ANGIOSPERMS: FERTILIZATION, EMBRYOLOGY AND SEED

(WITH DIAGRAMS)
Let us make an in-depth study of the fertilization, embryology and seed of
angiosperms. It is possible to get three types of seeds among both dicotyledons
and monocotyledons according to the final equilibrium.

They are: (1) Exalbuminous Seeds (2) Albuminous Seeds and (3) Perisperm
Seeds.

Pollination ends in a copious dusting of the stigma surface with pollen grains
(Fig. 414). The next stage is the germination of the pollen to form a pollen tube
wherein the male gametes are carried to the female gametophyte or the embryo
sac whose development is already complete. Fertilization or syngamy or
fecundation is the final phase of this biological process whereby the male and
the female gametes fuse with one another giving rise to the embryo.

The pollen grain, after being liberated from the anther is viable for a short or
long period. If it is transferred to the stigma during this period, it germinates.
The germination of the pollen may be helped by the fluid of the stigma. Pollens
may also be made to germinate in artificial culture media successfully.
Germination may begin almost immediately after pollination as in sugarcane
(Saccharum) Sorghum or it may take several hours or even days.

The pollen absorbs liquid from the moist surface of the stigma and the intine
comes out through one of the germ pores in the form of a single unbranched
tube (Fig. 409). Abnormally, however, several pollen tubes may develop out of
the same pollen as in some Malvaceae, Cucurbitaceae, etc., or, the single pollen
tube may be branched.

The pollen tube grows easily through the stigmatic papillae (Fig. 415) and then
passes into the tissues of the style. The length of the stylar tissue which the
pollen tube has to traverse may vary from nil in sessile stigmas to some 50 cm
in the maize plants provided with silk (Fig. 416). Styles are solid or hollow.

The centre may be filled with a longitudinal transmitting tissue or, it may be a
canal full of mucilage. The pollen tube may grow through this or through the
tissue of the stylar wall. Usually, only the distal portion of a long pollen tube
c6r»tains living protoplasm and, as the nuclei within it passes forward, callose
plugs are left in the empty portions behind them.

When the pollen tube (now with the male gametes) reaches the ovary its course
is to find out an ovule and to reach the embryo sac. The pollen tube usually
enters the ovule (orthotropous or anatropous) through the micropyle. This is
known as porogamy (Fig. 417). But, in some cases the pollen tube is found to
enter through the chalaza. This latter method, known as chalazogamy (Fig.
418), is found in Casuarinaceae, Betulaceae, Fagaceae and other members of
Archichlamydeae.
Formerly, much phylogenetic importance used to be attached to this character
but, now it seems that the phenomenon is more of physiological importance as
the same species may show both types of behaviour. In some cases (e.g.,
Cucurbita) the pollen tube enters the embryo sac piercing the integuments. This
is termed mesogamy.

In the passage of the pollen tube towards the micropyle, it has to pass through
some empty space. Sometimes the pollen tube is directed in its growth through
this distance by an obturator which forms a sort of a plug on the micropyle and
through which the pollen tube may grow. The obturator is usually of placental
origin.

Fertilization:
On, penetrating the nucellus, whether through the micropyle, or, by a round
about way through the chalaza, or, by piercing the integuments, the pollen tube
penetrates the embryo sac and enters the egg apparatus. Ultimately, the tip of
the pollen tube bursts and both the gametes are discharged. Probably the
synergids play little or no role in this act. One of these gametes fuses with the
egg cell of the egg apparatus.

The other gamete fuses with the secondary fusion nucleus. This behaviour of
the two male gametes is termed double fertilization which was first observed by
Nawaschin (1898) in Lilium and Fritilaria species. As a result of the first
fertilization the oospore cell is formed which is the mother cell of the embryo
and is a diploid cell containing 2n complement of chromosomes whereas the
microspore and all nuclei of male and female gametophytes are haploid with n
complement of chromosomes.

The secondary fusion nucleus of the embryo sac, however, normally becomes
2n at the time of the fusion undergone by it. (Exceptions to this have already
been mentioned in connection with female gametophyte development). So, the
fusion of this nucleus with the second male gamete is triple fusion and the
resultant nucleus is triploid or 3n. This is the first nucleus of the endosperm.

Embryogeny and Endosperm Formation:

In the post-fertilizational changes within the ovule, the embryo and the
endosperm are seen to develop simultaneously (Fig. 420). The oospore forms
the embryo while the fertilized fusion nucleus (product of triple fusion)
develops the endosperm. The other nuclei or cells within the embryo sac
(synergids, antipodal cells) may behave differently but are destined to
disorganise sooner or later.
Development of the Endosperm:
The Angiospermic endosperm, except in special cases, is a triploid (3n) tissue as
it is a product of triple fusion involving double fertilization. It is, thus, distinct
from the endosperm of Gymnosperms and heterosporous Pteridophytes where
the endosperm is a simple haploid (n) tissue of the gametophyte not involving
any complication like polar fusion or fertilization.

The real function of the endosperm is supplying nutrition to the growing

embryo.

There are at least two families, Orchidaceae and Podostemonaceae, where the
product of double fertilization soon disintegrates and endosperm development is
completely suppressed.

In other plants, three types of endosperm development, nuclear, cellular

and helobial, are usual:
(1) Nuclear Type:
This is the common type of endosperm development. Here, the endosperm
nucleus gives rise to a number of free nuclei (Figs. 421 a, 424) which remain in
the peripheral layer of embryo sac cytoplasm surrounding a large central
vacuole. The nuclei, usually, become cut off by cell walls at a later stage if they
be not already absorbed by the growing embryo. Such cell wall formation
usually begins from the basal periphery.

The cells soon organise into an endosperm tissue and goes on increasing further.
Development of the nuclear endosperm in Capsella bursa-pastoris (Fig. 424) is
described later. In some cases the central vacuole may not be filled up even in
the mature seed. This is seen in the palms. In coconut, the central cavity full of
coconut water is the original embryo sac vacuole while the nuclei around it
form the peripheral endosperm kernel.

(2) Cellular Type:

In a number of plants belonging to Annonaceae, Gentianaceae, etc., and in
Adoxa, Peperomia, Villarsia (Fig. 421B), etc., division of the original
endosperm nucleus is immediately followed by wall formation so that the
endosperm is cellular from the beginning although some cells may enclose more
than one nucleus.

(3) Helobial Type:

Among members of Helobiales (e.g., Vallisneria, Eremurus, Limnophyton, etc.)
there is a type of endosperm development which is intermediate between the
nuclear and the cellular types. Here, a partition wall develops between the two
nuclei resulting from the first division of the endosperm nucleus so that the
embryo sac is divided into two compartments.

A large number of free nuclei is now developed in the upper chamber while the
lower nucleus forms a few of them or may not divide at all (Fig. 421C).

Development of the Embryo:

Side by side with the development of the endosperm, the zygote or oospore is
developing the embryo. Both dicotyledons and monocotyledons begin embryo
development in the same way but, there is considerable difference in later
differentiation. In all Angiosperms the zygote (Fig. 422A) divides to develop a.
two-celled proembryo (Fig. 422B). In most plants the first wall between the two
cells is transverse while only in a few cases the first wall is more or less vertical
(Piperad type, Fig. 423A1).
Of the two cells the one near the micropyle is termed the basal cell while the
one pointing towards the centre of the embryo sac is called the terminal cell.
The basal cell usually forms the suspensor and may or may not contribute
towards the future embryo in whose development the terminal cell takes the
main part.

Johansen (1950) recognizes six types of embryo development among the

Angiosperms according to the manner of differentiation of the basal and
terminal cells:
Although there is no essential difference between the embryogeny of
monocotyledons and that of dicotyledons in the early stages, later
differentiation makes the two embryos so different that the development of
two such embryos are described separately below:
(1) Typical development of dicotyledonous embryo and endosperm:
The classical example of dicotyledonous embryo development is the plant
Capsella bursa-pastoris of Cruciferae. The ovule is campylotropous so that the
embryo sac and the later developed endosperm as well as embryo are
horseshoe-shaped.

Moreover the micropyle being pointed downwards, the embryo looks upside
down. Embryo development here is of the typical Onagrad or Crucifer type
described above.

The oospore (Fig. 424A) divides by a transverse wall forming a large basal cell
and a smaller terminal cell (Fig. 424B). The basal cell now divides transversely
while the terminal cell divides by a vertical wall developing a │ -shaped 4-
celled proembryo (Fig. 424B).
Next, the second basal cell quickly divides by a number of transverse walls
giving rise to a row of cells called the suspensor (Fig. 424C). The lowest
(micropylar) cell of the suspensor remains disproportionately large. As the
suspensor increases in length, it pushes down the terminal embryonal cells
deeper into the embryo sac. The embryonal cell, meanwhile, divides by three
walls at right angles to one another giving rise to eight cells or octants (Fig.
424C).

This is called the embryonal mass. The lowest cell of the suspensor is called the
hypo physis. The embryonal mass, along with the hypophysis, divides further
(Fig. 424 D & E).

Ultimately, the four terminal octants form the two cotyledons, the four
micropylar octants form the hypocotyl and the core of the radicle while the
hypophysis forms the cortex and the epidermis of the radicle as well as the root-
cap (Fig. 424 F & G). In the final stage (Fig. 424H) we find the mature embryo
with the plumule developed also out of the four terminal octants. The suspensor
gradually withers as the radicle is developed.

As the embryo is developing, the endosperm also keeps pace. Endosperm

development, in this case, is strictly nuclear. The endosperm mother nucleus
rapidly gives rise to a number of free nuclei which are arranged in the
cytoplasm lining the periphery of the embryo sac while the centre is occupied
by a big vacuole (Fig. 424D).

Gradually, cell wall formation begins from the periphery (Fig. 424F). The
antipodal cells, in this, case, form a tissue which is short-living. The endosperm
supplies nutrition to the embryo mainly through the suspensor. As the embryo
of Capsella increases, the endosperm presses upon and sucks in plenty of food
material from the nucellus which is soon consumed.

In the final stage (Fig. 424H) the nucellus has been completely consumed and
most of the endosperm has also been consumed by the embryo. In the mature
seed even this remnant of endosperm disappears so that the seed becomes
exalbuminous.
(2) Typical Monocotyledonous Embryo:
There is no essential difference between the embryogeny of monocotyledons
and that of dicotyledons. The endosperm develops in the same way but, as a
monocotyledonous embryo develops a single cotyledon instead of two, there is
some difference in later differentiation.

Luzula forsteri of Juncaceae may be taken as a type for monocotyledonous

embryogeny. Here also the development is of the Onagrad or Crucifer type. The
oospore divides by a transverse wall to form a large basal cell a and a’ smaller
terminal cell b (Fig. 425A).
The terminal cell b divides into two by a longitudinal wall and the basal
micropylar large cell a cuts off a cell a’ by a transverse wall developing a │ -
shaped 4-celled proembryo (Fig. 425B).
The two cells b now divide into four quadrants by another wall at right angles to
the first, the cell a’ divides into two by a longitudinal wall and the cell a cuts off
a second cell a” by a transverse wall (Fig. 425C). The cell acts as the
suspensor— while all the other cells take part in the formation of the embryo.
The quadrants of b divide further giving rise to a tissue of which the lower half
b (l) is destined to give rise to the lower part of the single cotyledon and the
upper half b'(l’) to the upper part of the cotyledon, the hypocotyl with the core
of the radicle and, later, the plumule.

The cell a’ on the top of it, forms a tissue which develops the upper part of the
radical while a” develops the root-cap tip (Fig. 425 D, E, & F).

Apomixis:
Apomixis is a term coined by Winkler to signify any asexual method of
propagation not involving the normal production of embryo by fertilization.
This definition includes even propagation by bulbils. But, following the usual
practice, here a number of cases are being discussed which involve seed or
embryo formation without fertilization.

Parthenogenesis means development of an embryo directly from an egg cell or a

male gamete. Apogamy means the development of a sporophyte (i.e., embryo)
out of any gametophytic cell without the union of gametes. Apospory means the
formation of a gametophyte (i.e., embryo sac or pollen) on a sporophyte without
any reduction division.
Parthenogenesis, therefore, is a type of apogamy. Parthenogenesis of a normal
haploid egg into an embryo has been noticed in Solanum nigrum, etc. This
embryo and the resultant plant are haploid. This is termed haploid
parthenogenesis. Haploid plants are usually sterile. Cases are also known (as in
different species of Nicotiana and Crepis) where such embryos actually
developed from male gametes. A second type of parthenogenesis, called diploid
parthenogenesis, has also been noticed.

In these cases, the embryo sac develops from a diploid, i.e., sporophytic cell of
the ovule without any reduction division (i.e., by apospory) so that the embryo
sac and all cells within it are diploid. The diploid egg of this embryo sac
develops a diploid embryo. This has been noticed in Taraxacum albidum and
other plants.

Apogamy of other types also has been noticed. Any other haploid cell of a
normal gametophyte (e.g., synergid or antipodal cell) may develop into an
embryo. The resultant plant also is haploid as has been noticed in the orchid
Orchis. On the other hand, if a synergid or an antipodal cell of a Taraxacum
type embryo sac (i.e., diploid developed aposporously) develops into a diploid
embryo, it will be a case of diploid apogamy.

Apospory, i.e., development of a gametophyte on the sporophyte without

reduction division as seen in Taraxacum albidum has also been noticed in a few
other plants like Eupatorium glandolusum, several species of Hieracium, etc.

An embryo may also develop directly from any diploid sporophytic cell (e.g.,
cells of nucellus, integument, etc.). This may be considered as vegetative
growth of the Category of bulbils although they are not always so simple. This
is sometimes called adventive embryony or sporophytic budding.

Polyembryony:
Polyembryony or the development of several embryos within the same ovule is
quite common among Gymnosperms but rare among Angiosperms. This is often
found in Citrus species (lemons and oranges —Fig. 426).

In these, the additional embryos are, usually, of nucellar origin, as discussed

under adventive embryony above. Such additional embryos may also originate
from the cleavage or splitting of the zygote (Cymbidium bicolor) or proembryo
as seen in Erythronium americanum. Additional embryos are known to develop
by epidermal budding of the first embryo (Nicotiana rustica) or from a
filamentous proembryo (Nymphaea advena).

More commonly, two embryos are found in a seed of which one is normal, and
the other is apomictic, the latter being usually haploid. Such haploid-diploid
twin embryos are known in many cultivated plants like rice, linseed, wheat,
potato, chilli, cotton, etc.

N.B. Johansen is of opinion that the term polyembryony should be strictly

applied only when all the embryos are developed from the zygote, the
proembryo, or by budding of the first embryo. The haploid-diploid embryos or
such apomictic embryos are, thus excluded.

The Seed:
When the embryo is developed fully, the ovule or the megasporangium becomes
the seed. With increase in the size of the ovule the integuments slowly dry up
and form the protective seed coat, the outer integument forming the testa and
the inner the tegmen.

Simultaneously, the internal organs also lose moisture and all food substances
are transformed into insoluble reserve forms. Physiological activities are
reduced to a minimum until the whole structure becomes dormant and almost
dry when we call it seed. The seed is, therefore, a mature integumented
megasporangium.

It has been seen that the developing seed contains a growing embryo within a
growing endosperm, and the whole is contained within the nucellus. As the
embryo increases, the endosperm decreases and the growth of both decreases
the nucellus. Finally, an equilibrium is reached when seed development is
complete.

According to the final equilibrium, it is possible to get three types of seeds

among both dicotyledons and monocotyledons:
A. If the embryo completely fills up the ovule consuming both endosperm and
nucellus, the seed is exalbuminous, there being no endosperm (Fig. 427). In
such a seed food for the embryo is kept stored in the cotyledons.
B. If the endosperm be not completely consumed the embryo remains
comparatively small and a large quantity of stored food (oily, mealy, horny,
etc.) remains within the endosperm which the embryo could not consume but
would consume in the germinating stage. Such a seed is albuminous (Fig. 428).
C. If even the nucellus is not consumed fully, it remains in the seed as a layer of
perisperm (Fig. 429) over and above the endosperm and the embryo. This is the
real nature of the perisperm as has been described in a number of
monocotyledonous and dicotyledonous seeds.

Aril and Caruncle:

Alter fertilization, an outgrowth looking like a third integument is sometimes
seen to develop from the funicular (i.e., hilar) end. In litchi (Litchi chinensis)
such a juicy fleshy outgrowth develops within the pericarp.

This (Fig. 430) is called the aril. In nutmeg (Myristica fragrans of

Myristicaceae), such-a red-coloured, dry, leathery and branched aril (Fig. 431)
forms the valuable spice ‘mace’ of commerce. There is a pulpy aril in Garcinia
cowa and Garcinia mangostana.
In castor and some other Euphorbiaceae, a smaller outgrowth develops near the
micropylar end. This is called the caruncle (Fig. 35). In pansy (Viola) such a
caruncle develops at the hilum.