EMBRYOGENESIS: Dicots,

Monocots, and Gymnosperms

Bio 176
 Embryogenesis
• Embryogenesis is the process by which the embryo
forms and develops.
– It starts with the fertilization of the egg cell (ovum) by a sperm cell.
– Once fertilized, the ovum is referred to as a zygote, a
single diploid cell.
– The zygote undergoes mitotic divisions and cellular differentiation,
leading to development of a multicellular embryo.

Guppy fish embryo Day 12 chicken embryo

 Plant Embryogenesis
• It is the process that produces a plant embryo from a fertilized
ovule by asymmetric cell division and the differentiation of
undifferentiated cells into tissues and organs.

• It occurs during seed development, when the single-

celled zygote undergoes a programmed pattern of DNA
replication then mitotic cell division resulting in a mature embryo.

• A similar process continues during the plant's

life within the meristems of the stems and roots.
• Therefore, embryogenesis in plants, is
concerned primarily with establishing the
basic shoot-root body pattern of the plant
and accumulating food reserves that will be
used by the germinating seedling after a period
of embryonic dormancy within the seed.
• A major problem in plant development is to unravel the
mechanisms operating during embryogenesis that
enable a plant to specify its body plan and tissue
differentiation patterns.
– Especially, a detailed understanding of the events that
govern plant embryo formation.
– One obstacle in achieving this goal is the location of
embryos within the plant and their relative inaccessibility
to experimental manipulation.
In flowering plants, reproductive processes occur within
floral organs.
• Because egg cell formation, fertilization, and
embryogenesis occur within the pistil, it has been
difficult to dissect the major events that take place
during the early stages of higher plant development.
– Now, it is possible to isolate plant eggs and fertilize them in
vitro in order to investigate the initial events of plant
embryogenesis.
– Genetic approaches have been used to identify genes
required for various embryogenic processes, including
pattern formation.
• Embryogenesis occurs naturally as a result of sexual
fertilization and the formation of the zygotic
embryos.
• The embryo along with other cells from the mother
plant develops into the seed, which, after germination,
grows into a new plant.
• Generally, embryogenesis is divided into two
(2) phases/processes:

– The first involves morphogenetic events which form

the basic cellular pattern for the development of
the shoot-root body and the primary tissue layers;
• It also programs the regions of meristematic
tissue formation.
– The second phase, or postembryonic development,
involves the maturation of cells, which involves cell
growth and the storage of macromolecules (such as
oils, starches, and proteins).
• These macromolecules are required as a 'food and energy
supply' during germination and seedling growth.
• Embryogenesis involves cell growth and division, cell
differentiation and programmed cellular death.
• The zygotic embryo is formed following
double fertilization of the ovule, giving rise to two
distinct structures:
– the plant embryo and
– the endosperm
• Which together go on to develop into a seed.
 Sexual reproduction in plants
●The flowering plant life cycle
is divided into haploid and
diploid generations that are
dependent on each other.

● Flower parts undergo meiosis

to produce haploid products
(gametophytic)
— pollen grain (male
gametophyte) or
— embryo sac (female
gametophyte, contains the
egg cell)
• The pollen grain
contains two sperm cells,
whereas the embryo sac
contains a single egg.

• Other accessory cells

within the haploid male
and female gametophytes
help facilitate the
pollination and fertilization
processes.
 Pollination and Fertilization
• Two fertilization
events occur in
flowering plants.
– One sperm unites
with the egg cell to
produce a diploid
zygote (2n) and
initiate
embryogenesis.
 — The other sperm cell
unites with a specialized cell
(central cell), with 2 polar
nuclei, to form the triploid
endosperm (3n), which will
become a food source for the
developing plant embryo.

*Thus, double fertilization

occurs.
• Fertilization causes the
ovule, containing the embryo
and endosperm, to develop
into a seed and the ovary to
differentiate into a fruit,
which facilitates seed
dispersal.
The shoot-root body plan
• Following fertilization, the zygote undergoes an
asymmetrical cell division that gives rise to a:
– small apical cell (T, terminal cell), which becomes the
embryo
– large basal (B) cell (called the suspensor), which functions
to provide nutrients from the endosperm to the growing
embryo
*The product is an ovoid mass of tissue called the
proembryo.
Refer to Fig. 2
• Thus, plant embryogenesis can be divided into
three (3) phases in which distinct developmental
and physiological events occur:
(i) post fertilization- proembryo,
(ii) globular-heart transition, and
(iii) organ expansion and maturation
Refer to Fig. 2
• The asymmetric cleavage of the zygote results in the
formation of an embryo with a suspensor and embryo
proper that have distinct developmental fates.
– Cell lineages derived from the terminal cell and embryo proper will
specify the cotyledons, shoot meristem, hypocotyl region of the
embryonic axis, and part of the radicle, or embryonic root.
– By contrast, the large basal cell derived from the lower portion of the
zygote will divide and form a highly specialized, terminally differentiated
embryonic organ called the suspensor.

Refer to Fig. 2
• In Arabidopsis, the suspensor contains only 7 to 10
cells.
– The suspensor anchors the embryo proper to the
surrounding embryo sac and ovule tissue and serves as
a conduit for nutrients to be passed from the maternal
sporophyte into the developing proembryo (Fig. 2).
– The suspensor senesces after the heart stage and is not
a functional part of the embryo in the mature seed.
• In the globular stage, the embryo develops radial
patterning through a series of cell divisions, with the
outer layer of cells differentiating into the “protoderm”.
• The globular embryo can be thought of as two layers of
inner cells with distinct developmental fates:
– the apical layer will go on to produce cotyledons and
shoot meristem, while the lower layer produces
the hypocotyl and root meristem
Refer to Fig. 2
• Embryonic organs and tissue-types differentiate
during the globular-heart transition phase.
• Two (2) critical events must occur after the embryo
proper forms:
(i) regions along the longitudinal apical-basal axis must
differentiate from each other and generate embryonic
organ systems, and
(ii) the three primordial tissue layers of the embryo
need to be specified.
Refer to Fig. 2
 • Subsequent cell differentiation events within the embryo
proper result in the production of an inner procambium
tissue layer and a middle layer of ground meristem
cells.

• A dramatic change in the morphology of the embryo

proper occurs just after the globular stage.

Refer to Fig. 2
• Bilateral symmetry is apparent from the heart stage;
provascular cells will also differentiate at this stage.
– The body plan and tissue layers of the mature embryo (and
postembryonic plant) have been established.
– Morphogenetic changes during this period are mediated by
differential cell division and expansion rates and by
asymmetric cleavages in different cell planes.
– No cell migration occurs, in contrast to the migration
events that take place in many types of animal embryos.
Refer to Fig. 2
• A major change in embryonic development occurs
during the organ expansion and maturation phase.
– A switch occurs during this period from a pattern
formation program to a storage product accumulation
program in order to prepare the young sporophyte for
embryonic dormancy and postembryonic development

*This is because embryogenesis terminates with

a dormancy period.
• In the organ expansion and maturation phase:
– The heart-shaped stage is followed by the torpedo-shaped stage, in
which elongation of the cotyledons and hypocotyl, as well as
extension of the vascular tissues, occurs.
– The cotyledons and axis increase in size dramatically as a result of
cell division and expansion events.
– Ground meristem cells within both these organs become highly
specialized and accumulate large amounts of storage proteins
and oils that will be utilized as a food source by the seedling after
germination.
Refer to Fig. 2
• At the end of the organ expansion and maturation
period, the embryo has reached its maximum size,
cells of the embryo and surrounding seed layers have
become dehydrated, metabolic activities have
ceased, and a period of embryonic dormancy within
the seed begins.
• Thus, embryogenesis terminates with a dormancy
period.
• In a dicot embryo,
the hypophysis, which
is the uppermost cell of
the suspensor,
differentiates to form
part of the root cap
called columella.
 Hormones Affect Embryo
Morphogenesis
• What physiological events cause the embryo proper to
initiate cotyledons and become bilaterally symmetric
during the globular-heart transition phase?
– Several experiments implicate a class of plant hormones, the
auxins, in this morphogenetic process.
– Auxins, such as indoleacetic acid (IAA), are involved in a
number of plant activities, including photo- and
gravitropism, apical dominance, and vascular cell
differentiation.
• Embryos in plants as diverse as bean and pine
synthesize auxins and transport them in a polarized,
basipetal direction (from the shoot meristem to root
tip) along the embryonic axis.

• The highest auxin levels occur at the globular stage of

embryogenesis.
• Agents that inhibit
polarized auxin transport
– Either:
• block the transition from
the globular to heart stage
completely or
• prevent the bilateral
initiation of cotyledons at
the top of the globular
embryo
• For example, zygotic embryos of the Indian mustard
(Brassica juncea), an Arabidopsis relative, fail to initiate
two laterally positioned cotyledons when treated with
auxin transport blockers in culture.
– A cotyledon-like organ does form, but as a collar-like ring
around the entire upper (apical) region of the embryo.
• In a similar study of Neuhaus et al.
(1998), they tested the effects of auxin
transport inhibitor N-(1-
naphthyl)thalamic acid (NPA) and an
antiauxin p-chlorophenoxyisobutyric
acid (PCIB), in vitro in zygotic
Brassica juncea embryos, which led
to a wide range of morphogenetic
alterations.
The Embryo Is Reprogrammed
Late in Embryogenesis
• How does the embryo prepare for dormancy and
postembryonic development?
• Late in embryogenesis a maturation program is
induced that is responsible for:
(i) synthesizing large amounts of storage products,
(ii) inducing water loss and a desiccated state,
(iii) preventing premature germination, and
(iv) establishing a state of dormancy

Refer to Table 1, slide 24

• Several specialized gene sets, such as those encoding
storage proteins and late embryo abundant (lea)
proteins, are activated transcriptionally during
maturation and then repressed (blocked) before
dormancy.
• These gene sets remain transcriptionally quiescent
(inactive) during seed germination when germination-
specific and postembryonic gene sets are transcribed.
• Abscisic acid maintains embryonic dormancy.
– The plant hormone abscisic acid (ABA) is involved in several
plant processes, including senescence, responses to
environmental stresses, growth inhibition, and
maintenance of a dormant state.
– Exogenous (externally applied) ABA prevents seed
germination as well as the precocious (early/advance)
germination of embryos in culture.
• In addition, Arabidopsis mutants that either cannot
synthesize ABA or fail to respond to ABA germinate
precociously.

• These data indicate that ABA prevents germination

while seeds are still dormant or present within siliques
(seed pod).
 Embryogenesis in Monocots
• The early divisions of the zygote in monocots follow the same
pattern as in dicots.
• However, in the Poaceae (grasses) family, which includes
wheat and the other cereals, the sequence and orientation of
later divisions in the proembryo are irregular, resulting in highly
complex mature embryos.
– The main feature of the cereal embryo is the development of an
absorptive organ known as the scutellum (considered equivalent to
the single cotyledon).
• The zygote or oospore elongates and then divides
transversely to form basal and terminal cells (Fig 2.31
A-B).
• The basal cell (towards micropylar end) produces a
large swollen, vesicular suspensor cell (Fig 2.31 C-D).
• The top cell after a series of divisions forms plumule
and a single cotyledon (Fig 2.31 E) .
• Cotyledon called scutellum, grows rapidly and pushes
the terminal plumule to one side (Fig 2.31 F-G).
– The plumule comes to lie in a depression.
• The middle cell, after many divisions forms hypocotyl
and radicle (Fig 2.31 F-G).
– It also adds a few cells to the suspensor.

*In some cereals both plumule and radicle get covered by

sheaths developed from scutellum called coleoptile and
coleorrhiza, respectively (Fig 2.31 H).
• Other organs of the embryo for which there are no
counterparts in the dicot embryo are:

– a sheath like tissue covering the root (coleorrhiza),

• On one side of the coleorrhiza there is also a small, flaplike out
growth called the epiblast.
• Epiblast represents rudiments of second cotyledon.
– a tissue that covers the shoot (coleoptile), and
– an internode known as the mesocotyl.
Embryogenesis in Gymnosperms
• Gymnosperms, like all vascular plants,
have a sporophyte-dominant life
cycle, which means they spend most
of their life cycle with diploid cells, while
the gametophyte (gamete-bearing
phase) is relatively short-lived.

• Two spore types, microspores and

megaspores, are typically produced in
pollen cones or ovulate cones,
respectively.
• During pollination, pollen grains are physically transferred
between plants from the pollen cone to the ovule.
• Pollen is usually moved by wind or insects.
• The pollen grain germinates and grows into the ovule, penetrating the
female gametophyte and eventually fertilizing an egg nucleus.
• There may be more than one egg fertilized, but only one zygote
develops into an embryo.
• It may be an extended time, such as one year or longer, between
pollination and subsequent fertilization.
• Once a zygote is formed, further development leads to a seed, which
consists of an embryo surrounded usually by nutritive tissue and a
seed coat.
• The nutritive tissue of "gymnosperm" seeds is derived from the female
gametophyte.
 Pollination and Fertilization

• Whole grains enter each ovule through a microscopic gap in the ovule
coat (integument) called the micropyle.
• The pollen grains mature further inside the ovule and produce sperm cells.
 Pollination and fertilization in Ginkgo
• Two main modes of
fertilization are found in
gymnosperms:
– Cycads and Ginkgo have
motile sperm that swim
directly to the egg inside the
ovule.
– Conifers and gnetophytes
have sperm with
no flagella that are moved
along a pollen tube to the
egg.

Ginkgo biloba and the cycads are the only

living seed plants with motile sperm cells.
 Ginkgo pollination

Fertilization does not occur immediately after

pollination. It only takes place in fall. Inside the
Pollination droplet is present on the archegonia egg cells are maturing. During the months
micropyle at the tip of the integument. following pollination, the pollen forms two sperms in the
pollen chamber, absorbing water and nutrients from the
female tissue - a nucellus.

The pollen chamber and the space

around the tentpole unite into a bigger The pollen tube ruptures and the Ginkgo
archegonial chamber. This fluid could sperms swim upwards in "the sea" toward the
be regarded as "the sea" in the seed. eggs.
Figure 1 (a-d). Fertilization and embryo development in the gymnosperm Picea
(a conifer).

(a) Fertilization of the two female nuclei, each in a separate archegonium at the
base of each scale of a cone. Left, fertilization; Right, after first mitotic division.
(b–d) Each nucleus then undergoes mitotic divisions to form free nuclei, followed
by cell wall formation, resulting in 16-celled proembryos; only the front set of
nuclei and cells is visible (development of one of the fertilized eggs is shown; the
other will abort or undergo incomplete development and degenerate).
Figure 1 (e-h). Fertilization and embryo development in the gymnosperm Picea.

(e–f) Three-tiered mature terminal embryo with subterminal suspensor cells which
elongate to form the primary suspensor.
(g–h) Embryonal mass divides to form the mature embryo with one to several
cotyledons, which varies between Picea species from 4 to 15.
Conifer Seed Germination
and Dormancy
Conifer Seed Germination and Dormancy
• A mature pine seed contains an uncurved embryo with many
cotyledons.
– The embryo is embedded in a nutritional tissue which is the
megagametophyte (n) tissue.
• This is haploid maternal tissue and is by some researchers called
primary endosperm.
• The diploid nucellar tissue is usually obliterated.
• The diploid integument tissue develops into the seed coat.
• The ovulate pine cone becomes woody as it matures.
• The scales grow and remain closed while the cone matures.
 References Cited
• Suárez, María F., and Peter V. Bozhkov. 2008. Plant embryogenesis. Totowa, NJ: Humana
Press. p 3.
• Goldberg, Robert B., de Paiva, Genaro, and Ramin Yadegari. 2012. Plant Embryogenesis:
Zygote to Seed. www.sciencemag.org.
• http://lifeofplant.blogspot.com/2011/12/angiosperm-plant-formation.html
• http://www.yourarticlelibrary.com/biology/embryo-in-flowering-plants-structure-types-and-
development/11808
• Current topics in developmental biology; v. 67. 2005. [S.l.]: Elsevier Academic Press. p 135.
• Hadfi, K., Speth, V. and G. Neuhaus. 1998. Auxin-induced developmental patterns in
Brassica juncea embryos. Development 125, 879-887pp.
• https://www.researchgate.net/publication/279350635_Seeds/figures?lo=1
• http://courses.washington.edu/bot113/summer/WebReadings/gymnospermPlants/gymnosp
erms.html
• https://en.wikipedia.org/wiki/Gymnosperm
• http://www.seedbiology.de/evolution.asp