Plant Embryogenesis
(Zygotic and Somatic)
John J Harada, University of California, Davis, California, USA
Mark F Belmonte, University of Manitoba, Winnipeg, Manitoba, Canada
Raymond W Kwong, University of California, Davis, California, USA
Based in part on the previous version of this Encyclopedia of Life Sciences
(ELS) article, Plant Embryogenesis (Zygotic and Somatic) by John J
Harada and Raymond W Kwong.
Zygotic embryogenesis in higher plants describes the
developmental period in which the zygote undergoes a
series of differentiation events, leading to the formation
of a mature embryo. Establishment of the major embry-
onic organs and shoot and root apical meristems occur
though partitioning events along the apical–basal axis,
and many of these events are guided by the hormone
auxin. Formation of the three embryonic tissue systems
occurs along a radial axis perpendicular to the apical–
basal axis. The mature zygotic embryo is generally
developmentally arrested, metabolically quiescent and
enclosed within maternal tissues of the seed. Somatic cells
can be induced to divert from their normal fate and
develop into embryos in a process termed somatic
embryogenesis. Auxin and other plant hormones appear
to play critical roles in inducing embryogenic com-
petence. Zygotic and somatic embryogenesis represent
parallel developmental programs in which cells acquire
embryogenic cell fate and develop into mature embryos.
Introduction
Plants alternate between the haploid gametophytic and the
diploid sporophytic generations during their life cycles.
The sporophyte produces the megaspores and microspores
that give rise to the female and male gametophytes,
respectively. Egg and sperm cells made in the gametophytes
ELS subject area: Plant Science
How to cite:
Harada, John J; Belmonte, Mark F; and Kwong, Raymond W (October
2010) Plant Embryogenesis (Zygotic and Somatic). In: Encyclopedia of
Life Sciences (ELS). John Wiley & Sons, Ltd: Chichester.
DOI: 10.1002/9780470015902.a0002042.pub2
ENCYCLOPEDIA OF LIFE SCIENCES & 2010, John Wiley & Sons, Ltd. www.els.net
Introductory article
. Introduction
Article Contents
. Description of Embryogenesis
. Establishing the Plant Body Plan
. Somatic Embryogenesis
. Summary
Online posting date: 18th October 2010
combine to form the zygote, the initial stage of the sporo-
phytic generation. See also: Gametophyte and Sporophyte
Embryogenesis traditionally describes the develop-
mental period in which the zygote undergoes a series of
differentiation events, resulting in the formation of a
mature embryo with cotyledons, shoot apical meristem
(SAM), hypocotyl, root, root apical meristem (RAM) and
three embryonic tissue systems. Many facets of higher
plant embryogenesis are novel. For example, two fertil-
isation events are required for reproduction: one produces
the zygote and the other generates the endosperm, an
extraembryonic tissue system that is essential for zygotic
embryo development and that serves a nutritive role for the
embryo/seedling. Another unique aspect is that higher
plant embryogenesis can be divided conceptually into two
distinct phases. The body plan of the plant is established
during the early morphogenesis phase. In the late matur-
ation phase, the embryo acquires the ability to withstand
desiccation and accumulates storage reserves used to
nourish the developing seedling. This mode of embryo-
genesis differs substantially from that of lower plants that
do not make seeds and do not undergo the maturation
phase. Finally, plants are totipotent, that is, plant cells can
change their fate and be induced to regenerate a fully dif-
ferentiated adult organism. A variety of cell types, includ-
ing zygotic cells, somatic cells, microspore cells, nucellar
cells and unfertilised eggs, can undergo embryogenesis
although it is not known to what extent initiation of
embryo development through these different pathways
overlap. See also: Apical Meristems; Meristems
In this article, we discuss selected examples to illustrate
the processes that occur during embryogenesis in higher
plants. Although there is diversity in the details, the con-
ceptual processes underlying embryogenesis are likely to
apply broadly.
Description of Embryogenesis
Zygote formation
The ability of higher plants to make seeds has provided
many selective advantages to seed plants as compared with
1
 Plant Embryogenesis (Zygotic and Somatic)
nonvascular plants and seedless vascular plants (Steeves,
1983). Fertilisation occurs within the female gametophyte
that is surrounded by maternal tissue allowing this repro-
ductive step to occur in nonaqueous environments.
Development within the confines of maternal tissues also
affords some level of protection to female gametophytes
and embryos. In addition, the seed serves as an efficient
dispersal unit in which the embryo can remain dormant
until conditions for germination and seedling survival are
favourable. See also: Gametophyte and Sporophyte; Plant
Reproduction; Seed Germination and Reserve Mobiliza-
tion; Seeds
The multicellular female gametophyte, or embryo sac,
develops within the ovule, the progenitor of the seed. Figure
1a shows a female gametophyte of the Polygonum type that
consists of six haploid cells – one egg cell, two synergid cells
and three antipodal cells – and one diploid central cell in
which the two polar nuclei have fused. After pollination,
the male gametophyte forms a pollen tube that grows into
the ovule micropyle and penetrates one synergid cell of the
female gametophyte. Double fertilisation occurs following
the release of two sperm cells into the female gametophyte.
One sperm cell migrates to the egg cell, fuses with its plasma
membrane and its nucleus undergoes karyogamy with the
egg cell nucleus to form the zygote. The other sperm cell
fuses with the central cell to produce the triploid endo-
sperm mother cell. The developing endosperm undergoes
an initial period of nuclear divisions without cytokinesis,
producing a syncytium of nuclear–cytoplasmic domains.
Cellularisation of the endosperm occurs later in develop-
ment. In many plants such as beans, the endosperm can be
absorbed largely or completely by the developing embryo.
Alternately, the endosperm persists in the seed as a storage
organ and serves as a nutrient source for the developing
seedling as occurs in cereals. See also: Carpels; Endosperm
Development; Gametogenesis; Stamen and Pollen
Development
Morphogenesis phase of embryo
development
Higher plants display a diversity of embryo development
patterns. For example, six classifications of embryogenesis
have been defined based on the sequences of early cell
division, although the early cleavages of some plants such
as cotton are so variable that they cannot be classified in
these groups (Natesh and Rau, 1984). Therefore, there
is no universal pattern of embryo morphogenesis. As an
example, we describe embryogenesis characteristic of the
dicotyledonous crucifers Arabidopsis, Brassica and Cap-
sella bursa-pastoris.
Figure 1b and c show that in crucifers, the first division of
the zygote is transverse and asymmetric, generating cells of
different fates. The small apical cell gives rise to the embryo
proper with the exception of a part of the root meristem,
and the basal cell yields the suspensor and the other root
meristem components. However, this first division of the
zygote in other plants can be oblique, symmetrical or
2 ENCYCLOPEDIA OF LIFE SCIENCES & 2010, John Wiley & Sons, Ltd. www.els.net
longitudinal, indicating that a transverse, unequal division
is not a requirement for plant embryo formation (Natesh
and Rau, 1984).
The apical cell initially undergoes two longitudinal div-
isions to produce the four-cell embryo proper (Figure 1d).
The first transverse division of the embryo proper produces
the octant-stage embryo with an upper and lower tier of
cells, as shown in Figure 1e. The division plane created by
this cleavage, designated the O’ line, has been implicated to
serve as a boundary between two distinct embryonic
regions, the apical and the central domains (Mayer et al.,
1991). The next divisions of the octant-stage embryo occur
parallel to the embryo surface and form the protoderm, the
epidermis precursor that is the first morphologically
detectable embryonic tissue (Figure 1f). This set of cleavages
sets off outer from interior cells and establishes the globu-
lar-stage embryo, often described as having a radially
symmetrical morphology (Figure 1g).
The basal cell undergoes a series of transverse divisions
to produce the hypophysis and the suspensor (Figure 1e).
The hypophysis is the uppermost derivative of the basal cell
that, in many plants, serves as the precursor of the quies-
cent centre (QC) of the RAM and the central root cap cells.
Thus, the embryonic RAM is derived from descendants of
both the apical and the basal cells. The suspensor is
ephemeral and performs a structural role by pushing the
embryo proper into the nutrient-rich endosperm. It is also
thought to serve as a conduit for the transport of nutrients
and growth factors to the developing embryo. Although
crucifers possess a single file of 6–11 suspensor cells, there
are substantial variations in the number and layers of
suspensor cells in other plants (Kawashima and Goldberg,
2010). The suspensor usually degenerates at late stages of
embryo development.
In dicots, emergence of the first discernible organs, the
two cotyledons, generates a triangular, transition-stage
embryo (Figure 1h). This dramatic change in embryo
morphology is often described as a change from radial to
bilateral symmetry. Concomitant with the emergence of
cotyledon organs, embryonic tissue systems become
defined. The middle ground meristem and the innermost
procambium, the vascular tissue precursor, become dis-
tinguishable as a result of longitudinal divisions in the
interior of the embryo. Formation of the cotyledons reveals
the second major embryonic organ system, the axis. Con-
tinued cell divisions of the cotyledon primordial result in
the formation of a heart-stage embryo.
Major events of morphogenesis involve establishment of
the apical meristems (Figure 1i). Following embryogenesis,
stems, leaves and, later, floral organs are generated from
the SAM. By the late heart stage, the SAM consists of one
or two outer cell layers of tunica and an inner layer of
corpus. Functionally, it is divided longitudinally into three
zones, central, peripheral and rib. Primordia that produce
the first true leaves are initiated on the flanks of the SAM
during late embryogenesis. The embryonic RAM, which
consists of a layer of stem cells that surround the QC, a
group of cells that divide only rarely, becomes active in
 Plant Embryogenesis (Zygotic and Somatic)

Figure 1 The ovule and representative stages of embryo development. (a) Ovule, (b) zygote, (c) two-cell embryo, (d) two- or four-cell embryo proper,
(e) octant-stage embryo, (f) 16-cell embryo proper, (g) globular-stage embryo proper, (h) transition-stage embryo proper, (i) torpedo-stage embryo proper,
(j) bent cotyledon-stage embryo and (k) mature-stage embryo. Abbreviations: Ac, apical cell; An, antipodal cells; Ax, axis; Bc, basal cell; Cc, central cell; Ch,
chalazal region; Co, cotyledons; Ec, egg cell; Fg, female gametophyte; Fu, funiculus; Gm, ground meristem; Hy, hypophysis; Ii, inner integument; Mi,
micropyl region; O’, O’ line; Oi, outer integument; Pc, procambium; Pd, protoderm; Pn, polar nuclei; RAM, root apical meristem; Rc, root cap; SAM, shoot
apical meristem; Su, suspensor and Sy, synergid cells.

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 Plant Embryogenesis (Zygotic and Somatic)
heart-stage embryos. Different initials give rise to the
procambium, the ground meristem cells that will become
the cortex and endodermis, the epidermis and lateral root
cap, and the central root cap (Jiang and Feldman, 2005).
See also: Apical Meristems
Maturation phase
By the torpedo stage, most cell divisions and events asso-
ciated with embryonic tissue and organ formation are
complete, and morphological development has essentially
arrested (Figure 1i). During the maturation phase of
embryogenesis, cellular processes occur that are associated
with preparing the embryo and seed for desiccation,
metabolic quiescence and germination. First, macro-
molecular reserves, including storage proteins, storage
lipids and carbohydrates, accumulate and are sequestered
in discrete storage organelles within the embryo or endo-
sperm. These reserves are used primarily after germination
as a nutrient source for growth until the seedling becomes
photosynthetically active. Cell expansion that accounts for
embryo growth during the maturation phase occurs largely
as a result of reserve accumulation. Second, the embryo
gradually acquires the ability to withstand the stresses of
desiccation imposed by water loss at the end of the mat-
uration phase. The precise mechanisms involved in con-
ferring desiccation tolerance are not well understood, but
carbohydrates and a class of late embryogenesis abundant
(LEA) proteins have been implicated in this process
(Angelovici et al., 2010). Third, germination appears to be
actively suppressed during the maturation phase. Inhib-
ition of germination is mediated initially by the hormone
abscisic acid (ABA) and later by restricted water uptake
(Bewley, 1997). The onset of desiccation signals the end of
embryogenesis and induces a period of metabolic quies-
cence. See also: Dormancy in Plants; Plant Storage
Products (Carbohydrates, Oils and Proteins); Seed Ger-
mination and Reserve Mobilization; Seeds
Figure 2 Apical–basal embryonic domains. Three embryonic domains, apical, central and basal, can be discerned in the octant-stage embryo. The apical
domain gives rise to the shoot apical meristem and most of the cotyledons. The central domain gives rise to part of the cotyledons, the hypocotyl, the root
and most of the root apical meristem. The basal domain gives rise to part of the root apical meristem. Abbreviation: Su, suspensor.
4 ENCYCLOPEDIA OF LIFE SCIENCES & 2010, John Wiley & Sons, Ltd. www.els.net
Establishing the Plant Body Plan
In conceptual terms, plant embryo formation represents a
series of partitioning events in which organs and tissues are
formed from larger domains. The shoot–root axis consti-
tutes the major apical–basal axis of the plant body. During
embryogenesis, the body plan of the plant is elaborated
along this axis, producing the SAM, cotyledons, hypo-
cotyl, root and RAM. Tissue formation results from seg-
mentation events that occur radially from this axis.
See also: Phenotypic and Developmental Plasticity in
Plants; Positional Information in Plant Development
Apical–basal patterning
Specification of embryonic domains
An early event in morphogenesis appears to be compart-
mentalisation of the embryo into three distinct domains
along its apical–basal axis (Figure 2). The apical domain is
postulated to give rise to the SAM and most of the coty-
ledons, the central domain forms part of the cotyledons, the
hypocotyl, the root and the root meristem initials, and the
basal domain produces the QC of the RAM and central
root cap initials (Jurgens, 2001). The domain hypothesis
originated from analyses of Arabidopsis mutants with
defects in seedling morphology that were interpreted to
result from the absence of specific embryonic domains
(Mayer et al., 1991). gurke (gk) mutant seedlings that do
not have cotyledons and exhibit hypocotyl defects were
interpreted to lack the apical and, possibly, central
domains. The central domain appears to be absent in fackel
mutants in which cotyledons of seedlings are directly
attached to roots. monopteros (mp) mutants lack roots and
hypocotyls and are considered to have a central domain
deletion and a defective basal domain. Finally, gnom (gn)
mutants appear to consist of a ball of cells and were
interpreted to represent deletions of the apical and basal
domains.
 Plant Embryogenesis (Zygotic and Somatic)

The existence of ‘transcriptional territories’ support the
apical–basal domain hypothesis (Goldberg et al., 1994).
Transcriptional territories are defined by the spatial
activities of genes expressed during embryogenesis. For
example, the promoter of the soybean Kunitz trypsin
inhibitor gene is active in the micropylar region of a
globular-stage embryo, corresponding to the basal
domain. By contrast, the soybean seed lectin gene pro-
moter is active in a ring of equatorial cells around the
middle of a globular-stage embryo encompassing the pro-
toderm and ground meristem but not the procambium.
Thus, this promoter is active in a region of the central
domain. Similarly, expression of the Arabidopsis genes
WUSCHEL-RELATED HOMEOBOX2 (WOX2) and
CUP-SHAPED COTYLEDON3 (CUC3) is limited to the
upper tier of octant- and globular-stage embryos corres-
ponding to the apical domain (Haecker et al., 2004; Vroe-
men et al., 2003). By contrast, PLETHORA1 (PLT1)
initially marks a region of the central domain (Aida et al.,
2004). Thus, transcriptional territories appear to corres-
pond to genetically defined apical, central and basal
domains.
Embryonic domain specification appears to occur early
in embryogenesis. Phenotypic consequences of the
domain-deletion mutations can be traced to the earliest
stages of embryo development. For example, the first div-
ision of the gn mutant zygote is abnormal in that the
cleavage is symmetrical or oblique rather than asymmetric
and transverse as in wild type (Mayer and Jurgens, 1993).
mp embryo development deviates from wild type initially at
the octant stage when mutant embryos possess four rather
than two tiers of cells (Berleth and Jurgens, 1993). Differ-
ential gene expression patterns also provide support for
early compartmentalisation of the embryo. At the two-cell
embryo stage, a number of Arabidopsis genes including
AtMERISTEM L1 LAYER (AtML1), PINFORMED1
(PIN1), MP, BODENLOS (BDL) and WOX2 are active
predominantly in the apical cells, whereas PIN7, WOX8
and WOX9 are expressed primarily in the basal cell
(Weijers and Jurgens, 2005).
Role of auxin in apical–basal patterning
Regulated transport of the hormone auxin is implicated to
play critical roles in several major patterning events during
plant development, including embryogenesis (Vanneste
and Friml, 2009). From the two-cell embryo stage until the
early globular stage, auxin is transported from basal to
apical regions of the embryo, creating an auxin maximum
in the embryo proper, as shown in Figure 3 (Friml et al.,
2003). Auxin transport is mediated primarily through the
activities of the PIN family of auxin efflux carriers. For
example, localisation of PIN7 at the apical (upper) end of
the basal cell in a two-cell Arabidopsis embryo and its

Figure 3 Auxin flux during early embryo development of Arabidopsis thaliana. From the two-cell embryo stage to the pre-globular stage, PIN-FORMED7
(PIN7, purple) localises to the apical membrane of suspensor cells mediating auxin transport (green arrow) from the suspensor into the embryo proper,
where an auxin maximum (blue) is created. By the globular stage, PIN1 and PIN7 localise to the basal membranes and establish apical-to-basal auxin flux.
The activity of PIN1 localises to the basal membrane of the procambial cells. The activities of these transporters establish an auxin maximum in the
hypophysis and the uppermost cells of the suspensor of the globular-stage embryo (dark blue). The hypophysis then divides asymmetrically to eventually
form the quiescent centre of the embryo. Adapted from Jenik et al. (2007) with permission of Annual Reviews.

ENCYCLOPEDIA OF LIFE SCIENCES & 2010, John Wiley & Sons, Ltd. www.els.net 5
 Plant Embryogenesis (Zygotic and Somatic)
descendants appears to create the auxin maximum in the
apical cell. At the globular stage, however, apical-to-basal
auxin transport that is maintained throughout the
remainder of the plant life cycle is initiated through redis-
tribution of PIN carriers. PIN7 is relocalised to the basal
end of basal cell derivatives, and PIN1, which does not
display polar localisation in the embryo proper until the
globular stage, becomes restricted to the basal end of
procambial cells. The newly localised PIN efflux carriers
appear to cause a redistribution of the auxin maximum to
the hypophysis, the uppermost cell of the suspensor, and its
derivatives. Later in embryogenesis, auxin maxima form at
the site of incipient cotyledon primordial as a result of
relocalisation of PIN1. Thus, intracellular location of PIN
efflux carriers determines sites of auxin maxima during
embryo development. See also: Polar Auxin Transport
The importance of auxin flux in embryo development is
indicated by defects caused by a mutation affecting PIN
localisation. gn mutants have striking defects in apical–
basal patterning, including an abnormal division plane of
the zygote and the apparent loss of apical and basal
domains (Mayer and Jurgens, 1993). GN is an adenosyl
ribosylation factor guanine nucleotide exchange factor
that is required for the recycling and polar localisation of
PIN proteins (Steinmann et al., 1999). Thus, polar auxin
transport and the creation of auxin maxima are dependent
on GN-mediated cellular processes. The phosphorylation
status of PIN proteins determines their localisation to
apical or basal plasma membranes, which is controlled by
the serine-threonine kinase, PINOID (PID), and PRO-
TEIN PHOSPHATASE2A, respectively. Mutation of PID
causes defects in cotyledon separation during embryo-
genesis, emphasising the importance of proper PIN local-
isation. The following section highlights the roles of auxin
signalling in apical–basal patterning of the embryo.
See also: Auxin
Establishment of embryo domains
The basal domain derives from the hypophysis and gives
rise to the QC and central root cap cells. The auxin max-
imum generated by apical-to-basal transport is required for
the formation of the hypophysis cell and its subsequent
specification as QC cells. Two genes required for hyp-
ophysis and RAM formation, MP and BDL, encode
transcription factors involved in auxin signalling, and they
may affect transcription of PIN genes and the establish-
ment of the auxin maximum within the embryo. Both
genes, however, are expressed in procambial cells in the
central domain bordering the hypophysis, suggesting
intercellular signalling plays a key role in RAM formation
(Weijers et al., 2005). Auxin signalling may also play a role
in root formation by actively suppressing processes that
occur normally in the apical domain. TOPLESS (TPL)
encodes a transcriptional corepressor of auxin-regulated
genes (Szemenyei et al., 2008). A semidominant mutation
that inactivates the function of TPL and its family mem-
bers results in conditional root formation from the apical
6 ENCYCLOPEDIA OF LIFE SCIENCES & 2010, John Wiley & Sons, Ltd. www.els.net
domain, and, therefore, an embryo with two opposing
roots. Thus, auxin-dependent pathways play key roles in
establishment of the basal domain.
Auxin-independent pathways are also important for
domain specification, and in many cases, they intersect with
auxin-mediated pathways. For example, specification of
QC cell identity is thought to be controlled through
mechanisms dependent and independent of auxin. Speci-
fication of QC cell fate likely results from the auxin-induced
expression of the PLT family of transcription factors that
regulate root stem cell fate (Aida et al., 2004). On the
contrary, an auxin-independent network involving
SHORTROOT (SHR) and SCARECROW (SCR) is also
required for QC specification and RAM organi-
sation (Jiang and Feldman, 2005). See also: Root Apical
Meristems
Another example of the interplay between auxin-
dependent and auxin-independent pathways involves the
WOX genes that constitute a family of transcription factors
with important roles in early patterning events of the
embryo. WOX8 and WOX9 are expressed collectively in
the basal cell and its descendents and in the lower tier of the
embryo proper during early embryonic stages. Inactivation
of these two genes results in a severe reduction in PIN1
activity, the absence of the auxin maximum in the hypo-
physeal cell and defects in both embryo proper and sus-
pensor development (Breuninger et al., 2008). WOX genes
are also required for patterning of the apical domain.
WOX2, which is expressed in the zygote and the apical cell
of the two-cell embryo proper, appears to act redundantly
with WOX1 and WOX3/PRESSED FLOWER for estab-
lishment and maintenance of the protoderm (Breuninger
et al., 2008). Moreover, genetic studies showed interaction
between WOX2 and WOX8 with MP, suggesting the auxin-
independent WOX pathway converges with auxin signal-
ling pathways in controlling embryo patterning.
The apical domain originates from the upper tier of the
octant-stage embryo and gives rise to the SAM and most of
the cotyledons. Although GK appears to be essential for
partitioning of the apical domain, it is not known how this
gene encoding acetyl-coenzyme A carboxylase, an enzyme
involved most commonly in fatty acid biosynthesis, is
required for domain establishment (Baud et al., 2003).
Formation of the SAM occurs through a series of
compartmentalisation events within the apical region.
WUSCHEL, a homeodomain transcription factor
required for SAM formation, is initially expressed at the
16-cell stage in the upper tier of the embryo proper (Mayer
et al., 1998). As these cells divide asymmetrically, WUS
expression becomes restricted to the innermost cells. WUS
is required for expression of CLAVATA genes, and col-
lectively these genes establish the stem cell niche and
regulates SAM homeostasis (Carles and Fletcher, 2003). A
second partitioning pathway involves the formation of a
boundary around the SAM and the two cotyledons
by the CUC transcription factors (Koyama et al., 2007).
CUC gene expression in the apical domain early
in embryogenesis activates the expression of

SHOOTMERISTEMLESS (STM), a homeodomain
transcription factor that promotes SAM formation. STM
then downregulates CUC genes, restricting CUC expres-
sion to a domain surrounding the SAM.
Cell fate determination in radial patterning
The embryo consists of three primary tissue systems:
the outer protoderm, the middle ground meristem and the
inner procambium that are arrayed radially from the
apical–basal axis as shown in Figure 4. Clonal analyses and
cell ablation studies show that although the precise series of
cell divisions that occur early in embryogenesis give the
appearance of lineage-dependent specification of cell fate,
positional information plays a predominant role in speci-
fying cell identity. Consistent with this idea, Arabidopsis
fass (fs) mutant embryos do not undergo normal patterns
of cell division, and they produce misshapen seedlings
(Mayer et al., 1991). Yet, mature fs mutant embryos have
the full complement of functional tissues, suggesting that
consistent cell division patterns are not critical in estab-
lishing the radial pattern.
The outer protoderm layer appears to be specified
coincident with cell divisions parallel to the surface of the
octant-stage embryo. Markers for protoderm cell fate such
as AtML1, PROTODERMAL FACTOR1 (PDF1) and
PDF2 become restricted to the outermost layer and even-
tually the embryo (Abe et al., 2003; Lu et al., 1996). They
are expressed in the developing epidermis during post-
embryonic development, but they are not required for root
epidermis development. See also: Epidermis: Outer Cell
Layer of the Plant
The embryonic root consists of concentric rings of
endodermis, cortex and protoderm surrounding the pro-
cambium (Figure 4). The former two arise from an asym-
metric division of the ground meristem initial and involves
two GRAS (gibberellic acid insensitive (GAI), repressor of
GAI (RGA) and scarecrow (SCR))-type transcription
factors, SHR and SCR (Jiang and Feldman, 2005). SHR is
synthesised in the procambium and transported into the
adjacent ground tissue where it activates the SCR gene.
SCR promotes division of the ground tissue initial and
specification of endodermis identity.
Figure 4 Radial patterning of embryonic tissues. The three embryonic
tissue systems are the protoderm, the ground meristem and the
procambium. In the embryonic root, the ground meristem gives rise to the
cortex and the endodermis.
ENCYCLOPEDIA OF LIFE SCIENCES & 2010, John Wiley & Sons, Ltd. www.els.net
Plant Embryogenesis (Zygotic and Somatic)
Somatic Embryogenesis
Plants can make embryos in various ways. Zygotes formed
from egg cells as a result of fertilisation undergo zygotic
embryogenesis. Somatic cells can be induced to form dip-
loid embryos (Yang and Zhang, 2010). Microspore cells
can be diverted from their normal pathway of pollen
development to make haploid embryos (Maraschin et al.,
2006). Various cells of the ovule can undergo adventitious
or parthenogenic embryogenesis in a suite of processes that
give rise to asexual seeds known as apomixes (Koltunow
and Grossniklaus, 2003). These forms of embryogenesis,
including somatic embryogenesis, are examples of the
totipotency of plants, that is, the ability of plant cells to
form a fully differentiated organism. Studies of these
alternative pathways of embryogenesis may provide clues
about the mechanisms that induce embryonic develop-
ment. See also: Plant Cell Culture
Somatic cells must usually be induced to form embryos,
although somatic embryos can sometimes form naturally
on leaf margins of plants such as the Mother of Thousands
(Kalanchoe daigremontiana). Somatic embryos are most
commonly generated by treating cells with auxin anal-
ogues, such as 2,4-dichlorophenoxyacetic acid and some-
times cytokinins, followed by culture of the cells in
hormone-free medium (Yang and Zhang, 2010). Thus,
induction is considered a bipartite process in which cells
treated with auxin acquire the competence to undergo
somatic embryogenesis, but they are only able to differen-
tiate into somatic embryos in the absence of this plant
growth regulator. However, hormone treatments are not a
requirement for somatic embryogenesis. Somatic cells can
be induced to undergo embryogenesis through various
procedures that include, but are not limited to, exposure to
hormones, heat or pH shock and treatment with various
chemicals (Ikeda-Iwai et al., 2003). Thus, there does not
appear to be one universal signal capable of inducing
embryogenic cell fate. However, the precise mechanisms by
which cells acquire competence in response to hormone or
other treatments are not known.
Despite the important practical applications of somatic
embryogenesis in the plant sciences, much remains
to be learned about the cellular processes that underlie
somatic embryogenesis. The development of somatic
embryos appears to follow pathways similar to zygotic
embryos. Mutations responsible for defects in zy-
gotic embryo development cause similar anomalies in
somatic embryo development (Mordhorst et al., 2002).
Thus, major questions focus on the mechanisms that cause
cells to change their fate and become embryogenic.
Significant progress has been made in the past 15 years. We
highlight recent advances in understanding the mech-
anisms controlling somatic embryogenesis in Arabi-
dopsis. The knowledge gained from this model system
should provide the necessary starting point in under-
standing embryogenic competence and somatic embryo
development in other plants, including economically
important crop species.
7
 Plant Embryogenesis (Zygotic and Somatic)
Embryogenic competence
What are the endogenous factors that promote the com-
petence of cells to undergo somatic embryogenesis?
Although somatic embryos can form from cells in many
plant organs, immature zygotic embryos are generally the
preferred source of material for producing embryogenic
cell cultures. For example, Arabidopsis maturation-phase
embryos show the highest induction frequency of somatic
embryogenesis (Gaj et al., 2005). Thus, embryonic cells
appear to be more competent to undergo somatic
embryogenesis. Stem cells or undifferentiated cells also
appear to have higher competence for somatic embryo-
genesis. Mutant seedlings with enlarged SAMs, such as
primordia timing, clavata1 and clavata3, exhibit a higher
frequency of somatic embryo formation following auxin
treatment than wild-type seedlings (Mordhorst et al.,
1998). Consistent with this finding, overexpression of
WUS, which normally promotes stem cell identity in the
SAM, induces somatic embryogenesis in the absence of
auxin (Zuo et al., 2002). Overexpression of BABY BOOM,
a gene implicated in promoting RAM stem cell identity,
also is sufficient to induce somatic embryogenesis (Boutilier
et al., 2002). Thus, genes that enhance apical meristem
formation or stem cell identity may confer embryogenic
competence with and without exogenous auxin treatment.
Many studies have focused on characterising the cellular
processes that occur as cells acquire competence for som-
atic embryogenesis. Carrot cells have long been a model
system for plant somatic embryogenesis (Steward et al.,
1958). Embryos can form from single cells, and imaging
studies suggest that morphological shape is not a consistent
indicator of embryogenic competence. However, two
molecular markers of competent cells have been identified.
Expression of the somatic embryo receptor-like kinase
(SERK) gene correlates with the acquisition of competence
(Schmidt et al., 1996). A second marker is a cell wall antigen
found on the surface of competent cells that interacts with
the JIM8 (John Innes monoclonal 8) monoclonal antibody
(McCabe et al., 1997). Other molecular markers of somatic
embryogenesis, such as LEAFY COTYLEDON1 (LEC1),
FUSCA3 (FUS3), ABA INSENSITIVE3 (ABI3) and LEA
gene expression, are associated with both somatic and
zygotic embryogenesis (Braybrook and Harada, 2008).
Therefore, it is not clear if these genes are specific markers
of embryogenic competence or of embryo development.
Somatic embryogenesis can also be induced through the
ectopic expression of genes encoding transcription factors
that accumulate primarily during embryo development,
LEC1 (Lotan et al., 1998), LEC2 (Stone et al., 2001, 2008)
and MYB115 and 118 (Wang et al., 2009). These genes are
sufficient to induce a cellular environment that promotes
somatic embryogenesis. All of these transcription factors
are implicated to play regulatory roles during the matur-
ation phase of zygotic embryogenesis. Together with the
observation that cells from maturation-phase em-
bryos undergo somatic embryogenesis at a high frequency,
these findings suggest that cellular processes that
8 ENCYCLOPEDIA OF LIFE SCIENCES & 2010, John Wiley & Sons, Ltd. www.els.net
occur during maturation may be associated with embryo-
genic competence.
Clues about the potential mechanisms by which these
transcription factors promote somatic embryogenesis come
from analyses of genes regulated by the LEC2 transcription
factor, which is a prolific inducer of somatic embryogenesis
(Stone et al., 2001, 2008). Induction of LEC2 activity results
in the rapid activation of genes encoding key enzymes
involved in auxin biosynthesis, YUCCA2 and YUCCA4.
Auxin has also been shown to interact genetically with LEC1.
Thus, competence for somatic embryogenesis through
ectopic expression of LEC transcription factors may occur
through auxin. Although the central role for auxin in embryo
development is well established, we do not know how
embryogenic cells become hormone autonomous. Further
studies will help establish the genetic mechanisms underlying
this critical step in embryo development. See also: Auxin
An additional insight into the mechanisms controlling
embryogenic competence was obtained through studies
of the Arabidopsis PICKLE (PKL) gene, that encodes a
CHD3 (chromodomain helicase DNA-binding 3) chro-
matin-remodelling factor (Ogas et al., 1999). Roots of pkl
mutant seedlings display embryonic characteristics, such as
oil bodies, and excised pkl roots produce somatic embryos
in hormone-free culture. Unlike wild type, postembryonic
pkl mutant roots accumulate LEC1 and LEC2 messenger
ribonucleic acid (mRNA), suggesting that the LEC genes
may confer embryogenic competence to pkl mutant roots.
The Pkl2 mutant root phenotype can be suppressed by the
hormone giberellic acid (GA), suggesting that the wild-type
PKL gene may serve a role in a GA signalling pathway to
repress the embryogenic competence of postembryonic
roots and allow vegetative development to occur. The
LEC2 transcription factor activates AGAMOUS-LIKE15,
which in turn activates the GA2ox6 gene encoding a GA-
inactivating enzyme (Braybrook and Harada, 2008).
Therefore, the LEC transcription factors may promote
embryogenic competence indirectly through its potential
effects in repressing GA levels. These findings are consist-
ent with studies that show exogenous GA suppresses
somatic embryogenesis (Wang et al., 2004). The hormone
ABA generally has effects on plant growth and develop-
ment that are antagonistic to those of GA. Mutations in
several ABA signalling genes drastically affect somatic
embryogenesis in Arabidopsis (Gaj et al., 2006). In Datura,
applications of ABA promotes somatic embryogenesis,
perhaps through co-ordinated down regulation of the GA
pathway (Kikuchi et al., 2006). Thus, several plant hor-
mones may be involved in conferring embryogenic com-
petence to somatic cells. See also: Abscisic Acid (ABA);
Plant Growth Factors and Receptors; Regulatory Genes in
Plant Development: MADS
Summary
Embryogenesis in seed development has long been recog-
nised as a critical period of the higher plant life cycle.

Beginning with one of two fertilisation events that result in
the formation of the zygote, the embryo develops within the
confines of the maternal tissue until it becomes meta-
bolically and developmentally arrested at the end of
embryogenesis. The SAM and RAM and embryonic organ
systems appear to arise through partitioning events along
the apical–basal axis, as suggested by the existence of
genetically defined morphological domains and transcrip-
tional territories. Embryonic tissue systems are formed
through compartmentalisation along the radial axis.
Localisation of the hormone auxin plays many critical roles
in establishing the body plan of the plant during embryo
development.
Somatic embryogenesis is an example of the ability of
plant cells to alter their cell fates to become embryogenic
and has been used to address long-standing questions
about plant cell determinacy. Hormone treatments are
most commonly used to induce somatic embryogenesis,
although somatic embryos can arise naturally or as a
consequence of treatments that do not involve hormones.
Studies with Arabidopsis transcription factors that are
sufficient to induce somatic embryogenesis in the absence
of exogenous hormone treatment emphasise the likely
importance of endogenous hormones that cause cells to
acquire embryogenic cell fate. Taken together, our under-
standing of plant embryo development should convert into
practical procedures with the goal to improve our under-
standing of zygotic and somatic embryogenesis.
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