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SEMINAR-IV
ON
BIOCHEMICAL AND MOLECULAR
MECHANISMS OF SEED DEVELOPMENT

PRESENTATION BY
GAJENDRA KHIDRAPURE
III Ph.D. Scholar, Dept. SST, UASR
(ID No. PHD12AGR2019)
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CONTENT
Introduction
Stages of seed development
Biochemical mechanisms
Molecular mechanisms
Integrated overview of seed development
Analysis of seed development through advanced techniques
Case studies
Conclusion
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Introduction

Parts of developing seed

Events/mechanisms of seed development

1. Fertilization and fusion of male and female gametes
2. Cell division
3. Cell enlargement
4. Dry matter accumulation
5. m RNA synthesis and accumulation
6. Synthesis and accumulation of starch, protein, lipids etc.
7. Increases in fresh and dry weight
8. Desiccation tolerance
9. Maturation and drying
10. Detachment/dispersion from mother plant
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(Desai et al., 1982)

Stages of seed development

(Jones, 1927)

Stage I: Histodifferentiation

Stage II: Cell expansion

Funiculus is the connection b/n the mother plant and ovule

Stage III Maturation drying

(Jones, 1927)

A- All the seed parts are

differentiated. Scutellum acts as a
conduct for exchange of storage
food reserves b/n the endosperm
and embryo

B- Cotyledons and distinct shoot

and root meristem is fully formed
Hence, suspensor and the basal
cell begin to disappears
B

Mature stage of monocot (A) and dicot (B) seed

10

(Black, 1994 )

Molecular and biochemical mechanisms

Molecular mechanisms (Expression of specific gene)

12

(Goldberg et al., 1989)

Trait specific gene expression during seed development

13

(Goldberg et al., 1989)

Mechanism of protein synthesis under the control of gene

during seed development
14

(Goldberg et al., 1989)

Photosynthetic products are the primary source for further

operation of biosynthetic pathways
(Goldberg et al., 1989)
15

Conversion and utilization of accumulates in developing seeds

16

(Goldberg et al., 1989)

Accumulation of storage protein, carbohydrates and FA

17

(Goldberg et al., 1989)

Average (%) composition: Major storage tissue

18

(Black, 1994 )

Changes in nutrition:
Cereals low in lysine
Legumes low in sulphur amino acids
Oil seeds (degree of unsaturated oil content)

New compounds; detergents and lubricants

Golden rice and beta-carotene

The cell expansion stage ends when the seeds reach physiological
maturity; maximum dry weight
(www. plant physiol., 2014)
19

20

An integrated overview of seed development in Arabidopsis

A-Structure of
the seed at 4
DAF
H

I
J

Fig. Different changes during seed development observed

under microscopy

B-Triangular
stage at 5 DAF
C & D- Early &
Late heart stage
at 5 & 6 DAF
E- Torpedo
stage at 7 DAF
F- Upturned-U
stage at 8 DAF

G- Mature cotyledon stage at 10 DAF; H- Formation of integument

at 10 DAF
21
I-

Excised stained embryo at 10 DAF;

J- KI staining (Sebastien
of starchet after
al., 2002)

-Dry weight
-Fresh weight
-Water content

Fig. Growth parameters of developing seeds

E.E.M.- Early Embryo Morphogenesis
22

(Sebastien et al., 2002)

23

Major physiological, biochemical and

molecular mechanism

Fig. Nutrient status in the seed and major physiological,

biochemical and molecular events affecting seed development.
(Sebastien et al. 2002)

Significance of pollen tube growth for seed development in Arabidopsis

A- Germinating pollen on stigmatic surface

B- Growth of pollen tube
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Fig. Phenotypic analysis

(Singh et al., 2002 )

DNA and RNA Synthesis during the cell expansion phase of

cotyledon development in Vicia faba L.

25

(Adele and Whitfeld, 1973)

26

(Adele and Whitfeld, 1973)

Carbohydrate metabolism in the developing and maturing wheat seeds

27

Fig. Patterns of carbohydrates () accumulation in

developing and maturing seeds
(Black et al., 1996)

Protein synthesis and accumulation in developing

endosperm of wheat
Membranes were isolated from whole
cell extracts of endosperm and
incubated with 5 mM ADP-glucose
buffer for 1 min at 25 C.
Membrane proteins were separated by
10% SDS-PAGE Each lane was loaded
with 25 mg protein and Stain the
protein.
Photolabelled protein visualized by
autoradiography.

Fig. Changes in protein content during wheat endosperm

development
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(Emes et al., 2003)

Mechanisms associated with biochemical changes during

seed development in Pines
Vanildo, S., Tiago, S., Balbuena1, C., Santa C., Eny, I., Floh, M.,
Guerra and Walter, H., 2004

Plant Growth Regulation, 44: 147156.

29

(Vanildo et al., 2004)

IAA increased continually

from globular stage reaching
maximum at
cotyledonary
stage whereas lowest in mature
seed

Fig. IAA and ABA levels in

megagametophytes
containing
developing embryos during seed
development

Highest ABA content occurred

at globular stage, followed by
a continuous reduction until
stabilization at the precotyledonary stage whereas
lowest in mature seed

(1) Globular (2) torpedo (3) pre-cotyledonary (4) cotyledonary (5) mature
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(Vanildo et al., 2004)

IAA and tryptophan is considered as the main precursor for

biosynthesis of different hormones, enzymes and storage materials at
initial phase of seed development i.e., during establishment of embryo
Endogenous ABA plays a role in the stimulation of specific storage
proteins
Probably, the increase in starch content during seed development was
due to triacylglycerols breakdown in the megagametophytes and
subsequent carbohydrate metabolism

31

(Vanildo et al., 2004)

Protein and starch content

progressively
increased
during seed development,
reaching their maximum
values at the mature stage
The increase in DM during
seed development is a
result of the synthesis and
deposition of storage
substances
Fig. Protein, starch and DM percentage in
megagametophytes containing developing
embryos during seed development
(1) Globular (2) torpedo (3) pre-cotyledonary (4) cotyledonary (5) mature
32

(Vanildo et al., 2004)

INTERACTION OF GA WITH OTHER HORMONES

Fig. Gibberellins (GA) interacts positively and negatively with other

plant hormones throughout the plants life cycle
33
(David and Naomi, 2005)

Fig. Interactions between GA and ABA. ABA suppresses GA

responses through DELLA protein. Interactions mediated by changes
in protein activity or stability; mediated by specific gene expression
34

(Shani et al., 2006)

Genetic analysis as a tool to investigate the mechanisms

underlying seed development in maize
A

Biochemical and molecular mechanisms

indicate that both embryo and endosperm
formation require a distinct series of events
leading to the acquisition of a proper structure
From this study it is clear that the progression of
Presenceand
of aendosperm
proper endosperm
is required
embryo
development
relies for
on
successful
embryo
development
the
interaction
between
them
Greater seed size with its accompanying
increase in stored reserves is considered as the
most significant trait

A- Embryo

B- Endosperm

Note: Safranin-fast green staining of a longitudinal section

of an embryo (A) and endosperm (B) at 14 DAP in maize.
fluorescein fluorescence images
35

(Gabriella et al.,

Embryo and endosperm are genetically identical, except for ploidy level
Embryo (2n)
Endosperm (3n)
So the ratio b/n the (M:P) contribution differs in each compartment

From this study, evidence was obtained that what is critical for
proper seed development is not the overall ploidy level, but the relative
contribution from the male and female parents
2: 1 ratio (M:P) genomes is necessary for endosperm development.
This mechanism as a consequence, for proper seed development

36

(Gabriella et al.,

ARABIDOPSIS SEED DEVELOPMENT IS ASSOCIATED WITH TEMPORALLY

DISTINCT METABOLIC SWITCHES
Aaron, F., Ruthie, A., Hadar, L., Itzhak, O., Ewa, U., Alisdair, R., Fernie and Gad, G., (2006)

Plant Physiol. 142: 839854

37

Fig A. Decreasing in metabolites during the period

of reserve accumulation in Arabidopsis seeds
38

Observation on 14 and 17 DAF

(Aaron et al.,2006)

Fig B. Increasing in metabolites during the

desiccation period of Arabidopsis seeds
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Observation on 17 DAF

(Aaron et al., 2006)

Large scale identification and quantitative profiling of

phosphoproteins expressed during seed filling in oilseed rape
Ganesh, K., Umar Agrawal and Jay, J., 2006

Molecular & Cellular Proteomics , 5.11: 2044-2059

40

Identification and quantification of phosphoproteins during seed filling in Brasicca

2-D gels

Fig. Schematic depiction of experimental design for quantitative

expression analysis and database construction
(Ganesh et al. 2006)
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42

Fig.Reference map of phosphoproteins expressed during seed

development in Brasicca (Ganesh et al. 2006)

Fig. Dynamic variations of

phosphoprotein profiles

43

Fig. Brasicca seed

phosphoproteins belong to 10
major functional classes
(Ganesh et al. 2006)

44

Genomic profiling to identify genes required for soybean seed

development
A-Globular stage
B and C- Globular
Molecular &
Biochemical

stage before (B) and

after (C) capturing

ent, Endothelium;

LCM (Laser Capture

ep, embryo proper;

Micro dissection)

epd, epidermis;
es, endosperm;
hi, hilum;
ii, innerintegument;

image
D- Functional
category distribution

oi, outer integument;

s, suspensor

(Brandon et al., 2007)

Secondary metabolites in seed defence mechanisms

(Richard and Roger, 1994)
New Phytol., 127: 617-633

(Richard and Roger, 1994)

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Many secondary metabolites found in seeds have a
role in defence against herbivores; pests and pathogens

Defence may involve anti-feedant activity, toxicity or

acting as precursors to physical defence systems
Eg. Aromatic amino acids, Alkaloids, Phenolics,
Phytoalexins etc.

Cinnamic acid and Salicylic acid are acts as a signals

for secondary metabolites production during seed
development of many plant sp.

(Richard and Roger, 1994)

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In addition to defence against pest and pathogen, these

compounds may be involved in protection in seed storage,
protection from UV, protection against osmotic and other
environmental

stresses

during

germination

and

also

allelopathic interactions with other plants etc.

Fig. Some of the secondary products and other metabolites derived
from Shikimic and aromatic amino acids and their biosynthetic
relationship during seed development
(Richard and Roger,

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Do legume storage proteins play a role in

defending seeds against pest?..
(Mauricio et al., 2012)
American Society of Plant Biologists, 124:
516-522

48

(Mauricio et al., 2012)

Resistant and susceptible of seeds associate with chitin containing

of bruchids midgut
High affinity for chitin to vicilins content of cotyledons of
resistant seeds possibly hampers absorption of nutrients in midgut
In addition, hydrolysis of vicilins is reduced due to the low level of
proteolytic activity found in the midgut of C. maculatus
Schematic representation of how storage proteins play a role in defending seeds

Takeningested
together
these vicilin
factors
seem
to account
larval
Bruchids
cowpea
from
resistant
seedsfor
have
not mortality
ofsurvive
this insect
feeding on resistant cowpea seeds
because
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(Mauricio et al., 2012)

Legume seed storage proteins that have been synthesized during

seed development are acts as defense proteins against bruchid

Legumin and vicilin in developing seeds are characterized as

metabolically inactive but act as defense seed proteins under stress

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CONCLUSION

Seed development study has become exciting

due to the availability of new and sophisticated
techniques

i.e.

proteomics,

genomics,

QTL

mapping, marker technology and RNA profiling

using Gene Chip arrays etc.

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PATH AHEAD

Little is known about the molecular and genetic mechanisms

responsible for seed development
Genes have to identify particularly that are unique to seed
development, compartment and seed filling
Suitable genes (conserved motifs) have to identify for better and
complete understanding of seed development mechanisms in detail
Conclusive evidence linking these mechanisms with hybrid failure
remains patchy
However mechanisms of seed development is becoming clearer but
still lot of scope is there to improve upon
52

Thank you

References:
Aaron, F., Ruthie, A., Hadar, L., Itzhak, O., Ewa, U., Alisdair, R., Fernie and Gad, G., 2006,
Arabidopsis seed development and germination is associated with temporally distinct
metabolic
switches. Plant Physiol., 142: 839854.
David, W. and Naomi, O., 2007, Mechanisms of cross talk between gibberellins and other
hormones. Plant Physiol., 144: 12401246.
Gabriella, C., Giuseppe, G. and Silvana, D., 2005, Genetic analysis as a tool to investigate the
molecular mechanisms underlying seed development in maize. Annals of Bot., 96: 353362.
Mauricio, P., Isabel, R., Gerhardt, M., Fatima, G. and Jose, X., 2012, Do storage proteins play a
role in defending seeds. American Soci. Plant Biologists, 124: 516-522.
Nicolas, G., Bradford, K., Come, D. and Pritchard, H., 2002, The biology of seeds recent research
advances. Proceedings of seventh international workshop on seed, Salamanca, Spain,
pp. 11-24.
Sebastien, B., Jean, P., Martine, M., Loic, L. and Christine, R., 2002, An integrated overview of
seed development in Arabidopsis thaliana. Plant Physiol. Biochem., 40:151160.
Shani, E., Yanai, O. and Ori, N., 2006, The role of hormones in seed development. Curr. Opin. Plant
Biol., 9: 484489.
Steve, W., Sarah, E. and Sheldon, C., 1971, Seeds: biology, development and ecology. CABI publication,
North America. pp. 102-120.