Hoogenboom Jennifer 202201 MSC

Horticultural Management and Environment Control Strategies for Cannabis
(Cannabis sativa L.) Cultivation
by
Jennifer Hoogenboom
A Thesis
presented to
The University of Guelph
In partial fulfilment of requirements

for the degree of
Master of Science
in
Environmental Science
Guelph, Ontario, Canada
© Jennifer Hoogenboom, January 2022

ABSTRACT
HORTICULTURAL MANAGEMENT AND ENVIRONMENT CONTROL STRATEGIES

FOR CANNABIS (CANNABIS SATIVA L.) CULTIVATION
Jennifer Hoogenboom Advisors:
University of Guelph, 2022 Dr. Mike A. Dixon
Dr. Thomas Graham
The objective of this project was to establish horticultural management recommendations for
each growth stage of cannabis with respect to growing medium, light quality and flux,
temperature, and CO2 supplementation. A peat-based potting mix produced longer roots
compared to peat plugs; however, rockwool media resulted in 97% rooting success. There was
no effect of light quality on root dry mass; however, the cuttings under red light exhibited
significantly more and longer roots. Low light acclimated plants exhibited contrasting Net
Carbon Exchange Rate (NCER) responses to 15 light spectral qualities; no differences in NCER
were observed between 15-32°C. NCER responses to light and CO2 concentration indicated
optimal light intensity of 410-960 µmol m-2s-1 and ~1600 ppm CO2 for stock, 21 DAP vegetative
and 35 DAP flowering plants. Plants in a peat-based mixture with slow-release fertilizer
exhibited higher floral yield than organic mixtures.

iii
ACKNOWLEDGEMENTS
This work was supported by the Natural Sciences and Engineering Research Council of
Canada (NSERC) and ABcann Medicinals Incorporated (now Vivo Cannabis Inc.).
First, I’d like to sincerely thank my advisors, Dr. Mike Dixon and Dr. Thomas Graham,
for this unique and fantastic opportunity. My time at the facility is an experience that I greatly
appreciate. The lessons I’m taking away extend far beyond skills at a lab bench. I was given the
space to grow independently while consistently being supported throughout my time in Napanee.
Thank you to Dr. Youbin Zheng for his knowledge and advice on my advisory committee. I
would also like to express immense gratitude to Dr. Mike Stasiak; his humility and generosity of
his time and expertise ensured that his contributions to experimental design and technical support
were always available in quick succession. Special thanks to the students in the CESRF lab for
their input and support during our online meetings and before the final defence. Thank you to
Ping Zheng for the nutrient analysis; it was a significant endeavour due to the large sample size.
Thank you to Theresa Rondeau-Vuk for your input during lab meetings and for ensuring that all
administrative work ran smoothly. To employees of ABcann, you have all provided me with
support and assistance to shadow daily operations, set up and run my experiments. Thanks
mainly, in no specific order, to Charlotte Presley, Christopher Gauvreau, Graham Wise, Ian
Langback, James Adams, Jeffrey Ray, Kelly Conboy, Kim Warner, Leiann Ford, Matt Vine,
Matthew Christie, Nate Boldrick, Ryan Benvenuti, Michael Keegan and Ryan Ford for their
contributions and support towards my research. To my loved ones, especially my parents (Pieter
and Dina), thank you so much for supporting me throughout my research, I would not have
gotten through it without your encouragement.
iv
TABLE OF CONTENTS
Abstract ........................................................................................................................................... ii
Acknowledgements ........................................................................................................................ iii
Table of Contents ........................................................................................................................... iv
List of Tables ................................................................................................................................ vii
List of Figures .............................................................................................................................. viii
List of Abbreviations ...................................................................................................................... x
List of Appendices ......................................................................................................................... xi
Chapter 1: Literature Review .......................................................................................................... 1
1.1 Cannabis physiology .................................................................................................................... 1
1.2 Historical barriers in Canada ........................................................................................................ 2
1.3 Cultivation .................................................................................................................................... 3
1.3.1 Indoor controlled environment ............................................................................................................... 4
1.3.2 Propagation ............................................................................................................................................ 6
1.3.3 Growing media and fertilizer ................................................................................................................. 7
1.3.4 Net carbon exchange rate ....................................................................................................................... 9
1.4 Light ........................................................................................................................................... 10
1.4.1 Photosynthetic pigments ...................................................................................................................... 13
1.4.2 Photoreceptor pigments ........................................................................................................................ 13
1.4.3 Horticultural research on light ............................................................................................................. 14
1.5 Objectives ................................................................................................................................... 18
1.6 Hypotheses ................................................................................................................................. 19
Chapter 2: Methods ....................................................................................................................... 20
2.1 Site and Growth Chamber Description ...................................................................................... 20
2.2 Experiment 1 – Effect of elevated CO₂ and light intensity, on whole plant net carbon exchange
rate during the vegetative stage ............................................................................................................... 21
2.2.1 Plant material ....................................................................................................................................... 22
2.2.2 Gas exchange measurements................................................................................................................ 23
2.2.3 Harvest ................................................................................................................................................. 24
2.2.4 Statistical analysis ................................................................................................................................ 24
2.3 Experiment 2a – The effect of growing media on root development during propagation ......... 25
2.3.1 Plant material ....................................................................................................................................... 25
2.3.2 Harvest ................................................................................................................................................. 26
2.4 Experiment 2b – The effect of spectral quality of light on root development during propagation
.................................................................................................................................................... 27
2.4.1 Plant material ....................................................................................................................................... 27
2.4.2 Propagation .......................................................................................................................................... 28
2.4.3 Light ..................................................................................................................................................... 29
2.4.4 Rooting performance measurements .................................................................................................... 30
v

2.5 Experiment 3 – Growing media considerations for container-grown production ...................... 31
2.5.1 Propagation .......................................................................................................................................... 32
2.5.2 Transplant and Fertilization ................................................................................................................. 33
2.5.3 Measurements ...................................................................................................................................... 35
2.5.4 Harvest ................................................................................................................................................. 35
2.6 Experiment 4 – Effects of CO₂ concentration, temperature, light intensity and spectral
composition on whole plant net carbon exchange rate for early vegetative and flowering stages ......... 36
2.6.1 Propagation .......................................................................................................................................... 36
2.6.2 Transplant............................................................................................................................................. 37
2.6.3 Gas Exchange ....................................................................................................................................... 38
Chapter 3: Results ......................................................................................................................... 40
3.1.1 CO₂ response ........................................................................................................................................ 41
3.1.2 Light response ...................................................................................................................................... 42
.................................................................................................................................................... 44
3.4.1 Rooting response .................................................................................................................................. 45
3.4.2 Growing media pH and EC .................................................................................................................. 45
3.4.3 Growth response................................................................................................................................... 48
3.5 Experiment 4 – Effect of CO₂, light intensity, temperature and spectral composition on whole
plant net carbon exchange rate for early vegetative and flowering stages .............................................. 50
3.5.1 CO₂ response ........................................................................................................................................ 50
3.5.2 Light response ...................................................................................................................................... 52
3.5.3 Temperature response .......................................................................................................................... 54
3.5.4 Light spectra response .......................................................................................................................... 56
Chapter 4: Discussion ................................................................................................................... 57
.................................................................................................................................................... 63
4.5 Experiment 4 – Effects of CO₂ concentration, temperature, light intensity and spectral
composition on whole plant net carbon exchange rate for early vegetative and flowering stages ......... 68
4.6 Summary .................................................................................................................................... 72
Chapter 5: Conclusion................................................................................................................... 77
Literature Cited ............................................................................................................................. 78
vi
Appendices .................................................................................................................................... 98
Appendix I ............................................................................................................................................... 98
Appendix II.............................................................................................................................................. 99
Appendix III .......................................................................................................................................... 101
Appendix IV .......................................................................................................................................... 103
Appendix V ........................................................................................................................................... 104
Appendix VI .......................................................................................................................................... 106
vii
LIST OF TABLES
Table 1. Growth chamber environmental parameters for stock and cuttings during the
experiment. Abbreviations: D= Day, N=Night, DAP= Days after planting. ................................ 26
Table 2. Growth chamber environmental parameters for stock and propagation stages during the
experiment. Abbreviations: DAP: Days after planting, D: Day, N: Night. .................................. 28
Table 3. Growth chamber environmental parameters for stock and propagation growth stages of
cannabis during the experiment. Abbreviations = D: Day, N: Night, DAP: Days after planting . 33
Table 4. Growth chamber environmental parameters for vegetative and flowering growth stages
during the experiment. Abbreviations = D: Day, N: Night, DAP: Days after planting ................ 34
Table 5. Controlled environment chamber schedules for propagation, vegetative and flowering
growth stages during the experiment. Abbreviations: DAP: Days after planting, D = Day, N =
Night. ............................................................................................................................................ 38
Table 6. Root length of the three longest roots in propagated vegetative cuttings in two different
mediums, peat plug and potting mix at 14 days after planting ..................................................... 44
Table 7. Effect of light quality treatments on the average number of roots and root lengths of the
five longest roots 14 days after planting ....................................................................................... 45
Table 8. The average (n=4) pH of three growing medium treatments, Recipe 1, Recipe 2 and the
peat based mixture obtained with the pour-through method with applications of RO water at 26-
96 DAP.......................................................................................................................................... 46
Table 9. The average (n=4) EC of three growing medium treatments, Recipe 1, Recipe 2 and
peat based mixture at 26-96 DAP obtained with the pour-through method using RO water ....... 47
Table 10. Evaluation of dry weight of shoot, leaf and flower at harvest of three growing medium
treatments ...................................................................................................................................... 49
viii
LIST OF FIGURES
Figure 1. Schematic illustration of the experimental PGC Flex and PS1000 chambers. The PGC
Flex was divided into two sections (one or two-tier), totaling nine sub-sections. There were a
total of 8 chambers in a research lab located in the Vivo facility. ................................................ 21
Figure 2. PS1000 growth chamber arrangement of one experimental unit during acclimatation.
Chamber was equipped with variable spectrum LED arrays. ....................................................... 22
Figure 3. Normalized spectra for full spectrum in PS1000 chamber. Spectral distribution was
measured using a Li-180 spectrometer (LI-COR Biosciences., Lincoln, USA). .......................... 23
Figure 4. Schematic and example of a replicate of the planting density and arrangement of the
12 cuttings in a propagation tray before placement in a growth chamber. ................................... 29
Figure 5. Normalized spectra for seven channel LED lights (Intravision Light Systems, Inc.,
Toronto, ON, Canada). Each fixture had four rows and contained dimmable spectrum channels
for green (530 nm), red (660 nm), blue (440 nm) and white (5650 K), which were individually
controlled with a fully automated system (Argus Controls., Surrey, British Columbia, Canada).
Spectral distribution was measured using a Li-180 spectrometer (LI-COR Biosciences., Lincoln,
USA). ............................................................................................................................................ 30
Figure 6. Arrangement of treatments (Soilless mixture, Recipe 1 and Recipe 2) and growth
chamber replicates. Each box represents a growth chamber with randomly arranged sub-samples.
....................................................................................................................................................... 34
Figure 7. CO₂ concentration (ppm) of vegetative plants in the PS1000 chambers measured over
24 hours. During the dark period due to respiration, CO₂ accumulated and was drawn down.
When lights were on. CO₂ was added to adhere to the setpoints controlled by Argus (Argus
Controls., Surrey, British Columbia, Canada) and to offset photosynthetic uptake. Values
represented as means ± SEM (n = 9). ........................................................................................... 40
Figure 8. NCER response of vegetative plants in the PS1000 chambers over a range of CO₂
concentrations from 480, 800, 1000, 1200, 1600 and 1980 ppm. NCER was measured at 178
µmol m-2s-1, 55% relative humidity and 25°C. Values represented as means ± SEM (n=9, R2=
0.9996). ......................................................................................................................................... 41
Figure 9. NCER response of vegetative plants in the PS1000 chambers over a range of light
intensities from 115, 215, 410, 500, 685, 775 and 860 μmol m-2s-1. NCER was measured at 520
ppm CO₂, 55% relative humidity and 25°C. Values represented as means ± SEM (n=9, R2 =
0.9967) .......................................................................................................................................... 42
Figure 10. A) Example of a tray in a PGC Flex chamber, each chamber subsection contained
three trays B) Example of root formation for a subsample of each growing medium a) Peat plug
b) Potting mix c) Water d) Rockwool cubes ................................................................................. 43
ix
Figure 11. Image of the adventitious root formation of a subsample grown under each light
spectrum a) White b) Red c) Blue d) R:B e) Green at 14 DAP in the PGC Flex chambers ......... 44
Figure 12. Nutrient delivery system pH and EC measurements of four replicates over a range of
26-96 DAP of the Recipe 1, Recipe 2 and the peat based mixture. Solid lines represent the pH,
and the dotted line represents the EC measurements corresponding to the colours in the legend.
Reported values are means ± SEM. .............................................................................................. 48
Figure 13. Cannabis experimental plants at 104 DAP in a) Soilless growing medium b) Recipe 1
growing medium c) Recipe 2 growing medium ........................................................................... 49
Figure 14. A) NCER response for a 21 DAP vegetative plant (n = 3, R= 0.9999) over a range of
CO₂ concentrations at 605, 815, 1000, 1200, 1600 and 1975 at light intensity of 100 μmol m-2s-1.
B) NCER response for a 35 DAP flowering plant (n = 4, R = 0.9983) over a range of CO₂
concentrations from 545, 805, 1000, 1200, 1600 and 1975 at 140 μmol m-2s-1. .......................... 51
Figure 15. NCER response for a 21 DAP vegetative plant and 35 DAP flowering over a range of
light intensities from 128, 241, 459, 564, 767, 865 and 960 μmol m-2s-1 at 25°C and 520 ppm. A)
21 DAP vegetative plant (n = 4, R= 0.9938) at 65% humidity. B) 35 DAP flowering plant (n = 4,
R = 0.9948) at 60% humidity. ....................................................................................................... 53
Figure 16. Temperature NCER response at increasing intervals of 15, 18, 23, 26, 29, 32 °C for a
21 DAP vegetative plant and 35 DAP flowering plant at 520 ppm. Points represent mean ± SE
values (n = 4). A) 21 DAP vegetative plant at 65% humidity, 100 μmol m-2s-1 and 520 ppm. B)
35 DAP flowering plant at 65% humidity, 140 μmol m-2s-1 and 520 ppm. .................................. 55
Figure 17. NCER response at different light qualities for a 21 DAP vegetative plant at 250 (μmol
m-2s-1). Grey bars represent mean ± SE values (n=3). .................................................................. 56
Figure 18. NCER response at different light qualities for a 35 DAP flowering plant at 250 (μmol
m-2s-1). Grey bars represent mean ± SE values (n=4). .................................................................. 57
x
LIST OF ABBREVIATIONS
CESRF – Controlled Environment Systems Research Facility

CBC – Cannabichromene
CBD – Cannabidiol
CBG – Cannabigerol
CBN – Cannabinol
CO2 – Carbon Dioxide
CP – Compensation Point
FR – Far-Red
GPP – Geranyl pyrophosphate
HVAC – Heating, Ventilation and Air Conditioning
LED – Light Emitting Diode
MVA – Mevalonate
NCER – Net Carbon Exchange Rate
PPFD – Photosynthetic Photon Flux Density
R:B – Red:Blue
RH – Relative humidity
RO – Reverse Osmosis
THC – Tetrahydrocannabinol
UV – Ultraviolet
VPD – Vapor Pressure Deficit
xi
LIST OF APPENDICES
Appendix I – Technical summary of “PS1000” chambers (a version of the Guelph Bluebox

Chamber) growth chamber
Appendix II – Technical summary of “PGC Flex” growth chambers
Appendix III – Light intensity maps of White, Blue, Red, R:B (1:1) and Green for Experiment
2b
Appendix IV – Analysis report using saturated medium extract method to remove water-soluble
nutrients of the potting mix (ProMix HP Mycorrhizae)
Appendix V – Light intensity maps of R:B (4:1), White, White + FR, White + ultraviolet (UV),
R:B (1:4) for the case study
Appendix VI – Case Study – The influence of spectral quality on NCER, nutrient uptake and
morphology
1
Chapter 1: Literature Review
1.1 Cannabis physiology
Cannabis (Cannabis sativa L.) is an annual, herbaceous and monotypic genus originating
from Asia, specifically in modern day India and China (McPartland et al., 2019; Ren et al., 2019;
Small et al., 1976). It has been cultivated for over 10,000 years and is primarily grown for three
post-harvest products: grain, fibre and phytochemicals, notably phytocannabinoids and
terpenoids (Russo, 2007; Zhang et al., 2018). In the literature, cannabis can also be identified as
hemp, dagga, bhang, locoweed, marihuana, marijuana, hashish, or hasheesh. Cannabis is a
naturally dioecious species with sexual dimorphism occurring during flower development, but
can also differentiate into hermaphroditic inflorescences spontaneously (Moliterni et al., 2004;
Rana et al., 2015). Cannabis is a wind-pollinated (Small & Antle, 2003), primarily a short-day
plant (Spitzer-Rimon et al., 2019; Tournois, 1912), requiring an average of ~12 hours of
darkness to initiate flowering. Day-neutral cultivars have the exception of flowering without
being governed by a specific photoperiod (Hall et al., 2012). The production cycle can be
separated into three different categories: propagation, vegetative and flowering. It is typical to
have a separate room designated for each growth phase due to the different photoperiod and
environmental demands. The end product of the production cycle can be based on the final yield,
but also the chemical composition of the product as it is of interest to producers and consumers.
Cultivars are bred to provide different products, which has led to a diverse range of
genetic, morphological and biochemical variation. The morphological characteristics can vary
drastically among cultivars, heights can range from 0.2-6.0 m, palmate leaves can contain 3-13+
leaflets, leaflets can be broad or narrow, and there are multiple distinctions with branching, stem
quality, and inflorescence density (Jin et al., 2021; Small et al., 1976; Zhang et al., 2018). A
systematic approach to characterizing cultivars is through their dry inflorescence
phytocannabinoid ratios, namely tetrahydrocannabinol (THC) and cannabidiol (CBD), which can
be assigned into one of three chemotypes (Fischedick et al., 2010; Small & Beckstead, 1973).
Chemotype I or drug-type cultivars are dioecious, determined by greater than 0.3% THCA and
less than 0.5% CBD. Chemotype II cultivars have intermediate quantities of both THCA (more
2
than 0.3%) and CBD (more than 0.5%). Chemotype III or fibre-type cultivars are typically
monecious, are characterized by less than 0.3% THCA and greater than 0.5% CBD. Chemotype
II and III are selectively bred for fibre, oil and grain production (Small & Beckstead, 1973).
Medical research has focused on the minor phytocannabinoids for potential therapeutic
application (Gonçalves et al., 2019); this research has led to the discovery of two new
chemotypes. Like type III varietals, Chemotype IV contains less than 0.001% THC, less than
0.5% CBD, and CBG greater than 0.03% (Fournier et al., 1987). Chemotype V refers to varietals
that exhibit little to no phytocannabinoid content (Mandolino & Carboni, 2004). This thesis
focuses on studying chemotype I, the drug type cannabis, made available for recreational and
medicinal Canadian consumers in 2018.
1.2 Historical barriers in Canada
Cannabis is a plant that has been both honoured and outlawed throughout history.
Cannabis is considered one of the oldest crops distributed worldwide (Schultes et al., 1974).
Research available today has focused primarily on pharmacological properties, cannabis use
policy, hemp field production, addiction and potential harms (Amaducci et al., 2008, O'Grady,
2020) but have significantly lacked research of large-scale quality cannabis production. Cannabis
was introduced to America in 1545 (Small et al., 1975). However, the earliest known evidence of
the first hemp crop grown in Canada was planted in Port Royal, now known as Nova Scotia, in
1606 (Abel, 1982). Despite cannabis being a multipurpose crop, it was primarily grown for fibre
for several hundred years until 1938, when The Canadian Opium and Narcotics Act made
cannabis cultivation illegal (Cherney & Small, 2016). This effectively eliminated the cannabis
industry until 1994, when Canada began issuing licenses to industries for research, paving the
way in 1998 to permit commercial cultivation under the control of Health Canada (Cherney &
Small, 2016). The last two decades have led to immense policy changes within the Canadian
cannabis industry for recreational and medical markets. Following suit, in 2001, the Canadian
Medicinal Marijuana Access Regulations granted legal access to cannabis for medical patients
with authorization to grow. The legal status of cannabis production had radically shifted in
Canada with the Canadian Senate enactment of the Cannabis Act, Bill C-45. This created a
landmark moment, as Canada had become the second country in the world in 2018 under federal
3
jurisdiction to obtain this approval. It is estimated to be a 22.6 billion dollar industry in Canada
(Deloitte, 2016).
The versatility of this multi-purpose crop has been demonstrated by its uses among
different types of consumers. The clinical utility of cannabis has been researched and shown to
be of positive value in patients who cannot benefit from traditional medicines or can use it in
conjunction with pain management (Vigil et al., 2017; Whiting et al., 2015). These studies
include epilepsy (Silvestro et al., 2019), post-traumatic stress disorder (Bonn-Miller et al., 2014),
psoriasis, multiple sclerosis and Parkinson’s disease (Kindred et al., 2017). In conjunction with
the recent legalization, it is of increasing importance to cannabis producers to identify the best
methodology in producing quality flowers for both recreational and medicinal consumers.
1.3 Cultivation
Standard technologies in agriculture are being studied on cannabis, however, the optimal
growing procedures of every cultivar are not available. Although there are advantages to outdoor
production, the significant benefit being low production costs and low energy utilization, there
are also many challenges. The main challenge to commercial outdoor cannabis cultivation is
reduced environmental control. With outdoor production, the grower has less control over
growth factors such as length of the growing season, photoperiod (daylength), light quality
(spectral distribution) and light quantity (intensity). Additionally, vastly more energy comes
from natural sunlight than from any artificial light source. Wavelengths can range from
approximately 250 - 2500 nm, with about 8% light composition containing harsh ultraviolet
radiation (Hollósy, 2002). The wavelengths of UV radiation (100-400 nm) are divided into three
subcategories, UV-A (315-400 nm), UV-B (280-315 nm) and UV-C (100-280 nm). Sunlight
reaching the surface of the earth is comprised of about 95% UV-A and 5% UV-B. This could
damage plant DNA, as UV-B causes reactive oxygen species to actively scavenge enzymes from
the plant (Carvalho & Folta, 2014), however plants have found a way to adapt to UV-B
minimizing harmful transmittance. Exposure to UV-B stimulates production of enzymes to repair
damage and accumulate phenolic compounds that absorb damaging wavelengths of light (Santin
et al., 2018).
4
By growing under sunlight, growers are limited to the natural spectra that the sun is
outputting, with no way to adjust between growth stages to optimize specific metabolic
processes. This was elucidated in a study conducted by Livadariu et al., 2018, where cannabis
seeds were improved by illumination with LEDs by influencing the success rate of seeds and
total fresh weight. Results indicated that cannabis seeds grown with LEDs are of higher quality
than when grown under sunlight. Natural canopies do not create homogenous environments due
to daily and seasonal fluctuations in light composition. Data presented in Hemphill et al. (1980)
study showed that cannabinoid content in 12 cultivars analyzed from different geographical
regions contained different concentrations. In another study, Zhang et al. 2018 analyzed 52
cannabis cultivars that exhibited distinct latitudinal gradients patterns, differences likely due to
climatic heterogeneity and geographical isolation. Medical producers require high quality,
uniform yield and the production of certain phytocannabinoid compounds or ratios between the
different phytocannabinoids. Due to the need for standardization of medicine, outdoor cultivation
is not sufficient for medicinal products. This indicates that the optimal growing environment for
cannabis would be in an indoor controlled environment system where it would be possible to
take advantage of a light source that provides a consistent light composition.
1.3.1 Indoor controlled environment
Indoor production can control the significant variables that affect plant growth and
development to obtain consistent and uniform yield. This is important for producing certain
phytocannabinoids or a ratio of specific phytocannabinoids throughout the canopy and between
production cycles. The practice of indoor cultivation is driven by the pursuit of security for such
an important crop. In addition, there are advantages, including reducing the risk of cross-
pollination, effective management of pests or diseases, having a year-round growing season and
producing synchronized floral induction (Magagnini et al., 2018). Furthermore, under stable
environment conditions, producers in Canada can have up to six harvests within a year,
suggesting that indoor crop facilities are far more productive (Magagnini et al., 2018; Niu &
Masabni, 2018).
Large-scale cultivation of indoor cannabis is an energy-intensive proposition. The

legalization of cannabis in Canada has caused a substantial increase in energy demand.
5
Depending on local conditions, operational costs can represent half of the wholesale value of the
finished product (Mills, 2012). In California, a state that has had recreational use legal for four
years, annual energy expenditure for cannabis production amounts to ~3% (Mills, 2012). Factors
influencing energy consumption in indoor facilities required for successful growth include the
facility type, lighting fixture, carbon dioxide input, irrigation strategies, heating, ventilation and
air conditioning (HVAC). Of these, lighting represents 38% of the total electricity consumed,
followed by ventilation and humidification (30%) (Mehboob et al., 2020). A challenge that
indoor producers have to mitigate is how to balance the optimal growing conditions for specific
cultivars and the production costs.
Characterizing the photosynthetic response to environmental parameters can determine

the most cost-effective parameters. Indoor systems can control temperature, humidity, air
exchange, carbon dioxide, light quality and light intensity to provide a programmed recipe
optimal for the net carbon exchange rate (NCER) for a wide range of crops (Benke & Tomkins,
2017). Research in growth chambers is an invaluable tool in allowing researchers to characterize
a plant. Growth chamber studies of whole-plant cannabis and environment interaction provide
better baseline data to be applied in closed environments. This is evident when data is
extrapolated from studies that focus on light properties identified in purified compounds. The
observations investigated in Moss & Loomis, 1952, resulted in underestimating the absorbance
of green light (Shimadzu et al., 2019). Studies like these will not extend well to single leaf data
or whole plants and highlight the advantage of whole-plant studies.
The two main environment variables that producers can manipulate to optimize growth
are light and CO₂. Adjusting lighting applications or CO₂ input instead of changing genetics or
retrofitting an HVAC system is a more feasible approach for producers (Hall et al., 2012). Yield
increases have been shown to be correlated with elevated CO₂; however, each cultivar has an
optimal level that depends on the transpiration and respiration interactions in the plant (Bugbee
et al., 1994). In a study conducted by Chandra et al., 2011, it was elucidated that four different
cultivars grown under elevated CO₂ conditions (545 and 700 ppm) increased photosynthetic rates
by an average of 44%. In another study, the effect of photosynthetic photon flux densities from
500-2000 µmol m−2 s−1 observed the optimum light intensity to be 1500 µmol m−2 s−1 (Chandra et
6
al., 2008). It is essential to identify the minimum CO₂ and light to avoid inadvertent plant stress,
as studies have shown that long-term enrichment can be associated with feedback inhibition of
photosynthesis (Bugbee et al., 1994).
1.3.2 Propagation
The selection of cannabis globally has generated a unique arena of genetics, and it is
essential to preserve the integrity of genetics over generations. Cannabis consumption relies on
uniform whole flower development to ensure accurate and reliable medical products are being
provided. Unlike products used for extraction, which can utilize post-harvest technology to
change the chemical profile, whole flower products do not have this advantage.
It is of particular importance to have a healthy propagation phase as that determines the

future success of the plant. Adequate relative humidity, temperature, light and airflow are
environmental parameters that should be carefully considered for quality root formation. Another
primary consideration is the propagation technique which can be selected between three options.
Cannabis can be propagated by seed (Jovičić et al., 2019), stem cuttings (Caplan et al., 2018) and
in-vitro propagation (Piunno et al., 2019). Planting seeds allows producers to obtain new genetics
however it does have some disadvantages; if it is non-feminized, the crop output has the
disadvantage of requiring staff to identify and cull males to prevent cross-pollination. Feminized
and non-feminized seeds offer the burden of non-uniform plants resulting in inconsistencies in
cannabinoid profile and yield in the final product. Clonal propagation through stem cuttings is a
method that production facilities commonly use for herbaceous and woody plants; despite being
laborious, the final output is uniform and consistent plants between generations. It allows
producers to bypass the juvenile phase of cannabis, reaching maturity faster and preserving the
genetic characteristics (Chandra et al., 2020). Clonal propagation involves taking a stem cutting
containing one or more nodal segments and leaves from a mother plant, known as the stock
plant, and causing it to regenerate into a new plant. Stock plants are maintained in a vegetative
stage for their extended periods and are typically kept in a separate room from production plants.
Modifications in lighting quality, lighting intensity, and CO₂ can alter the morphology of a stock
plant to influence the maintenance of the stock plant (i.e. low light intensity reduces defoliation
frequency), number and size of clones (Campbell et al., 2019b; Swarts et al., 2018). In-vitro
7
propagation is a valuable tool to complement conventional breeding technology. In-vitro

propagation can be used to preserve elite genetics or plant regeneration to form whole plants.
The rate and success of stem cutting establishment depends on genetics, the efficiency of
operations, stock plant size and stem architecture. Other factors can include the endogenous
auxin in the cutting (Fogaça & Fett-Neto, 2005), use of synthetic auxin (indole butyric acid,
IBA), stem diameter, age of the cutting, leaf area (Aminah et al., 1997), number of leaves or
position of the cutting (Caplan et al., 2018). Caplan et al., 2018 study was the first scientific
paper to evaluate the influence of the success of a stem cutting based on leaf number (2-3), tip
removal, location of the cutting from the stock plant and rooting hormone. This study identified
that utilization of IBA, non-removal of leaf tips, apical or basal cuttings from a stock plant and
cuttings with three leaves increased the rooting success.
1.3.3 Growing media and fertilizer
In propagation and production, numerous growth media are available to be used,

categorized into organic, soilless, and soil. An optimal indoor growing medium would have high
water and air holding capacity (Zheng et al., 2007), anchor the plant appropriately, supply
sufficient solutes for nutrient uptake and have the ability to be reused or recycled properly. Soil
contains non-sterile inorganic particles, organic matter and can vary in fertility and porosity.
Soilless cultivation is defined as plant production without the use of soil and can be categorized
into hydroponics, aquaponics, aeroponics and soilless substrate culture (Maluin et al. 2021).
Soilless substrate culture is characterized by the use of sterile inert organic substances, including
peat moss (Michel, 2010), compost pine bark, and coir (Caplan et al., 2019) or inorganic
substances including limestone, sand, perlite and vermiculite (Zheng, 2019). Typically, soilless
substrate culture is a mixture of organic and inorganic substances. Further, it can use media
surfactants that can aid in the wetting and movement of water to the root zone. Aquaponics
utilizes the symbiotic relationship between plants and fish whereby the nutrient rich aquaculture
water is fed to hydroponically growing crops. Aeroponics is defined by plants being suspended
in the air and spraying them in the form of a mist with a nutrient solution.
8
Hydroponics involves growing plants in an aqueous nutrient solution with the roots fully
exposed or placed in an inert medium such as glass beads, clay aggregates, or rockwool (Bussell
& McKennie, 2004). Compared to hydroponics, soil and organic systems have a large buffering
capacity, and are easier to set up due to the lack of constant aeration, water circulation, electricity
and nutrients. Rockwool is a popular option in hydroponic systems since the material is
chemically inert, has a high water retention capacity, high air porosity and is available in many
different sizes and shapes (Bussell & McKennie, 2004). Lastly, organic growing media are
comprised of soil or organic soilless mixtures, it differs by the regulated use of specific soil
conditioners (i.e, manure, compost, or mulch) and organic fertilizers. Each of these growing
media has varying physical and chemical properties, requiring different fertilization and
irrigation strategies throughout a production cycle (Nemati et al., 2021; Zheng, 2020). In indoor
environments, these growing media are typically controlled with automatic irrigation methods
and control systems. There is minimal scientific research available on optimizing propagation of
cannabis in controlled environments with growing media (Caplan et al., 2018; Casanoa, 2019;
Chandra et al., 2020; Potter, 2009). However, this has been thoroughly investigated amongst
other species (Alam, 2020; Isfendiyaroglu et al., 2009; Rajkumar et al., 2017; Tate & Page,
2018; Vaughn et al., 2021).
The massive production of synthetic fertilizers used in soil and soilless growing media
have high costs for agriculture and negative consequences that can lead to environmental
pollution, nutrient leaching into water systems, nutrient loss through volatilization, low nutrient
use efficiency and crop productivity decline (Liu et al., 2014; Zheng, 2018). When considering
strategies for cost-effective production, the additional labour due to multiple fertilizer
applications required throughout a production cycle and the potential buildup of toxic chemicals
in the media or irrigation system should be considered. For producers, synthetic fertilizer offers
accurate concentrations and is flexible to changes throughout the production cycle, allowing
producers to tackle nutrient deficiencies with a fast-acting formula. In addition, when stored
under proper conditions, these products can last for a very long time. Despite the benefits, there
is a significant concern that many production facilities are taking part in the overuse of
fertilization (Clark & Zheng, 2017) and underutilizing the potential that the combination of soil
or soilless medium and slow-release fertilizer, or fertile soil has to offer.
9
Slow-release fertilizer (or controlled-release fertilizer) releases nutrients over a period of

generally 1-3 months through microbial decomposition or through applications of water by
breaking down a physical barrier (Lawrencia et al., 2021; Trenkel, 2013). Research has shown
that the use of fertilizer consistently results in increasing overall biomass and yield. Studies often
show an increased growth response to fertilization in hydroponic solutions (Bevan et al., 2021)
and soil (Clark & Zheng, 2014; Coffman & Gentner, 1977). However, if the soil fertility is
optimal, any additional fertilizer input provides no benefit to the overall yield of the plant.
1.3.4 Net carbon exchange rate
Plants will acclimate to environment conditions through changes in photosynthesis,

physiology and metabolism to optimize plant productivity (Herrmann et al., 2019; Walters,
2005). Acclimation can occur under short-term conditions, from second to minute responses or
have long-term responses to fluctuating conditions (Morales & Kaiser, 2020). NCER response
graphs describe the relationship between the NCER (µmol CO2 m−2 s−1) and a specific
environmental parameter. NCER is an important metric, as it plays a significant role in dry
biomass accumulation. NCER is defined as the rate of CO₂ uptake in net photosynthesis minus
the CO₂ evolution in net respiration (including microbial respiration in non-sterile systems) and
is considered a predictor of growth rate as it estimates net C gain (Dutton et al., 1988).
To understand the growth metrics of a plant cultivar, placing the plant under controlled
environmental parameters to obtain response graphs is a method widely used on many other
plant species in research. Cannabis (Chandra et al., 2008, 2011; Rodriguez-Morrison et al.,
2021a) and other C3 plant species show a response profile that exhibits a rectangular hyperbolic
model to elevated CO₂ (Allen & Prasad, 2004), light intensity (Evans et al., 1993; Herrmann et
al., 2020; Kanemasu & Hiebsch, 1975; Lobo et al., 2013; Rivera-Méndez & Romero, 2017; Zhou
et al., 2020) and light quality (Li et al., 2020). Closed system gas exchange measurements are
most often conducted using small portable leaf-level systems or whole plant sealed chambers of
known volume. A whole plant completely isolated in a growth chamber of known volume can
provide more accurate gas exchange measurements compared to leaf level measurements. This
10
was observed by Rodriguez-Morrison (2020a) who noted that differing physiological ages of
tissues and location in the canopy can reduce the leaf responses to light intensity.
The NCER generally follows a curve where photosynthetic uptake overtakes respiration
(i.e. compensation point) and increases linearly until the curve starts to plateau (Taíz and Zeiger,
2006). Plants exhibit a wide range of responses to increasing PPFD (Demmig-Adams & Adams,
1992), temperature, humidity, CO2 and light quality. NCER response graph results from a plant
being exposed to incremental increases of a specific environmental parameter. It’s crucial that
when assessing a particular environmental factor, all the other environmental parameters are
maintained under controlled conditions, as they can affect the NCER and prevent reliable
conclusions.
The response graphs can derive the compensation point (CP), saturation zone (i.e.
plateau), max NCER, saturation point and a dark respiration rate (Herrmann et al., 2020). Under
dark conditions, there is no accumulation of C, CO2 emitted is due to dark respiration
(mitochondrial respiration) given off by the plant (Taíz and Zeiger, 2006). For the case of high
light acclimated crops, it is typical to see photosynthetic rate maxima occur at light intensities
above 600 µmol m−2 s−1. Beyond saturation, the plant will enact mechanisms to dissipate the
excess energy and initiate photoprotective processes (Demmig-Adams & Adams, 1992). The
performance of a cannabis plant under steady-state environment conditions allows producers and
researchers to determine the best practices that they could apply to their production facility,
depending on their goals and system limits.
1.4 Light
From the 1860s onwards there has been a constant stream of research on growing plants
under electric lights (Montoya et al., 2017). The era of electric lighting started with incandescent
lamps designed by Thomas Edison in 1879. Incandescent lamps were low in cost, generated heat,
emitted a greater proportion of red light, far-red and infrared radiation. Following this, Daniel
McFarlan Moore (1895) achieved success developing the first predecessor to fluorescent light
called the Moore Tube. The Moore tube was short-lived due to being expensive to replace, and
the other high intensity discharge lamps that used different gases and/or the pressure of such
11
gases in the bulb were more successful. High intensity discharge lights provided improved
electrical efficiency and longer operating life than incandescent lamps. In the 20th century, the
most popular and commercially successful high intensity discharge lamps were mercury vapor
(1901), metal halide (1912), and the modern fluorescent light (1926) (Gupta, 2017). In 1936, the
ﬁrst modern high-pressure discharge lamp was launched by Philips. These high-pressure lamps
had high electrical efficiencies, a high percentage of blue radiation, a long operating life, and
provided a relatively broad-spectrum light that was acceptable for a wide range of plant species.
These lamps have become popular for use in growth chambers and currently are the lamps of
choice for supplementing naturally lit greenhouses across much of the world. However, in
comparison to high-quality LEDs, traditional lighting cannot compete in the level of
performance. LEDs can have a lifetime of approximately 100k hours, while conventional
lighting typically has a lifetime of 10-20k hours (Bourget, 2008). LEDs have higher electrical
efficiencies, reducing the overall power consumption as well as lowering the cooling
requirements in growth rooms (Montoya et al., 2017).
As newer LED technology has become more cost-effective, the application has become
immensely popular, used as either sole-source lighting or supplemental in many different
production practices. The small size, customable setup (i.e. in series to form strings), efficient
energy utilization, durability, lower heat emission, and anti-static property have resulted in the
rapid deployment of LEDs in indoor growing environments (Benke & Tomkins, 2017; Massa et
al., 2008). In addition, LEDs can emit light of an intended spectrum (260 - 730 nm), allowing
specific wavelengths to be enriched, thus supplying the light quality essential for different
growth phases. The technology offers researchers the ability to study monochromatic light
qualities and various spectral combinations. All the experiments presented in this thesis take
advantage of LED technology.
Cannabis morphological and physiological state is determined by the light quality, light
quantity and photoperiod throughout the production period. A major limiting factor in plant
growth is light interception due to a plant’s system of receptors and signal transduction pathways
that generate a specific response for plant growth and development (Dixon et al., 1999). It has
been well established that narrow bandwidth illumination can alter specific responses, including
12
seed germination, de-etiolation, directional growth, organelle movement, flowering and

senescence (Carvalho & Folta, 2014). These effects are mediated by photosynthetic and
photoreceptor pigments, which upon being triggered with illumination will convert light into
biochemical energy or undergo a conformational change to be involved in signalling state
formation that leads to a specific physiological response (Kong & Okajima, 2016). There are
differences in responses to spectra among different species that can be identified to optimize
plant performance. Whether trends are consistent among cultivars remains a question. The
limited research available on appropriate light quality and intensity for optimal cannabis
production underscores the importance of studying the effect of LED technology.
Light intensity is the photosynthetic photon flux (PPF) density (PPFD), the quantity of
photons (μmol m-2s-1) that is used to drive photosynthesis. Light quality is represented by the
distribution of photosynthetically active radiation (PAR), the wavelengths of light between 400-
700 nm. McCree, (1971) generated a curve to provide a means to establish the optimal
wavelength selections and spectral ratios for different crops. The results obtained from the
McCree curve supported the optimal spectral range is between 400-700 nm, however recent
research provides evidence that photosynthetic photons extend beyond the range.
Physiologically, the curve describes leaf level response at low light intensities that can be
converted into chemical energy at the maximum photosynthetic efficiency. The curve coincided
with previous research that observed chlorophyll molecules isolated in an organic solvent absorb
blue and red wavelengths, indicative that these wavelengths are the most effective in
photosynthesis to excite electrons in chlorophyll. However, the curve also shows that the
wavelengths in between red and blue contribute positively. Although the McCree curve is widely
accepted, it contrasted with a prior study, conducted by Emerson, 1957, who showed that
supplementation of far red (>680 nm) with shorter wavelengths within the range of PAR
achieved higher photosynthetic rates compared to conditions in which plants were exposed to
isolated wavelengths; this suggested that the use of wavelengths with light greater than 680 nm
drive photosynthesis more efficiently than isolated wavelengths (Emerson & Rabinowitch,
1960). New research has determined that McCree, 1971 did not fully acknowledge that
wavelengths such as green (H. L. Smith et al., 2017; Terashima et al., 2009), UV (Lydon et al.,
1987; Rodriguez-morrison et al., 2021b) and far-red (Pettai et al., 2005; Zhen, 2020) can
13
effectively drive photosynthesis. The use of these spectra is still misrepresented in literature and
has been poorly used by the horticultural lighting industry in product development.
1.4.1 Photosynthetic pigments
The photosynthetic pigments are located in the chloroplasts and can be identified as
chlorophyll and carotenoids. In plants, chlorophyll a and chlorophyll b are the primary
photosynthetic pigments, absorbing wavelengths of light primarily in the blue and red regions
(Guidi et al., 2017). Chlorophyll a exhibits peak absorbance at 430 nm and 663 nm, while
chlorophyll b exhibits peak absorbance at 453 nm and 642 nm, as shown by the peaks in the
absorption spectra in an acetone solution (Eichhorn Bilodeau et al., 2019; Guidi et al., 2017).
Although chlorophyll a and chlorophyll b absorb light, chlorophyll a plays a unique and crucial
role in converting light energy to chemical energy. Chlorophyll b is considered an accessory
pigment because it is not necessary for photosynthesis to occur; however, it does help to expand
the absorption spectra of organisms. It acts indirectly in photosynthesis by transferring light it
absorbs to chlorophyll a.
Carotenoids are another critical group of accessory pigments that absorb violet and blue-
green light (400-500 nm). In photosynthesis, carotenoids help capture light, but they also have an
essential role in getting rid of excess light energy (Ouzounis & Alle, 2015). When a leaf is
exposed to the sun, it receives a large amount of energy. If that energy is not handled correctly, it
can damage the photosynthetic mechanisms. Carotenoids in chloroplasts help absorb the excess
energy and dissipate it as heat (Ouzounis & Alle, 2015). It is also worth noting that non-
photosynthetic pigments such as anthocyanins and associated flavonoids influence the coloration
of leaf tissue due to their optical properties and can serve as a photo-protectant from ultra-violet
light (Albert et al., 2009).
1.4.2 Photoreceptor pigments
Photoreceptor pigments will mediate responses to light in the UV to far-red spectra. The
physiological and morphological responses are determined by a particular wavelength absorbed
by the plant. These receptors can be divided into five major classes of photoreceptors; these
include the UV-A/blue light-sensing cryptochromes, phototropins and Zeitlupe family members;
14
red/far-red light sensing phytochromes and UV-B sensing receptor UVR8. Blue light (390-500
nm) and UV-A (315-400 nm) captured by cryptochrome plays a role in initiating hypocotyl de-
etiolation, internode shortening (Cope & Bugbee, 2013), photoperiodic control of floral
initiation, phototropic response in which plants bend toward the light, opening of the stomata and
decreasing leaf expansion (Benke & Tomkins, 2017; Briggs & Christie, 2002; Cashmore et al.,
1999). Phototropin is responsible for light detection (phototropism), stomatal opening and light-
induced chloroplast movement. Red light and far-red (600-750 nm) are mediated by
phytochrome and can exist in two photo reversible forms dependent upon the photon flux density
(Kong & Okajima, 2016; H. Smith, 2000). This receptor mediates seed germination, de-
etiolation, shade avoidance (Fiorucci & Fankhauser, 2017), and photo-morphogenic response
(Whitelam et al., 1998). UVR8 is responsible for UV-B (280-315 nm) induced responses
including photomorphogenesis and accumulation of flavonoids (Heijde & Ulm, 2012).
Predictable outcomes are similar amongst various species with different LED spectra
(Landi et al., 2020). Numerous studies have indicated that cryptochrome and phytochrome can
mediate elongation of stems; however, blue light is more effective in suppression in a range of
species (Appelgren, 1991; Cosgrove, 1981; Hoenecke et al., 1992; Kigel & Cosgrove, 1991).
Under blue and red LEDs, lettuce has been shown to obtain higher yields than white light.
Monochromatic blue light spectra illuminated on lettuce displayed higher growth, leaf area and
biomass, while monochromatic red promotes leaf elongation, lower carotenoid levels and lower
growth rates (Carvalho & Folta, 2014). In combination, blue and red light induces higher
carotenoid levels (Carvalho & Folta, 2014). Therefore, it can be said that the differential effects
of blue and red light, either in isolation or combination, result in altered phenotypes in lettuce
(Massa et al. 2008).
1.4.3 Horticultural research on light
Light protocols are currently being developed to identify the best light quantity
(Rodriguez-morrison et al., 2021a) and quality for commercial growth for individual cultivars.
Ideal light conditions are dependent upon the production facilities’ goals, so optimal light quality
or quantity may be adjusted over time accordingly. For example, if a production company is
focused on aesthetics, this may have a different optimal plant growth protocol than if a company
15
focuses on phytocannabinoid yield. Lighting conditions also need to be balanced with the
practicality of light sensitivity for employees working in the production facility. Additionally,
producers need to be profitable, so another consideration is that blue LEDs contain more energy
than red light. As a result, the red LEDs are less expensive to purchase and have less power
consumption. Specific spectra may result in more positive physiological responses, but in
comparison to broad-spectrum, it may be difficult for employees to identify pests, pathogens and
acknowledge signs of plant stress in the form of discoloured plant tissue (Hawley et al., 2018).
LEDs can be modified into various functional forms to provide different overhead light
or inter-canopy lighting to irradiate plants within the canopy (Massa et al., 2008; Morrow, 2008).
Numerous studies have shown that utilizing the photosynthetic capacity in the lower canopy can
increase productivity by 23-87% (Stasiak et al., 1998). It has been well established in cannabis
that active compounds extracted from inflorescences positioned near the top are more
concentrated than lower in the canopy due to differences in photon flux (Namdar et al., 2018). A
study conducted by Hawley et al., 2018 evaluated the influence of sole-source lighting compared
to RGB and RB sub-canopy lighting with cannabis plants. This study indicated that the value of
additional LEDs in the lower canopy increased the yield, concentration of THC and overall
terpene content.
Traditional systems do not yield the same output compared to LED lighting systems, as
indicated by research with other crops (Hernandez et al., 2020) and select studies conducted on
cannabis (Amrein et al., 2020; Campbell et al., 2019b; Magagnini et al., 2018; Namdar et al.,
2019). Magagnini et al., 2018 evaluated three different light sources (∼450 μmol⋅m−2 s−1), HPS
and two different LEDs to find that total phytocannabinoid content had matured faster, produced
26-38% more THC and 29-35% more CBD in the LED treatments. A recent study design
conducted by Amrein et al., 2020 (∼520 μmol⋅m−2 s−1) observed an increase in CBD and yield
under LEDs. Plants under HPS treatment had longer internodes, therefore taller plants, and
higher total biomass dry weight than the LED. In another study, researchers utilized 4:1 blue and
red LED fixtures versus conventional fluorescent and high-pressure sodium (HPS) lamps in
different growth stages to assess the biological activity (Namdar et al., 2019). Results paralleled
the Magagnini et al., 2018 study, as plants that were vegetative or flowered under LED lights
16
were more compact and had a higher amount of secondary metabolites overall. In a recent study,
dry flower yield was higher under two LED treatments compared to the ceramic metal halide
lamp light which exhibited a higher biomass allocation in the main stem and leaves (Reichel et
al., 2021). Similarly, Wei et al. (2021) demonstrated two LED R:B light treatments obtained
greater height, stem diameter, leaf numbers, cannabinoid content and yield compared to ceramic
metal halide grown plants. A similar response was detected for cannabis by Danziger &
Bernstein (2021), where higher levels of CBGA were measured under the LED compared to HPS
lighting. However, within the same study, CBDA and THCA varied among three cultivars,
demonstrating genotypic variability.
Using LED technology allows researchers to understand how plants respond to individual
wavelengths and how each wavelength can influence morphology. Recent research by
Westmoreland et al., 2021 investigated the effect of blue photon fraction on floral yield and
potency of hemp. The study elucidated that as the blue photons increased from 4-20%, there was
an overall decrease in yield. Another published paper observed that cannabis grown under LED
broad-spectrum was 23% taller and had 20% more leaf biomass than blue-red (1.0) in six
different cultivars (Lalge et al., 2017). Early research on cannabis has indicated that light quality
affects the phytocannabinoid content measured in young leaves (Mahlberg & Hemphill, 1983)
and whole flowers (Lydon et al., 1987). Mahlberg & Hemphill, 1983 identified that leaves from
plants grown in daylight, filtered red, green and blue light could influence the ratio of CBD to
THC accumulation. The authors concluded that THC content did not differ significantly under
shaded daylight, filtered red, filtered blue and daylight control, however THC content in filtered
green and darkness was lower. In contrast, CBD content grown in filtered blue and dark was
undetectable but was present in filtered green and daylight. Hemp seedlings under different light
spectra developed higher leaf length and width under R:B and R:B + FR light; higher leaf
number and nodes under a full spectrum with additional far-red and UV; higher shoots under
R:B, RGB + FR, and R:B + FR and UV (Jahirul et al. 2021). Livadariu et al., 2018 demonstrated
that the use of green light increases the total yield, antioxidant capacity, and THC content and
blue light causes an increase in protein, flavonoids and polyphenols in seedlings. These studies
indicate that an optimized light spectrum throughout all growth stages can significantly improve
the value and quality of cannabis.
17
The correlation of the ratio of ultraviolet (UV) light and increasing THC production is
based on evidence of Small & Beckstead, 1973, who provided a phytocannabinoid profile of
uniform plants grown outdoors in multiple regions. The UV-B irradiance level from each region
was established and it was evident that higher levels of THC were observed from areas with
higher levels of UV-B. Early research from Lydon et al., 1987 reported no significant
physiological or morphological effect among UV-B treatments; however, the content of THC
increased in both leaves and flowers in drug-type plants. However, this is in contrast with
Rodriguez-Morrison et al. (2021b) who found that increasing UV-B radiation throughout an
entire production cycle altered the secondary metabolite profile such that there was no
enhancement of THC or CBD, yet there was a reduction of terpene concentration and suppresses
cannabis yield in two cultivars. This suggests that UV-B radiation applied throughout an entire
production cycle induced morphological changes that provided no commercial benefit. Fairbairn
& Liebmann, 1974 evaluated no lighting influences on phytocannabinoid production by
comparing greenhouse-grown plants supplemented with UV or HPS lamps. In another study,
cannabis leaves were exposed to UV-C radiation to evaluate metabolite modifications (Marti et
al., 2014). It was evident that there were significant increases in dehydrostilbenes,
glycerophospholipids and cinnamic acid amides. Fairbairn & Liebmann, 1974 and Mary &
Crombie, 1977 also assessed the effect of the absence of light in mature plants. Both studies
found a parallel increase of dry weight and cannabinoid content for plants grown under an
artificial light source.
Research has indicated that increasing light intensity will positively impact floral yield
(Chandra et al., 2008; Eaves et al., 2020; Hollósy, 2002) and floral biochemical composition
(Potter & Duncombe, 2012; Vanhove et al., 2011). Chandra et al., 2008 elucidated that a specific
high yielding Mexican cultivar optimum for photosynthesis was 25-30°C and 1500 μmol m-2s-1
at 350 ppm and 55% relative humidity. In another study conducted by Chandra et al., 2015,
researchers found an increasing trend in net photosynthesis with light intensity up to 2000 μmol
m-2s-1 in four cultivars at 350 ppm, which added to the previous report that it may not be a
cultivar specific trend. Eaves et al., 2020 had a similar experimental design and identified that
yields increased linearly up to 1500 μmol m-2s-1 at 1100 ppm and saw no yield differences
18
among six different broad LED lighting spectra. Vanhove et al., 2011 study identified that four
different cultivars grown under 600 W lights obtained a higher yield and THC content than
plants grown under 400 W. Vanhove et al., 2011 and Potter & Duncombe, 2012 both identified
that the larger yield cultivars correlated with higher irradiance levels exhibited a higher
concentration of total phytocannabinoid content. More recently, leaf level photosynthesis of
flowering plants was analyzed; it was observed that dry inflorescence yield increased linearly
with increasing light intensity up to 1800 µmol m−2 s−1 (Rodriguez-Morrison et al., 2021).
Compare this to Moher at al. (2021) who determined that 600 μmol m-2s-1 was optimal for
improved plant morphology and 900 μmol m-2s-1 obtained higher above ground dry weight.
1.5 Objectives
The main areas of focus in this thesis were growing media, propagation and photobiology.
Combining the results of the experiments will provide growers and researchers detailed
information on the effects of environment conditions on morphology, net carbon exchange rate
(NCER), and/or yield to quantify these effects. The main objectives were:
1) Measure NCER response of stock plants exposed to a range of CO2 concentrations (420-
2020 ppm) at a constant light intensity and exposed to a range of light intensities (115-
859 µmol m-2s-1) at a constant CO2 concentration (Experiment 1)
2) Evaluate the growth of cannabis vegetative cuttings in four growth media based on root
and shoot development (Experiment 2a)
3) Evaluate the response of vegetative cuttings under five different light spectra of LED
lighting based on root and shoot development (Experiment 2b)
4) Evaluate the growth of cannabis in a peat-based mixture with slow-release fertilizer and
organic soil mixtures irrigated with RO water throughout an entire production cycle
based on morphology and yield (Experiment 3)
5) Measure NCER response of a 21 DAP vegetative and 35 DAP flowering plant exposed to
a range of CO2 concentrations (545-1975 ppm) at a constant light intensity and a range of
light intensities (128-961 µmol m-2s-1) at a constant CO2 concentration (Experiment 4)
19
6) Measure NCER response of a 21 DAP vegetative and 35 DAP flowering plant exposed to
a range of temperature (15-32°C) and various light qualities at a constant light intensity
and CO2 concentration (Experiment 4)
1.6 Hypotheses
There were nine main hypotheses for the experiments conducted:
Experiment 1
H01: At a constant CO2 concentration, increasing light intensity does not increase stock
plant NCER
HA1: At a constant CO2 concentration, increasing light intensity increases stock plant
NCER
H02: At a constant light intensity, increasing CO2 concentration does not increase stock
plant NCER
HA2: At a constant light intensity increasing CO2 concentration increases stock plant
NCER
Experiment 2a
H03: Growing medium does not increase cannabis root growth during cutting
establishment
HA3: Growing medium does increase cannabis root growth during cutting establishment
Experiment 2b
H04: Light quality does not increase cannabis root growth during cutting establishment
HA4: Light quality does increase cannabis root growth during cutting establishment
Experiment 3
H05: Growing medium does not increase growth and development
HA5: Growing medium does increase growth and development
Experiment 4
H06: At low light levels, increasing CO2 does not increase NCER
HA6: At low light levels, increasing CO2 increases NCER
H07: At a constant CO2 concentration, increasing light intensity does not increase NCER
20
HA7: At a constant CO2 concentration, increasing light intensity increases NCER

H08: At low light levels, increasing temperature does not increase NCER
HA8: At low light levels, increasing temperature increases NCER
H09: At a constant CO2 and light intensity, light quality does not increase NCER
HA9: At a constant CO2 and light intensity, light quality increases NCER
Chapter 2: Methods
2.1 Site and Growth Chamber Description
The experiments were conducted at Vivo Cannabis Inc, a federally licenced cannabis
production facility (Napanee, ON, Canada). The research lab contained three plant growth
chambers (PGC Flex, Conviron, Winnipeg, Manitoba, Canada) and five custom whole
plant/small canopy-scale net CO2 assimilation rate measurement chambers (PS1000, Controlled
Environment Ltd., Winnipeg, Manitoba, Canada). These PS1000 chambers (Appendix I)
controlled environment variables precisely while in situ full canopy NCER was measured. The
PGC flex chambers (Appendix II) were divided with two sections which were one-tier or two-
tier; the chambers were configured to total nine sub-sections amongst the three chambers, as
illustrated in Figure 1. Each sub-section was an isolated unit that could be configured with
different light spectra or intensities.
21
Figure 1. Schematic illustration of the experimental PGC Flex and PS1000 chambers. The
PGC Flex was divided into two sections (one or two-tier), totaling nine sub-sections. There
was a total of 8 chambers in a research lab located in the Vivo facility.
CO2 concentration was measured by the LI-COR Li-820 CO₂ gas analyzer and controlled
with a Sierra 810 Mass-Trak Flow (Sierra Instruments, Monterey, California). The multi-point
closure, hermetic seal and aluminum interior in all chambers ensured that the plants were sealed
in a closed system. Environment parameters were controlled by ARGUS Control systems
(ARGUS Control Systems Ltd., Surrey, BC, Canada). Each chamber was fully automated and
contained water-cooled seven channel LED lights that range from 365-780 nm (Intravision Light
Systems, Inc., Toronto, ON, Canada). The independently controlled spectral peaks available
were UV-A (λmax = 365 and 380 nm), Blue (λmax = 448 nm), White (5650 K), Green (λmax = 568
nm), Red (λmax = 655 nm) and Far-red (λmax = 735 nm).
2.2 Experiment 1 – Effect of elevated CO₂ and light intensity, on whole plant
net carbon exchange rate during the vegetative stage
The objective of this study was to evaluate a vegetatively growing stock plant under
fluctuating light intensities (115 to 859 µmol m-2s-1) and CO2 (420 to 2020 ppm) conditions to
obtain the NCER response. This study utilized whole plants in a closed system growth chamber.
22
The results of this study will assist the indoor cannabis industry in determining how much CO2
concentration and PPFD is suitable for this cannabis cultivar in the vegetative growth stage.
2.2.1 Plant material
Vegetative plants were obtained from the stock room at 123 days old (4 plants in Trial
One) and 153 days old (5 plants in Trial Two). All plants appeared to have similar height and
foliage densities. One plant was selected and transferred into a PS1000 chamber (Figure 2).
Figure 2. PS1000 growth chamber arrangement of one experimental unit during acclimatation.
Chamber was equipped with variable spectrum LED arrays.
Plants were in 101.6 mm x 101.6 mm x 101.6 mm rockwool blocks (Grodan.,

Roermond, The Netherlands). Chambers were set at 55% relative humidity, 520 ppm CO₂,
25°C day temperature, 23°C night temperature and acclimated for eight days before testing.
Light (178 µmol m-2s-1, 18-hour photoperiod) at plant canopy level was measured with an
Apogee MQ-500 quantum meter 600 mm from the light source and set as a full spectrum
illustrated in Figure 3. Plants were fertigated as needed using Dual Fuel 1 (5N-0P-1K) and
Dual Fuel 2 (0N-3P-6K) (Green Planet., BC, Canada).
23
Figure 3. Normalized spectra for full spectrum in PS1000 chamber. Spectral distribution was
measured using a Li-180 spectrometer (LI-COR Biosciences., Lincoln, USA).
2.2.2 Gas exchange measurements
To study the NCER whole-plant response of vegetative Cannabis sativa (cv. Wappa),
plants were exposed under different PPFD and CO₂ levels to determine the optimal
environmental parameters for NCER. Plants in trial one were exposed to a range of CO₂
concentrations (420, 820, 1020, 1220, 1620, 2020 ppm) controlled at 178 µmol m-2s-1, 55%
humidity and 25°C temperature. Thereafter, plants were exposed to a range of PPFD (115, 215,
410, 504, 596, 686, 774, 859 µmol m-2s-1) controlled at 520 ppm CO₂, 55% humidity and 25°C.
Plants in trial two were exposed to the same environmental scenarios but in reverse order (PPFD
followed by CO2). All parameters for PPFD and CO₂ were first recorded at the lowest value and
then subsequently increased over a period of 48 hours. Each sequential setpoint was scheduled
for 2-3 hours during the 18-hour photoperiod for a total of 12 hours (CO2) and 16 hours (PPFD).
24
2.2.3 Harvest
At harvest, stems were cut at the base of the stem just above the mineral wool block and
all leaves were separated from the stems. Leaf area was measured using an LI-3100C Area Meter
(LI-COR Biosciences, Lincoln, USA). The fresh and dry weight of stems and leaves were
measured using a Sartorius ENTRIS 6202-1S Precision Balance (Sartorius, Göttingen,
Germany). Fresh samples were placed in an incubator oven at 61°C for five days (Fisher
Scientific IsoTemp 500 Series., Toronto, Canada). Samples were re-evaluated to note if constant
mass was obtained for dry weight analysis.
2.2.4 Statistical analysis
This experiment was conducted twice, in the first trial there were four replicates and in
the second trial, there were five replicates. The trials occurred in two separate time periods,
however, the singular plant within the chamber was considered as a replicate since the plants
were exposed to the same environmental parameters between chambers. To account for chamber
effects, the PPFD and CO2 response curves were conducted in reverse order between trials and
the first 15 minutes after exposure to a new environmental setpoint were negated. Values
presented here are the mean of nine replicates ± SE. A curve was fitted to the data using
Microsoft Excel 2019. The efflux formula (Stasiak., 2019) follows a similar theory in a reported
study (Dossa et al., 2015) in a constant volume chamber.
The NCER is calculated using the following formula:
△ 𝐶𝑂2 ∗ 𝑛
𝑁𝐶𝐸𝑅 (𝑢𝑚𝑜𝑙 𝐶𝑂2 𝑚−2 𝑠 −1 ) = (1)
1 𝑚𝑖𝑛
𝑃𝑛 ∗ ∗ −1
60 𝑠
Where:
𝑃𝑉
• n = 𝑅𝑇 (mols)
• V = chamber volume (litres)

• R = ideal gas constant (0.082 atm*L mole-1 K-1)
• P = atmospheric pressure (0.98 atm)
25
• T = constant temperature (296.15 Kelvin)

• △ 𝐶𝑂2 is the change in concentration per unit time (ppm min-1)
• 𝑃𝑛 factor (leaf area of subsamples in m2)
To obtain the NCER response, the formula was divided by a negative value to obtain a
positive relation. Leaf area measurements normalized the NCER. The continuous lines are best
fit 3rd-degree polynomials. The NCER responses were analyzed using repeated-measures
analysis of variance with multiple comparison tests to determine treatment effects.
2.3 Experiment 2a – The effect of growing media on root development

during propagation
The following experiments (2a and 2b) studied two different environment variables
affecting propagation strategies. Improving management of these environment variables can
increase root growth and provide the possibility to expedite production cycles. This experiment
will describe the effect of four different growing media (rockwool, fertilized water, peat plugs
and a potting mixture) on the shoot and root biomass/formation using vegetative clonal
propagation. The results of this study would be used to identify the optimal growing media for
propagation to obtain successful rooting and quality root formation.
Clonally propagated plant cuttings were placed in a chamber set to ‘white’ light (5650 K)
and environment conditions listed in Table 1 for 14 days. Thirty-six Cannabis sativa (cv.
‘Wappa’) cuttings from stock plants were established in four sub-sections of the PGC Flex plant
growth chambers (Conviron, Winnipeg, Manitoba, Canada), totalling 144 cuttings. Cuttings were
excised from stock plants and taken at a length of ~100 mm cut at 45° with 3-4 leaves. Thirty-six
cuttings were equally distributed in one of four propagation media, namely (1) 38.1 mm diameter
x 38.1 mm tall JIFFY peat pellets (Jiffy Pot., Brandon, Manitoba, Canada), (2) 38.1 mm x 38.1
mm x 38.1 mm rockwool cubes (Grodan., Roermond, the Netherlands), (3) nutrient solution, and
(4) ProMix HP Mycorrhizae soilless mixture (Premer Tech Horticulture, Rivière-du-Loup, QC,
26
Canada) in a 76.2 mm diameter x 76.2 mm tall JIFFY peat pot (Jiffy Pot., Brandon, Manitoba,
Canada).
All cuttings were presoaked or irrigated with CloneX (Green Planet., British Columbia,
Canada; 1.0N-0.4P-1.0K) solution at the recommended concentration of 5 ml/L (pH: 6.0, EC:
1.0) and dipped 15 mm in stim root #2 (Plant-Prod., Brampton, Ontario) before placement in the
growing medium. Cuttings in nutrient solution were placed in a plastic cup that supported the
clone with saran wrap.
Table 1. Growth chamber environmental parameters for stock and cuttings during the
experiment. Abbreviations: D= Day, N=Night, DAP= Days after planting.
Stock Cuttings
Temperature (°C) 24 D 23 N 25
Humidity (%) 60 7 DAP 85
4 DAP 75
3 DAP 65
Carbon Dioxide (ppm) 450 450
PPFD (μmol m s )-2 -1
225 80
Photoperiod (hours) 18 18
2.3.2 Harvest
Plants were harvested at 14 days. Stems were cut at the growing medium interface; all
leaves were removed and leaf area was measured using an LI-3100C area meter (LI-COR
Biosciences., Lincoln, USA). Stem diameter, nodes, root lengths of the three longest roots,
height and number of leaves were measured. The fresh and dry weight of roots, stems and leaves
were measured using a Sartorius ENTRIS 6202-1S Precision Balance (Sartorius, Göttingen,
Germany). To calculate the root dry weight in the rockwool treatment, the weight of the dried
growing medium containing the roots was subtracted from the weight of the dried growing
medium before planting the cuttings. Plants were dried in an incubator oven at 61°C (Fisher
Scientific IsoTemp 500 Series., Toronto, Canada) for two weeks.
27
The experiment was a randomized complete block design with one factor: growing
medium. The experiment included four replications with 36 subsamples in total (9 subsamples
per replicate). Data were analyzed using RStudio 1.4.1717 (R Core Team, 2020) at a Type 1
error rate of <0.05. Outliers were evaluated and removed where appropriate. Block was treated
as a random effect used to account for the random variation between chambers. The Tukeys HSD
test compared treatments using the statistical model: Y = 𝜇 + block + trmt + e. The residuals
were tested for independent observations, normality and equality of variance with plots and
Shapiro Wilks test.
2.4 Experiment 2b – The effect of spectral quality of light on root

development during propagation
The spectral composition of light has a significant influence on plant growth and
morphology. To uncover the mechanisms of cannabis development, photobiology research can
help researchers understand which wavelengths are most relevant to cannabis growth and
development in the propagation stage. The following experiment evaluated the influence of five
different LED light spectra (red, blue, green, white and a combination of red and blue (R:B), on
the shoot and root biomass/formation using vegetative clonal propagation. The results of this
study may provide the optimal light spectra for cannabis propagation in a controlled
environment.
Stock plants were grown in 152.4 mm x 152.4 mm x 152.4 mm rockwool blocks

(Grodan., Roermond, the Netherlands) and maintained in the PGC Flex growth chambers for the
duration of this study. Stock plants were replaced once throughout the experiment with younger
plants. Feeding solutions for stock plants were made with Dual Fuel 1 (5.0N-0P-1.0K), Dual
Fuel 2 (0N-3.0P-6.0K) and ProCal (2.0N-0P-0K) (Green Planet., British Columbia, Canada).
Environment parameters are listed in Table 2.
28
Table 2. Growth chamber environmental parameters for stock and propagation stages
during the experiment. Abbreviations: DAP: Days after planting, D: Day, N: Night.
Stock Propagation
Temperature (°C) 26 ± 2 D 23 ± 2 N 25
7 DAP 85
Humidity (%) 55 ± 5 4 DAP 75
3 DAP 65
Carbon Dioxide (ppm) 450 450
PPFD (μmol m-2s-1) 175-215 80
Photoperiod (hours) 18 18
2.4.2 Propagation
Twelve clones per replication, 240 cuttings in total, were excised from stock plants and
taken at a length of ~100 mm cut at a 45° with 2-3 leaves and a minimum of 3 nodes. Cuttings
were grown in 38.1 mm x 38.1 mm x 38.1 mm rockwool cubes (Grodan., Roermond, the
Netherlands) and presoaked in CloneX (Green Planet., British Columbia, Canada; 1.0N-0.4P-
1.0K) solution at the recommended concentration of 5 ml/L. Cuttings were dipped 15 mm in stim
root #2 (Plant-Prod., Brampton, Ontario) before placement in rockwool cubes. Cuttings were
evenly spaced in a 78-cell propagation tray (Mondi, Vancouver, BC) (Figure 4). Trays were
randomly arranged in the growth chambers and set to environment parameters listed in Table 2.
Cuttings were irrigated every other day with CloneX at the recommended concentration of 5
ml/L (pH: 5.65, EC: 1.00).
29
Figure 4. Schematic and example of a replicate of the planting density and arrangement of the
12 cuttings in a propagation tray before placement in a growth chamber.
2.4.3 Light
Chambers were adjusted to five different light spectra, comprising combinations of

red (λmax = 660 nm), blue (λmax = 440 nm), white (5650K) and green (λmax = 530 nm).
Normalized spectra of lights in each chamber are illustrated in Figure 5. The combination
utilized was R:B (1:1), white, green, blue, and red. Heat maps of each light intensity are
available in Appendix III. Due to the limitations of the LED lights, neutral density screens were
used to obtain lower PPFD targets.
30
Toronto, ON, Canada). Each fixture had four rows and contained dimmable spectrum channels
for green (530 nm), red (660 nm), blue (440 nm) and white (5650 K), which were individually
controlled with a fully automated system (Argus Controls., Surrey, British Columbia, Canada).
Spectral distribution was measured using a Li-180 spectrometer (LI-COR Biosciences., Lincoln,
USA).
2.4.4 Rooting performance measurements
Plants were harvested at 14 days. Stems were cut at growing medium interface; all leaves
were removed and leaf area was measured using LI-3100C area meter (LI-COR Biosciences.,
Lincoln, USA). The rockwool material was carefully picked out with forceps to obtain the roots.
The number of roots and root lengths of the five longest roots were measured. Plant materials
were dried in an incubator oven at 61°C (Fisher Scientific IsoTemp 500 Series., Toronto,
Canada) for two weeks. The dry weights of roots, stems and leaves were measured using a
Sartorius ENTRIS 6202-1S Precision Balance (Sartorius, Göttingen, Germany).
31
The experiment was an incomplete block design with one factor: light quality. The
experiment included four replications per factor with 48 subsamples, totalling 240 subsamples.
Outliers were evaluated and removed where appropriate. Eleven of the samples, dispersed
amongst all treatments and blocks, were measured below the detection limit of the Sartorius
ENTRIS 6202-1S Precision Balance (Sartorius, Göttingen, Germany). Due to the limitation of
the scale measuring below 0.01, to account for these samples without negating them they were
converted to a value of 0.001. Data were analyzed using RStudio 1.4.1717 (R Core Team, 2020)
at a Type 1 error rate of <0.05. A one-way mixed model ANOVA was used to determine the
effects of light quality on growth attributes including the dry weight of stem and roots, leaf area,
root number and the length of the three longest roots. Block was treated as a random effect used
to account for the random variation between chambers. Treatments were compared with Tukeys
HSD test using the statistical model: Y = 𝜇 + block + trmt + e. The residuals were tested for
independent observations, normality and equality of variance with plots and the Shapiro Wilks
test.
2.5 Experiment 3 – Growing media considerations for container-grown

production
The following experiment will describe the effect of reduced labor input of fertilizer
application throughout an entire production cycle. Reducing operations costs in commercial
cannabis production facilities is essential for cannabis cultivators to remain competitive. One
way to accomplish this would be to decrease manual labour with the automated irrigation
system. Drip irrigation systems with water-soluble fertilizer require daily to weekly maintenance
to limit complications. These complications include biofilm buildup, algae, insoluble salt, or
sediment that can form in the filters, lines or emitters. Cannabis production regulations prevents
the use of biocides, therefore growers are more limited in options to combat these issues. These
complications can result in sections of a growing room not receiving equal irrigation distribution.
A possible solution to excessive maintenance is to implement a reverse osmosis (RO)

water treatment for the irrigation system and avoid fertilizer inputs and water quality issues. The
32
lack of nutrient reservoir tanks would create more available space for plant production. In
addition, an ideal system wouldn’t require any changes in fertilizer input throughout an entire
production period. This can be accomplished in two ways: slow-release fertilizer or a high-
quality soil mixture that can fertilize the plant for the entire production cycle. This study aimed
to evaluate the suitability of the application of RO water into a peat based growing medium
amended with slow-release fertilizer versus an organic soil medium.
2.5.1 Propagation
Sixty Cannabis sativa (cv. ‘Wappa’) plants were clonally propagated in the PGC Flex
plant growth chambers (Conviron, Winnipeg, Manitoba, Canada), from stock plants maintained
under environment conditions presented in Table 3. Cuttings were excised from stock plants
with 2-3 leaves at a length of ~80 mm cut at a 45° angle. Thirty Jiffy peat pellets and cuttings
(Jiffy Pot., Brandon, Manitoba, Canada) were presoaked in aloe water (BuildASoil., Montrose,
Colorado) at a concentration of 1.3 ml/L for four hours. Rockwool cubes (Grodan., Roermond,
the Netherlands; 38.1 mm x 38.1 mm x 38.1 mm) were presoaked in CloneX (Green Planet.,
British Columbia, Canada; 1.0N-0.4P-1.0K) solution at the recommended concentration of 5
ml/L (pH: 5.70, EC: 0.60). Thirty cuttings were dipped 15 mm in stim root #2 (Plant-Prod.,
Brampton, Ontario) before placement in rockwool cubes. Each set of 30 clones was contained in
separate 78 cell trays in a propagation tray (Mondi., Vancouver, BC; 546.1 mm x 244.5 mm x
63.5 mm) with a transparent propagation dome. Growing media were irrigated as needed using
aloe water for the peat medium and CloneX solution for the rockwool medium.
33
Table 3. Growth chamber environmental parameters for stock and propagation growth stages of
cannabis during the experiment. Abbreviations = D: Day, N: Night, DAP: Days after planting
Stock Plants Cuttings

Temperature (°C) 24 D 23 N 26
Humidity (%) 60 80 70 (7 DAP) 60 (14 DAP)
Carbon Dioxide 450 450
(ppm)
PPFD (μmol m-2s-1) 225 80
PAR White R:B (3:2)
Photoperiod 18 18
2.5.2 Transplant and Fertilization
Sixteen rooted cuttings (~110 mm height) were transplanted 22 DAP into round 11.36 L
blow-molded black pots (279.4 mm diameter x 241.3 mm height) with one plant per pot, plugs
were placed at a depth of 50 mm. Eight rockwool cuttings were placed in growing medium (1),
four peat medium cuttings were placed in growing medium (2) and four peat medium cuttings
were placed in growing medium (3). Treatments were arranged as illustrated in Figure 6. Pots
were filled with one of three growing media, (1) ProMix HP Mycorrhizae (Premer Tech
Horticulture, Rivière-du-Loup, QC, Canada), (2) Recipe #1 or (3) Recipe #2 (Greenstar Plant
Products Inc., Langlwt, B.C., Canada). Analysis report of (1) Promix peat based soilless mixture
is available in Appendix IV. Promix soilless medium contained 65-75% peat moss, perlite,
limestone, a wetting agent, and mycorrhizae. Recipe #1 contained 60% peat moss, 25% perlite,
and 15% compost and Recipe #2 contained 50% peat moss, 33% perlite and 17% compost.
Recipes 1 and 2 rooted cuttings were dipped in mycorrhizal inoculant (Premier Tech Ltd.,
Canada) before placement in soil. Soilless mixture pots were amended with osmocote pellets at
the recommended ratio of 5.62g/kg. Growing media were irrigated as needed using reverse
osmosis (RO) water. All chambers were set to ‘white’ light (5650 K) under vegetative
environment conditions in a step-wise program presented in Table 4. Plants were switched to a
flowering schedule 46 DAP for eight weeks.
34
Table 4. Growth chamber environmental parameters for vegetative and flowering growth stages
during the experiment. Abbreviations = D: Day, N: Night, DAP: Days after planting
Vegetative Flowering Drying

Temperature (°C) 25 D 23 N 23 D 21 N (51 DAP) 18
19 N (61 DAP)
17 N (67 DAP)
Humidity (%) 70 55 50
Carbon Dioxide 500 500 500
(ppm)
PPFD (μmol m-2s-1) 150 250 400 600 0
(21 DAP) (35 DAP) (48 DAP) (61 DAP)
Photoperiod 18 12 0
Figure 6. Arrangement of treatments (Soilless mixture, Recipe 1 and Recipe 2) and growth
chamber replicates. Each box represents a growth chamber with randomly arranged sub-
samples.
35
2.5.3 Measurements
Rooting success at 13 DAP was measured with a binomial scale as the percentage of
cuttings that formed adventitious roots in each treatment. Growing medium pH and EC were
measured biweekly using the pour-through method. Pour-through solutions were measured using
a HI9814 pH/EC/TDS/Temperature portable meter (Hanna Instruments, Woonsocket, R.I., USA)
for pH and EC measurements <6 mS cm-1. Due to limitations of the conductivity range, a
LAQUA Twin EC Meter (HORIBA Advanced Techno Co., Ltd., Kyoto, Japan) was used when
EC measurements were >6 mS cm-1.
Plant height was measured as the vertical distance from the soil surface to the apical tip
of the plant. Length and width were measured at the widest part of the plant perpendicular to the
stem. Growth index was calculated as height x length x width x 300-1 (Ruter, 1992). Number of
leaves (33-41 DAP) and nodes (41-82 DAP) was measured at the vegetative and flowering
growth stages respectively.
2.5.4 Harvest
Plants were harvested at 104 DAP. Stems were cut at growing medium interface. All
leaves were removed and flowers were spread on a seven-layer mesh drying rack and dehydrated
for 18 days under drying environment conditions presented in Table 4 according to Vivo
standard operating procedures. The dry weight of roots, flowers and leaves was measured using a
Sartorius ENTRIS 6202-1S Precision Balance (Sartorius, Göttingen, Germany). Samples were
re-evaluated until constant mass was obtained. The dry weight of flower was determined by the
moisture content using a halogen moisture analyzer HE73 (Mettler Toledo., Mississauga,
Canada) when the moisture content was <13%.
The experiment was a completely randomized design with one factor: medium type. Data
were analyzed using RStudio 1.4.1717 (R Core Team, 2020) at a Type 1 error rate of <0.05. A
one-way mixed model ANOVA was used to determine the effects of growth medium on growth
36
attributes, including pH, EC, growth index, the number of leaves/nodes and dry weight of stem,
leaves and flowers. Block was treated as a random effect used to account for the random
variation between chambers. Treatments were compared with Tukeys HSD test using the
statistical model: Y = 𝜇 + block + trmt + e. The residuals were tested for independent
observations, normality and equality of variance with plots and Shapiro Wilks test.
2.6 Experiment 4 – Effects of CO₂ concentration, temperature, light intensity

and spectral composition on whole plant net carbon exchange rate for
early vegetative and flowering stages
The performance of a cannabis plant under steady-state environment conditions allows

producers and researchers to determine the best growth metrics that they could apply to their
production facility, depending on their goals and system limits. The following section assesses
the low light acclimated NCER response at different growth stages of cannabis. NCER
measurements of whole plants as either 21 DAP vegetative or 35 DAP flowering plants were
obtained in two experiments. In the first experiment, plants were exposed to a range of CO2
concentrations (420 to 2020 ppm) at a constant light intensity; thereafter, the same plants were
exposed to a range of light intensities (115 to 859 µmol m-2s-1) at a constant CO2 concentration in
sequential steps. In the second experiment, plants were exposed to a range of temperatures (15 to
32°C) in sequential steps while holding light and CO2 levels constant. The same plants were then
exposed to various light spectra (B, R:B (4:1), R:B (1:1), R, W, W+Fr, W+UV, R:B (1:4), R+Fr,
B+UV, R+UV, R:B (1:1) + Fr, B+Fr, W+B, W+R) at a constant light intensity and CO2
concentration. The results of this study could be used to provide the initial environmental
parameters for a full production experiment and determine the optimal spectra, temperature, light
intensity and CO2 concentration for a 21 DAP vegetative and 35 DAP flowering plant.
2.6.1 Propagation
Two sets of 53 Cannabis sativa (cv. Wappa) cuttings were excised 38 days apart from
stock plants and taken at a length of ~100 mm cut at a 45° with 2-3 leaves. One set of cuttings
was used to measure the effect of light intensity on NCER, and the second set of clone cuttings
was used to evaluate the impact of temperature and light spectra. Cuttings were placed in PGC
37
Flex plant growth chambers (Conviron, Winnipeg, Manitoba, Canada) under broad-spectrum
light as a pre-conditioning treatment. Cuttings were grown in 38.1 mm x 38.1 mm x 38.1 mm
rockwool plugs (Grodan., Roermond, the Netherlands) presoaked in CloneX (Green Planet.,
British Columbia, Canada; 1.0N-0.4P-1.0K) solution at the recommended concentration of 5
ml/L. Twenty-one cuttings were evenly spaced in a 78-cell propagation tray (Mondi, Vancouver,
BC) for a total of three trays in the PGC Flex chamber. Plants were irrigated every other day
with CloneX (pH: 5.65, EC: 1.00) at the recommended concentration of 5 ml/L.
2.6.2 Transplant
Propagated plants were transplanted into 101.6 mm x 101.6 mm x 101.6 mm rockwool

blocks 14 DAP (Grodan., Roermond, the Netherlands). Each plant was placed in the PGC flex
chamber and moved around randomly after four days. The plants remained in the PGC Flex
chamber under vegetative conditions (Table 5) for one week. Transplanted plants were top
irrigated with 500 ml every two days using Dual Fuel 1 and Dual Fuel 2 (Green Planet., British
Columbia, Canada; pH: 5.65, EC: 1.00) at the recommended concentration of 0.5 ml/L. Thirty
vegetative plants were randomly selected and transferred into five PS1000 chambers, totalling six
subsamples per chamber, under vegetative environmental parameters (Table 5) to acclimate for
24 hours before NCER measurements. The remaining plants were switched to the flowering
environment conditions (Table 5) for one week in the PGC Flex plant growth chambers to initiate
flowering. Flowering plants were top irrigated with 500ml every two days using 1 ml/L Dual Fuel
½ and 5 ml/L ProCal (Green Planet., British Columbia, Canada; pH: 6.0, EC = 1.30). After a
week, five flowering plants, one plant per chamber, were randomly selected and transferred to the
PS1000 chambers to acclimate for one week before NCER measurements.
38
Table 5. Controlled environment chamber schedules for propagation, vegetative and flowering
growth stages during the experiment. Abbreviations: DAP: Days after planting, D = Day, N =
Night.
Propagation Vegetative Flowering

Temperature (°C) 25 25 25 D 23 N
7 DAP 85 65 60
Humidity (%) 4 DAP 75
3 DAP 65
Carbon Dioxide (ppm) 450 500 500
PPFD (μmol m-2s-1) 80 100 140
Photoperiod (hours) 18 18 12
2.6.3 Gas Exchange
To study the NCER whole-plant response of 21 DAP vegetative and 35 DAP flowering
plants, cannabis plants were exposed to: varying light intensities and CO2 concentrations
(experiment 1); varying light spectra and temperature (experiment 2) to determine the optimal
environmental parameters for NCER. In the first experiment, 21 DAP vegetative plants were
exposed to CO2 concentrations (605, 815, 1000, 1200, 1600 and 1975 ppm) at a light intensity of
100 μmol m-2s-1, 65% humidity and 25°C. 35 DAP flowering plants were exposed to CO2
concentrations (545, 805, 1000, 1200, 1600 and 1975 ppm) at a light intensity of 140 μmol m-2s-
1, 60% humidity and 25°C. Thereafter, 21 DAP vegetative plants and 35 DAP flowering plants
were exposed to sequentially increasing light intensities (128, 241, 459, 564, 767, 865 and 961
μmol m-2s-1) at 520 ppm CO2, 65% humidity (vegetative), 60% humidity (flowering) and 25°C.
All parameters for PPFD and CO₂ were first recorded at the lowest value and then subsequently
increased over two days. Each light and CO2 setpoint was scheduled during an 18-hour
photoperiod for two hours. In the second experiment, 21 DAP vegetative and 35 DAP flowering
plants were exposed to a range of temperature (15, 18, 23, 26, 29, 32 °C) and light spectra
comprising combinations of red (R), blue (B), white (W), UV, far-red (FR) and green (G). The
combinations for the vegetative and flowering plants were B, R:B (4:1), R:B (1:1), R, W, W+Fr,
39
W+UV, R:B (1:4), R+Fr, B+UV, R+UV, R:B (1:1) + Fr, B+Fr, W+B, W+R set at 250 μmol m-
2s-1. Responses of NCER to temperature and light spectra were collected over a period of three
days. The temperature was first recorded at the lowest value and then the temperature was
subsequently increased over a period of nine hours on the first day set at 520 ppm CO2, 65%
humidity (vegetative) 60% humidity (flowering). Light spectra data were collected over the next
two days with eight spectra on the second day and seven spectra on the third day set at 520 ppm
CO2, 60% humidity and 23°C. Temperature and light spectra conditions were scheduled for 1.5
hours and 1 hour respectively.
Reference section 2.2.3 for harvest protocol. A rectangular hyperbola model for the light
intensity, temperature and CO₂ response was applied to the data using Microsoft Excel 2019. The
experiments occurred in two separate time periods. Each chamber was considered a replicate
within each experiment since the plants were exposed to the same environmental parameters
between chambers. To account for chamber effects the first 15 minutes after exposure to a new
environmental setpoint were negated. Values presented here are the mean of 3-4 replicates ±
SE. A best fit curve was applied to the data using Microsoft Excel 2019. The formula (Stasiak.,
2019) follows a similar theory described in Dossa et al., 2015. Refer to equation (1) in section
2.2.4.
To obtain the NCER response, the formula was divided by a negative value to obtain a
positive curve. Leaf area measurements normalized the NCER. The continuous lines are best fit
3rd degree polynomials. Statistical analysis for the temperature and light quality data were
analyzed using RStudio 1.4.1717 (R Core Team, 2020). One-way mixed model ANOVA were
performed to determine the effect of temperature and light spectra on the NCER. Tukeys HSD
test was applied for all multiple comparison tests with a significance level of P < 0.05. The
residuals of the analyses were tested for normality and equality of variance using The Shapiro-
Wilk test. The light and CO₂ NCER responses were analyzed using repeated-measures analysis
of variance with multiple comparison tests to determine treatment effects.
40
Chapter 3: Results
This study examined the effect of light and CO₂ on the NCER of vegetative stock
cannabis plants. The mature leaf area ranged from 0.39 – 0.66 m2 and the dry weight of stem and
leaves ranged from 37-66 grams. Figure 7 depicts a typical example of a growth chamber's
fluctuations of CO₂ concentration in a 24-hour cycle. Variation was less than 8% from the
programmed setpoints. Supplementation of CO₂ came from an external source within the facility
to compensate for photosynthetic uptake.
Figure 7. CO₂ concentration (ppm) of vegetative plants in the PS1000 chambers measured
over 24 hours. During the dark period due to respiration, CO₂ accumulated and was drawn
down. When lights were on. CO₂ was added to adhere to the setpoints controlled by Argus
(Argus Controls., Surrey, British Columbia, Canada) and to offset photosynthetic uptake.
Values represented as means ± SEM (n = 9).
41
3.1.1 CO₂ response
Net carbon exchange rate response with increasing CO2 concentration is illustrated in
Figure 8. The NCER became CO₂ saturated at approximately 1980 ppm, with a maximum
NCER of 3.48 μmol CO₂ m-2s-1. The respiration rate was -1.12 µmol CO₂ m-2s-1. As the CO2
concentration increased, the NCER increased linearly up to 900 ppm and then diverged from the
linear response to an asymptotic stage until it reached the maximum point. Between each CO₂
concentration setpoint of 480, 800, 1000, 1200, 1600 and 2000 there was a 39.15%, 11.03%,
7.88%, 8.89% and 1.46% increase in NCER respectively. There was a 50.51% increase in NCER
when doubling the concentration of CO₂ from 480 to 1000 ppm.
Figure 8. NCER response of vegetative plants in the PS1000 chambers over a range of CO₂
concentrations from 480, 800, 1000, 1200, 1600 and 1980 ppm. NCER was measured at 178
µmol m-2s-1, 55% relative humidity and 25°C. Values represented as means ± SEM (n=9, R2=
0.9996).
42
3.1.2 Light response
Net carbon exchange rate response with increasing light intensity is illustrated in
Figure 9. The NCER maximum was 6.03 μmol CO₂ m-2s-1 and occurred at ~ 686 µmol m-2s-1.
The CP was estimated using the best fit 3rd degree polynomials when the NCER was zero. The
rate of respiration was -0.38 µmol CO₂ m-2s-1 and the light CP was 28.6 µmol m-2s-1. As the
PPFD increased, the NCER increased initially linearly (up to 400 µmol m-2s-1) and then began
decreasing beyond the maximum value of 6.03 μmol CO₂ m-2s-1. The delta NCER between
each light intensity, 115, 215, 410, 500, 686, 774, 859, was 168.52%, 65.17%, 12.73%,
11.48%, -2.49%, -0.68% respectively. There was a 21.71% increase in NCER observed when
doubling the light intensity from 410 to 859 µmol m-2s-1.
Figure 9. NCER response of vegetative plants in the PS1000 chambers over a range of light
intensities from 115, 215, 410, 500, 685, 775 and 860 μmol m-2s-1. NCER was measured at
520 ppm CO₂, 55% relative humidity and 25°C. Values represented as means ± SEM (n=9,
R2 = 0.9967)
43

during propagation
The rooting success was measured on a binomial scale in which success was determined
by the presence of any visible adventitious roots (Figure 10) present at harvest. The clonally
propagated plants in the nutrient solution had a 0% success rate, whereas the rockwool method,
peat plug and potting mix had a 97%, 80% and 80% success rate respectively.
Figure 10. A) Example of a tray in a PGC Flex chamber, each chamber subsection contained
three trays B) Example of root formation for a subsample of each growing medium a) Peat
plug b) Potting mix c) Water d) Rockwool cubes
No differences (p < 0.05) were observed in stem and root dry weight, nodes, stem
diameter, height, and a number of leaves. Root lengths of the three longest roots were measured
in Table 6. There was an effect on root length 1 (p = 0.0002), root length 2 (p = 0.0014) and root
length 3 (p = 0.0016). The root length of the first longest root in the potting mix was 114%
greater than the peat plug. Root lengths 2 and 3 were 107% and 115% longer, respectively.
44
Table 6. Root length of the three longest roots in propagated vegetative cuttings in two different
mediums, peat plug and potting mix at 14 days after planting
Treatment Root length 1 (mm) Root length 2 (mm) Root length 3 (mm)
Peat plug 63.0 ± 8.7 44.3 ± 5.9 35.7 ± 5.1
Potting mix 135.2 ± 22.9 *** 91.8 ± 13.7 ** 76.9 ± 12.1 **
Data are means ± SEM (n=4). Data followed by an asterisk are significantly different (p > 0.05) according
to one-way analysis of variance test (ANOVA) with multiple comparisons test, estimated marginal means
with a Tukey-Kramer adjustment
*, **, *** represents p<0.05, 0.01, and 0.001 respectively

Figure 11 illustrates a comparison of light treatments of propagated plants. Based on

observations, the root formation between all treatments was similar. There were no differences
between leaf area, stem and root dry weight of rooted cuttings amongst all light quality
treatments.
Figure 11. Image of the adventitious root formation of a subsample grown under each light
spectrum a) White b) Red c) Blue d) R:B e) Green at 14 DAP in the PGC Flex chambers
Root number and root lengths for the longest five roots were analyzed to determine the
overall effects of light spectral treatments (Table 7). The number of roots did not differ;
however, the red treatment had a significantly larger quantity of roots compared to the RB
treatment by 41.67%. The longest and 2nd longest rooting lengths had no statistically significant
45
differences between the white and green light and all treatments. There were differences in the
first longest root between the red treatment compared to the blue and RB treatment by an
increase of 36.99% and 37.93%, respectively, and between the second-longest root by an
increase of 15.81% and 10.98%, respectively. There were no statistically significant differences
between the root length of the 3-5 longest roots between all treatments.
Table 7. Effect of light quality treatments on the average number of roots and root lengths of the
five longest roots 14 days after planting
Light Root Root length Root length Root length Root length Root length
quality Number 1 (mm) 2 (mm) 3 (mm) 4 (mm) 5 (mm)
White 15 ± 0.88 ab 30.0 ± 0.9 ab 26.1 ± 1.0 ab 23.9 ± 0.9 a 21.8 ± 0.9 a 18.8 ± 0.9 a
Blue 15 ± 0.78 ab 29.2 ± 0.8 a 25.3 ± 0.6 a 23.3 ± 0.6 a 21.3 ± 0.6 a 18.5 ± 0.6 a
Red 17 ± 1.35 a 40.0 ± 1.2 b 29.3 ± 1.0 b 26.1 ± 1.1 a 23.7 ± 1.0 a 21.5 ± 1.0 a
RB 12 ± 0.75 b 29.0 ± 1.0 a 26.4 ± 0.8 a 24.2 ± 0.9 a 21.8 ± 0.8 a 19.9 ± 0.9 a
Green 15 ± 0.72 ab 30.7 ± 0.8 ab 28.4 ± 0.6 ab 26.4 ± 0.6 a 24.3 ± 0.6 a 22.2 ± 0.6 a
Data are means ± SEM; Data followed by the same letter are not significantly different (p < 0.05)
according to analysis of variance (ANOVA) with multiple comparisons, estimated marginal means with a
Tukey-Kramer adjustment

production
3.4.1 Rooting response
Rooting success was determined based on a binomial scale by the presence of

adventitious roots; the plant that was clonally propagated in the peat plug had a 53% success
rate, whereas the rockwool method had a 96% success rate.
3.4.2 Growing media pH and EC
The pH (Table 8) and EC (9) are described and illustrated (Figure 12) based on the
DAP. The overall pH in the vegetative stage (26-41 DAP) ranged from 4.99-5.72 and in the
flowering stage (54-96 DAP) it ranged from 4.90-5.45. Looking at the specific mediums, the pH
46
in the vegetative stage for Recipe 1 ranged from 5.12 -5.25, for Recipe 2 the pH ranged from
4.99-5.00 and for the peat-based mixture the pH ranged from 5.64-5.72. The pH in the flowering
stage for Recipe 1 ranged from 4.94-5.14, Recipe 2 ranged from 4.90-5.27 and peat based ranged
from 5.25-5.45.
Overall, at 26 DAP, the peat-based mixture pH was 11.72% and 14.40% higher than
Recipe 1 and 2, respectively; at 41 DAP, the peat-based mixture pH was 13.03% higher than
Recipe 2. For the rest of the production period (54-96 DAP), the pH levels amongst all
treatments were not significantly different.
Table 8. The average (n=4) pH of three growing medium treatments, Recipe 1, Recipe 2 and the
peat based mixture obtained with the pour-through method with applications of RO water at 26-
96 DAP
DAP Recipe 1 Recipe 2 Peat based mixture

26 5.12 ± 0.12 a 5.00 ± 0.03 a 5.72 ± 0.09 b
41 5.25 ± 0.13 ab 4.99 ± 0.23 a 5.64 ± 0.12 b
54 5.14 ± 0.07 a 5.27 ± 0.04 a 5.38 ± 0.11 a
68 4.99 ± 0.19 a 4.97 ± 0.22 a 5.25 ± 0.05 a
82 4.94 ± 0.16 a 4.90 ± 0.22 a 5.34 ± 0.08 a
96 5.02 ± 0.17 a 4.99 ± 0.18 a 5.45 ± 0.09 a
Reported values are means ± SEM (n=4, 4-8 subsamples) from 26-96 DAP. Lettering indicates statistical
comparisons (p < 0.05) within a time point, not between time points. Tukey – Kramer levels across all
treatments are shown.
The overall EC in the vegetative stage (26-41 DAP) ranged from 2.66-6.67 mS cm-1 and
in the flowering stage (54-96 DAP), the EC ranged from 1.68-5.04 mS cm-1. More specifically,
the EC in the vegetative stage for Recipe 1 ranged from 4.25-6.07 mS cm-1, for Recipe 2 the EC
ranged from 6.08-6.67 mS cm-1, and for peat-based mixture the EC ranged from 2.66-4.53 mS
cm-1. In the flowering stage, Recipe 1 ranged from 2.09-5.04 mS cm-1, Recipe 2 ranged from
3.07-4.64 mS cm-1 and peat-based mixture ranged from 1.68-4.50 mS cm-1. At 26 DAP, the peat-
47
based mixture EC was 56.18% and 60.12% lower than Recipe 1 and 2, respectively. The EC
values amongst all treatments and between 41-96 DAP were not statistically significant.
Table 9. The average (n=4) EC of three growing medium treatments, Recipe 1, Recipe 2 and
peat based mixture at 26-96 DAP obtained with the pour-through method using RO water
DAP Recipe 1 Recipe 2 Peat based mixture

26 6.07 ± 0.08 a 6.67 ± 0.27 a 2.66 ± 0.23 b
41 4.25 ± 0.53 a 6.08 ± 0.27 a 4.53 ± 0.29 a
54 5.04 ± 1.04 a 4.64 ± 1.05 a 4.50 ± 0.68 a
68 3.16 ± 0.19 a 3.54 ± 0.50 a 2.57 ± 0.45 a
82 2.80 ± 0.34 a 3.08 ± 0.41 a 2.61 ± 0.53 a
96 2.09 ± 0.36 a 3.07 ± 0.43 a 1.68 ± 0.26 a
Reported values are means ± SEM (n=4) from 26-96 DAP. Recipes 1 and 2 contained four subsamples,
and the peat-based mixture contained eight subsamples. Lettering indicates statistical comparisons (p <
0.05) within a time point, not between time points. Tukey – Kramer levels across all treatments are
shown.
48
Figure 12. Nutrient delivery system pH and EC measurements of four replicates over a range
of 26-96 DAP of the Recipe 1, Recipe 2 and the peat based mixture. Solid lines represent the
pH, and the dotted line represents the EC measurements corresponding to the colours in the
legend. Reported values are means ± SEM.
3.4.3 Growth response
There were no differences in the growth index, the quantity of leaves, number of nodes,
shoot and leaf dry weight (Table 10) throughout the production period. The peat-based mixture
flower weight was 129.51% and 87.85% significantly larger than Recipe 1 and Recipe 2,
respectively (Table 10).
49
Table 10. Evaluation of dry weight of shoot, leaf and flower at harvest of three growing medium
treatments
Treatment Shoot Weight (g) Leaf Weight (g) Flower Weight (g)
Peat based mixture 9.18 ± 1.08 a 32.56 ± 2.82 a 44.41 ± 2.82 a
Recipe 1 7.26 ± 1.44 a 23.75 ± 3.85 a 19.35 ± 2.40 b
Recipe 2 8.07 ± 1.80 a 27.60 ± 3.37 a 23.64 ± 1.84 b
Data are means ± SEM (n=4). Recipes 1 and 2 contained four subsamples, and the peat based mixture
contained eight subsamples. Data followed by the same letter are not significantly different (p < 0.05)
according to analysis of variance (ANOVA) with multiple comparisons, estimated marginal means with a
Tukey-Kramer adjustment
Initially, the peat based growing medium plants were visibly smaller, but all leaves
appeared healthy exhibiting a dark green. This study showed visual signs of nutrient disorders
shortly following the flowering stage, the organic recipes both had chlorotic tissue throughout
the whole canopy while the peat-based mixture exhibited dark green leaves (Figure 13).
Figure 13. Cannabis experimental plants at 104 DAP in a) Soilless growing medium b) Recipe
1 growing medium c) Recipe 2 growing medium
50
3.5 Experiment 4 – Effect of CO₂, light intensity, temperature and spectral

composition on whole plant net carbon exchange rate for early vegetative
and flowering stages
3.5.1 CO₂ response
Photosynthetic response with increasing CO2 concentration is illustrated in Figure 14.

The NCER for the 21 DAP vegetative plant became CO₂ saturated at approximately 1980 ppm,
and the maximum NCER was 2.40 ± 0.13 μmol CO₂ m-2s-1. The rate of respiration was -1.19 ±
0.02 µmol CO₂ m-2s-1. Doubling of CO2 concentration between 814 and 1607 ppm observed an
NCER increase of 298.11%. The NCER for the 35 DAP flowering plant became CO₂ saturated at
approximately 2020 ppm, and the maximum NCER was 3.07 ± 0.50 μmol CO₂ m-2s-1. The rate of
respiration was -0.88 ± 0.08 µmol CO₂ m-2s-1. Doubling of CO2 concentration between 814 and
1607 ppm observed an NCER increase of 107.94%. The two growth stages NCER response were
compared between programmed CO₂ concentrations with repeated measures and post-Hoc
comparison test showed no significant differences.
51
Figure 14. A) NCER response for a 21 DAP vegetative plant (n = 3, R2= 0.9999) over a range
of CO₂ concentrations at 605, 815, 1000, 1200, 1600 and 1975 at light intensity of 100 μmol
m-2s-1. B) NCER response for a 35 DAP flowering plant (n = 4, R2 = 0.9983) over a range of
CO₂ concentrations from 545, 805, 1000, 1200, 1600 and 1975 at 140 μmol m-2s-1.
52
3.5.2 Light response
Photosynthetic responses with increasing light intensity are illustrated in Figure 15. The
NCER for the 21 DAP vegetative plant became light-saturated at 564 µmol m-2s-1, and the
maximum NCER was 5.94 ± 0.48 μmol m-2s-1. The rate of respiration was -0.52 ± 0.26 µmol
CO₂ m-2s-1 and the light CP based on the trendline was estimated to be ~14.90 µmol m-2s-1. As
the PPFD increased, the NCER initially increased linearly (up to 400 µmol m-2s-1) and then
plateaued until it reached the maximum value. Between 128 and 459 μmol m-2s-1 there was an
NCER increase of 105.88%.
The NCER for the 35 DAP vegetative plant became light-saturated at 559 µmol m-2s-1,
and the maximum NCER was 6.39 μmol CO₂ m-2s-1. The respiration rate was -0.77 µmol CO₂ m-
2s-1 and the light CP was estimated to be 21.70 µmol m-2s-1. As the PPFD increased, the NCER
initially increased linearly (up to ~400 µmol m-2s-1) and then plateaued after it reached the
maximum value. Between 128 and 459 μmol m-2s-1, there was an NCER increase of 116.84%.
The two growth stages NCER response were compared between light intensities with repeated
measures and post-Hoc comparison test showed no significant differences.
53
Figure 15. NCER response for a 21 DAP vegetative plant and 35 DAP flowering over a range
of light intensities from 128, 241, 459, 564, 767, 865 and 960 μmol m-2s-1 at 25°C and 520
ppm. A) 21 DAP vegetative plant (n = 4, R2= 0.9938) at 65% humidity. B) 35 DAP flowering
plant (n = 4, R2 = 0.9948) at 60% humidity.
54
3.5.3 Temperature response
The effect of temperature on the vegetative plants is illustrated in Figure 16. A one-way
mixed effects ANOVA was performed to compare the effect of temperature on the NCER. The
analysis revealed no statistically significant difference in average NCER with respect to
temperature in the 21 DAP vegetative and 35 DAP flowering plant.
55
Figure 16. Temperature NCER response at increasing intervals of 15, 18, 23, 26, 29, 32 °C for
a 21 DAP vegetative plant and 35 DAP flowering plant at 520 ppm. Points represent mean ±
SE values (n = 4). A) 21 DAP vegetative plant at 65% humidity, 100 μmol m-2s-1 and 520
ppm. B) 35 DAP flowering plant at 65% humidity, 140 μmol m-2s-1 and 520 ppm.
56
3.5.4 Light spectra response
The light quality NCER response of a 21 DAP vegetative plant is represented in Figure
17. Significant differences were represented by the different letters above each bar in the graph
and it showed a lot of variation between light spectral combinations. No statistical differences
were observed between R:Fr, R:B (4:1), R:B (1:4), W+Fr, W+UV, R+UV, RB(1.0)+Fr and W+R
spectral combinations. Treatment R+Fr obtained the highest NCER (4.67 ± 0.10 (μmol CO₂ m-2s-
1) response and was statistically significant from B, B+UV, B+Fr, W, R:B (4:1), W+B and R, by
an increase of 73.93%, 55.21%, 50%, 48.5%, 47.52%, 40.57% and 32.64% respectively. The
lowest NCER was exhibited with blue light (2.57 ± 0.31 μmol CO₂ m-2s-1). The spectral
combinations that exhibited no difference with any other treatment were R:B (4:1) and R:B (1:4).
Figure 17. NCER response at different light qualities for a 21 DAP vegetative plant at 250 (μmol
m-2s-1). Grey bars represent mean ± SE values (n=3).
The light quality response curve of a 35 DAP flowering plant is illustrated in Figure 18.
Significant differences were represented by the different letters above each bar in the graph and
it showed a lot of variation between light spectral combinations. No statistically significant
57
differences with W+R, B, R:B (4:1), RB (1.0), R, W+Fr, R+Fr, R+UV and RB (1.0)+Fr spectral
combinations. Treatment W+R obtained the highest NCER (4.99 ± 0.21 μmol CO₂ m-2s-1) and
was statistically different from B+Fr, W+UV, W+B, R:B (1:4), W+UV, white by an increase of
41.76%, 34.86%, 30.97%, 28.61%, 24.13% and 23.51% respectively. The lowest NCER was
exhibited with B+UV (3.70 ± 0.27 μmol CO₂ m-2s-1). The spectral combinations that exhibited no
difference with any other treatment were B, RB(1:1), and R.
Figure 18. NCER response at different light qualities for a 35 DAP flowering plant at 250 (μmol
m-2s-1). Grey bars represent mean ± SE values (n=4).
Chapter 4: Discussion
To test the hypotheses (HO1 and HO2), this study deployed whole plants in a closed
system growth chamber while modifying CO₂ and light intensity to obtain the NCER. The NCER
response curve provided a way to predict the growth of cannabis over a range of parameters
58
(Leonardos et al., 1994). In the literature, reports of NCER of cannabis at different PPFDs and
CO₂ concentrations are limited. The NCER has been extensively researched with other plant
species acclimated to different environment conditions (Massa et al., 2015). In recent papers,
Chandra et al., 2008 and Chandra et al., 2011a assessed the leaf-level photosynthetic response of
cannabis to variations in photosynthetic photon flux densities and carbon dioxide. These studies
determined that 1500 μmol CO₂ m-2s-1 and 700-750 ppm CO₂ was the most efficient
environmental condition and that this was consistent amongst different cultivars. To identify the
optimal environment conditions, the vegetative stage was chosen as a starting point to allow the
plant to be at its optimal growth potential before transitioning into flowering or being used as
stock plants for vegetative propagation. Plants were acclimated to the stock room environmental
parameters in the production facility and the environmental setpoints were replicated in the
growth chamber. The setpoints were subjectively low compared to the levels of light penetration
and CO₂ that a cannabis plant can withstand (Chandra et al., 2017). Although these conditions
limit the plant growth rate, this allows producers to keep up with plant maintenance of the stock
room with limited staff available.
The graphical illustration in Figure 7 exhibits extensive data collected over 24 hours.
This low variation (8%) from the setpoints indicates that short-term studies are more practical.
They are less likely to have significant cyclic variation during the duration of the experiment
from mechanical failure. The carbon dioxide increased during the night due to mitochondrial
respiration and was subsequently utilized by plants through photosynthesis when the lights
turned on. Mid-day spike was observed due to human entry into the chamber; this spike was also
evident in the study conducted by Romer (2012). The cyclic CO₂ concentrations represented the
balance between the injection rate into the chamber, the lag time between the gas analyzer and
the on-off valve and the photosynthetic uptake (Bernier et al., 1994; Bunce, 2020).
Environmental properties that can influence NCER measurements in a growth chamber

include irradiance and CO₂ and were illustrated in Figure 8 and Figure 9. There are multiple
factors (temperature, VPD, air quality, fertilizer) that can influence photosynthesis. For the
purposes of this experiment these environment conditions were held constant, while the primary
parameters of interest, light intensity and CO₂ concentration, were manipulated. In certain
59
scenarios, the absorption of too much light or CO₂ will stimulate the plant to initiate special
mechanisms to mitigate the excess through long-term developmental and short-term responses.
Mechanisms to deal with excess photons include non-photochemical quenching, savaging of
toxic photoproducts and targeting molecules for oxidative degradation; mechanisms to deal with
excess exposure to CO2 include charges to hormones levels, formation of non-structural
carbohydrates and increase in phenolic compounds. However, there are ultimately limits to
acclimation under these conditions (Taíz and Zeiger, 2006).
Short-term responses to high light conditions involve the dissipation of excess light
through the use of carotenoids. Long-term responses involve changes in leaf anatomy to
maximize light absorption by being perpendicular to the overhead light source or to minimize
absorption by changes in leaf orientation to reduce heating and water loss. Additionally, plants
can exhibit smaller and thicker leaves under high light conditions due to the additional levels of
palisade cells and allow chloroplasts to rearrange parallel to the light source (Moher at al., 2021;
Rodriguez-morrison et al., 2021). Low light intensities will enable plants to have thinner and
broader leaves to allow the movement of chloroplasts to a greater surface area (Higa & Wada,
2015). The leaves absorb approximately 85-90% of spectra emitted; the remainder is reflected at
the leaf surface or transmitted through the leaves to the lower canopy (Guidi et al., 2017; Taíz
and Zeiger, 2006). The value of adding additional light via an increase in light intensity can
directly influence quantum efficiency. Quantum efficiency measures how much CO₂ can be fixed
for every additional photon of light added, given if CO₂ is not limited.
The observations in the light curve indicated a proportional increase in NCER,

demonstrating initially a linear relationship followed by an asymptomatic response between light
intensity and NCER (Figure 9). Prior to the lights turning on, the plant does not undergo the
process of photosynthesis due to mitochondrial respiration, hence the NCER is below 0. When
the NCER is above 0, the linear relationship generated is because photosynthesis was light-
limited at those light levels. Beyond the maximum NCER, there was no advantage of additional
PPFD to the NCER since the plant was CO₂ limited. Literature has demonstrated that had the
plant been acclimated under high light conditions, the CP would have been higher since the
respiration rates would be higher so it would take more photosynthetic processes to bring the
60
exchange rate to a net value of 0 (Taíz and Zeiger, 2006). Additionally, the curve would be
situated higher in the graph as the NCER would have increased as exhibited in Rodriguez-
morrison et al (2021a) study.
When doubling of the CO₂ concentration in a controlled environment, the NCER

typically increases by 30-50% in C3 plant species (Ainsworth et al., 2002; Allen & Prasad,
2004). This is evident in the graph (Figure 8) when doubling the concentration from ~500 to
1000 ppm increased NCER by 50%. The observations also indicated that the rate of fixation had
not formed a plateau, so the true threshold had not been reached. The entry of CO₂ into the
leaves is based on the concentration gradient between the carboxylation site and ambient air. At
high CO₂ concentration, the plateau, photosynthesis is likely limited by the Calvin cycle to
regenerate RuBP (Onoda et al., 2005).
The results proved the alternative hypothesis to be true for both the light and CO2
concentration NCER response. At a constant CO2 concentration, increasing light intensity
increased stock plant NCER, and at a constant light intensity, increasing CO2 concentration
increased stock plant NCER. Ultimately, the optimal NCER for light warrants further
investigation based on the limitations of indoor production and goals; the optimal NCER under
varying CO2 concentration must be given further consideration due to the lack of inhibition.
Based on these figures, to obtain optimum NCER ~410-859 µmol m-2s-1 and ~1620-2020 ppm
are the desired setpoints under those environment conditions. To reduce energy consumption and
obtain environmental conditions that are not statistically different from the highest NCER, a light
of intensity of 410 µmol m-2s-1 and a CO2 concentration of 1620 ppm is optimal. This
phenomenon extrapolated over time will likely yield more biomass production if irrigation
strategies and fertilizer applications are not limiting as the plant water use, morphology and leaf
temperature changes.
The optimal NCER in terms of CO2 concentration was inconsistent with Chandra et al
(2011) and Chandra et al (2015) studies which observed optimal photosynthetic rates at 700-750
ppm. However, the optimal CO2 concentration was observed at the maximum CO2 concentration
measured, which does not conclude that concentrations above 750 ppm would achieve higher
61
rates. The optimum NCER in terms of light intensity was inconsistent with a long-term whole
plant study conducted on vegetative plants by Moher et al. (2021) that achieved desirable
morphological characteristics at 600 µmol m-2s-1 and 15% higher dry weight at 900 µmol m-2s-1.
Chandra et al. (2008) and Chandra et al. (2015) observed maximum leaf level photosynthesis and
yields at ~1500 µmol m-2s-1, however plants were acclimated at a higher light intensity of 700
µmol m-2s-1. Leaves 63 DAP acclimated at low light conditions (91 µmol m-2s-1) in the flowering
stage indicated saturating responses at ~600 µmol m-2s-1. However, plants grown at a light
intensity up to 1800 µmol m-2s-1 till harvest exhibited higher dry inflorescence yield and terpene
potency (Rodriguez-Morrison et al., 2021a). Moreover, this study exhibited temporal variability
in leaf level light intensity from weeks 1 to 9 of flowering plants, which shows the importance of
growing plants under a broad range of light intensities throughout a full production cycle. These
studies provide indications that the optimal light intensity exhibited in this study was not
commonly observed in typical production.

during propagation
This study was conducted to determine the impact of different growing media on root
biomass accumulation using vegetative clonal propagation. The results of this study could be
used to provide insight into the most successful growing medium in the propagation stage in a
controlled environment. Some limitations of this study resulted from the limited replications.
Issues arose with the rockwool treatment wherein dry root weights could not be resolved even
though this treatment exhibited the most successful root and shoot development and was used in
subsequent experiments as the propagation treatment of choice. The results could have been
confounded by the resolution of the scale, accumulated salts from the fertilizer or the
development of algae.
The water treatment failed to develop roots so those data were omitted from the study.
This does not reflect what occurs in other species when placing a stem cutting in water.
However, it is common to see rooting in anaerobic water take longer or cause poor rooting
(Hartmann et al., 2017). The clones did maintain turgidity, as observed in another study Grange
62
& Loach, 1983, but the results were possibly confounded by lack of time or aeration that may
have affected respiration.
Since the shoot system is already developed, propagation by stem cuttings produces new
adventitious root systems (Hartmann et al., 2017). Despite the shortcomings of incomplete
rockwool data collection, the rockwool plugs success rate indicated that it was the ideal growing
medium for cannabis propagation. This was based on the high rooting success (97%) and not on
the other morphological observations that included the dry weight of roots, root quantity and
rooting length. In another cannabis propagation study, rockwool significantly outperformed three
other soilless mixtures that differed by peat concentration in root number, length and mass in five
cultivars; the remaining three cultivars tested exhibited variability or insignificance amongst the
growth media (Campbell et al. 2021). Campbell et al. (2019) used peat-based plugs under
humidity domes and exhibited variability among cultivars in producing roots and the speed at
which they rooted. These studies suggested that rockwool is the optimal growing medium and
propagation of cannabis was influenced by cultivar selection.
The growth media that were successfully statistically analyzed included the peat plugs
and potting mix in a peat pot. They exhibited measurable morphological rooting behaviour and
were not skewed by methodology. Based on morphology, the clones did not show any signs of
evapotranspiration stress (wilting, curling of leaves, chlorosis). Growth attributes including
height, dry weight of stems, nodes, leaves and stem diameter were assessed to determine if the
clones were different from each other before placement in the respective media. There were no
significant differences between the cuttings morphology, so any differences observed in the root
zone were determined by the medium. In the results, there were no effects of media on the dry
weight of roots between the peat plugs and the potting mix. There was an effect on root length 1,
root length 2 and root length 3. This could indicate that the potting mix had a higher quality
medium or optimal aeration porosity and bulk density. However, the container of the potting mix
was double in size compared to the peat plug so the root zone had more volume to explore and
consequently, the amount of water and nutrients available would have been higher. The overall
root mass between the growing media was the same and it is predicted that the root surface area
would be smaller in the potting mix to accommodate the longer root structure and still obtain the
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same dry weight (Wu et al., 2016). Overall, results were consistent with the alternative
hypothesis that growth medium does increase cannabis root growth during cutting establishment.
Above ground morphological metrics are an important consideration when characterizing

the root zone. Stem diameter is an important variable to take into account when trying to obtain
successfully rooted cuttings. A study conducted by (Cockson & Barajas, 2019), showed that the
larger stem diameters (2.9-3.2 mm) had more root formation based on a root rating scale. This
could have been due to more carbohydrate reserves in the shoot that created a larger source-sink
relationship. A study conducted by Kathiravan et al., 2009 observed Jatropha curcas cuttings and
recorded better performance with the longest length (400 mm) in height. This could be due to the
smaller cuttings being more limited in auxin, carbohydrates or rooting co-factors for induction of
roots. Leaf quantity between 2-3 leaves has been shown not to influence rooting success rate but
exhibited 15% higher rooting quality with cuttings with three leaves (Caplan et al., 2018). The
number of nodes influences the root zone and is supported by studies (Mesay et al., 2015;
Shekhawat & Manokari, 2016) due to the hydraulic conductivity (Solikin, 2018). These studies
emphasize the importance of looking at these morphological characteristics as they can be
confounding in propagation studies.

The study’s objective was to evaluate the influence of five different light spectra on root
biomass using vegetative clonal propagation in rockwool cubes. The results of this study could
be used to provide a light recipe for cannabis propagation in a controlled environment. The
vegetative propagation technique demonstrated high rooting success (>99%) amongst all
treatments. This indicated that the cloning procedure employed by Caplan et al., 2018 under
LED lights and these environmental parameters could lead to successful rooting. In general, root
formation occurred after seven days, and by the 14th-day roots could be seen at the bottom of
most rockwool plugs, a typical occurrence exhibited in propagating vegetative clones (Chandra
et al., 2020).
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This experiment took place over several months and the stock plants were exchanged
midway for a new set obtained from the stock room onsite. Despite the exchange of stock plants
midway through the experiment, there were no differences in dry stem weight and leaf area due
to following the cloning procedure. As described in Section 4.2, it is important to reduce the
confounding variables exhibited from above-ground morphological characteristics. This
exchange of stock plants is typical in production facilities as producers will opt to replace stock
plants every 1-2 years to change the cultivars available or obtain new phenotypes that can
enhance production practices and final product quality. In the case of this study, the cultivar that
was replaced was of the same phenotype but was just younger. A preliminary experiment (data
not shown) suggested that the phenotype was an important factor in rooting response, as another
phenotype of cv. Wappa exhibited less rooting success and biomass based on physical
observations.
An analysis (data not shown) was performed to assess differences between replicates
between the stock plants and there were no significant differences to indicate that variability was
present. There are no studies available on how stock plants of the same cultivar but different ages
influence rooting success based on genetic or physiological differences; however, there is
evidence that the young cuttings will have a higher rooting success (61%) compared to old
shoots (23%) (Munoz-Gutierrez et al., 2009) and that different genotypes influence success and
speed of root formation (Campbell et al., 2019b). Collectively, these studies do not indicate
concern regarding the use of different stock plants for propagation. With cannabis propagation
studies, there remains a lack of empirical literature aimed at optimizing cannabis propagation
under different spectral qualities. Of the limited studies available, Campbell et al., 2019b found
that spectral quality can augment the plant structure in stock plants, influencing the height,
internode length, petiole length, fan leaf width and the number of meristems.
Research has reported that producers can use different light spectra to influence
adventitious root development and the spectral characteristics of light are altered with depth
under the growing medium surface. The absorption spectra of red/far-red light receptors known
as phytochromes, blue light receptors identified as cryptochromes and phototropins allow
coordinated signal transduction mechanisms that regulate photomorphogenesis. These
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photoreceptors are expressed in the roots and can regulate negative root phototropism, root
elongation, lateral rooting, root bending and inhibit rooting (Van Gelderen et al., 2018). Red
light has been reported to have increased root formation in Vaccinium corymbosum (Poudel et
al., 2008), Plectranthus scutellarioides (Cho et al., 2019); similarly, blue light has demonstrated
this observation in Kalanchoe blossfeldiana (Rasool et al., 2020), Symphyotrichum novi-belgii
(Schroeter-Zakrzewska & Kleiber, 2014); and white light has been reported in
Fragaria × ananassa (Zheng et al., 2019). In contrast, blue light has shown to inhibit root
growth and suppress lateral root elongation in Arabidopsis; while red light has demonstrated
promotion of root elongation, formation of lateral roots and regulation of root phototropic
responses (Mo et al., 2015). These studies show that light quality and root formation are
inconsistent amongst different species.
There were no significant differences in rooting biomass amongst all treatments in this
study. This confirms similar results from Navidad et al., 2020, in which there were no
differences observed in dry weight with Abies laciocarpa under HPS with 5% blue and HPS with
30% additional blue light. Additionally, in another study comparing the effect of red and blue
light, no differences were observed in the rooting percentage of hybrid franc under different
LEDs, however, longer roots were produced under fluorescent light with spectral distributions in
blue and red regions (Poudel et al., 2008). Within the same study, different LEDs affected
rooting percentages among two other species. However, this contrasted with Jahirul et al. 2021
study, which observed differences in root quality of cannabis seedlings under natural light, and
R:B spectra with additional far-red or far-red and UV. Thus, it is possible that this cultivar was
not sensitive to different light treatments in the propagation stage under the conditions of the
experiment. Despite the no differences in yield, there were differences observed amongst the root
quantity and root length of the first and second longest roots. Red light exhibited the longest root
lengths and quantity and was statistically the same as white and green. This indicates that blue
light, or shorter wavelengths, can inhibit root elongation by different synergistic interactions due
to the blue light receptors as observed in Arabidopsis. Thus, this study supported the null
hypothesis that light quality does not increase cannabis root growth during cutting establishment.
It is evident that light quality may influence root elongation through photomorphogenic action
but does not affect overall yield. This research has been significant in predicting suitable light
66
spectra applied to cannabis vegetative cuttings in an indoor controlled environment to obtain

proper root development. Based on the present study, the five lighting qualities are all practical
for obtaining proper root biomass.

production
Studies have investigated soilless and organic production methods (Burgel et al., 2020;
Caplan et al., 2017; Potter, 2014) with cannabis. However, an aspect that has not been addressed
is an RO-only application with a growing medium throughout an entire production cycle. If
proven to be a feasible approach, this methodology would allow for a low labour approach that is
more efficient than administering and re-adjusting liquid fertilizer throughout a production cycle.
Studies have shown that crops respond differently to slow-release fertilizer and growing media
(Agro & Zheng, 2014). This preliminary study aimed to evaluate the suitability of application of
RO water into peat based growing medium amended with slow-release fertilizer versus organic
soilless media throughout the entire production cycle after the propagation stage. Cannabis
producers lack access to scientific information regarding optimal growing medium recipes and if
the supply of nutrients can be limited to an application of controlled-release fertilizer or simply
using a fertile organic growing medium. Results were consistent with the alternative hypothesis
that growing medium does increase growth and development. In this study the best growing
medium was peat based with slow-release fertilizer added.
There is a consensus that nutrient assimilation and growth are dependent on the growth
medium and fertilizer application used (Maļceva et al., 2011). This was evident at the beginning
of the production phase since the propagation methods used for the peat-based mixture followed
the standard operating procedures for production. Using liquid fertilizer and rockwool cubes
demonstrated greater rooting success compared to the organic method using peat plugs
supplemented with aloe water. Rooted cuttings were selected to be transplanted as the organic
recipes would have had a delayed start or become necrotic if the unrooted and unhealthy clones
were randomly selected. The propagation methods of the organic clones would need to be
adjusted, either with a different medium, different environmental controls or a different type of
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cloning solution for higher rooting success. To maximize the root formation, further
considerations should be applied to the experimental design of the propagation in peat plugs
supplemented with aloe water.
The deficiencies in Figure 13 showed indications of nitrogen deficiency, but also may
have been a combination of symptoms of multiple deficiencies or toxicity (Cockson et al., 2019).
Leaf greenness has been positively correlated in literature with increased rates of slow-release
fertilizer (Chen et al., 2001), which was an observation exhibited in the peat-based growing
medium. The experimental design cannot quantify the relative contribution of the nutrient
availability within each medium; these factors should be studied in more detail in future
experiments. In the study, the pH levels were slightly low for cannabis plant growth (4.90-5.72),
and the low pH may have reduced Ca, Mg or caused Mn toxicity. The lower pH values exhibited
in the organic growing mediums may be due to higher NH4+ availability in the medium resulting
in NH4+ uptake and the release of protons from the roots (Caplan et al., 2019). The high ECs
seen in the media indicated over-fertilization relative to plant demand early in the production.
Similarly, this occurred in another study with osmocote applications that was observed to be due
to high NO3-N measurements. The EC measurements decreased over the production period in all
treatments as displayed in 9. This is a similar observation in multiple other studies with
controlled-release fertilizer (Agro & Zheng, 2014; Clark & Zheng, 2015, 2017). This suggests
that most nutrients available from the slow-release fertilizer and the organic growing media were
released and taken up by the plant.
Interestingly, despite the morphological observations’ indicative of nutrient deficiency

evident in the two organic medium recipes, there were no differences in the growth index,
quantity of leaves and the number of nodes throughout the production period. The growth index
decreased near harvest as it is normal to see changes in leaf morphology with leaves becoming
chlorotic, curling and falling off the plant as the flowering stage focuses all of its energy reserves
on flower production (Spitzer-Rimon et al., 2019). The lack of differences suggests that the
growing medium may not have influenced morphology.
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The significant differences in flower yield of the peat based mixture amended with slow-
release fertilizer is a result which Backer et al., 2019 attributed due to differences in nutrient
composition and timing of application as they exhibited a higher yield per W in a slow-release
fertilizer compared to a liquid nutrient regime. This contrasted with Caplan et al., 2019 where no
differences were observed in two coir-based organic growing media and a commercially
available peat-based growing medium. As well, Burgel et al., 2020 documented no differences in
floral yield in peat, peat substituted with 30% green fibre and coco coir fibre growing media. The
differences in floral yield can be contributed to the use of the slow-release fertilizer, however,
with reference to these studies which used fertilizers, it is likely explained by the use of a
different genotype that requires more nutrient uptake.
To satisfy the nutrient and water demands of a cannabis crop, the methods utilized in this
study obtained reasonable flower yields. Compared to other studies conducted with cannabis, a
study yielded ~58-110 g with the control plant under observations of a drought study (Stemeroff,
2017), 47-50 g observed with the different rates of organic fertilizer under ambient CO₂ (Caplan
et al., 2017) and another study looking at the effects of three different growing media yielded
3.84-9.19 grams (Burgel et al., 2020). Maturity to induction of flowers normally mature in 6 to 9
weeks (Chandra et al., 2020; Kazan & Lyons, 2015); in the case of this study it was
approximately 8.14 weeks indicating there was no delay in growth or alteration in flowering due
to suboptimal conditions. Overall, the peat-based mixture and organic media can be used for
cannabis production, however, based on the results of the experiment the peat based growing
medium is optimal to obtain a higher floral dry weight.
4.5 Experiment 4 – Effects of CO₂ concentration, temperature, light intensity

and spectral composition on whole plant net carbon exchange rate for
early vegetative and flowering stages
The objective of this experiment was to evaluate the influence of light intensity, CO 2
concentration, temperature and light spectra on 21 DAP vegetative plants and 35 DAP flowering
plants. The results of this study could be used to provide the initial environmental parameters for
a full production experiment and determine the optimal spectra, temperature, light intensity and
69
CO2 concentration for a 21 DAP vegetative and 35 DAP flowering plants. Growth chambers can
obtain a direct assessment of short-term responses of net carbon exchange rate under fluctuating
and reproducible environment conditions. It has been well established in the literature that
biomass production is correlated with these environmental factors. As exhibited in Section 4.1,
the light and CO₂ curves follow a similar pattern, obtaining a linear relationship between the
environmental variable and NCER followed by a plateau.
The light response curve of the 21 DAP vegetative and 35 DAP flowering plants is
represented in Figure 15. It was predicted that the plant would exhibit saturation to increasing
light intensity, therefore indicating that an optimum NCER can be obtained under the conditions
of the experiment. Despite the similarities between the two growth stages, these light curves
supported the alternative hypothesis that at a constant CO2 concentration, increasing light
intensity increases NCER. The CP and maximum NCER was higher in the 35 DAP flowering
plant however the repeated measures analysis revealed that the curves at different growth stages
were not statistically significant at each light intensity setpoint. This would suggest that the
NCER of a 21 DAP vegetative and 35 DAP flowering plant can be placed at the same light
intensity to obtain the optimal NCER. However, the lack of differentiation could be confounded
because plants that have acclimated under certain environment conditions will generally have a
higher NCER at the light intensity at which they were grown (Lanoue, 2016; Wilson & Cooper,
1969). These conditions were used to simulate light levels that were relevant to the stock plant
production but was most likely limiting its capacity as the optimal NCER was 459 μmol m-2s-1,
which is not a light intensity normally characterized in cannabis at these growth stages (Chandra
et al., 2008; Eaves et al., 2020; Moher et al., 2021; Rodriguez-morrison et al., 2021; Potter &
Duncombe, 2012; Vanhove et al., 2011). Results were inconsistent with Rodriguez-morrison et
al., 2021a study which exhibited optima leaf level NCER at ~600 μmol m-2s-1 under low light
conditions. In high light (1238 μmol m-2s-1), Rodriguez-morrison et al., 2021a showed that the
NCER is 50% higher than low light (91 μmol m-2s-1). It is predicted that had these plants been
acclimated under a higher PPFD, the light CP and maximum point would have been higher due
to the higher level of enzyme activity for carboxylation and electron transport (Taíz and Zeiger,
2006).
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The CO₂ response curve (Figure 14) increased rapidly at the initial setpoint and then
gradually became more stabilized. It was predicted that the plant would exhibit saturation to
increasing CO₂ concentration, however, the lack of a plateau indicated that the true maximum
was not obtained. It has been well established that doubling the CO₂ concentration can typically
increase the NCER by 30-50%, however depending on environment conditions plant NCER may
be as much as 75% (Ehleringer et al., 2005) to 90% (Parry, 2019). In contrast, in this experiment
when the CO₂ concentration doubled there was an increase of 298.11% and 107.94% in the
vegetative plant and flowering plant respectively. In both growth stages, the optimal NCER was
observed at 1600 ppm CO2. These results support the alternative hypothesis that at low light
levels, increasing CO2 increases plant NCER.
The photosynthetic responses to temperature are influenced by the stomatal conductance

(Chandra et al., 2008). The insignificant differences in the temperatures for both the 21 DAP
vegetative and 35 DAP flowering were unexpected and the null hypothesis (H8) could not be
rejected. Chandra et al., 2008 reported a maximum rate of photosynthesis at 30°C when
comparing plants grown between 20-40°C. When NCER is plotted as a function of temperature
under ambient CO2 concentration it is normal to see a characteristic bell shape when approaching
40°C (Taíz and Zeiger, 2006). The temperature went from a range of 15-35°C which may not
have been high enough to witness any degrading effects on the photosynthetic apparatus and due
to the low light levels optimal photosynthesis was not taking place. This confirms similar results
from Bazzaz et al., 1975 where there were no significant differences between 15-35°C from two
different cultivars in growth chambers compared to another cultivar which exhibited optimal
NCER at 25°C. Long-term production under the minimum (15°C) and maximum (35°C) ranges
would be interesting to see if morphological impacts lead to floral quality or yield issues.
Light spectral properties observed in cannabis have been shown to exhibit cultivar
dependent responses (Danziger & Bernstein, 2021). The results in this study supported the
alternative hypothesis that at a constant CO2 and light intensity, light quality increases NCER.
Certain wavelengths have been shown to activate photosynthesis, however, it has been well
researched that individual wavelengths are not as effective at driving a high rate of
photosynthesis compared to combinations of spectra or broad-spectrum (Goins et al., 1997;
71
Hogewoning et al., 2010). This was apparent in the 21 DAP vegetative plants as the highest
NCER was exhibited with R+Fr and was statistically the same as R:B (4:1), R:B (1:4), W+Fr,
W+UV, R+UV, RB(1.0)+Fr and W+R. However, this contrasted with the 35 DAP flowering
plants as they exhibited no differences with the highest NCER in W+Fr with monochromatic R
and B. This contrasts with literature, as red light is more effective at obtaining photosynthetic
efficiency than blue light. However, there is evidence that the NCER becomes weaker at low
light intensities (Liu & van Iersel, 2021).
The highest NCER obtained in both the vegetative and flowering plants was improved by
adding far-red in both lighting spectra, which is likely explained by the red to far red ratio. The
photoreceptor, phytochrome, plays a key role in the perception of the red to far red ratio. These
results suggest that the R:Fr ratio can increase the photosynthetic CO 2 assimilation by improving
the photosynthetic capacity. The value of additional UV-A is likely due to the blue light
photoreceptors, such as phototropin and cryptochromes, as they play a role in the UV-A
absorption (Chen et al., 2019). UV-A radiation has been considered to have photo inhibitory
effects; however, it is apparent that this light intensity and the short-term effect did not
downregulate the photosynthetic capacity.
Studies have indicated that the NCER would also be most efficient when plants are
illuminated with light of a similar spectral quality to which they were grown (Walters, 2005).
This was not apparent in the experiment as the plants were grown under white light and still
obtained similar NCER results with various light combinations. In addition, there were
significant differences in the response of cannabis at the two growth stages to different
wavelengths of light; however, there were some overlapping light qualities between the two
growth stages which include W+Fr, R:B (4:1), R+UV, RB(1.0)+Fr and W+R. The influence of
indoor lighting on photosynthesis does require further investigation to provide theoretical
guidelines.
There were some speculations in this experiment as to what is occurring on a

physiological level; however, it is important to remember that in the multitude of metabolic
reactions in photosynthetic carbon metabolism, cellular metabolism and photorespiration, there
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are many potential permutations of changes in the photosynthetic apparatus which could not be
addressed practically without further analysis (Zhu et al., 2010). In addition, leaf morphology
adjustments will change the cumulative NCER over a production period to acclimate to
conditions for optimal performance and fitness. Considering these points, under the conditions of
this experiment, the observations here indicated that beyond the maximum NCER, there was no
advantage of additional PPFD or any specific temperature setpoint. Furthermore, there would be
advantages to additional CO₂ concentration supplement as the plateau was not achieved and there
were many variabilities witnessed between different light qualities.
4.6 Summary
The experiments examined different horticultural management strategies at all growth stages
of one cultivar of cannabis (cv. Wappa). To fill the gaps in knowledge, five experiments were
performed using various controlled environment treatments in growth chambers under LED
lighting. Determining the growing protocol for a certain cultivar can increase the return on
investment for a production facility and improve plant productivity. Overall, finding the optimal
growing recipe for a cultivar is a complex process due to the dynamic interactions within the
plant and all the considerations of the environment that must be taken into account. For example,
changing one environmental parameter or irrigation strategy would influence or limit the yield or
NCER. Literature also shows variation among cultivars and phenotypes, indicating the
importance of executing research cycles on different cultivars to assess variation (Stack et al.,
2021). This means that the findings exhibited here cannot be applied to another cultivar with
anticipation of similar results. However, the results exhibited can be extrapolated with the same
cultivar to a large indoor growth space if the climate conditions are held precisely to replicate the
growth chambers.
One of the initial steps in cannabis cultivation is understanding if the growing media is
appropriate for a specific cultivar as it will affect the crop performance and management
(Campbell et al., 2019a). This directly relates to the water use efficiency, bulk density, air
porosity and moisture content of the growing media as certain cultivars have varying responses
to irrigation strategies (Stemeroff, 2017), highlighting the importance of cultivar selection for a
production facility that is designed to handle certain growing media. It is crucial to understand
73
how growing media are related to the success of the propagation stage as it directly translates to
how healthy the vegetative phase will be and thus the future success of the flowering period.
Rockwool is typically used in commercial facilities currently and had demonstrated the
highest success rate and root formation in Figure 10. This was further exhibited in Section 3.4 as
the rockwool cubes obtained a higher success rate in comparison to the organic propagation
strategy using peat plugs fertilized with aloe water. This observation was also exhibited in a
study, Campbell et al., 2021, where rockwool demonstrated higher rooting success in root
number, length and mass when compared between four soilless mixes amongst eight cultivars. A
container or inert medium provides a limited volume capacity for root development and water
storage (Burgel et al., 2020). This was exhibited in the findings, since the potting mx was in a
larger pot it obtained longer roots. This means that it will take longer for the cutting to expand its
root zone in the pot before transplant can occur, but it also means that the irrigation strategies
have to be more precise for optimal moisture conditions and aeration. Thus, it is difficult to
conclude which is the optimal growing medium between peat plugs and the potting mix,
however, it was evident that rockwool consistently obtained the highest success rate in rooting in
two experiments. Future research should consider propagation research with growing media
when placed under different environmental or pH/EC conditions. Future work should also
consider an aerated water treatment to see if it can be competitive with rockwool.
In both of these experiments (3.2 and 3.3), different morphological factors were analyzed
to determine if the clones were similar to each other to identify if the cloning procedure produced
identical vegetative cuttings. This was determined by measurements of the shoot dry weight, leaf
area, nodes, height, quantity of leaves and nodes. Results showed that the cloning procedure
successfully produced identical clones, so the dependent variable is not confounded. As
discussed in Section 3.3, the results supported the alternative hypothesis (H4) for root
length/quantity but not with dry weight which was unexpected due to the role that light spectra
plays in plant growth. Plants have interaction effects between light pathways and photoreceptors
that affect the sugar and plant growth regulator production. Based on the results, a full spectrum
(5650K) is recommended to allow producers to inspect the crop for pest and disease
management. The lack of difference in mass between the spectral qualities tested in the
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experiment may be due to the cultivar. Future experiments with different ratios of R:B and
different spectral qualities with multiple cultivars would determine if this is cultivar specific.
The results in Section 3.4 indicated that the peat-based mixture was the optimal growing
medium. Interestingly, this is the first study conducted with cannabis using a granular slow-
release fertilizer. These findings provided insight into the beneficial aspects that slow-release
fertilizer can have on yield and morphology. It is typical for production facilities to utilize liquid
nutrients because it allows for greater control and in general additional fertilizer input results in
greater biomass. When considering fertilizer options, the main focus in production facilities isn’t
necessarily about being the most cost-effective or environmentally friendly but about the
secondary metabolite profile and yield that can be achieved. The lack of fertilizer guidelines does
come at a cost since it is likely that producers are over-utilizing fertilizer causing significant
runoff (Shiponi & Bernstein, 2021). Future studies should look at more applications of slow-
release fertilizer with tissue nutrient samples to understand the correlations between yield,
nutrient input and morphology. In addition, future research should emphasize using slow-release
fertilizer that can last longer or multiple applications to provide an optimal fertilizer regime for
an entire production period. Lastly, the secondary metabolite profile should be considered as a
major deciding factor of a high-quality product.
Research has shown that cultivars display latitudinal distribution patterns correlated with
climatic factors (Hemphill et al., 1980; Zhang et al., 2018). This suggests that certain cultivars
thrive in specific environments that have different environmental setpoints. Generally, increased
NCER, is positively correlated with crop biomass production, floral yield potential and is
directly related to the environment conditions provided. This justifies research to investigate the
influence that each individual environmental condition can have on the NCER. A whole plant
isolated in a growth chamber of known volume can provide precise gas exchange measurements
that can accurately translate to large indoor production spaces. This was investigated in
experiments 1 and 4.
As described in Chapter 4.1 and Chapter 4.5, NCER responses in plants can predict plant
productivity. In the two experiments, measurements of NCER of cannabis at different light
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intensities, spectral qualities, temperature and CO2 were obtained at three different growth
phases (21 DAP vegetative, 123-153 DAP vegetative stock plant, 35 DAP flowering plant) under
LED lighting. It is important to note that vegetative plants are not typically maintained for 123-
153 days as the grower will want to switch to the flowering period to obtain frequent harvests.
However, if the growing conditions are optimal, stock plants can be maintained for a long time
for vegetative propagation or preservation of genetics, depending on the goals of the production
facility. The illustration in Figure 7 demonstrated that whole plant photosynthesis showed see-
saw patterns and had little deviations from the setpoints. Due to this, obtaining averages of
NCER normalized with leaf area from the data can be viewed as reliable.
It was unexpected that the 35 DAP flowering plant exchanged more carbon than the stock
plant in the light intensity NCER response graph. Typically, in angiosperms, there are tradeoffs
with reproduction and fitness. The higher NCER was likely because the flowering plant was
smaller and contained less leaf biomass which allowed for anatomical structure and arrangement
of leaves to obtain greater light penetration. This contrasted with the CO2 NCER response graph,
as the stock plant exhibited a higher NCER than the vegetative and flowering plants. The
complex process of photosynthesis is influenced by numerous variables, with the anatomy,
position and angle of leaves being a large factor. The stock plants contained greater leaf area
which is a major contributing factor to gas exchange. Additional leaf surface area allowed the
plant to have a greater capacity to exploit CO2 for NCER gains. The shape and presence of
additional stomata likely played a role in the stock plant’s ability to exchange more carbon with
additional CO2 supplementation. In addition, the doubling of CO2 observed in experiment 1 was
50%, which is a commonly exhibited phenomenon. However, in section 3.5 when the CO₂
concentration doubled there was an increase of 298.11% and 107.94% in the vegetative and
flowering plants respectively. This suggests that the smaller plant architecture was CO2 limited
in the acclimation stage. Overall, it supports the conclusion that NCER response studies are
complex as there are many factors to take into account.
It is noteworthy that in the temperature NCER response graph there were no significant
differences between all temperatures tested (15-32°C). However, since the plants were
acclimated under low light conditions the NCER was affected. In the light quality response
76
graph, it was anticipated that R:B and white spectral combinations with or without additional FR
and UV would likely be effective in driving NCER processes. Although, the fact that
monochromatic light sources (R and B) exhibited statistically insignificant results with these
spectra was unexpected in the 35 DAP flowering. It is likely probable that the benefits of spectral
combinations compared to monochromatic are only evident at higher light intensities in a
reproductive plant. Studies conducted on improved experimental design with plants tested under
higher light intensities may support this theory. Overall, any spectral combinations that obtained
statistically the same NCER suggest that any of the options would give equivalent dry matter
yields in a long-term study. Future studies should assess other growth stages weekly to see which
light qualities are best for vegetative versus reproductive stages and whether there are significant
differences between vegetative and reproductive stages.
Many studies have been conducted to determine the NCER response of leaves and whole
plants to determine the optimal environmental parameters. However, most of these studies have
examined the short-term or leaf-level response to a different environmental setpoint. The long-
term whole plant response may exhibit differences in gas exchange due to the different
morphological characteristics of the plant, including leaf thickness and leaf area, or be due to
changes in the metabolic reactions. These differences would be interesting to see the long-term
responses to compare between different growth stages. Limitations in the research facility
constrained a full analysis of long-term development when exposed to different light qualities.
However, the case study in Appendix VI proposed a methodology that should be considered in
future experiments. An important consideration is having enough subsamples to harvest
throughout production to normalize the NCER response. Another critical factor is the influence
of spectral quality on the secondary metabolite profile. Furthermore, these studies can compare
different chemotypes or cultivars within the same chemotype to reflect typical production.
This work provided valuable research considerations for horticultural management that
can be used in future experiments. Many different combinations of acclimation strategies can be
applied for NCER measurements (i.e. high light, low light), short term versus long term NCER
response, light qualities and growing media can be used to continue research conducted with this
particular cultivar.
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Chapter 5: Conclusion
The primary objective of this thesis was to assess different horticultural management
strategies in cannabis cultivation by addressing growing media selection and root development in
the propagation stage and using measurements of NCER in response to light, temperature and
CO2 as the plant developed. Standardization and optimization of cannabis horticultural
techniques are required to obtain consistent yields and quality for medicinal and recreational
consumers.
Experiments 2 and 3 concluded that rockwool was the optimal growing medium based on
the success rate. The methodology used in the second experiment provided insight into the
implications of obtaining rootzone data among different media. In experiment 2a, aerated water
treatment and a balance with a higher resolution would have benefitted the methodology to
reduce the statistical limitations in the rockwool and water treatment. There were no significant
differences in morphology between the potting mix and peat plug treatments (nodes, height,
leaves, stem diameter, dry weight of stem and roots), but differences were observed in the
potting mix obtaining longer root length of the three longest roots. Experiment 2b exhibited no
differences in root dry weight under five light spectra qualities, red, blue, green, R:B (1:1) and
white. However, the cuttings under red light produced a greater mass and longer roots when
looking at the two longest roots compared to the R:B and blue cuttings. In the third experiment,
the cultivation of cannabis in a peat-based mixture exhibited a higher floral yield compared to an
organic soil medium . However, there was no impact on growth attributes or the pH and EC of
the nutrient delivery system. The results indicated that the peat based growing medium with
slow-release fertilizer was suitable for the production of indoor container-grown cannabis to
obtain a higher floral yield.
Experiments one and four indicated that plants adjust their physiology to acclimate to
environment conditions and optimize the NCER. The difference in NCER performance is a
criteria for producers to predict the growth performance of cannabis and obtain the most cost-
effective environment conditions. The optimal NCER light conditions for the stock plants, 21
DAP and 35 DAP, were 410, 459 and 459 μmol m-2s-1 respectively. Among all experiments,
78
plants exhibited an optimal CO2 response at ~1600 ppm. It was interesting that temperature
resulted with no significant NCER differences between 15-32°C on young vegetative and
flowering plants but it should be noted that the light level was low and chosen to replicate stock
plant growing conditions. It indicated that short-term NCER response of low light acclimated
plants to temperature was not informative for understanding plant responses to temperature. In
addition, 21 DAP vegetative plants, 35 DAP flowering plants and stock plants can handle a high
level of PPFD and CO₂ concentrations for gas exchange processes. Lastly, W+Fr, R:B (4:1),
R+UV, RB(1.0)+Fr and W+R were the optimal light qualities that could be applied between the
two growth stages.
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Appendices
Appendix I
Controlled Environment Systems Research Facility PS1000 model technical summary.
Controlled Environment Systems Research Facility

Guelph BlueBox model PS1000
The PS1000 plant growth chamber is the sixth generation of whole plant photosynthesis systems
to be designed at the University of Guelph’s Controlled Environment Systems Research Facility
(CESRF). These growth chambers are capable of high-resolution measurement of whole plant
photosynthesis and evapotranspiration. They are used as a precision tool to better study plant
physiological responses in response to manipulation of multiple variables including:
Temperature Nutrients
Humidity Plant water status
Carbon dioxide Insect predation
Oxygen Pathogen application/response
Light (quantity, quality) Chemical application (pesticide, bio control, fertilizer)
Technical Specifications
· 1600 Watt water-cooled multispectral and

programmable LED lighting system with available UV
(368 & 380nm), blue (448nm), white (5650K), green
(568nm), red (655nm) and far red (735nm) irradiation
· Passive pressure compensation to improve system
closure
· Carbon dioxide enrichment from 0 – 10,000 ppm
· Continuous CO2 (0-20,000ppm) data recording
· Temperature control range from 15°C - 35°C +/-
0.5°C
· VPD control from 0.2 - 1.5 kPa
· Variable speed air flow with bottom up distribution
· Available active venting system to reduce O2 and
ethylene buildup during long term studies
· Integrated Argus Control System - full data graphing
and recording of all sensors and actuators
· Made of primarily non-off gassing inert materials
· 1200 x 600 x 800 mm (HxDxW) growing volume can
accommodate a wide variety of crops
Issue 01 Revision 01 2018/04/02

99
Appendix II
Controlled Environment Systems Research Facility PGC Flex model technical summary.
100
101
Appendix III
Light intensity mapping was produced for the PGC Flex two-tier chambers for white,
blue (B), red (R), R:B (1:1), green. Light maps were produced by measuring the PPFD
(μmol/m2s) using a Li-180 spectroradiometer (Li-Cor Biosciences., Lincoln, NE, USA) from the
top of the plant, or ~450 mm from the light source.
62.76 67.14 65.58 64.87 67.3 65.57 60.67

69.94 75.12 75.39 76.86 74.39 72.59 68.73
72.73 76.85 77.51 77.94 76.98 74.18 70.48
67.57 72.74 74.7 73.46 74.58 72.08 68.64
Figure 1. Heat map of PPFD distribution of the white (5650 K) spectra in a PGC Flex
Chamber
65.03 70.64 71.22 70.77 70.71 69.35 65.71

70.82 77.88 79.21 79.7 77.63 75.96 71.38
74.49 79.45 80.65 80.84 79.33 77.56 72.59
71.92 74.59 77.14 78.02 74.9 73.78 69.06
Figure 2. Heat map of PPFD distribution of the blue (448 nm) spectrum in a PGC Flex
Chamber
65.95 69.9 68.87 66.76 69.41 66.92 63.93
71.91 75.46 77.59 77.01 76.09 74.61 70.18
74.94 78.52 80.54 80.04 78.85 75.51 71.56
71.51 74.57 76.92 75.36 75.1 71.97 69.64
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Figure 3. Heat map of PPFD distribution of the red (655 nm) spectrum in a PGC Flex
Chamber
66.65 70.37 67.87 67.38 69.25 68.56 64.02

72 76.52 78.02 77.35 76.76 75.24 70.78
75.32 79.15 80.77 80.68 78.94 77.28 71.51
71.89 75.39 77.45 76.68 74.96 73.3 69.25
Figure 4. Heat map of PPFD distribution of the red:blue spectra in a PGC Flex Chamber
68.2 70.96 69.96 69.64 71.22 69.86 65.53

73.53 77.41 78.69 78 76.6 74.84 70.39
74.71 77.63 79.39 78.95 78.05 75.21 71.2
70.94 73.95 75.05 73.06 72.58 70.86 67.17
Figure 5. Heat map of PPFD distribution of the green spectrum in a PGC Flex Chamber
103
Appendix IV
Table 1. Analysis report conducted by Premier Tech Horticulture using saturated medium
extract method for the ProMix HP Mycorrhizae (Premer Tech Horticulture, Rivière-du-Loup,
QC, Canada) potting mix. Media analysis indicates the fertility status at the time of production.
Chemical Properties
pH (1:3) 5.03
EC (mmhos/cm) 1.74
Inc 7d pH 6.11
pH 5.04
EC corr (mmhos/cm) 2.44
N-NO3 (ml/L) 207
P-PO4 (ml/L) 67
K (ml/L) 109
Ca (ml/L) 286
S-SO4 (ml/L) 104
Mg (ml/L) 46
Na (ml/L) 32
Cl (ml/L) 15
Fe (ml/L) 4.46
Cu (ml/L) 0.14
Zn (ml/L) 0.37
Mn (ml/L) 2.17
Physical Properties
Weight of litre (g) 172.4
Humidity (%) 49.3
Dry bulk density (g/L) 87.4
Direct wettability (h:mm:ss) 0:00:12
104
Appendix V
Light intensity mapping was produced for the PS1000 chambers for R:B (4:1), white,
white + far-red (FR), white + ultraviolet (UV), R:B (1:4). Light maps were produced by
measuring the PPFD (μmol/m2s) using an Li-180 spectroradiometer (Li-Cor Biosciences.,
Lincoln, NE, USA) from the top of the plant. Normalized spectra available in Appendix VI
(Figure 2).
492.80 414.60 538.00 419.30 481.90

482.50 466.50 538.10 491.00 463.90
444.70 510.00 546.10 496.90 482.80
404.20 534.10 537.30 504.30 512.00
Figure 1. Heat map of PPFD distribution of the R:B (4:1) spectra in the PS1000 chamber.
Measurements were taken 960 mm from the light source.
525.70 536.50 527.70 518.20 498.80

529.40 531.40 531.00 522.20 494.80
518.70 533.30 523.00 510.80 488.80
505.60 510.00 513.70 500.80 480.30
Figure 2. Heat map of PPFD distribution of the broad spectra in the PS1000 chamber.
105
527.40 538.60 534.60 515.70 505.10

532.20 537.20 535.60 526.40 508.80
523.10 527.10 527.00 518.30 500.70
500.30 513.90 510.00 489.20 475.90
Figure 3. Heat map of PPFD distribution of the broad spectra + additional far-red in the
PS1000 chamber. Measurements were taken 778 mm from the light source.
529.50 539.80 534.20 524.70 508.30

532.00 536.90 534.60 528.60 507.20
522.60 528.20 522.60 518.40 501.80
498.30 510.00 514.90 504.80 484.40
Figure 4. Heat map of PPFD distribution of the broad spectra + additional ultraviolet light in
the PS1000 chamber. Measurements were taken 778 mm from the light source.
525.70 536.50 527.70 518.20 498.80

529.40 531.40 531.00 522.20 494.80
518.70 533.30 523.00 510.80 488.80
505.60 510.00 513.70 500.80 480.30
Figure 5. Heat map of PPFD distribution of the R:B (1:4) spectra in the PS1000 chamber.
106
Appendix VI
Case Study – The influence of spectral quality on NCER, nutrient uptake and morphology
Methods
The following case study evaluated the influence of five different LED light spectra (R:B
4:1, R:B 1:4, White, White + FR, White + UV) on whole plant morphology and NCER. There
were two objectives of this case study 1) investigate the nutrient uptake to evaluate a cannabis
plant throughout an entire production cycle at weekly intervals 2) evaluate the NCER in response
to increasing light intensity and carbon dioxide concentration at biweekly intervals. Results of
this case study would reveal the long-term response of plants when exposed to a specific light
spectra throughout an entire production cycle.
Chamber design
Experiments were conducted in five custom PS1000 controlled environment growth

chambers. Each chamber contained a two-tier shelving unit that had a Precision Balance
MS32001L/03 (Mettler Toledo., Mississauga, Canada). On the scale there was a 37.85 L plastic
bin that had connections to a 20L reservoir and a drain into the bucket, creating an ebb and flow
system, the experimental setup illustrated in Figure . The experiment was 86 days in duration;
14 days in propagation, 23 days in vegetative and 49 days in flowering.
107
A) B)
Figure 1. A) PS1000 growth chamber arrangement of one experimental unit during vegetative
growth stage. Chamber was equipped with variable spectrum LED arrays; each chamber was
designated to a specific light spectrum. Nutrient delivery system was set up as an ebb and flow
which was measured and exchanged weekly. B) Each box represented a container, or
experimental unit. Chamber had 12 clones till 14 DAP, 6 plants till 28 DAP and 2 plants for
the remaining duration of the experiment.
Experimental design
Sixty Cannabis sativa (cv. Wappa) cuttings were excised and taken at a length of ~100
mm cut at a 45° with 2-3 full expanded leaves. Twelve cuttings were placed in each PS1000
chamber under broad spectrum light at 50 μmolm-2s-1, 25°C day, temperature, 23°C night
temperature, 90% humidity and CO₂ = 500 ppm. Propagated plants were grown in rockwool
plugs (Grodan., Roermond, the Netherlands; 38.1 mm x 38.1 mm x 38.1 mm) that were
presoaked in CloneX (Green Planet., British Columbia, Canada; 1.0N-0.4P-1.0K; pH: 5.50,
EC: 0.95) solution at the recommended concentration of 5.0 ml/L. Cuttings were dipped 15 mm
in stim root #2 (Plant-Prod., Brampton, Ontario) before placement in rockwool plugs. Cuttings
were evenly placed in a 78-cell tray (Grodan., Roermond, the Netherlands) under a humidity
dome. Cuttings were irrigated twice a day for 10 minutes with CloneX (Green Planet., British
Columbia, Canada). At 7 DAP, light conditions were adjusted to 120 μmolm-2s-1 and 80%
108
humidity. Chambers were adjusted to five different light spectra, comprising combinations of
ultraviolet (λmax = 365 nm), red (λmax = 660 nm), blue (λmax = 440 nm) white (5650K) and far-
red (λmax = 735 nm). The combination utilized was R:B (4:1), R:B (1:4), white, white + UV and
white + FR. Normalized spectra of Intravision lights in each chamber are illustrated in Figure.
Heat maps of each light intensity are available in Appendix V.
Toronto, ON, Canada). Each fixture had four rows with one fixture and contained dimmable
spectrum channels for R:B (4:1) (660 and 440 nm), White (5650 K) + FR (735 nm), R:B (1:4)
(660 and 440 nm nm), White (5650 K) and White (5650 K) + UV (365 nm) which were
individually controlled with a fully automated control system (Argus Controls., Surrey, British
Columbia, Canada). Spectral distribution was measured using a Li-180 spectrometer (LI-COR
Biosciences., Lincoln, USA).
Propagated plants were transplanted into rockwool blocks (152.4 mm x 152.4 mm x

152.4 mm) 14 DAP. Rockwool blocks were covered with black and white poly film to prevent
algae growth. At 16 DAP humidity was adjusted to 75%, at 22 DAP light intensity was adjusted
109
to 300 μmol m-2s-1 and at 27 DAP light intensity was adjusted to 500 μmolm-2s-1. At 37 DAP,
plants were switched to a 12-hour photoperiod and 1200 ppm. Approximately ~10 grams of dry
weight leaf material were removed from each plant at 73 DAP to ensure proper light penetration.
Nutrient delivery system
Nutrient samples were measured weekly from the reservoir, one sample prior to irrigation
and the second sample after ~7 days and are listed in Error! Reference source not found..
Hydroponic nutrient solution samples were measured and analyzed by a high-performance liquid
chromatograph (HPLC) to see differences before and after irrigation strategies. Nutrient
solutions were measured for H2PO4, Cl, NO2, NO3, SO4, Na, NH4, K, Mg and Ca. Daily, pH and
EC data were measured and adjusted to maintain 5.50 and 0.95 mS cm-1 for the first three days.
Following that the pH was adjusted accordingly, 5.75 set at 3 DAP, 5.85 set at 14 DAP, 5.75 set
at 36 DAP and 5.65 set at 59 DAP. EC was adjusted accordingly, 0.95 mS cm-1 was maintained
until 20 DAP, 1.25 mS cm-1 set at 21 DAP, 1.65 mS cm-1 set at 36 DAP, and 1.85 mS cm-1 until
86 DAP. For the vegetative growth stage Dual Fuel ½ and ProCal (Green Planet., British
Columbia, Canada) was used. For the flowering growth stage Dual Fuel ½, ProCal and
Massive, Rezin (Green Planet., British Columbia, Canada) was used. Cuttings were irrigated
twice a day for 10 minutes with CloneX (Green Planet., British Columbia, Canada) using the
ebb-flow design. At 20 DAP, plants were irrigated three times a day for 2 minutes. At 32 DAP,
plants were irrigated three times a day for 3 minutes.
Measurements
During the vegetative growth stage height, number of nodes, number of leaves, stem
diameter and length of the 3 internodes was recorded weekly. During the flowering growth stage
height, number of nodes and stem diameter were measured.
The photosynthetic response of each chamber was obtained biweekly under different
PPFD (115, 215, 410, 504, 596, 686, 774, 859 µmol m-2s-1) and CO₂ levels (420, 820, 1020,
1220, 1620, 2020) levels. NCER measurements corresponding to PPFD and CO₂ settings were
first recorded at the lowest value and these settings were subsequently increased over a period of
110
48 hours. To normalize the NCER, destructive measurements were obtained at 14 DAP (6 plants)
and 28 DAP (4 plants) for randomly selected plants in each chamber. Normalization data at 12
and 26 DAP were based on the destructive harvest obtained at 14 and 28 DAP respectively and
multiplied to obtain an estimate to account for the number of plants in the chamber.
Normalization data between 44-82 DAP was based on the leaf area at final harvest.
Harvest
At 14 DAP, 6 plants, and at 28 DAP, 4 plants were cut at the base of the rockwool cube,
all leaves were removed and leaf area was measured using a LI-3100C area meter (LI-COR
Biosciences., Lincoln, USA. Fresh weight and dry weight of stems and leaves were measured
and measured using a Sartorius ENTRIS 6202-1S Precision Balance (Sartorius, Göttingen,
Germany). Plant material was dried in an incubator oven at 61°C (Fisher Scientific IsoTemp 500
Series., Toronto, Canada) for 14 days. Dried samples were re-evaluated until constant mass was
obtained.
Fresh flowers were spread on a seven-layer mesh drying rack in the PS1000 chamber in
the dark at 18°C temperature, 50% humidity and a fan at high speed at 86 DAP. Dehydrated
flower tissue was measured for moisture content using a halogen moisture analyzer HE73
(Mettler Toledo., Mississauga, Canada). Five grams of dehydrated flower tissue from each
experimental unit were sent to RPC Science and Engineering (Fredericton, NB, Canada) for
cannabinoid and terpene analysis. Analysis was conducted on neutral cannabinoids THC, CBD,
CBN, CBG, and their acidic constituents. Terpene analysis included alpha pinene, beta pinene,
myrcene, limonene, terpinolene, caryophyllene, humulene, 3-carene, cis-ocimene, eucalyptol,
trans-ocimene, fenchol and borneol.
Results
The experiment was performed once due to limitations of the research facility. As such
this experiment was treated as a case study and limited conclusions can be made. Graphs and
tables are provided for the data measured.
CO₂ response
111
Figure 3. Average NCER response of a 12 DAP vegetative plant (n=1) over a range of CO₂
concentrations at 500, 750, 1000, 1200, 1600 and 1980 ppm at a light intensity of 120 μmol m-
2s-1
112
Figure 4. Average NCER response (n=1) at a light intensity of 500 μmol m-2s-1 a) 26 DAP
vegetative plant over a range of CO₂ concentrations from 650 to 1830 ppm b) 44 DAP
flowering plant at 820, 1000, 1200, 1600 and 1980 ppm c) 54 DAP flowering plant over a
range of CO₂ concentrations at 680, 810 970, 1300 and 1600 ppm d) 68 DAP flowering plant
over a range of CO₂ concentrations at 880, 1000, 1200, 1600 and 1980 ppm e) 82 DAP
flowering plant over a range of CO₂ concentrations at 910, 1000, 1200, 1600 and 1980 ppm
Light response
113
Figure 5. Average NCER response (n=1) at a CO₂ concentration of 500 ppm a) 13 DAP
vegetative plant over a range of light intensities from 0 to 860 μmol m-2s-1 b) 27 DAP
vegetative plant over a range of light intensities at 0, 190, 360, 450, 610, 690, 770 μmol m-2s-1
Figure 6. Average NCER response (n=1) at 27 DAP a) Vegetative plant over a range of light
intensities at 0, 90, 190, 360, 450, 610, 690, 770 μmol m-2s-1 at a CO₂ concentration of 1000
ppm b) Vegetative plant over a range of light intensities at 0, 90, 190, 360, 450, 610, 690, 770
μmol m-2s-1 at a CO₂ concentration of 1500 ppm
114
Figure 7. Average NCER response (n=1) at 1200 ppm over a range of light intensities 0, 100,
210, 400, 500, 680, 780, 860 μmol m-2s-1 a) 43 DAP flowering b) 55 DAP flowering plant c)
69 DAP flowering plant d) 83 DAP flowering plant
Potency
115
Figure 8. Horizontal bar graph of secondary metabolite concentration. Data were produced from
5.0 g of dehydrated bud (<13% moisture), sampled randomly from all five chambers a)
Cannabinoid concentration in percentage b) Terpene concentration in percentage
Nutrient delivery system

116
Figure 9. Electrical conductivity and pH measurements throughout the production period. Each
graph represents one experimental unit (n=1). Left axis are measurements for pH and the right
axis are measurements for EC.
117
Nutrient solution data taken weekly and analyzed by HPLC. Two samples were taken each week
and indicated as before and after to estimate nutrient absorption and accumulation.
Table 1. Hydroponic nutrient solution for R:B (1:4) spectra. Values represent one replication.
Samples were taken every ~7 days, with one sample at the beginning and the end of a 7-day
period. Abbreviations: DAP = Days after planting, B = Before, A = After.
DAP H2PO4 Cl NO2 NO3 SO4 Na NH4 K (mg/l) Mg Ca (mg/l)

(mg/l) (mg/l) (mg/l) (mg/l) (mg/l) (mg/l) (mg/l) (mg/l)
7B 55.332 5.24 25.488 401.892 14.896 10.604 5.06 141.52 19.816 92.548
7A 60.3 3.668 0 488.332 14.444 9.872 5.344 148.68 22.656 80.024
16 B 57.976 1.252 0.108 397.8 112.8 15.408 6.372 98.3 29.592 100.564
16 A 46.628 1.368 0 331.988 87.288 7.692 5.604 81.26 23.268 82.78
21 B 52.74 1.712 2.932 337.596 120.384 9.788 4.792 106.32 28.352 93.516
21 A 72.212 1.056 0 438.884 129.74 8.948 7.296 121.888 32.66 109
28 B 45.328 2.648 0 282.936 128.312 10.176 3.552 108.132 27.496 99.056
28 A 97.74 1.368 0 591.108 192.56 13.364 10.312 160 45.26 137.18
35 B 76.928 1.84 0 410.468 158.412 10.556 4.248 138.892 38.056 121.896
35 A 101.752 1.412 0.048 651.752 230.404 21.388 12.088 179.94 50.476 152.696
42 B 74.008 4.692 1.656 109.048 184.016 18.708 4.268 143.392 37.636 122.656
42 A 283.556 1.616 0.108 642.804 206.676 33.648 32.016 285.312 39.328 109.496
49 B 165.016 2.86 83.572 9.3 163.368 25.216 13.588 183.372 31.036 87.764
49 A 313.628 1.628 3.796 638.34 204.044 28.628 33.52 292.836 39.1 111.048
56 B 182.836 1.64 0.044 1.992 173.348 27.056 0.088 209.336 29.92 77.268
56 A 279.364 1.24 10.968 585.08 190.364 31.22 32.348 276.28 37.38 104.036
65 B 252.016 4.78 17.256 267.404 236.648 35.856 16.144 295.36 40.44 100.968
65 A 260.424 1.848 0.804 577.016 173.584 27.168 30.032 260.212 34.148 100.424
69 B 228.188 1.968 47.36 212.048 213 29.564 12.376 287.528 36.764 87.028
69 A 274.372 1.572 0.144 636.704 183.168 27.456 29.868 262.232 38.368 108.58
72 B 281.352 2.368 49.848 517.988 205.052 31.504 24.172 281.48 42.028 105.156
72 A 293.764 1.628 0 640.592 195.764 31.876 30.452 281.952 38.88 112.072
77 B 236.78 4.576 0.356 120.736 212.92 31.772 1.788 347.548 45.908 99.612
77 A 302.108 1.804 0 636.908 198 31.012 31.832 284.784 38.344 109.716
118
Table 2. Hydroponic nutrient solution for white spectra. Values represent one replication.

7B 56.764 6.58 0 414.732 12.928 9.932 2.888 138.072 18.848 70.988
7A 67.772 3.092 0.128 466.496 12.26 8.728 5.228 144.704 21.032 75.092
16 B 60.2 1.1 0 391.88 111.544 9.84 5.996 98 29.868 98.504
16 A 50.504 2.232 0.052 354.44 98.336 10.468 5.844 87.456 26.056 94.932
21 B 50.128 1.34 1.036 373.88 123.816 13.568 5.56 112.192 28.628 98.536
21 A 73.88 1.108 0.088 447.952 132.992 9.856 7.908 124.052 33.516 112.204
28 B 61.972 1.496 0.76 388.152 151.556 13.428 4.916 118.312 32.288 113.98
28 A 93.328 1.312 0 537.812 171.836 10.648 9.04 151.936 41.684 130.672
35 B 71.952 3.148 0.004 242.836 152.868 9.888 2.904 142.2 36.508 122.184
35 A 105.484 1.72 0 625.756 217.796 20.668 11.008 175.232 46.36 147.32
42 B 79.288 1.768 1.26 177.744 193.78 14.212 2.336 165.188 41.256 137.04
42 A 267.016 1.648 0.084 593.976 190.3 28.876 27.952 266.344 35.352 101.268
49 B 194.788 1.86 0.2 1.716 177.284 24.592 10.796 215.192 33.116 95.38
49 A 319.276 1.624 16.14 633.08 204.264 28.744 33.448 293.78 39.172 108.72
56 B 225.064 0.864 0.18 4.684 207.432 28.952 0.444 248.58 37.204 95.168
56 A 295.6 2.14 8.52 657.304 215.768 33.188 34.568 292.616 42.888 115.012
65 B 241.008 2.04 6.692 259.14 204.428 30.812 15.744 257.828 36.364 98.108
65 A 287.708 2.224 4.4 619.496 190.368 27.456 31.232 276.28 37.172 109.22
69 B 207.82 1.704 0 50.644 173.292 27.576 5.04 228.72 32.94 89.64
69 A 280.548 1.468 0.008 651.756 189.26 26.908 29.268 262.64 38.732 107.352
72 B 236.36 1.552 0 192.892 197.836 30.42 8.812 250.872 38.768 99.32
72 A 288.596 1.652 0.024 628.992 191.176 26.556 28.748 276.932 39.132 107.224
77 B 191.788 1.072 0.02 4.036 174.74 28.4 0.892 242.124 32.196 78.648
77 A 293.788 1.708 0.036 618.58 190.224 28.16 29.8 269.796 36.132 103.564
119
Table 3. Hydroponic nutrient solution for white spectra + FR. Values represent one
replication. Samples were taken every ~7 days, with one sample at the beginning and the end
of a 7-day period. Abbreviations: DAP = Days after planting, B = Before, A = After.

7B 54.224 3.368 0.196 438.424 14.804 11.384 3.996 140.228 20.312 77.736
7A 65.936 2.6 0.304 461.488 13.076 8.6 5.296 144.336 21.344 74.232
16 B 55.036 1.008 0.172 394.736 107.12 9.88 5.948 97.044 29.972 99.404
16 A 49.52 1.272 0.224 346.12 96.6 10.948 6.104 88.232 26.352 92.544
21 B 52.408 1.296 0.18 316.164 135.94 13.748 4.42 112.984 29.46 102.768
21 A 76.388 1.44 0.28 458.536 138.064 10.312 7.096 125.328 33.484 113.488
28 B 62.4 1.956 0 325.236 157.584 14.248 4.384 123.236 32.58 111.656
28 A 89.028 1.332 0.008 560.888 177.624 12.54 9.912 157.884 43.568 136.328
35 B 72.492 1.676 0.3 265.704 173.94 10.96 4.156 153.924 40.508 132.988
35 A 97.084 1.636 0.072 583.084 203.872 18.908 10.452 167.628 42.896 133.76
42 B 66.068 1.568 0.556 0.664 226.912 25.776 0.532 142.324 43.76 142.216
42 A 273.096 1.584 0.276 637.44 209.04 32.44 31.488 284.272 38.54 109.112
49 B 155.264 1.232 0.624 1.796 146.044 22.592 0.188 186.192 25.892 77.26
49 A 306.32 1.716 11.532 622.568 198.112 28.992 34.444 290.824 38.332 106.44
56 B 172.336 1.156 0.16 2.884 166.908 26.152 0.336 202.864 29.512 81.308
56 A 270.364 1.656 11.328 628.132 201.744 31.732 33.528 288.824 39.78 113.108
65 B 231.296 1.72 24.844 312.412 202.968 31.288 19.856 251.684 36.276 97.02
65 A 266.684 1.716 9.92 611.644 191.184 35.964 32.308 273.64 38.668 109.376
69 B 219.02 1.4 0.164 217.552 174.844 24.608 12.468 237.136 31.976 92.052
69 A 276.948 1.376 0.112 636.132 183.088 28.656 28.86 260.716 38.888 108.168
72 B 273.112 1.96 15.96 319.136 229.944 35.948 17.68 273.64 36.368 93.464
72 A 287.992 1.768 0 632.088 189.448 27.984 28.408 280.156 38.516 108.244
77 B 192.436 1.128 0 1.648 155.62 24.452 2.084 209.636 30.616 81.216
77 A 298.912 1.496 0.188 631.832 192.232 31.136 31.924 278.188 37.568 105.06
120
Table 4. Hydroponic nutrient solution for RB (0.5) spectra. Values represent one replication.

7B 53.824 3.444 0.532 453.744 14.732 10.212 4.3 143.748 20.852 77.308
7A 58.256 3.26 0.116 476.484 13.084 11.004 5.876 148.512 22.056 82.736
16 B 52.58 1.592 4.212 390.012 111.136 9.076 5.892 98.144 29.54 98.316
16 A 50.328 1.376 0 344.808 84.42 12.068 5.536 90.416 28.056 90.64
21 B 57.468 1.616 0.944 387.748 122.232 13.06 6.32 115.608 29.86 100.796
21 A 79.588 1.236 0.044 508.704 152.736 10.476 8.4 133.104 36.132 123.552
28 B 55.94 1.98 11.264 305.296 146.208 12.14 4.784 125.772 32.388 107.944
28 A 89.66 1.236 0 580.948 187.84 10.952 9.652 158.016 43.268 139.476
35 B 87.116 0.832 9.844 479.144 181.824 14.852 6.948 160.896 42.568 140.728
35 A 106.816 1.052 0 631.62 220.96 20.94 10.864 175.46 46.156 144.256
42 B 92.308 1.472 0.22 504.892 201.948 18.332 6.292 174.636 42.716 137.444
42 A 275.14 1.644 0.08 639.428 210.856 33.548 31.452 283.56 38.492 111.156
49 B 244.332 1.956 200.688 9.576 218.084 31.856 21.32 261.9 43.68 114.256
49 A 317.584 1.8 18.18 638.804 208.94 28.428 33.02 293.948 38.668 108.572
56 B 213.032 1.432 0 3.508 197.684 33.248 0 235.896 34.336 96.292
56 A 266.132 1.54 3.492 579.104 182.076 27.788 29.524 267.172 35.892 100.164
65 B 279.16 2.6 39.012 440.64 252.412 42.408 26.868 286.572 42.18 115.736
65 A 277.896 1.94 4.908 640.828 198.364 35.68 33.836 282.468 38.252 113.128
69 B 257.424 1.744 0.48 352.672 195.496 30.444 19.064 281.992 37.42 105.624
69 A 289.184 1.548 0.792 652.252 190.756 29.532 30.34 267.736 39.992 111.068
72 B 290.456 1.9 7.036 428.228 223.252 29.684 20.416 280.824 44.308 115.448
72 A 279.764 1.68 1.492 608.632 186.008 27.94 28.576 273.872 37.232 105.124
77 B 236.668 2.22 0 1.516 209.488 33.156 0.308 280.116 37.068 93.832
77 A 296.896 1.864 0.308 639.084 192.376 28.684 30.312 278.756 37.612 108.036
121
Table 5. Hydroponic nutrient solution for white spectra + UV. Values represent one
replication. Samples were taken every ~7 days, with one sample at the beginning and the end
of a 7-day period. Abbreviations: DAP = Days after planting, B = Before, A = After.

7B 47.908 2.996 0.088 404.66 12.996 8.412 3.752 130.616 19.108 71.74
7A 68.184 3.556 0 472.884 12.868 9.188 5.428 145.924 21.184 75.472
16 B 58.276 1.216 0.036 391.88 106.84 9.252 6.376 97.976 29.4 101.812
16 A 46.504 0.852 0.404 346.46 95.128 12.888 5.48 86.672 25.576 92.028
21 B 51.492 1.352 0.508 369.508 120.62 9.556 5.616 103.38 28.744 98.392
21 A 72.888 1.092 0 447.104 131.808 9.368 7.984 123.504 32.812 109.812
28 B 56.992 1.636 2.692 355.152 140.652 9.852 4.432 128.248 31.048 114.968
28 A 97.068 1.508 0 602.736 194.816 12.928 10.508 159.66 45.004 137.628
35 B 78.044 1.34 14.308 327.096 182.796 10.368 4.364 157.676 44.304 139.856
35 A 113.132 1.28 0.064 649.764 223.06 21.02 11.172 178.244 49.928 149.32
42 B 79.352 1.456 7.136 34.016 203.164 17.8 4.264 159.656 42.848 139.64
42 A 292.612 1.452 0 623.544 205.4 28.844 29.152 276.012 37.576 106.528
49 B 163.776 1.392 0 1.692 162.868 26.204 0.78 219.28 30.768 95.012
49 A 280.916 1.764 6.128 591.52 182.468 26.828 30.456 278.232 35.904 101.04
56 B 218.124 1.68 0.424 2.224 165.368 25.792 0 252.072 33.084 96.688
56 A 268.512 1.348 3.616 587.204 183.76 26.928 30.4 270.412 36.288 101.184
65 B 270.636 4.824 31.892 453.156 217.68 34.18 27.02 277.148 40.096 115.476
65 A 298.228 1.924 4.384 633.956 197.248 28.496 30.804 276.384 37.48 110.572
69 B 253.94 1.624 11.616 361.788 197.368 29.756 19.52 260.16 37.148 108.928
69 A 278.924 1.664 0 647.376 185.352 27.564 29.452 265.744 38.088 109.964
72 B 247.776 1.692 34.6 417.612 191.768 27.556 23.552 250.98 37.46 106.756
72 A 296.14 1.852 0 638.76 194.416 28.164 28.708 281.668 38.232 108.216
77 B 198.288 1.256 0 0 171.772 26.692 0 236.748 32.248 90.972
77 A 267.948 1.592 0.028 580.328 173.404 27.416 29.648 260.856 34.572 99.988
Growth parameters
122
Table 6. Averages of growth parameters, height, nodes, stem diameter, internode length (1-3),
and quantity of leaves measured throughout the vegetative period (7-35 DAP)
Stem 1st 2nd 3rd

Height
Treatment DAP Nodes diameter Internode Internode Internode Leaves
(mm)
(mm) (mm) (mm) (mm)
50.8 5 19.7 18.5 15.2 13.5 4
White 53.4 5 20.7 19.0 17.0 13.4 4
White+FR 7 57.8 5 20.7 19.8 17.7 14.3 4
R:B (4:1) 59.5 6 19.5 18.1 17.3 16.0 4
White+UV 54.0 5 20.1 19.9 18.4 14.0 4
R:B (1:4) 61.7 6 23.9 19.4 14.3 14.5 7
White 58.1 6 24.0 19.5 17.2 14.6 8
White+FR 14 72.8 7 26.4 19.3 18.3 15.3 9
R:B (4:1) 79.5 7 24.1 17.7 17.4 15.5 9
White+UV 77.9 7 27.4 19.9 18.6 14.2 10
R:B (1:4) 102.5 8 34.7 15.5 15.2 16.8 18
White 112.0 8 39.5 17.0 16.0 12.6 18
White+FR 21 141.2 9 41.9 20.7 17.7 15.8 19
R:B (4:1) 135.7 9 34.1 19.2 20.7 14.0 17
White+UV 129.0 9 36.7 18.0 17.2 12.0 21
R:B (1:4) 148.1 11 51.4 16.0 14.5 16.7 37
White 166.2 10 62.1 17.2 17.2 14.0 39
White+FR 28 198.2 11 60.0 20.0 17.5 15.8 37
R:B (4:1) 171.2 11 52.4 18.8 18.2 12.2 31
White+UV 193.0 11 58.5 18.0 18.3 13.5 42
R:B (1:4) 224.5 14 73.6 17.5 15.5 12.0 65
White 224.0 13 85.1 15.5 13.5 14.0 62
White+FR 35 273.0 13 79.9 15.5 21.0 13.5 67
R:B (4:1) 216.0 12 67.6 18.0 15.5 8.0 47
White+UV 235.5 13 65.1 21.5 16.5 12.5 56
123
Table 7. Averages of growth parameters, height, nodes and stem diameter of treatments
measured throughout the flowering period (42-63 DAP)
Treatment DAP Height (mm) Nodes Stem diameter (mm)

R:B (1:4) 330.0 14 8.70
White 341.0 14 8.81
White+FR 42 372.0 15 9.40
R:B (4:1) 295.5 14 8.16
White+UV 302.6 14 7.47
R:B (1:4) 424.0 15 9.39
White 406.5 16 11.28
White+FR 49 469.0 15 10.26
R:B (4:1) 422.0 15 8.91
White+UV 388.5 16 8.63
R:B (1:4) 470.0 17 9.39
White 402.0 15 9.44
White+FR 56 468.0 15 10.25
R:B (4:1) 436.5 14 10.85
White+UV 395.5 14 9.14
R:B (1:4) 468.5 16 9.37
White 416.5 15 10.36
White+FR 63 474.0 15 10.52
R:B (4:1) 451.0 15 9.77
White+UV 409.1 14 9.23
124
Table 8. Averages of growth parameters, height and stem diameter of treatments measured
throughout the flowering period (70-77 DAP)
Treatment DAP Height (mm) Stem diameter (mm)

R:B (1:4) 429.0 87.4
White 425.0 117.9
White+FR 70 479.5 96.2
R:B (4:1) 458.0 102.2
White+UV 413.0 85.7
R:B (1:4) 475.0 99.4
White 419.0 122.9
White+FR 77 476.7 128.9
R:B (4:1) 464.5 129.3
White+UV 414.0 123.0
Table 9. Averages of leaf area, dry weights of stem, leaf and flower measured at harvest
Treatment Leaf Area (mm2) Dry Weight Stem Dry Weight Flower Moisture (%)
and Leaf (g) (g)
R:B (1:4) 11758.0 17.06 32.93 11.88
White 16229.2 18.96 44.22 11.31
White+FR 17785.6 20.88 53.94 12.82
R:B (4:1) 14457.0 20.59 45.64 11.88
White+UV 11035.9 12.00 30.47 12.33
125
Figure 10. Cannabis experimental plants at 85 DAP a) R:B (1:4) b) White + UV c) R:B (4:1)
d) White e) White + FR
126
Discussion
Limitation of the research performed in the case study were that adequate replications
were not achieved, and therefore no statistical conclusions can be made from the data
measured. Casual inferences cannot be drawn and applied to indoor cannabis production
facilities nor to any growth chamber studies. However, the information gained from this case
study is an important first step at identifying the light metrics and nutrient uptake that should
be considered in future research. Future studies should continue this work in controlled
environments to understand propagation strategies, impact of different growing media, long-
term response in morphology, nutrient uptake, and physiology effects when placing plant
under different environment conditions.

Hoogenboom Jennifer 202201 MSC

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Horticultural Management and Environment Control Strategies for Cannabis

(Cannabis sativa L.) Cultivation

The University of Guelph

In partial fulfilment of requirements

Guelph, Ontario, Canada

© Jennifer Hoogenboom, January 2022

HORTICULTURAL MANAGEMENT AND ENVIRONMENT CONTROL STRATEGIES

Jennifer Hoogenboom Advisors:

University of Guelph, 2022 Dr. Mike A. Dixon

Dr. Thomas Graham

exhibited higher floral yield than organic mixtures.

2.4.5 Statistical analysis ................................................................................................................................ 31

CESRF – Controlled Environment Systems Research Facility

Appendix I – Technical summary of “PS1000” chambers (a version of the Guelph Bluebox

Appendix II – Technical summary of “PGC Flex” growth chambers

Chapter 1: Literature Review

1.1 Cannabis physiology

1.2 Historical barriers in Canada

1.3.1 Indoor controlled environment

Large-scale cultivation of indoor cannabis is an energy-intensive proposition. The

Characterizing the photosynthetic response to environmental parameters can determine

It is of particular importance to have a healthy propagation phase as that determines the

propagation is a valuable tool to complement conventional breeding technology. In-vitro

1.3.3 Growing media and fertilizer

In propagation and production, numerous growth media are available to be used,

Slow-release fertilizer (or controlled-release fertilizer) releases nutrients over a period of

1.3.4 Net carbon exchange rate

Plants will acclimate to environment conditions through changes in photosynthesis,

seed germination, de-etiolation, directional growth, organelle movement, flowering and

1.4.1 Photosynthetic pigments

1.4.2 Photoreceptor pigments

1.4.3 Horticultural research on light

There were nine main hypotheses for the experiments conducted:

HA7: At a constant CO2 concentration, increasing light intensity increases NCER

2.2.1 Plant material

Plants were in 101.6 mm x 101.6 mm x 101.6 mm rockwool blocks (Grodan.,

2.2.2 Gas exchange measurements

2.2.4 Statistical analysis

The NCER is calculated using the following formula:

• V = chamber volume (litres)

• T = constant temperature (296.15 Kelvin)

2.3 Experiment 2a – The effect of growing media on root development

2.3.1 Plant material

2.3.3 Statistical analysis

2.4 Experiment 2b – The effect of spectral quality of light on root

2.4.1 Plant material

Stock plants were grown in 152.4 mm x 152.4 mm x 152.4 mm rockwool blocks

Chambers were adjusted to five different light spectra, comprising combinations of

2.4.4 Rooting performance measurements

2.4.5 Statistical analysis

2.5 Experiment 3 – Growing media considerations for container-grown

A possible solution to excessive maintenance is to implement a reverse osmosis (RO)

Stock Plants Cuttings

2.5.2 Transplant and Fertilization

Vegetative Flowering Drying

2.5.5 Statistical analysis

2.6 Experiment 4 – Effects of CO₂ concentration, temperature, light intensity

The performance of a cannabis plant under steady-state environment conditions allows

Propagated plants were transplanted into 101.6 mm x 101.6 mm x 101.6 mm rockwool