LECTURE 3.

PLANT EMBRYOGENESIS

PLANT EMBRYOGENESIS
1. From fertilization until dormancy
One-cell zygote embryogenesismature embryo.
2. The basic body plan of the sporophyte is established
plan is repeated several times over and elaborated after dormancy is broken.
3. Challenges of embryogenesis are:
a. Form basic body plan: apical-basal (shoot-root) axis; radial patterning- produces three
tissue systems, axial patterning formation of cotyledons.
b. Form meristematic tissue for post-embryonic development of structure (leaves, roots,
flowers, etc.).
c. Establish a food reserve for the germinating embryo until it becomes autotrophic.

A. MORPHOGENETIC STAGES
1. Single CellThe zygote undergoes the first division-an asymmetric cell division
Result: to dissimilar cells- a smaller apical or terminal cell and a larger basal cell. The apical
cell receives most of the cytoplasm, and the basal cell receives the vacuole (Figure 9).
2. Polarity is established during this first asymmetric division. Most of the plant embryo
develops from the apical (terminal) cell.
3. Basal cell divides to form the suspensor. Cells of the suspensor divide transversely to form a
longitudinal file of cells.
Function of suspensor:
i) anchors the embryo to the endosperm
ii) nutrient conduit for the developing embryo

iii) The distal end of the suspensor is called the hypophysis contributes to gives rise to the root.
(The other suspensor cells divide to form a filamentous or spherical organ that degenerates later
in embryogenesis).
4. Two celled pro-embryo. The apical cell divides longitudinally to form two cells.
5. Four celled pro-embryo. The two cells each divide again longitudinally in a plane
perpendicular to the first division to form a quadrant or 4-celled filament (Figure 9).
6. Octant stage. These four cells then divide transversely to form an octant (Figure 9, 10). The
transverse walls of these 4 cells divides the embryo in half along what is known as the O' line.
This becomes a boundary between distinct domains of the embryo.
7. Globular stage. All cells then divide periclinally to form the first histologically distinct tissue,
the protoderm (Figure 11a). This stage is called the globular or dermatogen stage. The three
basic tissue systems (dermal, ground, and vascular) can be recognized at this point based on
characteristic cell division patterns.
8. Heart shaped stage. As cotyledons begin to form the globular shape of the embryo changes to
a heart-shaped appearance in dicots. In monocots, only a single cotyledon forms.

a. Transition to heart stage begins when two groups of cells divide periclinally causing bulges
that emerge as cotyledon lobes.
b. This represents a shift from radial to bilateral symmetry.
c. This also delineates the 2 main embryonic organ systems: the cotyledons and the shoot axis.
d. The root apical meristem (RAM) begins organizing at this stage.
e. The apical-most suspensor cell, called the hypophysis, becomes incorporated into the embryo
proper. Both the hypophysis and apical cell derivatives contribute to the formation of the RAM.
f. At this stage, procambium (vasculature) initials can first be discerned (Figure 11)
NB. Thus differentiation of most major tissue systems has begun by early heart stage.
9. Torpedo stage. Continued cell division, growth and differentiation lead to the late heart stage
and then to the torpedo stage.
a. By this point the suspensor is degenerating
b. And the shoot apical meristem (SAM) and root apical meristem (RAM) are established.

c. These meristems will give rise to the adult structures of the plant upon germination
10. Walking stick stage. Further growth of the cotyledons results in the walking stick stages. At
this point, embryogenesis is arrested, and the mature seed desiccates and remains dormant until
germination.

Figure 9. Diagrammatic representation of the different stages embryogenesis.

Figure 10. Stages of development in an angiosperm the embryo upto dormancy

B. MORPHOGENETIC CHANGES
1. Establishment of body plan.
2. Establishment of meristems
3. Establishment of food reserves; Dormancy
1. Establishment of body plan.
Two developmental patterns have been identified
a) Shoot-root axis determination.
1. Begins -asymmetric cell division to form terminal and basal cell.
2. Establishes polarity. The apical/terminal cell embryo proper while the basal cell
suspensor (more in lecture 4).

3. Hypophysis -distal end of the suspensor contributes to root formation. Rest degenerates

Function of the suspensor:

orients the embryo toward food source;

in angiosperms serve as a nutrient conduit for the developing embryo.

hormones

Evidence:
1. Isolate embryos with and without suspensor and culture them.
Result. Embryos without suspensor: do not survive at heart shape stage while embryos with a
suspensor are twice as likely to survive at the heart-shape stage.
Conclusion: need for a suspensor through the heart-shape stage in dicots (Yeung and Sussex
1979).
Possible reason: The suspensor may be a source of hormones.
Further evidence: In scarlet runner beans, younger embryos without a suspensor can survive in
culture if they are supplemented with gibberellic acid (Cionini et al. 1976).
b) Axial and Radial Symmetry.
1. Cell division and differentiation continue to give radial and axial and patterns. The cells
of the embryo proper divide in transverse and longitudinal planes to form a globular-stage
embryo with tiers of cells (Figure 11).
2. The emerging shape of the embryo depends on regulation of the patterns of cell division,
since the cells are not able to move and reshape the embryo (see lecture 4).
3. Cell division planes in the outer layer become restricted and this layer, called the
protoderm, becomes distinct.
4. Radial patterning emerges in the globular stage as the three tissue systems (dermal,
vascular, and ground) of the plant are initiated.
a. The dermal system forms from the protoderm and contributes to the outer
protective layers of the plant.
b. Ground tissue forms from the ground meristem and surrounds the developing
vascular tissue;
c. The vascular system forms from the procambium cells that differentiate in the
center of the globular embryo are for support and transport.

NB. The ground and vascular systems form independently.

Evidence Knolle mutants of Arabidopsis have their epidermal (L1) layer disrupted by
misoriented cell divisions from the early globular stage while the Keule mutants have bloated
and irregularly arranged epidermal cells recognizable at the globular stage. In both mutants the
inner tissue systems develop normally (Mayer et al. 1991).

1. Axial (bisexual) patterning.

Becomes evident after the cotyledons, the first leaves, begin to form at end of the
globular stage. Dicots have two cotyledons, which gives the embryo a heart-shaped
appearance as they form.

Hormones (specifically, auxins) may mediate the transition from radial to bilateral
symmetry (Liu et al. 1993). In monocots such as maize, only a single cotyledon emerges.

Function of cotyledons:
i. Become photosynthetic after germination and aid in nourishing the plant (although some never
emerge from the ground).
ii. Where food reserve in the endosperm is used up before germination, the cotyledons become
the nutrient source for the germinating seedling e.g. pea.
Cotyledons store food reserves such as starch, lipids, and proteins e.g. maize.

Figure 11. Radial and axial patterning. (a) Radial patterning in angiosperms begins in the
globular stage and results in the establishment of three tissue systems. (b) The axial pattern
(root-shoot axis) is established by the heart-shape stage. Adapted from
http://zygote.swarthmore.edu/phyto1.html

1.Establishment of meristems
a. The shoot apical meristem and root apical meristem formed: are clusters of embryonic cells
that persist in post-embryonic development and give rise to most of the sporophyte body.
b. The root meristem is partially derived from the hypophysis (the uppermost cell of the
suspensor) in some species. All other parts of the sporophyte body are derived from the embryo
proper. (More during shoot and root formation).
c. Genetic evidence indicates that the formation of the shoot and root meristem are regulated
independently.

Evidence from observations of mutants of Arabidopsis:

i. The shootmeristemless (STM) mutant in Arabidopsis, is able to form a root meristem but fails
to initiate a shoot meristem (Clark and Sheridan 1986; Barton and Poethig 1993).
ii.The STM gene is expressed in the late globular stage, before cotyledons form.
iii.STM's role appears to be to repress cell differentiation in the shoot apical meristem so that the
cells maintain their indeterminate state (Long et al. 1996).
iv.The shoot apical meristem will initiate leaves and ultimately the transition to reproductive
development after germination.
1.Establishment of food reserves; Dormancy

1.As the embryo reaches a maturation phase there is a shift from constructing the basic body plan
to creating a food reserve by accumulating storage carbohydrates, proteins, and lipids.
2.The high level of metabolic activity in the developing embryo is fueled by continuous input
from the parent plant into the ovule.
3.Eventually metabolism slows and the connection of the ovule (seed) to the ovary is severed by
the degeneration of the adjacent supporting sporophyte cells.
4.The seed loses water (dessication) and the integuments harden to form a tough seed coat and
Dormancy sets in ending embryogenesis. Embryo can persist in a dormant state for weeks or
years.
5.Maturation dormancy is due to precisely regulated program.
NB. The hormone abscisic acid is important in maintaining dormancy in many species.
Gibberellins, another class of hormones, are important in breaking dormancy.

References

Arabidopsis book (TAB) an electronic book published by ASPB at

http://www.aspb.org/publications/arabidopsis/toc.cfm
Graham, C.F., and Wareing, P.F. Eds. Developmental biology of plants and animals.
Blackwell Scientific Publications, Oxford.

Lecture 16

Embryogenesis

Overview
There are 3 major objectives for plant embryogenesis:
1. To establish the body plan (axis formation) and major organ systems of the
plant.
2. Establishing meristems (populations of cells that will continue postembryonic
growth).
3. To prepare the embryo for dormancy and subsequent germination. This involves
storage of compounds for nutrition of the germinating seedling, and acquiring
tolerance to desiccation.
Embryonic development can be divided into 3 overlapping stages:
1. A period of morphogenesis and histogenesis in which the major organ and
tissue systems of the plant are established. This period is characterized by a
high rate of cell division and differentiation.
2. A maturation phase characterized by cell expansion and the massive
accumulation of storage reserves. Fresh weight can increase 100 fold with little
to no cell division.
3. Acquisition of desiccation tolerance and the loss of water associated with the
onset of developmental arrest.

1. Establishment of the plant body.

Description of embryogenesis
Arabidopsis:
At the time of fertilization the egg cell is polarized, with a large vacuole toward the
micropylar end and the nucleus and most of the cytoplasm toward the chalazal
end. After fertilization, this polarity is maintained in the zygote. The first division

is perpendicular to the axis of polarity and is asymmetric. The apical cell receives
most of the cytoplasm, and the basal cell receives the vacuole. From this point on
the pattern of cell division varies among species.
The basal cell gives rise to the suspensor. Cells of the suspensor divide transversely
to form a longitudinal file of cells. The suspensor pushes the embryo proper into
the nutrient-rich endosperm and is also important for nutrient transport into the
developing embryo. Only the distal-most cell of the suspensor will contribute to
the embryo proper.
The apical cell divides longitudinally. Both cells then divide again longitudinally in a
plane perpendicular to the first division to form a quadrant. These four cells then
divide transversely to form an octant. The transverse walls of these 4 cells divides
the embryo in half along what is known as the O line. This becomes a boundary
between distinct domains of the embryo (see below). All cells then divide
periclinally to form the first histologically distinct tissue, the protoderm. This
stage is called the globular or dermatogen stage. Afterward, protodermal cells
divide predominantly anticlinally to form a distinct cell lineage. Inner cells divide
both anticlinally and periclinally cause growth as a mass. In the globular stage
embryo, there is expression of a lipid transfer protein gene specifically in the
protodermal cells. The distinct pattern of cell division and differential gene
expression are the first indications of histological differentiation.
Transition to heart stage begins when two groups of cells divide periclinally
causing bulges that emerge as cotyledon lobes. This represents a shift from radial
to bilateral symmetry. This also delineates the 2 main embryonic organ systems:
the cotyledons and the shoot axis. The root apical meristem (RAM) begins
organizing at this stage. The apical-most suspensor cell, called the hypophysis,
becomes incorporated into the embryo proper. Both the hypophysis and apical cell
derivatives contribute to the formation of the RAM. At this stage, procambium
(vasculature) initials can first be discerned. Thus differentiation of most major
tissue systems has begun by early heart stage. Continued cell division, growth and
differentiation lead to the late heart stage and then to the torpedo stage.
Differentiation of the shoot apical meristem (SAM) less obvious than the RAM. As the
cotyledon lobes begin to grow, a group of cells between them is seen to divide
infrequently. Later, the cotyledons and subtending tissues begin expressing genes
for storage proteins. The group of quiescent cells does not express these genes
and the lower boundary of the non-expressing cells in at a prominent cell wall
corresponding to the O line.
Maize:

After the first division, there is no set pattern of divisions. However apical cell
derivatives divide actively while basal cell derivatives (the suspensor) divide
infrequently. The suspensor is multiple cells thick. In the late proembryo stage a
protoderm differentiates, first over the apical region of the embryo, then
progressively downward. Like dicots, the protoderm divides anticlinally and
represents the first evidence of histodifferentiation.
At transition stage, a small wedge of cells becomes densely cytoplasmic and
begins rapidly dividing. These cells continue dividing and form a protuberance
that becomes the apical meristem. Internal to and basal to the developing
apical meristem, another group of cells assumes meristematic characteristics
(dense cytoplasm, rapid division). This becomes the root apical meristem.
Subsequently, procambial strands differentiate to connect the shoot and root
primordia and this defines the plant axis. A collar bulges up around the apical
meristem, but is not formed by the apex. This becomes the coleoptile which
ensheathes the shoot, protecting it as it pushes through the soil during
germination. As the embryo continues to grow, the apical meristem initiates
several leaf primordia (typically 6). This is in contrast to dicots which form no
leaf primordia prior to germination.
Axis Formation
Plant tissues contain an intrinsically fixed polarity. Stem segments will retain their
original apical-basal polarity no matter what the treatment. If the stem segment is
grafted in reverse orientation to a plant, roots will form at the original basal end.
The egg cell shows marked polarity prior to fertilization and this polarity is manifest
in the formation of the plant axis. Because the polarity is always present in the plant
tissues, it is difficult to study how the polarity is established. The in vitro fertilization
system described by Digonnet (Digonnet et al., 1997) has promise in this regard.
However most of the research to date has made use of the brown algae Fucus or
Pelvetia See the following for review: (Fowler and Quatrano, 1997).
Fucusmarine brown algae
zygotes free floating APOLAR
1st division is assymetricgives rise to a thallus cell and rhizoid
rhizoid gives rise to holdfast, thallus to fronds
direction of light determines the axis of polarityshade side becomes the rhizoid
axis determination is a stepwise process

1. axis formation: 4-10 hours after fertilization

requires a gradientmost convenient is light
shine directional light for 1 hour and then darkness or uniform lightaxis of
polarity forms according to the light direction :: treatment sufficient to induce
axis formation
shine directional light for 1 hour, then shine light from a different directionup
to 10 hours after fertilization, axis of polarity forms according to direction of
second pulse :: axis of polarity still labile
2. axis fixation : 10-12 hours after fertilization
changing direction of light will no longer change the axis of polarity
3. rhizoid formation : 14-18 hours after fert.
on side away from light
axis formation
1st detectable sign of asymmetry is an electrical current flowing inward at
presumptive rhiziod site
requires monovalent cation--medium containing only K + ions sufficient to support
axis formationits not clear if the electrical current is actually required for axis
formation;; its also not clear whether the ion is required for this current or
other events in axis formation or both
cortical clearinglocalized cortical vesicle exocytosishypothesized to
incorporate ion channels into the plasma membrane at presumptive rhizoid site
and contribute to the electrical current
requires microfilamentstreat zygotes with cytochalasinB during directional
light treatment, wash and place in dark, rhizoids form randomly relative to light
treatment
no detectable redistribution of microfilaments during axis formation
axis fixation
experimental approach--give zygotes directional light (light 1), then during axis
fixation period treat with inhibitors and give light from a different direction

(light 2)if inhibitor interfered with axis fixation, rhizoids will form according to
the second light direction, if not they will form according to the first.
requires microfilamentstreat zygotes with cytochalasinB, wash out and rhizoids
form according to second light direction
requires cell walldigest cell wall with enzymes to make protoplast
give light 1, remove wall during axis fixation, then no light 2rhizoid forms
according to light 1 therefore the cell wall is not required for axis formation
give light1, remove wall during axis fixation, then give light 2rhizoid forms
according to light 2therefore the cell wall is required for axis fixation

Involvement of auxin in embryo morphogenesis

Treatment of developing embryos with auxin polar transport inhibitors shows a
requirement for auxin polar transport, however, zygotic and somatic embryos
respond differently. Zygotic embryos treated at globular stage develop coyledons
in a fused cylinder. The embryos remain radially symmetrical thus auxin polar
transport is required to establish bilateral symmetry. Somatic embryos when
treated with the inhibitor fail to progress to the next stage of morphogenesis. Eg.
globular embryos continue spherical growth but no axis elongation or cotyledon
formation.
Positional information along the embryo axis
Torpedo stage embryos have recognizable cytological characteristics at different
points along their longitudinal axis. Torpedo stage embryos were transected at
various points along their longitudinal axis and allowed to regenerate root poles.
Regeneration involved the sequential production of cells with the correct
characteristics for their position along the axis. For example, embryos transected
near the apical pole first regenerated a hypocotyl, then a radicle and finally a
RAM. Thus regenerating cells respond to positional information that specifies
cellular characteristics along the longitudinal axis (Schiavone and Racusen,
1990).
Pattern mutants
The major pattern elements of the embryo include components of the longitudinal
axis, and the radial pattern of tissues found in stem cross sections. The
longitudinal elements include SAM, cotyledons, hypocotyl and root (including

RAM). The radial pattern consists of the epidermis, cortex (ground tissue) and
vasculature. A saturation mutagenesis for alterations in pattern revealed a small
group of genes controlling these elements. Reviewed by: (Jurgens et al., 1994).
Axial pattern:
gurke

apical deletion

fackel

central deletion

monopterous

basal deletion

gnom (emb30)

terminal deletion (both termini)

Radial pattern
knolle and keule

no epidermis

gurke deletes tissues that roughly correspond to those derived from cells above
the O line suggesting that the first transverse division of the embryo proper is
important for establishing the apical domain.
fackel mutants have severely shortened hypocotyls, suggesting the central
domain deletion, but also show abnormal numbers of cotyledons, shoot apicies
and other defects. The FACKEL gene is involved in the synthesis of a previously
unknown sterol compound (Jang et al., 2000; Schrick et al., 2000).
monopterous mutants lack a root and hypocotyl and show defects in vasculature.
MONOPTEROUS encodes a transcription factor that binds auxin-inducible gene
promoters and the mutants show defects in auxin polar transport (Hardtke and
Berleth, 1998).
gnom mutants (also called emb30) have abnormal cell division, expansion and
adhesion. The first cell division in the zygote is equal instead of assymetric.
GNOM encodes a protein related to a secretory yeast protein Sec7 which is
involved in golgi vesicle secretion (Shevell et al., 1994). It functions as an ARF
GEF (ADP-ribosylation factor G-protein Guanine exchange factor) that is required
for the polar localization of the auxin efflux carrier PIN1 (Steinmann et al., 1999).
Whether mislocalization of PIN1 is directly responsible for the pattern defects is
not clear.
knolle and keule mutants display abnormal cytokinesis leading to polyploidy and
multinucelate cells. Incomplete cytokinesis might prevent segregation of pattern
determinants between epidermal and internal cells. KNOLLE encodes a syntaxin

related protein (Lauber et al., 1997). Syntaxins are target membrane receptors
that interact with vesicle membrane proteins to promote secretory vesicle
membrane fusion. KEULE encodes a Sec1 protein that interacts with KNOLLE
(Assaad et al., 2001).
Segregation of cell wall determinants
Early Fucus embryo development resembles plant embryogenesis. The first cell
division results in a thallus cell that will form the shoot and a rhizoid cell that will
form the holdfast. If the rhizoid cell is completely ablated the thallus cell will
form a thallus only. If the thallus cell is completely ablated, the rhizoid will form a
rhizoid only. If protoplasts are made of the rhizoid cell, they will regenerate an
entire embryo indicating they still possess full totipotency. If the rhizoid cell is
ablated but fragments of rhizoid cell wall material remain attached to the apical
cell, apical cell derivatives that come in contact with the rhizoid wall material will
acquire a rhizoid fate. Similarly, if rhizoid derivatives contact wall material from
an ablated apical cell, they will acquire a thallus fate. This demonstrates the
presence of cell wall determinants in Fucus embryos (Berger et al., 1994).

Coupling of pattern formation, morphogenesis and differentiation

Several mutants that altered embryo shape without causing pattern deletions also
arose from the same pattern mutant screen discussed above. One of these
called fass causes a variety of morphological abnormalities, including variable
numbers of cotyledons, but contained all the basic elements of the axial and
radial patterns. This indicates that pattern formation and morphogenesis are
separable programs. fass mutants have abnormal patterns of cell division
indicating that the normal cell division orientation is not required for pattern
formation (Torres-Ruiz and Jurgens, 1994).
Another mutant called raspberry is arrested early in the globular embryo stage.
Accumulation cell type specific molecular markers indicated that tissue (cell)
differentiation proceeded beyond that normally seen at the stage of
morphogenesis at which the embryo was blocked. This indicates that cell
differentiation and embryo morphogenesis are controlled by separable programs
(Yadegari and al, 1994).
Somatic embryogenesis
Somatic cells can be induced to undergo embryogenesis. This was one of the
earliest and clearest demonstrations of totipotency, and therefore genomic
equivalence. Somatic tissue is induced to form callus. In the presence of auxin,

the callus will form pre-embryogenic masses (PEMs)small clumps of cells. Upon
removal of auxin the PEMs organize into a typical globular embryo which then
progresses through heart stage and torpedo stage. These embryos are the same
size and morphology as normal zygotic embryos. At the cotyledon stage, instead
of entering developmental arrest (like normal) the embryos initiate a shoot
meristem and seedling growth.
That embryo development can occur outside the environment of maternal tissue
indicates that the developmental program is intrinsic to the embryo per se. It
does not require maternal gene products, signals or spatial constraints for
normal morphogenesis. That is not to say the maternal environment is not
important because it is likely that the tissue culture conditions mimic some of
the conditions within maternal tissue.
The plant hormone, auxin, is required to induce competence of the PEMs to
undergo embryogenesis following removal of auxin. What the auxin or removal
from auxin actually does to induce embryo formation is unknown.
Embryonic development vs. seedling development (germination)
The plant hormone, abscisic acid (ABA) is intimately involved in controlling
embryogenesis and maturation. ABA deficient mutants show vivipary (ie. the
seeds germinate directly instead of going into dormancy following
embryogenesis. Furthermore, germination of dormant seeds can be inhibited by
exogenously applied ABA. Another hormone, gibberellic acid (GA) acts
antagonistically to ABA and promotes germination. GA deficient mutants
germinate poorly and exogenously applied GA can block dormancy and promote
vivipary.
Chemical environment of the endosperm: Raghavan and Torrey (1963) Late stage
embryos (cotyledon stage) placed in culture with a simple medium (inorganic
salts, vitamins and 2% sucrose) grew into normal seedlings with no dormancy
period. Early stage (globular-heart) embryos will not grow on simple medium.
They cease embryonic development and undergo cellular changes such as rapid
elongation and vacuolation associated with germination. Precise concentrations
of the plant hormones auxin and cytokinin allowed progression of embryonic
development. Alternatively, high osmoticum (10% sucrose or mannitol) allowed
embryonic development. Progressively lower concentrations of osmoticum are
required in later stages to complete embryogenesis. The osmoticum acts
independently of ABA because precocious germination of ABA deficient mutant
embryos can be inhibited.
Genetic control of developmental program: leafy cotyledons (lec) is a recessive
mutation that causes the cotyledons to acquire characteristics of leaves; they

have trichomes (hairs), leaf-like vasculature and lack storage proteins typical of
cotyledons. The mutant also displays vivipary and lacks desiccation tolerance.
The phenotype suggests that LEC normally regulates embryonic development
and in mutants, embryonic development is disrupted allowing the default
program of seedling growth to proceed. It also suggests homology between
cotyledons and leaves. This relationship would not otherwise be obvious because
leaves and cotyledons have very different ontogenies. Leaves are produced by
apical meristems whereas cotyledons arise prior to the meristem during
embryogenesis. Ectopic expression of the LEC gene in adult vegetative tissues
induces embryo formation (Lotan et al., 1998).
The suspensor
The suspensor is clearly involved in transport of nutrients to the embryo proper. In
some species the suspensor cells form haustoria or cell wall invaginations typical
of transfer cells. It is very metabolically active and thought to also provide
growth regulators to the embryo proper.
Removal of the embryo proper allows the suspensor to proliferate into a mass of
cells. In some plants, it will develop into another embryo. Thus the
developmental potential of the suspensor is greater than its fate and the
embryo proper is inhibitory to further growth.

2. Preparation for developmental arrest (embryo maturation).

Most cell division is complete by the beginning of the maturation phase of embryo
development, but the embryo can increase in size up to 100 fold. This is by cell
expansion and accompanies a massive accumulation of storage compounds. The
major storage compounds are proteins, starch and lipids. These storage
compounds are what give nutritional value to important crops such as cereals and
beans. They are also valuable for other uses such as production of vegetable oil
and starch which are used in a wide variety of ways ranging from cooking to
industrial lubricants and plastics. Therefore there is a huge economic interest in
seed storage compounds.
Accumulation of storage products
Storage proteins represent an important source of amino acids, nitrogen and
carbon for the germinating seedling. Storage protein mRNAs represent up to
20% of the total mRNA found in a maturation phase embryo. They are
synthesized on the RER and accumulate in the vacuole or as membrane bound
vesicles called protein bodies. The storage proteins are encoded by several

multigene families with up to 55 different genes coding for a given storage

protein. Synthesis is controlled at the transcriptional level, with a few regulatory
genes each controlling particular classes of storage proteins. An example is the
opaque2 gene of maize which codes for a transcription factor.
The regulation of starch and lipid accumulation, although no less important, is less
well understood. These compounds are produced by complex enzymatic
pathways. Each class of compound is a mixture of molecules with different chain
lengths, chain branching characteristics, levels of saturation and other chemical
modifications. Thus the synthesis of these compounds is much less straight
forward than storage proteins.
Acquisition of dessication tolerance
At the end of embryonic development, most seeds dehydrate to about 5%
moisture content. Such severe dehydration is lethal to most plant tissues and
embryos express a developmental program that allows them to survive. Two
problems faced by desiccated cells are high ionic concentrations and membrane
stresses. At such low moisture levels, solutes would tend to crystallize and
precipitate. Hydrophobic interactions with the aqueous solution are important for
maintaining the integrity of the lipid bilayer. With no aqueous phase, the
membrane becomes unstable and leaky.
A group of proteins called dehydrins are expressed in late maturation. The role for
these proteins in desiccation tolerance is supported by their induction by
drought stress in vegetative tissues and during desiccation of the resurrection
plant, one of the few plants that can tolerate desiccation of postembryonic
tissues. They are hypothesized to function in ion sequestration and in forming a
protective layer for stabilizing membranes.
Coupling of morphogenetic and maturation programs
Morphogenesis and maturation appear to be controlled by independent
developmental programs. In viviparous mutants which fail to undergo
maturation, morphogenesis is normal whereas other mutants arrested at various
stages of morphogenesis undergo normal maturation as evidenced by the
absence of necrosis following desiccation and the accumulation of storage
proteins.
Integration of these programs involves both hormonal mechanisms and genetic
programs. ABA is necessary to induce the expression of genes involved in
maturation and desiccation tolerance. ABA deficient viviparous mutants are
desiccation intolerant. An ABA independent genetic program is necessary to
confer ABA sensitivity to the embryo and mutants in this program show ABA

insensitive vivipary. The LEC gene, in which mutants both display seedling
instead of embyro morphological characteristics and bypass embryo maturation
are likely candidates for coordinating the two different programs.