Module 2

Plant Development
Learning objectives:
At the end of the module, you are expected to :
1. Understand the different processes and structures involved in plant embryonic
development
2. Learn how different plant parts derived from meristems.
3. Comprehend the flowering development and control of flowering in plants.

2.1. Embryonic Development

One general difference between plant and animal development is that most of
the development occurs not in the embryo but in the growing plant. Unlike an animal
embryo, the mature plant embryo inside a seed is not simply a smaller version of the
organism it will become. All the structures of the plants - shoots, roots, stalks, leaves
and flowers - are produced after the germination from localized groups of
undifferentiated cells knows as meristems. As such, two meristems are established in
the plant embryo: one at the tip of the root and the other at the tip of the shoot. Thus,
cells within the meristems can divide repeatedly and can potentially give rise to all plant
tissues and organs. This means that developmental patterning within the meristems to
produce structures continues throughout a plant’s life. Other key differences are as
follows:
1. Presence of rigid cell wall in plant cells therefore, there is no cell migration.
Instead, form is largely generated by differences in rates of cell division and by
division in different planes, followed by cell enlargement.
2. A complete, fertile plant can develop from a single differentiated somatic cells and
not just from a fertilized egg. This suggest that some differentiated plant cells
may retain totipotency (cells that can give rise to a new organism)

Plant embryos develop through several distinct stages

Like an animal zygote, the fertilized plant egg cell undergoes repeated division,
cell growth and differentiation to form a multicellular embryo. The first division of the
zygote is at the right angles to the long axis, dividing it into an apical cell and a basal
cell (Figure 2-1) establishing an initial polarity that is carried over into the apical-basal
polarity of the embryo and into the apical-basal axis of the plant. In many species, the
first zygotic division is unequal, wherein the basal cell is larger that the apical cell. As
such, the basal cell divides and gives rise to the suspensor, which attaches the embryo
to maternal tissue and is a source of nutrients. As such, the apical cell divides vertically
to form a two-celled proembryo, which will give rise to the rest of the embryo.
The next two division produce an eight-cell octant-stage embryo, which
develops into globular-stage embryo of around 32 cells (Figure 2-1). The embryo
elongates and the cotyledons (seed leaves) start to develop as wing-like structures at
one end, while an embryonic root forms at the other known as the heart stage.
Moreover, there are two groups of undifferentiated cells capable of continued division
are located at each end of this axis and these are known as apical meristems. The
meristem lying between the cotyledons gives rise to the shoot, while the one at the
opposite end of the axis, towards the end of the embryonic root will drive root growth at
germination. The region in between the embryonic root and the future shoot will become
the seedling stem or hypocotyl. Almost all above ground adult plant structures are
derived from the apical meristems. The main exception is radial growth in the stem,
which is most evident in woody plants, and which is produced by the cambium, a ring
of secondary meristematic tissue in the stem. After the embryo is mature, the apical
meristems remain quiescent until germination.

Figure 2-1. Pattern of cell division in embryonic plant development

Seedling structures can be traced back to groups of cells in the early embryo to
provide a fate map (Figure 2-2). In Arabidopsis, patterns of cleavage up to the 16-cell
stage are highly reproducible, and even at the octant stage, it is possible to make a fate
map for the major regions of the seedling along the apical-basal axis. The upper tier of
cells give rise to cotyledons and shoot meristems, the next tier is the origin of the
hypocotyl, and the bottom tier together with the region of the suspensor where it joins
the embryo will give rise to the shoot. At the heart stage, the fate map is clear

Figure 2-2. Fate map of the Arabidopsis embryo. The stereotyped pattern of cell division
in dicotyledon embryos means that at globular stage, it is already possible to map the
three region that will give rise to the cotyledons (dark green) and shoot meristem (red),
the hypocotyl (yellow), and the root meristem (purple) in the seedling. Scheres, B. et al.,
1994.

Furthermore, the radial pattern in the embryo comprises three (3) concentric
rings of tissue namely the outer epidermis, the ground tissue (cortex and
endodermis), and the vascular tissue at the center. This radial axis appears first at the
octant stage, when adaxial (central) and abaxial (peripheral) regions become
established. Subsequently, periclinal divisions occur when the plane of division is
parallel to the outer surface that give rise to the different ring of tissue whereas anticlinal
divisions happens when the plane of cell division is at right angles of the outer surface,
increasing the number of cells in each ring of tissue (Figure 2-3). At the 16-cell stage,
the epidermal layer (dermatogen) is established.
Figure 2-3. Periclinal and anticlinal divisions. Periclinal cell division are parallel to the
organ surface, whereas anticlinal division are at right angles to the surface

Gradients of the signal molecule auxin established the embryonic apical-basal

axis
The small, organic molecule auxin (indoleacetic acid or IAA) is one of the most
important and ubiquitous chemical signals in plant development and plant growth. It
causes changes in gene expression by promoting the ubiquitination and degradation of
transcriptional repressors known as Aux/IAA proteins. In the absence of auxin, these
bind to proteins called auxin-response factors (ARFs) to block the transcription of so-
called auxin-responsive gene. Auxin-stimulated degradation of Aux-IAA proteins frees
the auxin response factors, which can then activate, or in some cases repress, these
genes (Figure 2-4). Auxin-responsive genes include gene involved in specifying cell fate.
In some cases, auxin appears to be acting as a classical morphogen, forming a
concentration gradient and specifying different fates according to a cell’s position along
the gradient.

Figure 2-4. Auxin signaling pathway. In

the absence of auxin, AUX/IAA proteins
bind to and repress the activity of
transcription factors in the Auxin
Response Factor (ARF) family, which
bind TGTCTC-containing DNA
sequence elements in the promoters of
auxin-response gene. Auxin targets the
ubiquitin ligase complex SCF/TIR1 to
the AUX/IAA protein, causing it to be
ubiquitinated (Ub). This modification
targets AUX/IAA for degradation and
thus lifts the repression of the auxin-
responsive gene. Which can then be
transcribed.
Chapman and Estelle, 2009
 The earliest known function of auxin in Arabidopsis is in the very first stage of
embryogenesis, where it establishes the apical-basal axis. Immediately after the first
division of the zygote, auxin is actively transported from the basal cell by the auxin-
efflux protein PIN7, which is localized in the apical plasma membrane of the basal cell.
The auxin is required to specify the apical cell, which give to all the apical embryonic
structures such as shoot apical meristem and cotyledon. Through the subsequent cell
divisions, transport of auxin continues up through the suspensor cells and into the cell at
the base of the developing embryo until the globular-stage embryo of about 32 cells.
The apical cells of the embryo then start to produce auxin, and the direction of the auxin
transport is suddenly reversed. PIN7 proteins in the suspensor cells move to the basal
faces of the cells. Other PIN genes are activated, and the concerted actions of the PIN
proteins cause auxin from the apical region to be transported into the basal region of the
globular embryo. (Figure 2-5)

Figure 2-5. The role of auxin in patterning the early

embryo. Left panel : auxin produced in the original
basal cell accumulates in the two-celled proembryo
(green) through transport in the basal to apical
direction mediated by protein PIN7, which is located
in the apical membranes of the basal and suspensor
cells (purple arrows). Another PIN protein, PIN1,
transports auxin between the two cells of the
proembryo (red arrows). Cell division distributes
auxin into the developing embryo. Right panel; at
the globular stage, free auxin starts to be produced
at the apical pole and the direction of auxin transport
is reversed. PIN7 becomes localized in basal
membranes of the suspensor cells and the proteins
PIN1 and PIN 4 (orange arrows) transport auxin
from the apical region into most basal cell of the
embryo, known as the hypophysis, where it
accumulates

Plant somatic cells can give rise to embryos and seedlings

As gardeners well know, plants have amazing powers of regeneration. A new
plant can develop from a small piece of stem or root, or even from the cut edge of a leaf.
This reflects an important difference between the developmental potential of plant and
animal cells. In animals, with few exceptions, cell determination and differentiation are
irreversible. By contrast, many somatic plant cells remain totipotent. Cells from roots,
leaves, stems and even for some species, a single isolated protoplast (a cell from
which the cell wall has been removed by enzymatic treatment) can be grown in culture
and induced by treatment with the appropriate growth hormones to give rise to a new
plant (Figure 2-6). Plant cells proliferating in culture can give rise to cell clusters that
pass through a stage resembling normal embryonic development although the pattern
of cell divisions are not the same. These ‘embryoids’ can develop into seedlings. Plant
cells can also form callus, an apparently disorganized mass of cells that can form new
shoots or roots.

Figure 2-6. Cultured somatic cells from a mature plant can form an embryo and
regenerate into a new plant. The illustration shows the generation of a plant from single
cells. If a small piece of tissue from a plant stem or leaf is placed on a solid agar
medium containing the appropriate nutrients and growth hormones, the cells will start to
divide to form a disorganized mass of cells known as a callus. The callus cells are then
separated and grown in liquid culture. In suspension culture some of the callus cells
divide to form small cell clusters. These cell clusters can resemble the globular stage of
a dicotyledon embryo, including the heart-shaped and later stages to regenerate a
complete new plant.

2.2. Meristems

In plants, most of the adult structures are derived from just two region of the
embryo, the embryonic shoot and root meristems, which are maintained after
germination. The embryonic shoot meristem, for example, becomes the shoot apical
meristem of the growing plant, giving rise to all the stems, leaves and flowers. As the
shoot grows, lateral outgrowths from the meristem give rise to leaves and to side shoots.
In flowering shoots, the vegetative meristem becomes converted into one capable of
producing floral meristems that makes flower, not leaves. In Arabidopsis, the shoot
apical meristem, changes form a vegetative meristem that makes leaves around it in a
spiral pattern to an inflorescence meristem that then produces floral meristems thus,
flowers around it in a spiral pattern. The first stages of future organs are known as
primordia (singular primordium). Each primordium consists of a small number of
founder cells that produce the new structure by cell division and cell enlargement,
accompanied by differentiation.
There is usually a time delay between the initiation of two successive leaves in a
shoot apical meristem, and this results in a plant shoot being composed of repeated
modules. Each module consists of an internode (cells produced by the meristem
between successive leaf initiations), a node and its associated leaf, and an axillary
bud (Figure 2-7). The axillary bud itself contains a meristem, known as the lateral shoot
meristem, which forms at the base of the leaf, and can produce a side shoot when the
inhibitory influence of the main shoot tip is removed. Root growth is not so obviously
modular, but similar considerations apply, as new lateral meristems initiated behind the
root apical meristem give rise to lateral roots.

Figure 2-7. Plants shoots grow in a modular

fashion. The shoot apical meristem produces a
repeated basic structural module. The
vegetative shoot module typically consists of
internode, node, leaf and axillary bud (from
which a side branch may develop). Successive
modules are shown here in different shades of
green. As the plant grows, the internodes
behind the meristem lengthen and the leaves
expand.
Alberts et. al., 2002

A meristem contains a small, central zone of self-renewing stem cells

Shoot apical meristems are rarely more than 250 um in diameter in
angiosperms and contain a few hundred relatively small, undifferentiated cells
that are capable of cell division. Most of the cell divisions in normal plant
development occur within meristems, or soon after the cell leaves a meristem,
and much of a plant’s growth in size is due to cell elongation and enlargement.
Cells leave the periphery of the meristem to form organs such as leaves of
flowers and are replaced from s central zone of slowly diving, self-renewing stem
cells or initials at the tip of the meristem (Figure 2-8). Initials behave in the same
way as animal stem cell wherein they can divide to give one daughter that
remains a stem cell and one that loses the stem cell property. This daughter cell
continues to divide and its descendants are displaced towards the peripheral
zone of the meristem, where they become founder cells for a new organ or
internode, leave the meristem, and differentiate.

Figure 2-8. Organization of the

Arabidopsis shoot meristem. A
longitudinal section is shown. The
meristem has three main layer, L1, L2,
and an inner layer (yellow lines). Further
divided into a central zone (red), a rib
zone (yellow), and a peripheral zone
(blue). The stem cells or initials lie in the
central zone, while the peripheral zone
contains proliferating cells that will give
rise to leaves and side shoot. The rib
zone gives rise to the central tissue of
the plant stem

Meristem stem cells are maintained in the self-renewing state by cells

underlying the central zone that form the organizing center. As we shall see, it
is the microenvironment maintained by the organizing center that gives stem
cells their identity.

Root tissues are produced from Arabidopsis root apical meristems by a

highly stereotyped pattern of cell divisions
The organization of the tissues in the Arabidopsis root tip is shown in
Figure 2-9. The radial pattern comprises single layers of epidermal, cortical,
endodermal, and pericycle cells, with vascular tissue in the center (protophloem
and protoxylem). Root apical meristems resemble shoot apical meristems in
many ways and give rise to the root in a similar manner to shoot generation. But
there are some important difference between the two. One of which is shoot
meristem is at the extreme tip of the shoot, whereas the root meristem is covered
by a root cap (which is derived from one of the layers of the meristem). Another
is that there is no obvious segmental arrangement at the root tip resembling the
node-internode-leaf module.
 Figure 2-9. The structure of the root tip in Arabidopsis. Roots have a radial
organization in the center of the growing root tip is the future vascular tissue
(protoxylem and protophloem). This is surrounded by further tissue layers.

The root is set up early and a well-organized embryonic root can be identified in
the late heart-stage embryo (Figure 2.10). As such in the shoot meristem, a root
meristem is composed of an organizing center called the quiescent center in root, in
which the cells divide only very rarely, and which is surrounded by the stem-cell like
initials that give rise to the root tissue. A key function of the quiescent center is to
maintain the immediately adjacent initials in the stem-cell state and prevent them from
differentiating.
Figure 2-10. Fate map of root regions in the heart-stage of Arabidopsis embryo. The
root grows by the division of a set of initial cells. The root meristem comes from a small
number of cells in the heart-shaped embryo. Each tissue in the root is derived from the
division of a particular initial cell. At the center of the root system is a quiescent center,
which does not divide

Moreover, the role of auxin in root development continues into the adult plant.
Auxin in the root is transported out of cells via the PIN proteins, and enters adjacent
cells. It plays a key role in patterning the growth root. Modeling of auxin movement,
using the known distribution of PIN proteins in the membranes of the different cell types
has shown that PIN-directed flow can explain the formation and maintenance of a stable
auxin maximum at the quiescent center. Auxin is transported down the central vascular
tissue of the root tip, forming a concentration maximum at the quiescent center, and
then outwards and upwards through the outer layers of the root tip. As such, auxin is
also involved in the ability of plants to regenerate from a small piece of stem. In general,
roots form from the end of the stem that was originally closest to the root, whereas
shoots tends to develop from dormant buds at the end that was nearest to the shoot.

Root Hairs
Root hairs are formed from the epidermal cells at regular intervals around the
root, and this regularity is thought to be achieved by a combination of responses to
positional information and lateral inhibition by the movement of transcription factors
between cells. Files of cells that will make roots alternate with files of non-hair-
producing cells on the surface of the developing root. The importance of position is
shown by the fact that if an epidermal cell overlies a junction between two cortical cells,
it forms a root hair, whereas if it contacts just one cortical cell it does not. (Figure 2-11).
And if a cell changes its position in relations to the cortex, its fate will also change, from
potential hair forming cell to a non-hair-forming cell and vice versa. Most cell divisions in
the future epidermis are horizontal, increasing the number of cells per file, but
occasionally a vertical anticlinal division occurs, pushing one of the daughter cells into
and adjacent file. The daughter cell then assumes a fate corresponding to its new
position in relation to the adjacent cortical cells.

Figure 2-11. Organization of cell type in

the root epidermis. The epidermis is
composed of two type of cell:
trichoblasts (T), which will form root
hairs, and atrichoblasts (A), which will
not. Trichoblasts overlie the junction
between two cortical cells and
atrichoblasts are located over the outer
tangenital wall of cortical cells.

2.3 Flowering Development and Control of Flowering

Flowers contain the reproductive cells of higher plants and develop from the
shoot meristem. In most plants, the transition from a vegetative shoot meristem to a
floral meristem that produces a flower is largely under environmental control in line with
daylength and temperature being important determining factors. Moreover, flowers are
complex structures with their arrangement of floral organs including sepals, petals,
stamens, and carpels (Figure 2-12).
The conversion of a vegetative shoot meristem into one that makes flowers
involves the induction of so-called meristem identity gene. As such, the key regulator
of floral induction in Arabidopsis is the meristem identity gene LEAFY (LFY).

Organ identity of a flower

The individual parts of a flower each develop from a floral organ primordium
produced by the floral meristem. Unlike leaf primordia, which are all identical, the floral
organ primordia must each be given a correct identity and be patterned according to in.
An Arabidopsis flower has four (4) concentric whorls of structures (Figure 2-12), which
reflect the arrangement of the floral organ primordia in the meristem. The sepals (whorl
1) arise form the outermost ring of meristem tissue, and the petals (whorl 2) form a ring
of tissue lying immediately inside it. An inner ring of tissue gives rise to the male
reproductive organs - the stamens (whorl 3). The female reproductive organs - the
carpels (whorl 4) will develop from the center of the meristem. In a floral meristem of
Arabidopsis, there are 16 separate primordia, giving rise to a flower with four sepals,
four petals, six stamens and a pistil made up of two carpels (Figure 2-12).

Figure 2-12. Structure of an Arabidopsis flower. Left : Arabidopsis flowers are radially
symmetrical and have an outer ring of four identical green sepals, enclosing four
identical white petals, within which is a ring of six stamens, with two carpels in the
center. Right : Floral diagram of the Arabidopsis flower representing a cross section
taken in the plane indicated in the left diagram. This is a conventional representation of
the arrangement of the parts of the flower, showing the number of flower parts in each
whorl and their arrangement relative to each other .

Control of flowering
In Arabidopsis, Flowering is promoted by increasing daylength, which predicts
the end of winter and the onset of spring and summer (Figure 2-13). This behavior is
called photoperiodism. In some strains, flowering is also accelerated after the plant
has been exposed to a long period of cold temperature, a cue that winter has passed.
This phenomenon is known as vernalization. Grafting experiments have shown that
daylengths is sensed not by the shoot meristem itself, but by the leaves. When period
of continuous light reaches a certain length, a diffuse of flower-inducing signal is
produced that is transmitted through the phloem up to the shoot meristem. The pathway
that triggers flowering involves the plant’s circardian clock, the internal 24-timer that
causes many metabolic and physiological processes, including the expression of some
genes, to vary throughout the day. Moreover, expression of floral meristem identity
genes such as LEAFY throughout the plant is sufficient to confer a floral fate on lateral
shoot meristem thus, they develop as flowers.

Figure 2-13. Flowering can be controlled

by daylength and LEAFY expression. As
shown in the top panels, when wild-type
Arabidopsis is grown under long-day
conditions (left), few lateral shoots are
formed before the apical shoot meristem
begins to form floral meristems. When
grown under short-day conditions,
flowering is delayed and there are in
consequence more lateral shoots. The
gene LEAFY is normally expressed only
in inflorescence and floral meristems,
but if it is expressed throughout the
plant, (bottom panels), all shoot
meristems produced are converted to
floral meristems in both daylengths.

Furthermore, one of the genes regulated by the circardian clock is CONSTANS

(CO), which is a key gene controlling the onset of flowering and provides the link
between the plant’s daylength-sensing mechanisms plus the production of the flowering
signal. The expression of CO oscillates on a 24-hour cycle under the control of the
circadian clock, and its timing is such that peak CO expression occurs towards the end
of afternoon. This means that in longer days, peak expression occurs in the light,
whereas in short days, it will already be dark at this time. In the dark, the CO protein is
degraded and so the circardian control ensures that CO only accumulates to high
enough level to trigger the flowering pathway when light conditions are favorable (Figure
2-14)

Figure 2-14. The initiation of flowering is

under the dual control of daylength and the
circadian clock. The transcription factor
CONSTANS (CO) is required for
production of the flowering signal and is
expressed in leaves under the control of
the circadian clock. In short days,
expression of the CO gene peaks and the
protein is rapidly degraded. In long days,
peak expression occurs in the light, and
the CO protein accumulates.

CO is a transcription factor that activates a gene known as FLOWERING LOCUS

T (FT), producing the FT protein, which appears to act as the flowering signal. The FT
protein is thought to travel from leaf through the phloem to the shoot apical meristem,
where it acts in a complex with the transcription factor (FD), which is expressed in the
meristem, to turn on the expression of genes (AP1) that promote flowering (Figure 2-15).
If FT is activated in a single leaf, this is sufficient to induce flowering. Induction of
flowering also requires the downregulation of a set of floral repressor genes such as
FLOWERING LOCUS C (FLC). These suppress the transition from a vegetative to the
flowering state until the positive signals to flower are received. FLC encodes a protein
that binds to FT and suppresses its activity. After cold exposure, for example, FLC
activity is low, and the repression of FT be released.
Figure 2-15. Signals that initiate flowering in Arabidopsis. When the daylength becomes
longer after winter, the transcription factor CO accumulates in the leaf, which in turn
activates the transcription of FT in leaf phloem cells. The FT protein travels through he
phloem to the shoot apex. This interacts with the transcription factor FD to form a
complex, which acts together with the transcription factor LAEFY (whose own
expression in upregulated in the shoot apex by FT) to activate key floral meristem
identity genes such as AP1, which convert the vegetative meristem into one that will
produce floral meristems.

References

1. Campbell, S. (2013). Essential Biology. Singapore: Pearson Educ, Inc.

2. Hickman, et.al. (2014). Integrated Principles of Zoology. New York: McGrawHill
3. Wolpert, L., Beddington, R., Jessell, T., Lawrence, P., Meyerowitz, E, and Smith,
J. (2002) Principles of Development, Second Edition. Oxford University Press:
Oxford
4. Wolpert, L., Tickle, C., & Arias, A. M. (2015). Principles of Development. Oxford
University Press, USA.