Group Leader: Magcalas, Carlos Jose B.

Date Performed: 05/26/2021

Members: Marcial, Erin Mark Date Submitted:
Monterozo, Audrey Rose
Llanto, Ann Benadeth N.
Olave, Kyla Denise

NARRATIVE REPORT

SUBJECT: A Narrative Report on the BioMass Webinar about Photosynthesis Measurement in

Seaweeds led by Dr. Jayvee A. Saco

A webinar happened on May 21, 2021 and

discussed about photosynthesis measurement in
seaweeds. The webinar was attended by the BioMass
students of the Batangas State University. The speaker
for the Virtual Training on Photosynthesis Measurement
in Seaweeds was Dr. Jayvee A. Saco, a Balik Scientist
in DOST-PCAARRD, Center Head of VIP CORALS in
Lobo, Batangas State University, and Assistant
Professor at the College of Arts and Sciences and
College of Agriculture and Forestry in Batangas State
University. He first greeted the participants then
proceeded to give a little introduction on the topic.

Before proceeding on the discussion of

photosynthesis, he defined what are seaweeds. Then,
tackled the characteristics of seaweeds which are
marine macrobenthic algae and photo-auxotrophic.
Lastly, he tackled the habitat of the seaweeds which are
the shallow coastal areas.
 He also discussed the ecological importance of
seaweeds. Seaweeds are food for higher organisms,
shelter and nursing grounds, source of sediments,
stabilizer of the substrate, reef builder, and carbon
dioxide sink.

Aside from the ecological importance, seaweeds

also provide extensively to our economy. He asked the
participants to identify which item uses seaweeds. After
that, he said that all of the items use seaweed, even the
fabrics used.

In this part of the webinar, Dr. Saco briefly

discussed photosynthesis. In this process, seaweeds
need to utilize light energy in converting water and
carbon dioxide to oxygen and carbohydrates.
Photosynthesis helps the plants including seaweeds to
grow, thus, understanding the physiological mechanisms
controlled in the photosynthesis is very important for
their growth requirements to achieve efficient cultivation.
 Photosynthesis is carried out in a specialized
organelle called the chloroplast that consists of
photosynthetic pigment called chlorophyll that captures
light energy to convert to chemical energy.
Photosynthesis is divided into two processes, the light
dependent and light independent reactions. The light
absorption is held on Photosystem I and Photosystem II.
The light strikes at Photosystems simultaneously. When
the chlorophyll molecule absorbs light, the electron
obtained from water in PS II is energized into state and
produces the byproduct oxygen.

The hydrogen from the water is converted to hydrogen ion. Then, the excited electron is passed
onto the reaction center to a series of electron transport chains up to the PS I. The low energy electron in PS
I is reenergized by light and passed on to the reaction center. With the presence of hydrogen ions in the
energized electron, two sources of energy are produced (NADPH and ATP). This NADPH and ATP
produced in light reaction will be used in the light independent reaction for the conversion of carbon dioxide
to produce carbohydrates.

He added that when the chlorophyll absorbs

light energy, it undergoes one of the three things. First is
photosynthesis, de-excitation of excited chlorophyll
molecules produces excess energy in the form of heat
and emitted light as chlorophyll inflorescence.

There are two ways of measuring

photosynthesis as per mentioned by Dr. Saco. The first
one is using the Photosynthesis Irradiance (P-I) Curve.
This method is the common and traditional way of
measuring photosynthesis. This is done by measuring
their photosynthesis evolved, which is one of the
byproducts of photosynthesis. This type of method is
very effective and sufficient. The second method is
performed using Pulse Amplitude Modulation (PAM)
Fluorometry. This method is the most recent and
updated way of measuring photosynthesis.

In this method, the photosynthesis on the seaweeds is measured through their chlorophyll
fluorescence. This is a promising method since it involves measuring the actual functional unit of
photosynthesis which are the photosystems. However, we should know the basic principles of the method to
fully understand their mechanisms.
 Before discussing the specific procedures on
the photosynthetic measurements, Dr. Saco first
introduced the audience to the process of sample
collection. These steps are said to be vital and very
necessary in the study in order to prevent putting your
seaweed specimens under too much stress. The first
step in sample collection is the site reconnaissance. In
this procedure, you are tasked to check the condition of
the area of the targeted species to be collected. The
speaker gave emphasis on checking the substrate,
topography and physicochemical parameters of the area.
The factors stated serve as a basis on how species would respond from their ambient or immediate
environment.

Next, photo documentation and taking the

coordinates of the area is done. These parts are
important in order to make sure that you are choosing
the perfect location in finding your specimens. Dr. Saco
also said that this step could be a big help in checking
the photosynthetic response of the species.

The third step is the actual collection of the

samples. Researchers are advised to use scrapers or
their hand in gathering the specimens. If the seaweeds
are strongly attached to rocks, it is safer to use the
scraper. Dr. Saco also suggested using one plastic bag
per species in collecting the samples. As much as
possible, start sorting the sample on the site since it will
be harder for you to sort them in the laboratory.

For transporting samples, seaweeds are usually

stored in an airtight container with ice packs and ambient
water. Other bigger seaweeds like Sargassum can also
be used to maintain the temperature of the inside.
Following these guidelines could help you in keeping
your samples under good condition.
 Dr. Saco then proceeded with discussing the
required steps in preparing the sample for processing.
Firstly, he stated that the samples should be rid of any
epiphytes and debris, using filtered seawater or water
from the source where the sample was obtained.
Secondly, Dr. Saco explained that the sample should be
placed in the filtered water, in a location away from direct
sunlight, as it could cause further stress to the samples.
And lastly, Dr. Saco instructed that a hole, 8 mm in
diameter, should be punched to prepare the sample for
experimentation to induce healing.

Dr. Saco explained that using the

Photosynthesis-Irradiance or P-I curve for short, is one
of the best ways to measure the photosynthetic
response of the sample. The graph suggests that to
obtain the photosynthetic response, data which
correlates the apparent photosynthesis (measured as
the y-axis) and level of irradiance (measured as the x-
axis) should be used. The graph would begin at the light
compensation point, lc (the point where the minimum
level of irradiance would start to induce photosynthesis),
and rise up continuously until a certain threshold is
reached which would then be symbolized as Pmax (the
maximum photosynthetic rate).
A complete graph would also provide further information such as: the onset of light saturation lk,
light saturation ls, and initial slope a, which tells the efficiency of lower and higher levels of irradiation.

Upon explaining the prerequisites for the

experiment, Dr. Saco explained the actual setup of
measuring photosynthesis oxygen evolution, and the
functions of each part. The setup includes a Magnetic
Stirrer, Sample chamber, Black box, Neutral density
filters, a functioning Water flow-through system, an
Oxygen electrode, a light guide, and of course, a light
source. The function of the Water flow-through system is
to regulate the temperature, the light source controls the
light level, the density filters manage the light intensity,
the metal stirrer continuously stirs to obtain the accurate
measure of oxygen, and the oxygen electrode measures
the amount of oxygen saturation.

Dr. Saco also stated the importance of the prerequisites as a decrease in medium volume (0.6 ml)
and a decrease in the thallus area, would overall shorten the time of the incubation period (3 mins)
 After explaining the experiment procedures, Dr.
Saco then explained how the data obtained would be
processed. He also pointed out that in order to achieve
accurate data, changing the seawater per cycle, and
allowing the sample to respirate is necessary to ensure
that every cycle after the last one only varies with the
difference in light level provided. Dr. Saco elaborated
that in order to obtain the average respiration rate of the
sample, you need to get the slope intercept of each cycle
at 0% light level and gather the mean of the obtained
data. Similarly, to obtain the photosynthetic rate, you
need to measure the slope, but this time, measure the
ones generated at every specific light level.

Dr. Saco then proceed with the full discussion of

the second method in photosynthesis measurement. He
stated that the basic principle of PAM Fluorometry
technique is similar to signal modulation measurements
wherein the measurements is based on changes
amplitude driven by signal using light impulses through
time. In the graph, the y-axis is the fluorescence and x-
axis is time. So how does it work? On the far-right lower
side shows the PSII activity during the process. At dark
adapted state a weak measuring light releases
fluorescence, at this point, the fluorescence is still at
minimum level, but after some time when a very strong
and short pulse of saturating light has given and the
fluorescence level increases.

At this point, all reaction center are close, with this we can measure the maximum capacity of PSII.
At light adapted state, when the activating light is given the reaction centers are active and undergoes
photosynthesis. After some time, the fluorescence on the activating light decreases, but this time, when
another very strong short pulse is given the fluorescence level increases, from here we can measure the
efficiency of the light absorbed by PSII to be used for photosynthesis. When light is turned off, the
fluorescence decreases and the reaction center is open which gives of excess energy as heat, at this point,
we can measure the non – photochemical quenching coefficient.

Dr. Saco then showed the setup for PAM

Fluorometry technique, basically it is almost the same
setup as the P-I curve but without the presence of a
black-box. To keep it simple, he stated that the chamber
contains the seaweed sample with desired salinity of
water, then the fluorometer measures/controls different
sources of light (PSII and PSI activities carry out by
primary photochemistry of photosynthesis). The water
bath controls the water temperature. The stirring plate
performs continuous stirring to accurately measure the
photosynthetic rate.
 Dr. Saco showed one of his actual
measurement with one of his experiments. On the y-axis
is the fluorescence rate and on the x-axis is time in
seconds “s”. During the measurement, the measuring
light “ML” induces the fluorescence without the presence
of photosynthesis. Then the saturating light pulse “SP”
forces the photosynthetic process, thus, we can measure
photosynthetic reactions. Then the presence of active
light “AL” is the indication that the photosynthetic rate is
very active. Then there is also the far-red light “FR”, it is
the light source that relaxes the photosystem after active
photosynthesis
He also stated the important parameters in measuring PAM fluorometry response, from the picture
there is a “dark adapted state” & “light adapted state”. It is very important to have a “dark adapted state” to
know what specific conditions best fit the seaweed samples, from here we can measure the maximum
quantum of PSII. On the other hand, the “light adapted state”, from here we can measure the efficiency of
absorbed light by PSII. He added that the measurement parameters are highly dependent on what type of
seaweed species are to be used, so it is important to be specific.

Dr. Saco showed an actual representation of

the data using PAM fluorometry. To sum it up, the
growth and productivity of seaweeds are less active in
cold season and vice versa. The quantum yield
increases as the temperature rises. Lastly, Dr. Saco
shared a basic approach in studying seaweed which is
the “Ecophysiological technique”. This technique tackles
about on the effects of physiochemical parameters of
physiology of seaweeds and effects of biotic factors.

After discussion, the speaker was given a few questions and was able to answer all and satisfy the
audience with his answers. Dr. Saco was given a proper gratitude for his participation and a farewell by the
BioMass students.