Exercise 1

Introduction to Plant Embryology

1.1.Introduction
The embryology of angiosperms, a research area encompassing investigations on virtually
all the events relevant to sexual reproduction, has contributed much over the last century to
an understanding of the structural diversity in micro- and megasporogenesis. the
development of micro and megagametophytes, the fertilization, and the development of
embryo, endosperm and seed coat. At various times in the past, an attempt has been made
to compile accumulated data and to clarify embryological attributes for each family. Thus, the
embryology is at present incorporated in most general publications on angiosperm
systematics as one of the major sources of systematic characters. The systematic value of
embryological characters has often been persuasively argued. In so doing, studies have
usually demonstrated the utility of embryological information in clarifying the position of
certain problematic families.

Davis (1966) and Philipson (1974) have taken a broader view on assessing the systematic value
of embryological characters; in particular, they have attempted to show a restricted
occurrence of character states in angiosperms. The most illustrative evidence for consistent
occurrence of certain character states was presented by Dahlgren (1975) in his two-
dimensional map, the "Dahlgrenogram", a construct further refined by Gertrud Dahlgren in
the XIV International Botanical Congress at Berlin in 1987. Despite the increasing value of
embryological characters in angiosperm systematics, however, studies on embryology have
apparently become progressively inactive over the last two decades. Early in 1960s more than
150 articles on plant embryology were published a year, while early in 1980s, only 20-30
papers were published a year. This may have occurred in part because most of the major
easily accessible groups have already been studied, and in part because the time-consuming
and difficult tasks of embryology, largely based on microtome sections and optical
microscopy, are no longer fashionable and have been increasingly replaced by studies using
new technologies (e.g., electron microscopy and molecular techniques). However, the
information obtained by such new technologies is still extremely limited, and our knowledge
is still much far from sufficient to allow overall comparisons of any higher rank taxa. Further,
even where other evidence is not yet fully available or has failed to resolve disputed
systematic problems, embryological characters alone can often provide a sound basis for the
resolution of such problems.

1.2.Objectives:
After studying this unit, you will be able to:
1. define the term embryology.
2. know the developmental process in plant embryology.
3. know fertilization and post fertilization changes.

1.3.Life Cycle of Angiosperms

Angiosperms, have two phases in their life cycle a diploid sporophytic phase and a haploid
gametophytic phase. The sporophytic phase is the main phase of angiosperm's life cycle. At
maturity, the sporophyte produces flowers which bears fertile appendages stamens
(microsporophylls) and carpels (megasporophylls). The stamen has a proximal sterile part, the
filament, and a distal fertile part, the anther. Microsporangia present in anther contains
numerous pollen grains (microspores). The carpel has a proximal ovary which bears ovule, a
distal stigma to receive pollen grains and a sterile region style between stigma and ovary.
 Pollination is the transfer of pollen grains from anther to the stigma of flower. It occurs in
angiosperms either by sell-pollination or by cross pollination. It results in fertilization
fusion of egg (female gamete) present in embryo sac with male gametes present in pollen
tube.
In angiosperm, double fertilization and triple fusion is a unique phenomenon. Double
fertilization is fusion of one male gamete with egg cell and other male gamete with polar
nuclei is triple fusion because here three nuclei fuse together (one nucleus of male gamete
and two polar nuclei).

Double fertilization results in formation of zygote which develops into embryo. The
embryo may be dicotyledonous in dicot and monocotyledonous in monocots. The product
of triple fusion is primary endosperm cell which give rise to endosperm (cellular, nuclear
or helobial type). The endosperm is nutritive tissue and provides nutrition to the
developing embryo. The embryo develops into seed and ovary into fruits. The seed may
be endospermic, non endospermic or ex albuminous when endosperm is absent in it.
1.3.1. Microsporogenesis.
Typically, each anther has two lobes and each lobe has a pair of pollen sac or
microsporangia. Microspores or pollen grains are produced in microsporangia after
microsporogenesis. During microsporogenesis the nucleus of each microspore mother cell
or pollen mother cell (2n) undergo meiosis or reduction division, giving rise to four haploid
('n' number of chromosomes) microspores. Aggregates of four microspores are called
microspores tetrad. Based on the arrangement of spores, tetrad are classified as:
a.) Tetrahedral: Four microspores are arranged in tetrahedral manner.
b.) Isobilateral: The four microspores are arranged in one plane.
c.) T-Shaped: Out of the four microspore, two lies perpendicular to the others so
that tetrad has the shape of "T" .)
d.) Decussate: Microspores may be arranged in decussate manner. Sometimes
microspores do not separate from each other and remain stuck together in groups.
Such groups are called compound pollen grains eg. pollinium. In this all
microspores (pollens) in a pollen sac remain together to form a single mass called
pollinium.
Each pollen grain has an outer thick exine (decay-resistant outer coat) and inner
then intine (inner layer). The germination of pollen grain begins before pollination
and pollen grains have a generative cell and a tube cell. The generative cell further
divides mitotically to form two non-motile male gametes whereas tube cell forms
pollen tube. The pollen grain having male gametes with pollen tube represents the
mature male gametophyte.
1.3.2. Megasporogenesis
The ovule or megasporangium consists of nucellus and its protective coats the
integument. It remains attached to the placenta, on the inner wall of the ovary by a stalk
called funicle. The body of ovule attaches to the funicle at a point known as hilum.
An ovule has various parts like funicle, nucellus integument, micropyle, chalaza, hilum and
embryo sac. All parts of an ovule are sporophyte (2n) located in the nucellus, developed
from megaspore. The six main types of ovules e.g. orthotropous, anatropous,
campylotropous, amphitropous, hemainatropous and circinotropous.
The development of the megaspore within the ovule (megasporangium) is known as
megasporogenesis. During megasporogenesis the primary sporogenous cell
differentiated into nucellus functions as megaspore mother cell. It is the last cell of the
sporophytic generation and undergoes a reduction division (meiosis) to form four haploid
 megaspores. Out of four megaspores, three towards micropylar end degenerates and one
lowest remain functional. It represents first cell of female gametophyte and give rise
embryo sac.
The mature embryo sac has an egg (n) and two synergids (N) towards. micropylar end,
three antipodal cells (N), towards chalazal end and one secondary nucleus (2N) in the
center.

1.3.3. Pollination
The transfer of pollen grains from anthers to the stigma of a flower is called
pollination. The transfer of pollen grains from an anther to the stigma of the same flower
or to a flower on the same plant is known as self-pollination. For self-pollination flower
and anther and stigma of a flower ripe almost simultaneously must be bisexual. The
pollination of a flower by its own pollens is known as autogamy. The self- pollination
occurring between two different flowers present on the same plant is known as
geitonogamy.
The self-pollinated flower has homogamy and cleistogamy as an adaptation. In
homogamy, anther and stigma of a flower mature at the same time. In cleistogamy,
bisexual flower never open. Self-pollination is almost certain in a bisexual flower where
stamens and carpels mature at the same time. But the continued self-pollination
generation after generation results in weaker progeny.
The transfer of pollen grains from anther to the stigma of another flower is called cross-
pollination. For cross-pollination, flowers are mostly unisexual. Self- sterility, unisexuality,
dichogamy, heterostyly and herkogamy are various adaptation for cross- pollination.
Better seeds and healthier offspring are produced by cross-pollination. The process of
cross- pollination depends on various external agencies which may be abiotic or biotic.
The abiotic agents are nonliving agents such as air, water. The biotic agents include living
organisms such as insects, birds.
Pollination by wind is called anemophily
Pollination through water is called hydrophily
Pollination by bird is called ornithophily
Pollination by bat is called chiropterophily
Pollination by snail or slugs is called malacophilous
Pollination with the help of ants is called myrmecophily
Pollination by humans is artificial pollination or anthropophily
Cross-pollination results in healthier offspring, viable seeds and new varieties of crops.
1.3.4. Fertilization
In an embryo two sexual fusion occurs, one is syngamy (Le. fusion of one male
gamete with the egg) and another is triple fusion (ie fusion of other male gamete with
the polar nuclei or secondary nucleus) and therefore, the phenomenon is known as
double fertilization. This is a very unique phenomenon in angiosperms and was discovered
by S.G. Nawaschin in 1898. Once the pollen grain reaches the receptive stigma due to
pollination, it germinates producing a long slender pollen tube. Two male gametes and
the tube nucleus migrate into the pollen tube which now represents the mature male
gametophyte. Pollen tube penetrates the stigmatic tissue and pushes its way through the
style and then down the wall of the ovary. After arriving in the ovary, the pollen tube may
enter into the ovule through micropyle (porogamy), through chalazal end (chalazogamy),
 or through integuments (mesogamy). Irrespective of the route of the entry of the pollen
tube into the ovule, it always enters the embryo sac from the micropylar end. After
entering into the embryo sac, the tip of the pollen tube bursts and the two male gametes
are released. Out of the two male gametes one male gamete fuses with the egg nucleus
and result in the formation of zygote or oospore (2n) Second male gamete fuses with two
polar nuclei, resulting into primary endosperm nucleus (3n). This phenomenon is termed
as double fertilization.
1.3.5. Post Fertilization Development
Post fertilization development means development of the embryo and the
endosperm within the ovule. Both the development goes side by side in the ovule. The
endosperm (3N) develops from primary endosperm nucleus. This undergoes a series of
divisions and ultimately forms endosperm. Depending upon the mode of development,
three types of endosperms has been recognized. (1) nuclear type (2) cellular type (3)
helobial type. The endosperm is nutritive tissue, providing nutrition to the developing
embryo. With the development of endosperm, the oospore develops in the embryo side
by side. The development of embryo from diploid oospore is called embryogenesis.
1.3.6. Development of Dicot Embryo (Crucifer type)
1. Zygote (oospore) divides transversely. As a result of this a two-celled pro embryo is
formed
2. The larger basal cell at the micropylar end is called suspensor cell. The smaller one,
away from it termed as terminal cell or embryo cell.
3. The suspensor cell divides transversely a few times to produce a filamentous suspensor
of 6 to 10 cells. The suspensor helps in pushing the embryo in the endosperm.
4. The first cell of the suspensor (towards micropyle) becomes swollen and called
haustorium or vesicular cell.
5. The last cell of suspensor (towards embryo cell) is known as hypophysis. It forms radicle
and root cap.
6. The embryo cell undergoes two vertical divisions and one transverse division to form
quadrant and then octant stage. In octant, eight cells arranged in two tiers- epibasal
(terminal) and hypobasal (near the suspensor).
1.3.7. Development of Monocot Embryo
The early development of dicot and monocot embryos are similar uptooctant stage. Later
on differentiation starts.
1. The zygote or oospore elongates and then divides transversely to form basal and
terminal cells.
2. The basal cell (towards micropylar end) produces a large swollen, vesicular suspensor
cell. It may function as haustorium.
3. The terminal cell divides by another transverse wall to form two cells.
4. The top cell after a series of divisions forms plumule and a single cotyledon.
5. Cotyledon called scutellum, grows rapidly and pushes the terminal plumule to one side.
The plumule comes to lie in a depression.
6. The middle cell, after many divisions forms hypocotyl and radicle. It also adds a few
cells to the suspensor.
7. In some cereals both plumule and radicle get covered by sheaths developed from
scutellum called coleoptile and coleorhiza respectively.
1.3.8. Polyembryony and Apomixis
Polyembryony is defined as the development of several embryos within the same
ovule. It is common in gymnosperms but very rare in angiosperms. In angiosperms poly
embryo may be due to:
1. Cleavage of the pro embryo.
2. Embryos developing from cells of the embryo sac other than the egg.
3. More than one embryo sac in the same ovule.
4. Activation of some sporophytic cell of the ovule.
1.3.9. Apomixis, the phenomenon of substitution of sexual process by asexual methods. The
plants which show apomixis are called apomictic plants.
1. calyx It is the outer most whorl of the flower. It is composed of leaf like green sepals
2. corolla it is the second whorl of flower and consists of number of petals
3. androecium It is the third whorl of flower consisting of stamens. Stamen is the male
reproductive organ of flower. Each stamen is made up of filament and anther
4. gynoecium the last and fourth whorl of the flower consisting of pistil or carpel Pistil is
the female reproductive organ of the flower. Each pistil is composed of ovary, style and
stigma.