A DISSERTATION ENTITLED

SEED DIVERSITY STUDY AND GERMINATION OF SIX MEDICINAL

PLANTS.

FOR THE PARTIAL OF THE AWARD OF DEGREE OF MASTERS OF SCIENCE IN

BOTANY

SAVITRIBAI PHULE PUNE UNIVERSITY

SUBMITTED BY
MS. JADHAV ASHWINI LAHU
MSc Botany

UNDER THE GUIDANCE OF

Dr. D. N. MOKAT
PROFESSOR

DEPARTMENT OF BOTANY
SAVITRIBAI PHULE PUNE UNIVERSITY
PUNE-411007

APRIL 2022

1
 DECLARATION

I hereby declare that the project entiteled “SEED DIVERSITY STUDY AND
GERMINATION OF SIX MEDICINAL PLANT.” submitted to Savitribai Phule Pune
University by me for the partial fulfillment towords degree of Masters in Science [ Botany ].

Place : Pune
Date :

STUDENT
Ms. Jadhav Ashwini L.
Department of Botany ,
Savitribai Phule Pune University,
Pune.

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 ACKNOWLEDGEMENT

I would like to express my sincere gratitude to my guide Dr. D. N. Mokat, Department of

Botany Savitribai Phule Pune University, Pune for his dedicated efforts, intellectual guidance,
valuable advice in conducting project and preparation of this project report.
I am very thankful to Dr. N. Ade , Prof. & Head of department of Botany, Savitribai Phule
Pune University,for giving me an opportunity to work in the department of Botany,
University of Pune and for providing necessary laboratory facilities and encouragement.
I am very thankful to Miss. Tai Kharat for giving valuable guidance, encouragement and kind
suggestion during this project work.
I also thank my lab mate, Mr. Amit Mirje for his help during this project work.
I express my sincere gratitude to my parents for their ethical, moral and financial support.
Thank you for everything, Thank you everyone!

Miss. Jadhav Ashwini Lahu

M.Sc. II

3
 Content

Sr No. Chapters Page No.

i Certificate 2

ii Acknowledgement 4

1. Introduction 7

2. Review and literature 17

3. Material and Methods 20

4. Results and discussions 23

5. Conclusions 32

6. Reference 33

4
 Abbreviations Table
% Percentage
& And
°C Degree Celsius
µl microliter
APG Angiosperm Phylogeny Group
Cm Centrimeter
Conc. Concentration
Dia Diameter
DW Distilled water
et al. Et alia
GA3 Gibbrelic acid
gm Gram
H2O2 Hydrogen peroxide
Hrs Hours
i.e that is
Kno3 Potassium nitrate
Min Minutes
ml Mililitre
mm Millimetre
ppm Parts Per million
SPPU Savitribai Phule Pune University
temp Temperature
viz. namely
MS Murashige and Skoog Medium
IAA Indole acetic acid

5
 CHAPTER 1
Introduction
In the medicinal plant world, there is a lot of research on conservation and long-term use. A
medicinal plant that includes chemicals that can be utilized for medical uses, they are
substrates for the manufacture of effective pharmaceuticals from its one or more of its
components. Long before recorded history, plants are used for medical purposes. Food,
clothes, shelter, and medicine are all provided by plants. Traditional medicine has long relied
on medicinal plants.
In developing countries like India several plants based medicines also known as herbal
medicine are used. Ayurveda represents the oldest traditional medicines in the world, which
is recently getting an increasing scientific interest. Medicinal plants are used to treat many
different types of diseases in our daily lives. The validity of many herbal products has been
recognized by research in medicinal plants. Medicinal plants have been used in healthcare.
Global research on their efficacy has been done, with some of the findings leading to the
development of plant-based medicines. Traditional medicines, modern therapies, health
supplements, food supplements, herbal remedies, pharmaceutical intermediates and chemical
organization for synthetic drugs can all be identified in medicinal plants.
Different parts of the plant are used in Ayurveda systems of medicine for prevention and cure
of many illnesses and everyday ailments like common cold, headache, cough, cold, fever,
colic pain, sore throat, bronchitis, asthma, hepatic diseases, malarial fever, as an antidote for
snake bite and scorpion bite, headaches, night blindness, diarrhoea, skin diseases, tiredness
wound, arthritis, digestive disorders.
The aim of conservation is to support sustainable progress by protecting and using biological
sources in ways that do not reduce the world’s variety of genes and species or demolish
important habitats and ecosystems. In general, it involves activities as collection,
propagation, characterization, evaluation, disease indexing and elimination, storage and
distribution. The conservation of plant genetic resources is of two types, In-Situ & Ex-Situ
conservation.
Seed biology
Seed is an important cradle of plants life and keep the species in nature.

6
An embryonic plant is covered in a protective outer covering called a seed. They are product
of the ripened ovule, after fertilization by pollen and some growth within mother plant. The
embryo develops from zygote and seed coat from integuments of ovule.
The seeds are covered by the fruits. A true fruit is develops from only ovary but some fruits
are not form the ovary. False fruits are develops from some flowers part i.e. thalamus,
inflorescence and calyx, etc. Many plants seeds are used for their medicinal properties with or
without fruits.
Seed banks are a better way of storing the genetic diversity of many medicinal plants ex-situ
conservation. The botanic gardens are suggested to help conserve the biological and genetic
diversity of wild plant species.
The different type of seed shape viz., globose, globular, oblong, oval, obvoid, pyriform,
discoid, cuboid, cylindrical, oblong cylindrical, rounded, linear, biconvex, reniform, pinhed
– reniform, sectoroid, trapezoidal ellipsoid, rectangular, pyramidal, concavo-convex, plano-
convex, rhomboid, etc.
Germination:
Germination is the process of a seed, spore, or other reproductive body emerging after it has
been dormant for a period of time. Water is taken by the embryo during seed germination,
resulting in the rehydration and enlargement of the cells. Shortly after the beginning of water
uptake, the rate of respiration increases, and various metabolic processes, suspended or much
reduced during dormancy, restart.
Germination is considered complete when the radical (which becomes the primary root)
ruptures the root sheath and emerges from the seed. When the first true leaf develops, the
plant has developed into a "seedling". During germination, seedling growth and development
are completely reliant on the energy reserves stored in the seed.
Divide the number of healthy seedlings by the total number of seeds in the test and multiply
by 100 to get the germination percentage. The germination index (GI) is a measurement of
germination percentage and speed.
Seed Germination in Plants: The Most Important Factors (Internal and External
Factors):
 Water- Germination is impossible unless and until the seed is given access to an
external source of water.
 Oxygen
 Temperature

7
  Light.
Seed Dormancy:
i. Dormancy is the inability of seeds to germinate due to a lack of suitable conditions.

ii. Even in the availability of favourable conditions, a physical or physiological state of

viable seed that prevents germination Cause of seed dormancy:
1. Physiological
2. Undeveloped cotyledon
3. Immature embryo
4. Embryo dormancy
Methods of Breaking Seed Dormancy:
A. Scarification:
B. Temperature Treatments
C. Light Treatments
D. Treatments with growth regulators & other Chemicals.
The studied six medicinal plants are as follows:
Holoptelea integrifolia (Roxb.) Planch. Argyreia nervosa (Burm.f.) Bojer, Randia
dumetorum (Retz.) Poir. , Gymnema sylvestre (Retz.) R.Br. ex Sm., Abutilon indicum (L.)
Sweet, and Stereospermum chelonoides (L.f.) DC.
Scientific classification as per APG system:
Holoptelea Argyreia Randia Gymnema Abutilo Stereospermum
integrifolia (R nervosa dumetorum (R sylvestre (Retz n chelonoides (L.f.)
oxb.) Planch. (Burm.f.) etz.) Poir. .) R.Br. ex Sm. indicum (L.) DC.
Sweet
Bojer
Kingdom Plantae Plantae Plantae Plantae Plantae Plantae
Clade Angiosperms Angiosperm Angiosperms Angiosperms Angiosperm Angiosperms
Clade Eudicot Eudicot Eudicot Eudicot Eudicot Eudicot
Clade Rosids Asterids Asterids Asterids Rosids Asterids
Order Rosales Solanales Gentianales Gentianales Malvales Lamiales
Family Ulmaceae Convolvulaceae Rubiaceae Apocynaceae Malvaceae Bignoniaceae
Genus Holoptelea Argyreia Randia Gymnena Abutilon Stereospermum
Species H. integrifolia A.nervosa R. dumetorum G. sylvesre A. indicum S. chelonoides

8
Botanical description:
1) Holoptelea integrifolia (Roxb.) Planch.
Commom name - Indian Elm
Marathi - Vavla
Plant Description - Large deciduous trees, up to 25 m tall, with 6-8 mm thick whitish-
grey smooth bark, flame yellowish-grey with light brown band, and branchlets with
pubescence. Stipules are lateral and scarious, and the leaves are simple and alternating.
lateral nerves 6-9 pairs, pinnate, prominent, intercostae reticulate, prominent; lamina
ovate-oblong, apex acuminate, margin entire, distantly serrate when young, coriaceous,
glabrous above, appressed pubescent punctate below; petiole 5-10 Female flowers have
longer pedicels; ovary superior, compressed, 2-winged, 1-celled, ovule 1; style is bifid.
Flowers polygamous, appear before leaves, 5-8 mm across, greenish-purple; tepals 4 or 5,
free, anthers pubescent; Samara fruit, 3 cm diameter, orbicular, nerved wings, glabrous,
seed one.
Use - Holoptelea integrifolia Roxb. Indian Elm is a very valuable and well-known
traditional medicinal plant. Apart from the chemistry of plant components, significant
progress has been made in the biological activities and therapeutic applications of Indian
Elm over the last three decades.
2) Randia dumetorum (Retz.) Poir.
Synonym - Catunaregam spinosa (Thunb.) Tirveng.
Common name - Emetic nut tree
Marathi: Ghela, Wagatta, Gelphal
A large deciduous tree. Simple, obovate, wrinkled, glossy, pubescent leaves. Flowers
white, fragrant, solitary, on at the end of short branches. Fruits globose, berries with
longitudinal ribs, yellow when ripe. The seeds are numerous, compressed, and embedded
in a dark foul smelling pulp. Fruit is 1.8-4.5cm long, globose, crowned with persistent
calyx-limb, fruit contains numerous seeds, 0.4-0.6 cm long, compressed, smooth, brown
and very hard
Use - Fruit cures abscess, ulcers, inflammation, wounds, tumours, skin diseases and have
antibacterial activity. The pulp of fruit is believed by many practitioners to also have
anthelmintic properties and also used as an aborticide. Plants are used as a traditional
medicine in Ayurveda and fruits used in medicine as well as in food.
3) Argyreia nervosa (Burm.f.) Bojer

9
 An extensive woody climber, young shoots and branches covered by a silky white
pubescence. Leaves ovate-cordate, apiculate at tip, 7.5-30 cm in diam., glabrous
above, white tomentose beneath. Flowers rose-purple with 7.5-30 cm long white-
tomentose peduncles, born in axillary many-flowered cymes; bracts ovate-lanceolate,
acuminate, 3-4 cm long. Fruits globose, 2 cm in diam., apiculate, indehiscent.
Use – This plant has been studied pharmacologically for nootropic, aphrodisiac,
immunomodulatory antimicrobial, antioxidant, anti-inflammatory, anti-diarrheal anti-
hyperglycemic, hepatoprotective, antiviral, nematicidal, antiulcer, anticonvulsant,
analgesic, and central nervous system depressant properties.
4) Gymnema sylvestre (Retz.) R.Br. ex Sm.
Common name: Gurmar, Cowplant
Marathi: Bedakicha pala,
Gymnema is a gregarious woody climber with many branches that extends across the
top of large trees. The pubescence is present on young stems and branches.
Leaves are 3–5 cm long and up to 3 cm wide, ovate-elliptic, acute or shortly
acuminate, pubescent on both sides, with 6–13 mm long pubescent petioles; base
rounded or heart shaped. Umbellate cyme inflorescences produce flowers. The cup is
pubescent and nearly divided y to the base. Yellow corolla with thick, oval, and
recurved lobes, tube campanulate. Follicles are hard, lanceolate, and attenuated into a
beak, ranging up to 7.5 cm long and 1 cm wide.
Seeds are 1.3 cm long, narrowly ovoid–oblong, flat, and have a thin, broad, brown,
glabrous marginal wing.
Use - Gurmar is antiperiodic, astringent, hypoglycaemic, diuretic, stomachic, tonic,
and refrigerant. It is known to balance the excess sugar in the body and is utilized as a
glycosuria killer. It's also used to treatment for a variety of other urinary problems.
5) Abutilon indicum (L.) Sweet
Common name: Indian Mallow
Marathi: Petari
Indian Mallow is a velvety-pubescent shrub with circular-ovate or heart-shaped leaves
with roughly crenate-serrate, erect. The plant can grow to be 1-2 m tall. The leaves are
arranged alternately and have long stalks with velvety, delicate, pale hairs. Solitary
orange-yellow blooms with a diameter of 2-3 cm bloom in axils on long stalks of 4-7 cm.
The orange-yellow petals are triangular-obovate, 1 cm long or somewhat longer, and have

10
 stellate hairs on the staminal tube. The fruit is unusual in that it is circular in shape, with
11-20 radiating hairy carpels that are brown when dry, and each carpel is flattened and
boat shaped. Seeds are kidney shape. The plant is a weed that grows in disturbed areas.
Use- Extract of water-soaked dried seeds is used as purgative. Leaves are used as tonic.
Fever is treated with a root infusion.
It is useful in gout analgesic bleeding disorders, tuberculosis, ulcers and worms. It can be
used as Digestive, laxative, expectorant, diuretic, astringent, anti inflammatory,
anthelmintic and, healing.
6) Stereospermum chelonoides (L.f.) DC.
Synonyms: Bignonia chelonoides, Bignonia suaveolens, Stereospermum suaveolens
Common name: Fragrant Padri Tree
Marathi: Kalagori, Paadal
Deciduous trees up to 25 meters tall, with grey bark that exfoliates in flat scales.
Compound, imparipinnate, opposite, and estipulate leaves; Leaflets 5-11, opposite;
petiolules 10-20 mm, slender, pubescent; lamina 7.5-16 x 5-7.5 cm, elliptic or oval; base
oblique, acute or unequally rounded; rachis 150-450 mm long, pubescent, slender,
swollen at base; lateral nerves 6-8 pairs, pinnate, prominent, intercostae reticulate; apex
acuminate; margin entire, shiny and glabrous above and pubescent beneath, coriaceous;
apex acuminate; Flowers 18 mm long, bisexual, drooping panicles, pubescent; calyx
campanulate, hairy, lobes 3-5, short, broad; corolla 3-4 cm long, funnel shaped, pubescent
outside, lobes 5 unequal, rounded, crisped; stamens 4; didynamous included ststaminodes.
Ovary sessile, superior, 2-celled, syncarpous; ovules many; style slender; anther
divaricate; disc cupular, fleshy; stigma 2. seeds 3 cm long with a transverse grove,
funnel shaped, with a membranous wing with obtuse ends.
Use - indigestion, hiccups, vomiting, constipation, pain, fever, diabetes, liver problems,
and asthma. Skin wounds treated with Stereospermum. Stereospermum may help in
controlling blood sugar and cholesterol levels, as well as relieve pain, swelling, and
fevers, as well as protect the liver from toxins. It could help protect the brain from harm
caused by decreased blood flow or diseases like Parkinson's. Stereospermum seems to be
an antioxidant.

11
 Photo plate I
Plants

Holoptelea integrifolia (Roxb.) Planch. Abutilon indicum (L.) Sweet

12
Argyreia nervosa (Burm.f.) Bojer Gymnema sylvestre (Retz.) R.Br. ex Sm.

Stereospermum chelonoides (L.f.) DC Randia dumetorum (Retz.) Poir.

13
 Photo plate II
Fruits

Holoptelea integrifolia (Roxb.) Planch. Abutilon indicum (L.) Sweet

Argyreia nervosa (Burm.f.) Bojer Gymnema sylvestre (Retz.) R.Br. ex Sm.

Stereospermum chelonoides (L.f.) DC Randia dumetorum (Retz.) Poir.

14
 Photo plate III
Seeds

Holoptelea integrifolia (Roxb.) Planch. Abutilon indicum (L.) Sweet

Argyreia nervosa (Burm.f.) Bojer Gymnema sylvestre (Retz.) R.Br. ex Sm.

Stereospermum chelonoides (L.f.) DC Randia dumetorum (Retz.) Poir.

15
 Objectives

1. Study of morphology of six medicinal plants.

2. The objective of seed morphology is to provide a short description, based in
qualitative aspects. Fruits were studied carefully for exomorphic characteristic such as
morphometry, type, seed shape, size, colour and seed weight.
3. Study of seed biology with respect to seed type, viability, dormancy and germination
potential

16
 CHAPTER 2
Review of Literature

The review of literature has theoretical as well as practical significance in better

understanding of the previous research work and helpful to make effective work plans on
different aspects. The past work related to present investigation is briefly reviewed in this
chapter-
Several workers worked on medicinal plant and floristic study of Pune, Sanatapau (1993)
carried out “The flora of Khandala on the western ghats of India”. Kumar et al., (2013)
reported medicinal plant used in traditional health care practice in Bundelkhand region of
central India. They reported 100 plant species belongs to 43 genera and 50 families. Seed
morphology also has potential to be assessed as information that can lead to understanding of
certain characters that may be important in terms of phylogenetic analyses (Hufford, 1995).
Genetic diversity consists one of the main features which permits species to evolve and
hence, adapt to changing environments. Hence, a high level of genetic diversity in produced
seeds is necessary for the formation of adaptive future stands, for the ability to cope with the
challenges of global climate change, and also for defence against pollution and new
pathogens (Pakull et al., 2021).
In desert areas, it is well known that there is a massive consumption of seeds (Gutterman,
1994). Most studies have been essentially concerned with annual plant seeds (Morton, 1985;
Boeken & Gutterman, 1990), and studies of Cactus seeds have been carried out on some
Opuntia spp., mainly in the Chihuahuan Desert ‘Nopaleras’ (Janzen, 1971, 1986; Grenot &
Serrano et al.,1986).
The appearance of testa, tegmen, endosperm and cotyledons also constitute important
observations to be made while studying seed morphology. Pits and stipples may be seen over
the seed surface. Hilum and chalaza may also be seen. Size and surface features of seeds can
even vary widely even within a single sample (Barthet and Daun, 2011).
(H. Blum et al., 2006) The intention of the study is to improve the seed quality of medicinal
plants and herbs by using strategies with high practical applicability. They studied the seed-
treatment by physical and biological methods. They include Hot water treatment, electron
treatment and the vacuum-steam-treatment. A significant reduction of the infestation of
Mycocentrospora acerina on annual caraway Carum carvi var. annum was shown with a

17
temperature of 50 °C for 20 minutes (Reduction from 27 % to 0,7 %) temperature of 50 °C
for 10 minutes. They observe positive effects in the field trials. Seed treatment with low-
energy electrons Degree of efficiency tended was lower than with hot water treatment. Their
First positive results with Vacuum-Steam-Treatment of coriander were made. The
germination rate was certainly reduced massively by the treatment (especially with 80 °C / 1
min).
Plant propagation through seeds is the best method. There are many reports that seed with
physical dormancy can germinate through physical or chemical pre-treatments such as heat,
hot water, cold water, freezing treatments, GA3, etc. (Denton et al., 2013).
Mathew et al., (2011) investigated that the germination percentages of seeds of Ficus
racemosa, F. microcarpa, F. Religiosa and F. benghalensisare to be 5.0, 2.3, 27.7 and 82.0%,
respectively.
The highest germination rate was 2.94% was reported with GA3 200ppm pre-treatment
recorded by Madhushree et al., (2018). Srivastava et al., (2011), reported that the seeds of
Aconitum heterophyllum treated with IAA 500 ppm recorded high seed germination
(97.17%). Syzygium cuminii seeds of six treatments - control, water soaking for 24 hrs, hot
water (100ºC) for 5 sec, GA3 at 200 mg l-l for 10 min, KNO3 at 0.5 % for 10 min and thiourea
at 0.5 % for 10 min. seeds soaked in water for 24 hrs was found best seed germination
percentage (95.73 %) As compared to other treatments seeds soaked in water for 24 hrs was
found best seed germination percentage (95.73 %) was reported by Patil et al., (2018).
Andrographis paniculata seeds was pre-treated with 200 ppm GA 3 improved seed
germination (89%) reported by Sedeghi et al., (2010).
Khan et. al. [2011] reported that 70 to 90 % inhibition rate with treatment of 75% and 100%
wastewater on germination of wheat and rice.
Trivedi et al., (2014) reported that the Stereospemum suaveolens propagated through seeds
but this germination rate is very poor. This is very valuable medicinal species , now this
placed in threatened category. Wetness of one year old seed is high i.e.(15+-1.3) and current
year seed were low i.e. (81+-0.3) the fresh seed soaked in water before treatment. The study
were carried out with three treatments viz. Soil, coco peat and MS medium. They were
observed good germination rate in coco peat (68%), MS medium (66%) and low in soil (6%).
Awasti et al., (2015) worked on study of Steospermum tetragonum. They gave three
treatments on seeds with replications. They conclude that S. Tetragonum has very poor rate of

18
germination. The better result observed in the treatment of Distilled water other than Cow
dung slurry and Normal water.
Trivedi and Joshi (2015) reported that the cotyledonary node and nodal explants
Stereospermum suaveolens were cultured on WPM supplemented with individual BAP M,
Kn and TDZ. When micro-shoots were transferred to half strength WPM liquid medium,
rooting (100%) was observed.
Sutheesh et al., (2016) were studied on the influence of various organic treatments on
germination and seedling vigour in Sandal. They give 16 treatments to seeds which include
soaking in tap and hot water, acid scarification, GA 3, cow dung slurry and cow urine in diff.
conc. and duration. They obtained highest result in GA 3 (74.33) and lowest (44.33) H2SO4.
and also they observed that acid scarification and boiling water treatments decreased
germination percentage of the seed than control.

19
 CHAPTER 3
MATERIALS AND METHODS
The details of material used and method followed in the present investigation are described in
this chapter under the following subhead -
Collection of plant material:
Holoptelea integrifolia (Roxb.) Planch,. is collected from Dhamni Tal-Aambegaon , Dist-
Pune. Randia dumetorum, Argyreia nervosa, Abutilon indicum, Stereospermum
chelonoides collected from RCFC, of SPPU, Pune and Gymnema sylvestre collected from
botanical garden of botany department SPPU, Pune.
Identification:
Plants were identified by using Floras. The corrected botanical name and authentication were
reconfirmed by website like www.theplantlist.com, https://powo.science.kew.org/.
Material used for study –
1) Camera
2) Secateurs
3) Collection Bags
4) Field Notebook
5) Weighing Balance
6) Petri Plate
7) Beakers
8) Conical Flask
9) Filter Paper
10) Cotton
11) Alcohol
12) GA3 Solution – 100ppm, 200ppm, 300ppm,
13) H2O2
14) DW
15) Bavistin
16) KNO3
Seed germination
The fresh and matured seeds were collected. The dried seeds were stored in the
polythene zip pouch at room temperature. These seeds were used for the different

20
germination experiments. The effect of various seed pre-treatments were examined in
laboratory condition.
Propagation study of plants were completed by considering following parameters and
observation like fruit length, seed length, number of seed per fruit, number of seed in 100 gm,
germination percentages of seeds, plumule length with the help of thread and scale.
Germination test in vitro conditions-
Sterilization of the seeds:
Healthy seeds were washed for 10 min. under running tap water to remove any
adherent particles. The seeds were kept in 1% Bavistin solution, a broad spectrum fungicide,
for 10 min. and then washed in 5% Teepol, a liquid detergent for 5 min. The treated seeds
were then washed with distilled water for seven times to remove the traces of chemicals.
Then the seeds were dried using blotting paper. Seed germination experiments were carried
out in glass petriplates (100 x 17 mm). Each petri-plate was laden with double layer of
absorbent cotton at bottom and single layer of germinating paper moisturized with distilled
water. The 6 pre-treatments were selected by using previous literature data.
Pretreatments and germination evaluation:
The seed germination of each plants 10 seeds on each Petri plate within three replicates (90
seeds) with 6 pretreatments i.e. cold water, warm water, GA3 in different ppm, H2O2 in
different conc. and KNO3 different conc. for 24 Hrs for selected plant seeds. Petri-plates were
placed in normal room temperature at 27 °C. Moist condition was maintained by addition of
sterilized distilled water.
The different pretreatment of seeds are as follows:
1) Holoptelea integrifolia seeds were pretreated with GA3 100 PPM, 200 PPM, 300
PPM, hot water (40ºC) and cold water (27 °C) for 24 hrs.
2) Randia dumetorum seeds were pretreated with GA3 100 PPM, 200 PPM, 300 PPM,
hot water (40ºC) and cold water (27 °C) for 24 hrs..
3) Gymnema sylvestre seeds were pretreated with KNO3 0.2 %, 0.5%, hot water (40ºC)
and cold water (27 °C), GA3 100 PPM for 24 hrs.
4) Argyreia nervosa seeds were pre-treated with 0.5 µl and 1 µl H2O2 in 100 ml DW.
GA3 100 ppm, hot water (40ºC) and cold water (27 °C) treatments for 24 hrs of seeds.
5) Abutilon indicum seeds were treated with hot water and cold water (27°C) for 24 hrs.
6) Stereospermum chelonoides seeds treated with 30ºC, 40ºC, 50ºC, 60ºC, 70ºC temp.
of hot water for 24 hrs.

21
Preparation of GA3 Solutions:
Preparation of 100PPM GA3 Solution:
0.01 gm GA3 was weighed on weighing balance and then ethanol was added for dissolving
the GA3 and then 100 ml of water was added.
Preparation of 200 PPM GA3 Solution:
0.02 gm GA3 was weighed on weighing balance and then ethanol was added for dissolving
the GA3 and then 100 ml of water was added.
Preparation of 300 PPM GA3 Solution:
0.02 gm GA3 was weight on weighing balance and then ethanol was added dissolving the
GA3 and then 100 ml of water was added.
H2O2 treatment:
Preparation of 0.5µl: 0.5 µl H2O2 was added to 100 ml of water.
Preparation of 1 µl: 1 µl H2O2 was added to 100 ml of water.
The surface sterilized seeds of selected plant were immersed in 0.5µl H2O2 and 1 µl H2O2.
KNO3 pre-treatment
Preparation of KNO3 Solutions for
Preparation of 0.2% KNO3 solution: 0.02 gm KNO3 was added in 9.98ml of water
Preparation of 0.5% KNO3 solution: 0.05 gm KNO3 was added in 9.95 ml of water.
The surface sterilized seeds of selected plant were immersed in 0.2% KNO3 and 0.5% KNO3.
Control:
Seeds without pre-treatments were considered as control.
Method:
Selected seed source was placed in a Petri dish covered with filter paper. 90 seed (large
seeds) were counted and set to particular treatment. Selected seeds were spread in the filter
paper of the dish. Paper was moisten with enough water so that the seeds just barely float
(about 5ml). The dish was placed in dark. The number of seeds that germinate daily was
recorded.

22
 CHAPTER 4
Results and Discussions
Seed morphology is study on commercially medicinal important plants. To measure the fruit
type, fruit size, seed shape, length, colour, number of seed in 100gm respectively and
following result are observed.
Holoptelea integrifolia fruit size 2.5-3.5cm, fruit type samara, seed shape kidney, seed size
0.5-1.2cm no of seeds in 100 gm is 3000. Argyreia nervosa fruit size 1.2-2cm, berry, ovoid,
0.2-1cm, 1480. Randia dumetorum 1.8-4.5, berry 0.4-0.6cm, reniform, 3780. Gymnema
sylvestre 4-7cm, pod, ovoid, 1.3cm, 11700. Abutilon indicum 3-5cm, schizocarp, reniform,
0.2-0.8cm, 43000. Stereospermum chelonoides 35-40cm, legume, winged, 2-4cm, 4350.

Observation and observation table

Sr. Name of plant Fruit Fruit No of Seed Seed Seed No. Of Floweri
No size type seeds in Shape size colour seed in ng and
. 100gm 100gm fruiting
fruits
1 Holoptelea 2.5- Samara, 1490 Kidney 0.5- Whitish 3000 Februar
integrifolia (R 3.5 Dry fruit 1.2cm y -
oxb.) Planch. cm March
2 Argyreia 1.2cm Berry, 536 Ovoid – 0.2- Brown 1480 Decem
nervosa Globose concavo 1cm ber-
(Burm.f.) convex June
Bojer
3 Randia 1.8- berry 317 Reniform 0.4- Brown 3780 Decem
dumetorum ( 4.5cm 0.6cm ber -
Retz.) Poir. April
4 Gymnema 4 Pod 9600 Ovoid- 1.3cm Brown 11700 March-
sylvestre (Ret -7 obovoid June
z.) R.Br. ex
Sm
5 Abutilo 3- Schizoca 37000 Reniform 0.2- Black 43000 Septem
n indicum (L.) 5cm rp 0.8cm or dark ber -

23
 Sweet brown April
6 Stereospermu 35-40 Simple 750 Rectangul 2-4cm Whitish 4350 June-
m cm dry ar winged brown Septem
chelonoides ( dehiscent ber
L.f.) DC legume

In order to germinate the large no of seedlings of plants in short period of time, studied on
different pretreatment. Based on present study following result can be made:

Seed germination of Holoptelea integrifolia (Roxb.) Planch.:

Seed germination of H. integrifolia has 86.66% with GA3 200 PPM . GA3 100 and GA200
PPM has 84.44% germination, Hot water has 70% germination, cold water has 51.11%
germination, control has 21.11%. (Table no 1 and fig. no 1).
Table no 1 showing seed germination of holoptelea interifolia.
Sr. Treatments Total no of Germination Radical Plumule Root
No seed percentage (%) length length Shoot
germinated ratio
1 Hot water 63 70 4 7 0.5714
2 Cold water 46 51.11 4.2 6.8 0.6176
3 GA 100PPM 76 84.44 5.1 7.2 0.7083
4 GA 200PPM 78 86.66 5.1 6.9 0.7391
5 GA 300PPM 76 84.44 5.3 5.8 0.9137
6 control 19 21.11 2.3 2.6 0.8846

100 Seed Germination

90
80
70
60
Germination %

50
40
30
20
10
0
er er l
at at pm pm pm tro
W W 00p 00p 00p Co
n
t ld 1 2 3
Ho Co GA GA GA
Treatments

Figure 1 Showing seed germination of H. integrifolia

Seed germination of Stereospermum chelonoides (L.f.) DC.

24
S. chelonoides has 68.88 % germination at 40 ºC. At 30 ºC, 40 ºC, 50 ºC, 60 ºC, 70 ºC
germination was 43.33%, 68.88%, 47.77%, 57.77%, 53.33%, 53.33%, 43.33% respectively
(Table no 2 and fig. no 2).
Table no 2 showing seed germination of S. chelonoides.
Sr. Treatment Total no of Germination Radical Plumule Root
No s seed percentage (%) length length Shoot
Hot water germinated (cm) (cm) ratio
1 30 ºC 39 43.33 4.14 5.8 0.7137
2 40 ºC 62 68.88 6 6.1 0.9836
3 50 ºC 43 47.77 4 4.9 0.8163
4 60 ºC 52 57.77 4.9 4.29 1.1421
5 70 ºC 48 53.33 4.85 4.15 1.1686
6 control 39 43.33 4.84 5 0.968

80 Seed Germination
70
60
Germination %

50
40
30
20
10
0
30°C 40°C 50°C 60°C 70°C Control

Treatments

Figure 2 Showing germination% of S. chelonoides

Seed germination of Randia dumetorum (Retz.) Poir.

R. dumetorum has 100 % germination at GA3 100 and 300 PPM. GA3 200 PPM has 95.55
% germination, cold water has 96.66% germination, 86.66% germination with hot water
treatement.90% germination of control. (Table no 3 and fig. no 3).
Table no.3 showing seed germination of R. dumetorum.
Sr. Treatments Total no of Germination Radical Plumule Root
No seed percentage (%) length length Shoot
germinated ratio
1 Hot water 78 86.66 0.53 1.26 0.4206
2 Cold water 87 96.66 0.6 2.1 0.2857
3 GA 100PPM 90 100 0.48 2.15 0.2232
4 GA 200PPM 86 95.55 0.63 4.9 0.1285
5 GA 300PPM 90 100 0.84 3 0.28
6 control 81 90 0.46 2.8 0.1642

25
 105
Seed Germination
100

Germination %
95

90

85

80

75
Hot Water Cold Water GA 100ppm GA 200ppm GA 300ppm Control
Treatments

Figure 3 Showing germination % of R. dumetorum

Seed germination of Argyreia nervosa (Burm.f.) Bojer

A. nervosa has 30% germination with 0.5µl H2O2 . 1 µl H2O2 has 23.33 % germination, 19.99
% germination at hot water, 18.88% at GA3 100PPM , 4.44% germination of cold water, 0%
germination at control. (Table no 4 and fig. no 4).
Table no.4 showing seed germination of A. nervosa .
Sr. No Treatments Total no of Germination Radical Plumule Root
seed percentage length length Shoot
germinated (%) ratio
1 Hot water 18 19.99 1.6 2.8 0.5714
2 Cold water 4 4.44 2.2 3.5 0.6235
3 0.5µl H2O2 27 30 2.6 3.7 0.7027
4 1 µl H2O2 21 23.33 1.5 2.9 0.5172
5 GA 100PPM 17 18.88 1.4 3.1 0.4516
6 control 0 0 0 0 0
35
Seed Germination
30
25
Germination %

20
15
10
5
0
Hot Cold 0.5 μL 1 μL GA Control
Water Water H2O2 H2O2 100ppm
Treatments

Figure 4 germination % of A. nervosa

Seed germination of Gymnema sylvestre (Retz.) R.Br. ex Sm

G. sylvestre has 18.88 % germination at 0.2%KNO3.13.33% germination with 0.5%KNO3,

12.22% germination with hot water treatments, 6.66% germination with GA3 100PPM , 4.44

26
% germination at cold water treatments. In control 0% seed germination. (Table no 5 and fig.
no 5).

Table No. 5 showing seed germination of G. sylvestre .

Sr. Treatments Total no of Germination Radical Plumule Root

No seed percentage length length Shoot
germinated (%) ratio
1 Hot water 11 12.22 7.2 4.1 1.7560
2 Cold water 4 4.44 4.5 3.2 1.4062
3 GA 100PPM 6 6.66 6.3 4.6 0.1369
4 0.2% KNO3 17 18.88 7.1 3.9 1.8205
5 0.5% KNO3 12 13.33 6.9 5.4 1.277
6 control 0 0 0 0 0

Seed Germination
20
15
Germination %

10
5
0
... ... ... ... ... l
W W 10 2 % 5% n tro
t ld 0. 0. Co
Ho Co GA
Treatments

Figure 5 germination % of G. sylvestre

Seed germination of Abutilon indicum (L.) Sweet

A. indicum has 25.55% germination at hot water treatments. GA3 100 12.12 %
germination, GA200 14.14% germination, GA3 300 PPM 18.88 %
germination, 13.33% germination with cold water treatment, control with 0%.
(Table no 6 and fig. no 6).
Table No 6 showing seed germination of A. indicum.
Sr. Treatments Total no of Germination Radical Plumule Root Shoot
No seed percentage (%) length length ratio
germinated
1 Hot water 23 25.55 0.23 3.12 0.0737
2 Cold water 12 13.33 0.61 3.60 0.1694
3 GA 100PPM 11 12.22 0.63 2.42 0.2603
4 GA 200PPM 13 14.44 0.28 3.33 0.0840
5 GA 300PPM 17 18.88 0.19 2.63 0.0722
6 control 0 0 0 0 0

27
 30
Seed Germination
25
20

Germination %
15
10
5
0
er er l
at at pm pm pm ro
W W 0p 0p 0p ont
0 0 0 C
t ld 1 2 3
Ho Co GA Treatments
GA GA

Figure 6 Showing germination % of A. indicum

Photo plate IV

28
Holoptelea integrifolia

29
 Photo plate V
Randia dumetorum

30
 Photo plate VI
Stereospermum chelonoides

control 30 ºC

40 ºC 50 ºC

60 ºC 70 ºC

31
 Photo plate VII
Argyreia nervosa

32
 Photo plate VIII
Gymnema sylvestre

33
 CHAPTER 5
Conclusions

Holoptelea integrifolia plant fruit type samara, size 2.5-3.5cm and seeds kidney shaped, size
0.5-1.2cm. Total 3000 number of seeds in 100 gm.

Argyreia nervosa fruit berry, size 1.2-2cm and seeds ovoid, size 0.2-1cm as well as total 1480
number of seeds in 100 gm.

Randia dumetorum fruit berry, size 1.8-4.5; seeds reniform, size 0.4-0.6cm and total 3780
number of seeds in 100 gm.

Gymnema sylvestre pod size 4-7cm; seeds ovoid, 1.3cm and total number of seeds 11700 in
100 gm.

Abutilon indicum fruit schizocarp, size 3-5cm; seeds reniform, size 0.2-0.8cm, and total
number of seeds 43000 in 100 gm.

Stereospermum chelonoides fruit legume, size 35-40cm, seeds winged, size 2-4cm, and total
number of seeds 4350 in 100 gm.

In order to germinate the large no of seedlings of plants in short period of time, studied on
different pretreatment. Based on present study following conclusions can be made:

Seed germination of H. integrifolia has 86.66% with GA3 200 ppm so that concluded the
GA3 200 ppm treatment is best consequence on seed germination.

In A. nervosa has 30% germination with 0.5µl H2O2 i.e. 0.5µl H2O2 best germination
treatments.

R. dumetorum has 100 % germination at GA3 100 and 300 ppm. Result concluded the GA3
100 and 300 ppm most excellent treatments.

G. sylvestre (18.88 %) germination at 0.2%KNO3 and resulted that G. sylvestre seeds

germination at 0.2%KNO3.

S. chelonoides has (68.88 %) germination and A. indicum (25.55%) germination at 40 ºC hot

water treatment. So concluded that the most excellent outcome in 40 ºC hot water treatment of
seeds germinations of both plants.

34
 CHAPTER 6

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