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[From Developmental Biology, Eighth Edition, by Scott F. Gilbert, published by Sinauer Associates, Inc.]

An Overview of Plant Development

Susan R. Singer

Laurence McKinley Gould Professor of the Natural Sciences, Carleton College

20

THE DEVELOPMENTAL STRATEGIES OF PLANTS have evolved separately from those

of the animals over millions of years. The two kingdoms have many common- alities (and the land plants are sometimes referred to as “embryophytes,” call- ing attention to the significance of the embryo in their life histories), but some of the challenges and solutions found in plants are sufficiently unique to war- rant separate discussion in this chapter. What are the fundamental differences between development in animals and development in the land plants?

Plant cells do not migrate. Plant cells are trapped within rigid cellulose walls that generally prevent cell and tissue migration. Plants, like most metazoan animals, develop three basic tissue systems (dermal, ground, and vascular), but do not rely on gastrulation to establish this layered system of tissues. Plant development is highly regulated by the environment, a strate- gy that is adaptive for a stationary organism.

• Plants have sporic meiosis rather than gametic meiosis. That is, meiosis in plants produces spores, not gametes. Plant gametes are produced by mitotic divisions following meiosis.

• The life cycle of land plants (as well as many other plants) includes both diploid and haploid multicellular stages. This type of life cycle is referred to as alternation of generations and results in two different multicellular body plans over the life cycle of an individual.

• Plant germ cells are not set aside early in development. While this is also the case in several animal phyla, it is the case for all plants.

• Plants undergo extended morphogenesis. Clusters of actively dividing cells called meristems, which are similar to stem cells in animals, persist long after maturity. Meristems allow for iterative development and the formation of new structures throughout the life of the plant.

• Plants have tremendous developmental plasticity. Many plant cells are highly plastic. While cloning in animals also demonstrates plasticity, plants depend far more heavily on this developmental strategy. For example, if a shoot is grazed by herbivores, meristems in the leaf often grow out to replace the lost part. (This strategy has similarities to the regeneration seen in some animals.) Whole plants can be regenerated from some single cells. In addition, a plant’s form (including branching, height, and relative amounts of vegeta- tive and reproductive structures) is greatly influenced by environmental fac- tors such as light and temperature, and a wide range of morphologies can result from the same genotype (see Figure 2.15). The amazing level of plastici- ty found among the plants may help compensate for their lack of mobility.

The search for differences or fun- damental contrasts … has occupied many men’s minds, while the search for commonality of principle or essential similarities, has been pur- sued by few; the contrasts are apt to loom too large, great though they

may be. D’ARCY THOMPSON (1942)

Do not quench your

inspiration

and your imagination; do not become the slave of your model.

VINCENT VAN GOGH

not become the slave of your model. ” V INCENT VAN G OGH © 2010 Sinauer

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628 CHAPTER 20

Developmental mechanisms evolved independently in plants and animals. The last common ancestor of plants and animals was a single-celled eukaryote. Genome-level comparisons indicate that there is mini- mal homology between the genes and proteins used to establish body plans in plants and animals (Meyerowitz 2002). While both homeobox and MADS box genes were present in the last common ancestor of plants and ani- mals, the MADS box family controls major developmen- tal regulatory processes in plants, but not in animals.

Despite the major differences among many plants and ani- mals, developmental genetic studies are revealing some commonalities in the logic of their pattern formation, along with evolutionarily distinct solutions to the problem of cre- ating three-dimensional form from a single cell. The green plants include many organisms, from algae to flowering plants (angiosperms). Recent phylogenetic studies show a common lineage for all green plants, dis- tinct from the red and brown plants. While comparisons of developmental strategies among diverse plants is both fascinating and informative, this chapter focuses primari- ly on the flowering plants (angiosperms). The goal is to examine plant development within the larger context of developmental biology.

Gamete Production in Angiosperms

Plants have both multicellular haploid and multicellular diploid stages in their life cycles, and embryonic develop- ment is seen only in the diploid generation. The embryo, however, is produced by the fusion of gametes, which are formed only by the haploid generation. Understanding the relationship between the two generations is important in the study of plant development.

Gametophytes

Unlike animals, plants have multicellular haploid and mul- ticellular diploid stages in their life cycles (see Chapter 2). Gametes develop in the multicellular haploid gametophyte (from the Greek phyton, “plant”). Fertilization gives rise to

a multicellular diploid sporophyte, which produces hap-

loid spores via meiosis. This type of life cycle is called a haplodiplontic life cycle (Figure 20.1). It differs from the diplontic life cycle of animals, in which only the gametes are in the haploid state. In a haplodiplontic life cycle, gametes are not the direct result of a meiotic division. Diploid sporophyte cells under- go meiosis to produce haploid spores. Each spore goes through mitotic divisions to yield a multicellular, haploid gametophyte. There are two types of spores in angio- sperms. Megaspores produce female gametophytes, while microspores produce male gametophytes. Male and female gametophytes have distinct morphologies. Wind or mem-

Diploid generation Haploid generation MEIOSIS 1n Mitosis Path D Path A Path C Path B
Diploid generation
Haploid generation
MEIOSIS
1n
Mitosis
Path D
Path A
Path C
Path B
2n
1n
organism
organism
Mitosis
1n
2n
FERTILIZATION
Gametes
1n

FIGURE 20.1 Plants have haplodiplontic life cycles that involve mitotic divisions (resulting in multicellularity) in both the haploid and diploid generations (paths A and D). Most ani- mals are diplontic and undergo mitosis only in the diploid gen- eration (paths B and D). Multicellular organisms with haplontic life cycles follow paths A and C.

WEBSITE
WEBSITE

20.1

Plant life cycles. In plants there is an

evolutionary trend from sporophytes that are nutritionally dependent on autotrophic gameto- phytes to the opposite—gametophytes that are dependent on autotrophic sporophytes. This trend is exemplified by comparing the life cycles of mosses and ferns to that of angiosperms.

[Note: This Web topic is included at the end of this chapter.]

bers of the animal kingdom deliver the male gameto- phyte—pollen—to the female gametophyte. Mitotic divi- sions within the gametophytes are required to produce gametes. The diploid sporophyte results from the fusion of two gametes. Among land plants, the gametophytes and sporophytes of a species have distinct morphologies, and how a single genome can be used to create two unique morphologies is an intriguing puzzle. At first glance, angiosperms may appear to have diplon- tic life cycles because the gametophyte generation has been reduced to just a few cells (Figure 20.2). However, mitotic division follows meiosis in the sporophyte, resulting in a multicellular gametophyte, which produces eggs or sperm. All of this takes place in the organ that is characteristic of the angiosperms: the flower.

POLLEN The pollen grain is an extremely simple multicel- lular structure (Figure 20.3). The outer wall of the pollen grain, the exine, is composed of resistant material provid- ed by both the tapetum (sporophyte generation that pro- vides nourishment for developing pollen) and the microspore (gametophyte generation). The inner wall, the

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AN OVERVIEW OF PLANT DEVELOPMENT 629 Diploid sporophyte generation Haploid gametophyte generation Petals
AN OVERVIEW OF PLANT DEVELOPMENT
629
Diploid sporophyte generation
Haploid gametophyte generation
Petals
Microsporangium
Flower
Pollen
(2n)
Sporophyte (2n)
Microspores (1n)
Gametophyte
Anther
Carpel
MEIOSIS
Mitosis
Stamen
Filament
Ovary
Seed
Ovule
germination
Embryo sac
Megaspores (1n)
Gametophyte
Megasporangium
(2n)
Embryo
Endosperm
Embryo sac
FERTILIZATION
Pollen tube
Seed coat

FIGURE 20.2 Life cycle of an angiosperm, represented here by a pea plant (genus Pisum). The sporophyte is the dominant generation, but multicellular male and female gametophytes are produced within the flowers of the sporophyte. Cells of the microsporangium within the anther undergo meiosis to pro- duce microspores. Subsequent mitotic divisions are limited, but the end result is a multicellular pollen grain. Integuments and the ovary wall protect the megasporangium. Within the mega-

sporangium, meiosis yields four megaspores—three small and one large. Only the large megaspore survives to produce the female gametophtye (the embryo sac). Fertilization occurs when the male gametophyte (pollen) germinates and the pollen tube grows toward the embryo sac. The sporophyte gen- eration may be maintained in a dormant state, protected by the seed coat.

intine, is produced by the microspore. A mature pollen grain consists of two cells: a tube cell, and a generative cell within the tube cell. The nucleus of the tube cell guides pollen germination and the growth of the pollen tube after

the pollen lands on the stigma of a female gametophyte. The generative cell divides to produce two sperm. One of the two sperm will fuse with the egg cell to produce the next sporophyte generation. The second sperm will par-

(A)

next sporophyte generation. The second sperm will par- (A) (B) Exine Intine 1n 1n Tube cell

(B)

Exine

Intine

1n 1n
1n
1n

Tube cell

Tube cell

nucleus

Generative

cell

FIGURE 20.3 (A) Pollen grains have intricate surface patterns, as seen in this scanning electron micrograph of aster pollen. (B) A pollen grain consists of a cell within a cell. The generative cell will undergo division to produce two sperm cells. One will fertilize the egg, and the other will join with the polar nuclei, yielding the endosperm.

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630 CHAPTER 20 Stigma Style Ovule Megasporangium Ovary wall (eventual fruit) Integuments Megaspores Ovary 1n
630 CHAPTER 20
Stigma
Style
Ovule
Megasporangium
Ovary wall
(eventual fruit)
Integuments
Megaspores
Ovary
1n
Placenta
Micropyle
1n
1n
1n
Sepals
Placenta

FIGURE 20.4 The carpel consists of the stigma, the style, and an ovary containing one or more ovules. Each ovule contains megasporangia protected by two layers of integument cells. The megasporangia divide meiotically to produce haploid megaspores. All of the carpel is diploid except for the mega- spores, which divide mitotically to produce the embryo sac (the female gametophyte).

ticipate in the formation of the endosperm, a structure that provides nourishment for the embryo.

THE OVARY The angiosperm ovary is part of the carpel of

a flower, which gives rise to the female gametophyte (Fig-

ure 20.4). The carpel consists of the stigma (where the pollen lands), the style, and the ovary. Following fertiliza- tion, the ovary wall will develop into the fruit. This unique

angiosperm structure provides further protection for the developing embryo and also enhances seed dispersal by frugivores (fruit-eating animals). Within the ovary are one or more ovules attached by a placenta to the ovary wall. Fully developed ovules are called seeds. The ovule has one or two outer layers of cells, called the integuments. The integuments enclose the megaspo- rangium, which contains sporophyte cells that undergo meiosis to produce megaspores (see Figure 20.2). There is

a small opening in the integuments called the micropyle,

through which the pollen tube will grow. The integuments develop into the seed coat, a waterproof physical barrier that protects the embryo. When the mature embryo dis- perses from the parent plant, diploid sporophyte tissue accompanies the embryo in the form of the seed coat and the fruit. Within the ovule, meiosis and unequal cytokinesis yield four megaspores. The largest of these megaspores under- goes three mitotic divisions to produce the female game- tophyte, a seven-celled embryo sac with eight nuclei (Fig- ure 20.5). One of these cells is the egg. Two synergid cells

surround the egg and the pollen tube enters the embryo sac by penetrating one of the synergids. The central cell contains two or more polar nuclei, which will fuse with the second sperm nucleus and develop into the polyploid endosperm. Three antipodal cells form at the opposite end of the embryo sac from the synergids and degenerate before or during embryonic development. There is no known function for the antipodals. Genetic analyses of female gametophyte development in maize and Arabidop- sis* are providing insight into the regulation of the specif- ic steps in this process (Pagnussat et al. 2005).

Pollination

Pollination refers to the landing and subsequent germina- tion of the pollen on the stigma. Hence it involves an inter- action between the gametophytic generation of the male (the pollen) and the sporophytic generation of the female (the stigmatic surface of the carpel). Pollination can occur within a single perfect flower that contains both male and female gametophytes (self-fertilization), or pollen can land on a different flower on the same or a different plant. About 96 percent of flowering plant species produce male and female gametophytes on the same plant. However, about 25 percent of these produce two different types of flowers on the same plant, rather than perfect flowers. Staminate flowers lack carpels, while carpellate flow- ers lack stamens. Maize plants, for example, have stami- nate (tassel) and carpellate (ear) flowers on the same plant. Such species, which include the majority of the angio- sperms, are considered to be monoecious (Greek mono,

*A small weed in the mustard family, Arabidopsis is used as a model organism because of its very small genome.

Antipodal cells Polar nuclei Synergid Egg Integuments Micropyle (pollen entry point)
Antipodal cells
Polar nuclei
Synergid
Egg
Integuments
Micropyle
(pollen entry
point)

FIGURE 20.5 The embryo sac is the product of three mitotic divisions of the haploid megaspore; it comprises seven cells and eight haploid nuclei. The two polar nuclei in the central cell will fuse with the second sperm nucleus and produce the endo- sperm that will nourish the egg. The other six cells, including the egg, contain one haploid nucleus each.

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AN OVERVIEW OF PLANT DEVELOPMENT

631

“one”; oecos, “house”). The remaining 4 percent of species (e.g., willows, maples, and date palms*) produce stami- nate and carpellate flowers on separate plants. These species are considered to be dioecious (“two houses”). Only a few plant species have true sex chromosomes, yet they arose several times in flowering plant evolution (Charlesworth 2002). The terms “male” and “female” are most correctly applied only to the gametophyte genera- tion, not to the sporophyte (Cruden and Lloyd 1995).

SELF-INCOMPATIBILITY The arrival of a viable pollen grain on a receptive stigma does not guarantee fertilization. Interspecific incompatibility refers to the failure of pollen from one species to germinate and/or grow on the stigma of another species. Intraspecific incompatibility is incom- patibility that occurs within a species. Self-incompatibil- ity—incompatibility between the pollen and the stigmas of the same individual—is an example of intraspecific incompatibility (see Kao and Tsuikamoto 2004). Self-incom- patibility blocks fertilization between two genetically sim- ilar gametes, increasing the probability of new gene com- binations by promoting outcrossing (pollination by a different individual of the same species). Groups of close- ly related plants can contain a mix of self-compatible and self-incompatible species. Several different self-incompatibility systems have evolved. Recognition of self depends on the multiallelic self-incompatibility (S) locus (Nasrallah 2002). Gametophyt- ic self-incompatibility occurs when the S allele of the pollen grain matches either of the S alleles of the stigma (remem- ber that the stigma is part of the diploid sporophyte gen- eration, which has two S alleles, while a single pollen grain carries one S allele). In this case, the pollen tube begins

*The discovery that plants had sexes was important to the economy of date palms in the ancient Near East over two thousand years ago. Since only the female trees bore fruit, date farmers planted just a few male trees, then hand-pollinated the many female trees. This practice greatly increased the fruit yield per acre, and such pollina- tion events became associated with spring fertility festivals.

developing but stops before reaching the micropyle (Fig- ure 20.6A). Sporophytic self-incompatibility occurs when one of the two S alleles of the pollen-producing sporophyte (not the gametophyte) matches one of the S alleles of the stigma (Figure 20.6B). Most likely, sporophyte contribu- tions to the pollen exine are responsible for this type of self- incompatibility. The S locus consists of several physically linked genes that regulate recognition and rejection of pollen. An S gene has been cloned that codes for an RNase (called S RNase) that is sufficient, in the gametophytically self-incompati- ble petunia pistil, to recognize and reject self-pollen (Lee et al. 1994). The pollen component of gametophytic self- incompatibility in the petunia, SLF (S-locus, F-box), is an F-box gene within the S locus (Sijacic et al. 2004). A different, more rapid gametophytic response to self- incompatibility has been investigated in poppies, a rela- tive of the more basal flowering plants. Calcium ions accu- mulate in the tip of the pollen tubes, where open calcium channels are concentrated (Jaffe et al. 1975; Trewavas and Malho 1998). There is direct evidence that pollen tube growth in the poppy is regulated by a slow-moving Ca 2+ wave controlled by the phosphoinositide signaling path- way (Figure 20.7; Franklin-Tong et al. 1996). Ca 2+ influx occurs at both the tip of the pollen tube and on the shanks. Altered calcium influx is observed when the pollen tube is self-incompatible with the style, which leads to F-actin depolymerization, destabilization of the pollen cytoskele- ton, and cessation of pollen tube growth (Franklin-Tong et al. 2002; Franklin-Tong and Franklin 2003). The incompat- ible pollen tube then undergoes programmed cell death (Thomas and Franklin-Tong 2004). In sporophytic self-incompatibility, a ligand on the pollen is thought to bind to a membrane-bound kinase receptor in the stigma, starting a signaling process that

Members of the F-box family of genes share a common “F-box domain” for binding transcription factors. Although some F-box genes have been found in other eukaryote groups, most of these genes are unique to the plants.

(A)–Gametophytic self-incompatibility (B)–Sporophytic self-incompatibility Pollen Pollen S1 S2 S1 S2 S1/S2
(A)–Gametophytic self-incompatibility
(B)–Sporophytic self-incompatibility
Pollen
Pollen
S1
S2
S1
S2
S1/S2
S2/S3
S2/S3
S1/S2
S2/S3
S2/S3
Stamen
Carpel
Carpel
Stamen
Carpel
Carpel

FIGURE 20.6 Self-incompatibility. S1, S2, and S3 are different alleles of the self-incompatibility (S) locus. (A) Plants with gametophytic self-incom- patibility reject pollen only when the genotype of the pollen (i.e., the gametophyte) matches either one of the carpel’s two alleles. (B) In sporo- phytic self-incompatibility, the geno- type of the pollen parent (i.e., the sporophyte), not just that of the hap- loid pollen grain, can trigger an incompatibility response.

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632 CHAPTER 20 Embryo Ovule sac Callose plug Pollen tube Sperm cells Tube nucleus Ca
632 CHAPTER 20
Embryo
Ovule
sac
Callose
plug
Pollen
tube
Sperm
cells
Tube
nucleus
Ca 2+ channels
D
ir
ec
tio
n
o f
Ca
2+
wa
ve

FIGURE 20.7 Calcium and pollen tube tip growth. After com- patible pollen germinates, the pollen tube grows toward the micropyle. Waves of calcium ions play a key role in this growth of the tube. (After Franklin-Tong et al. 1996.)

leads to pollen rejection. In Brassica, one of the genes of the S locus encodes a transmembrane serine-threonine kinase (SRK) that functions in the epidermis of the stigma and binds a cysteine-rich peptide (SCR) from the pollen (Fig- ure 20.8; Kachroo et al. 2001). There are numerous examples of plant populations that have switched from self-incompatible to self-fertilizing sys- tems. Changes in the S locus, specifically the SKR and SCR genes, could account for these evolutionary changes. The Nasrallahs (2002) created self-incompatible Arabidopsis thaliana plants (which are normally self-compatible) by introducing the SKR and SCR genes that encode self-recog- nizing proteins from A. lyrata (a self-incompatible species). This experiment demonstrates that A. thaliana still has all of the downstream components of the signal cascade that can lead to pollen degradation. The mechanism of pollen degradation is unclear, but appears to be highly specific.

FIGURE 20.8 Receptor-ligand self-recognition is the key to self-incompatibility in Brassicas. Allelic variability in both the SRK and SRC genes leads to a variety of possible combinations of lig- and and receptor proteins. Unlike the common self-recognition systems of animals, including immunity and mating, self-incom- patibility results from the binding of SRK and SRC proteins of self (from allelic S loci) rather than nonself. (After Nasrallah 2002; photographs courtesy of J. Nasrallah.)

POLLEN GERMINATION If the pollen and the stigma are com- patible, the pollen takes up water (hydrates) and the pollen tube emerges. The pollen tube grows down the style of the carpel toward the micropyle (Figure 20.9). The tube nucle- us and the sperm cells are kept at the growing tip by bands of callose (a complex carbohydrate). It is possible that this may be an exception to the “plant cells do not move” rule, as the generative cell(s) appear to move forward via adhe- sive molecules (Lord 2000). Pollen tube growth is quite slow in gymnosperms (up to a year), while in some angiosperms the tube can grow as rapidly as 1 cm per hour. Genetic approaches have been useful in investigating how the growing pollen tube is guided toward unfertil-

ized ovules. In Arabidopsis, the pollen tube appears to be

guided by a long-distance signal from the ovule (Hul- skamp et al. 1995; Wilhelmi and Preuss 1999). Analysis of pollen tube growth in ovule mutants of Arabidopsis indi- cates that the haploid embryo sac is particularly important in the long-range guidance of pollen tube growth. Mutants

(A) Self-pollination

(self-self)

(B) Cross-pollination

(self-nonself)

S1

å å S1
å
å
S1
S2 S2
S2
S2
(B) Cross-pollination (self-nonself) S1 å å S1 S2 S2 ç × ç × S1 S3 S2
ç × ç × S1 S3 S2 S4 Pollen coat Stigma cell wall SRK-SCR NO
ç ×
ç ×
S1
S3
S2
S4
Pollen coat
Stigma cell wall
SRK-SCR
NO
binding
SRK-SCR
binding
SRK
NO SRK
activation
activation

Pollen tube inhibition

SRK NO SRK activation activation Pollen tube inhibition Pollen tube growth © 2010 Sinauer Associates, Inc.

Pollen tube growth

activation Pollen tube inhibition Pollen tube growth © 2010 Sinauer Associates, Inc. This material cannot be

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AN OVERVIEW OF PLANT DEVELOPMENT

633

(A)

AN OVERVIEW OF PLANT DEVELOPMENT 6 3 3 (A) (B) Pollen tubes Tube cell nucleus Sperm

(B)

Pollen tubes Tube cell nucleus Sperm nuclei
Pollen tubes
Tube cell
nucleus
Sperm nuclei

Style cells

Pollen

tube

FIGURE 20.9 Pollen tube germination. (A) Scanning electron micrograph of an Arabidopsis pollen tube en route to the ovule for fertilization. (B) Lily pollen tubes grown in vivo and removed from the ovary. Each green strand is an individual pollen tube and contains two sperm nuclei (bright blue stain) and a fainter (lighter blue) tube cell nucleus. Note the huge number of pollen tubes, all “racing” to fertilize a single egg. (Photographs courtesy of E. Lord.)

which nourishes the developing embryo. This second event is not true fertilization in the sense of male and female gametes undergoing syngamy (fusion)—that is, it does not result in a zygote, but in nutritionally supportive endo- derm. When you eat popcorn, you are actually eating “popped” endosperm. The other accessory cells in the embryo sac degenerate after fertilization. The zygote of the angiosperm produces only a single embryo.* Double fertilization, first identified a century ago, is generally restricted to the angiosperms, but it has also been found in the gymnosperm genera Ephedra and Gne- tum, although no endosperm forms. Friedman (1998) has suggested that endosperm may have evolved from a sec- ond zygote “sacrificed” as a food supply in an early gym- nosperm lineage with double fertilization. Investigations of Amborella, the most closely related extant relative of the basal angiosperm (Figure 20.10A), is providing information on the evolutionary origin of the endosperm (Brown 1999). It is probable that the first angiosperm had a four-nucleus embryo sac (Williams and Friedman 2002, 2004). The critical cell to consider is the central cell, which is fertilized by the second sperm to cre- ate the endosperm. In eight-nuclei embryo sacs, there are seven cells. The central cell contains two nuclei and, when fertilized, produces a triploid endosperm. In Nuphar, a basal angiosperm, the embryo sac consists of four nuclei, and the central cell has a single nucleus that, when fertil- ized, develops into a 2n endosperm (Figure 20.10B). The 2n endosperm provides convincing evidence that the four- celled embryo sac in Nuphar does not result from the degra- dation of four nuclei. If other cells had degraded, a 3n endosperm would be predicted.

*The gymnosperm zygote, on the other hand, produces two or more embryos after cell division begins, by a process known as cleavage embryogenesis.

with defective sporophyte tissue in the ovule but a normal haploid embryo sac appear to stimulate normal pollen tube development. While the evidence points primarily to the role of the gametophyte generation in pollen tube guidance, diploid cells may make some contribution. The Arabidopsis gene POP2 encodes an enzyme that degrades γ-amino butyric acid (GABA) and establishes a gradient of GABA in the style up to the micropyle (Palanivelu et al. 2003). The pop2 mutant has misguided pollen tube growth, presumably because there is no GABA gradient. POP2 is expressed in both the pollen and the style, which may explain why wild-type pollen tubes find their way to the micropyle when the style has a pop2 genotype. The wild-type enzyme in the pollen tube may degrade GABA and create a suffi- cient gradient to guide itself to the micropyle. As the final step in pollen guidance, the two synergid cells in the embryo sac may attract the pollen tube. In Tore- nia fournieri (wishbone flower), the embryo sac protrudes from the micropyle and can be cultured. In vitro, it can attract a pollen tube. Higashiyama and colleagues (2001) used a laser beam to destroy individual cells in the embryo sac and then tested whether or not pollen tubes were still attracted to the embryo sac. A single synergid was suffi- cient to guide pollen tubes; however, when both synergids were destroyed, pollen tubes were not attracted to the sac.

Fertilization

The growing pollen tube enters the embryo sac through the micropyle and grows through one of the synergids. The two sperm cells are released, and a double fertilization event occurs (see Southworth 1996). One sperm cell fuses with the egg, producing the zygote that will develop into the sporophyte. The second sperm cell fuses with the bi- or multinucleate central cell, giving rise to the endosperm,

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634 CHAPTER 20

FIGURE 20.10 Ancestral angiosperms. (A) Amborella trichopoda. This plant is more closely related to the first angiosperm than any other extant species. (B) Ancestral angiosperms prob- ably had 2n endosperms. The embryo sac of the basal angiosperm Nuphar (yel- low water lily) has a single nucleus in its central cell , which when fertilized will produce a 2n endosperm. DAPI staining shows that the DNA content is 1n , not n. Because this is a section of tissue, the egg cell is hidden behind the two syn- ergids and is shown in the insert. (A photograph courtesy of Sandra K. Floyd; B photograph courtesy of William Freidman.)

(A)

(B)

Synergids

photograph courtesy of William Freidman.) (A) (B) Synergids Egg Fertilization is not an absolute prerequisite for
photograph courtesy of William Freidman.) (A) (B) Synergids Egg Fertilization is not an absolute prerequisite for
photograph courtesy of William Freidman.) (A) (B) Synergids Egg Fertilization is not an absolute prerequisite for
photograph courtesy of William Freidman.) (A) (B) Synergids Egg Fertilization is not an absolute prerequisite for

Egg

Fertilization is not an absolute prerequisite for angi- osperm embryonic development (Mogie 1992). Embryos can form within embryo sacs from haploid eggs and from cells that did not divide meiotically. This phenomenon is called apomixis (Greek, “without mixing”), and results in viable seeds. The viability of the resulting haploid sporo- phytes indicates that ploidy alone does not account for the morphological distinctions between the gametophyte and the sporophyte. Embryos can also develop from cultured sporophytic tissue. These embryos develop with no associ- ated endosperm, and they lack a seed coat.

Embryonic Development

Embryogenesis

In plants, the term embryogenesis covers development from the time of fertilization until dormancy occurs. The basic body plan of the sporophyte is established during embryo- genesis; however, this plan is reiterated and elaborated after dormancy is broken. The major challenges of plant embryogenesis are:

To establish the basic body plan. Radial patterning pro- duces three tissue systems (dermal, ground, and vascu- lar), and axial patterning establishes the apical-basal (shoot-root) axis. To set aside meristematic tissue for postembryonic elab- oration of the body structure (leaves, roots, flowers, etc.). •To establish an accessible food reserve for the germinating embryo until the embryo becomes autotrophic.

Embryogenesis is similar in all angiosperms in terms of the establishment of the basic body plan. There are differences in pattern elaboration, however, including differences in the precision of cell division patterns, the extent of endosperm development, cotyledon development, and the extent of shoot meristem development (Esau 1977; Steeves and Sussex 1989; Johri et al. 1992).

Central cell

Maternal effect

genes play a key role in establishing embryonic patterns in animals (see, for example, the discussion of Drosophila in Chapter 9). The extent of extrazygotic gene involvement in plant embryogenesis is an open question, complicated by at least three potential sources of influence: sporophytic tissue, gametophytic tissue, and the polyploid endosperm.

All of these tissues are in close association with the egg/ zygote (Ray 1998). Endosperm development could also be affected by maternal genes. Sporophytic and gametophyt- ic maternal effect genes have been identified in Arabidop- sis, and it is probable that the endosperm genome influ- ences the zygote as well. The first maternal effect gene identified, SHORT INTEGUMENTS 1 (SIN1), must be expressed in the sporo- phyte for normal embryonic development to occur (Ray et al. 1996). Two transcription factors (FBP7 and FBP11) are needed in the petunia sporophyte for normal endosperm development (Columbo et al. 1997). A female gametophyt- ic maternal effect gene, MEDEA,* has protein domains sim- ilar to those of a Drosophila maternal effect gene (Gross- niklaus et al. 1998). Curiously, MEDEA is in the Polycomb gene group (see Chapter 9), whose products alter chro- matin, directly or indirectly, and affect transcription. MEDEA affects an imprinted gene (see Chapter 5) that is

MATERNAL EFFECTS IN EARLY EMBRYOGENESIS

*Another name from Greek mythology, after Euripides’ Medea, who killed her own children.

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AN OVERVIEW OF PLANT DEVELOPMENT

635

(A)
(A)
(B) Direction of light
(B)
Direction of light

FIGURE 20.11 Axis formation in the brown alga Pelvetia compressa. (A) An F- actin patch (orange) is first formed at the point of sperm entry; the blue spot marks the sperm pronucleus. (B) Later, light was shone in the direction of the arrow. The sperm-induced axis was overridden, and an F-actin patch formed on the dark side, where the rhizoid will later form. (Pho- tographs courtesy of W. Hables.)

expressed by the female gametophyte and by maternally inherited alleles in the zygote, but not by paternally inher- ited alleles (Vielle-Calzada et al. 1999). The significance of maternal effect genes in establishing the sporophyte body plan has been highlighted by Pagnussat and co-workers’ (2005) screen of 130 female gametophytic mutants. Near- ly half the mutations were in a maternal gene, further implicating the female gametophyte or maternal genome in embryo development.

Polarity is estab-

lished in the first cell division following fertilization. Because angiosperm embryos are deeply embedded in multiple layers of tissue, the establishment of polarity is also investigated in brown algae, a model system with external fertilization (Belanger and Quatrano 2000; Brown- lee 2004). The zygotes of these plants are independent of other tissues and are amenable to manipulation. The ini- tial cell division results in one smaller cell, which will form

FIRST ASYMMETRIC DIVISION: BROWN ALGAE

8 hours after fertilization

the rhizoid (root homologue) and anchor the rest of the plant, and one larger cell, which gives rise to the thallus (the main body of the sporophyte). The point of sperm entry fixes the position of the rhizoid end of the apical- basal axis. This axis is perpendicular to the plane of the first cell division. F-actin accumulates at the rhizoid pole (Figure 20.11A; Kropf et al. 1999). However, light or grav- ity can override this fixing of the axis and establish a new position for cell division (Figure 20.11B; Alessa and Kropf 1999). Once the apical-basal axis is established, secretory vesicles are targeted to the rhizoid pole of the zygote (Fig- ure 20.12). These vesicles contain material for rhizoid out- growth, with a cell wall of distinct macromolecular com-

FIGURE 20.12 Asymmetrical cell division in brown algae. Time course from 8 to 25 hours after fertilization, showing algal cells stained with a vital membrane dye to visualize secretory vesicles, which appear first, and the cell plate, which begins to appear about halfway through this sequence. (Photographs courtesy of K. Belanger.)

this sequence. (Photographs courtesy of K. Belanger.) 25 hours after fertilization Secretory vesicles Cell plate
this sequence. (Photographs courtesy of K. Belanger.) 25 hours after fertilization Secretory vesicles Cell plate
this sequence. (Photographs courtesy of K. Belanger.) 25 hours after fertilization Secretory vesicles Cell plate

25 hours after fertilization

courtesy of K. Belanger.) 25 hours after fertilization Secretory vesicles Cell plate © 2010 Sinauer Associates,
courtesy of K. Belanger.) 25 hours after fertilization Secretory vesicles Cell plate © 2010 Sinauer Associates,
courtesy of K. Belanger.) 25 hours after fertilization Secretory vesicles Cell plate © 2010 Sinauer Associates,
courtesy of K. Belanger.) 25 hours after fertilization Secretory vesicles Cell plate © 2010 Sinauer Associates,
courtesy of K. Belanger.) 25 hours after fertilization Secretory vesicles Cell plate © 2010 Sinauer Associates,

Secretory

vesicles

Cell

plate

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636 CHAPTER 20

Root Shoot Cotyledons meristem meristem Embryo Terminal cell 2-Cell embryo Suspensor Suspensor Zygote Basal
Root
Shoot
Cotyledons
meristem
meristem
Embryo
Terminal cell
2-Cell embryo
Suspensor
Suspensor
Zygote
Basal cell
Hypophysis
FIGURE 20.13 Angiosperm embryogenesis. A representative
dicot is shown; a monocot would develop only a single cotyle-
don. The embryo proper forms from the terminal cell; the basal
cell divides to form the suspensor, which will degenerate as
development progresses. The point of interface between the
suspensor and the embryo is the hypophysis. While there are
basic patterns of embryogenesis in angiosperms, there is
tremendous morphological variation among species.
Globular stage
Heart stage
Torpedo stage
embryo
embryo
embryo
position. Targeted secretion may also help orient the first
plane of cell division. Maintenance of rhizoid versus thal-
lus fate early in development depends on information in
the cell walls (Brownlee and Berger 1995). Such cell wall
information also appears to be important in angiosperms
(see Scheres and Benfey 1999).
FIRST ASYMMETRIC DIVISION: ANGIOSPERMS
The basic body plan
of the angiosperm laid down during embryogenesis also
begins with an asymmetrical* cell division, giving rise to
a terminal cell and a basal cell (Figure 20.13). The terminal
cell gives rise to the embryo proper. The basal cell forms
closest to the micropyle and gives rise to the suspensor.
The hypophysis is found at the interface between the sus-
pensor and the embryo proper. In some species it gives rise
to a portion of the root cells. (The suspensor cells divide to
form a filamentous or spherical organ that degenerates
later in embryogenesis.) In both gymnosperms and
angiosperms, the suspensor orients the absorptive surface
of the embryo toward its food source; in angiosperms, it
also appears to serve as a nutrient conduit for the devel-
oping embryo.
The study of embryo mutants in maize and Arabidopsis
has been particularly helpful in sorting out the different
developmental pathways of embryos and suspensors.
Investigations of suspensor mutants (sus1, sus2, and rasp-
berry1) of Arabidopsis have provided genetic evidence that
the suspensor has the capacity to develop embryo-like
structures (Figure 20.14A,B; Schwartz et al. 1994; Yadegari
et al. 1994). In these mutants, abnormalities in the embryo
proper appear prior to suspensor abnormalities. † Earlier
experiments in which the embryo proper was removed
also demonstrated that suspensors could develop like
embryos (Haccius 1963). A signal from the embryo prop-
er to the suspensor may be important in maintaining sus-
pensor identity and blocking the development of the sus-
pensor as an embryo. Molecular analyses of these and
other genes are providing insight into the mechanisms of
communication between the suspensor and the embryo
proper (Figure 20.14C).
The SUS1 gene has been renamed DCL1 (DICER-LIKE1)
because its predicted protein sequence is structurally like
that of Dicer in Drosophila melanogaster and DCR-1 in
Caenorhabditis elegans (Schauer et al. 2002). These proteins
may control the translation of developmentally important
mRNAs. This is an exciting discovery that will lead to a bet-
ter understanding of the regulation of development beyond
the level of transcriptional control. Intriguingly, DCL1 has
several alleles that were originally assumed to be complete-
ly different genes regulating very different developmental
processes in plants. DCL1 alleles include sin1 alleles. These
mutants affect ovule development (discussed in the next
section) and the transition from vegetative to reproductive
development (discussed later in the chapter). The carpel fac-
tory (caf1) allele of DCL1 causes indeterminancy in floral
meristems leading to extra whorls of carpels. Extrapolat-
ing from Drosophila Dicer function, DCL1 protein may be
involved in cleaving small, noncoding RNAs into even
smaller, 21- to 25-nucleotide, single-stranded RNA prod-
ucts that could cleave to mRNAs and affect translation.
Many questions about the role of microRNAs as possi-
ble developmental signals are arising from the work being
done on DCL1 alleles and on leaf asymmetry genes, which
will be discussed later (Kidner and Martienssen 2005).

*Asymmetrical cell division is also important in later angiosperm development, including the formation of guard cells of leaf stomata and of different cell types in the ground and vascular tissue systems.

Another intriguing characteristic of these mutants is that cell differ- entiation occurs in the absence of morphogenesis. Thus, cell differen- tiation and morphogenesis can be uncoupled in plant development.

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AN OVERVIEW OF PLANT DEVELOPMENT

637

(A)

AN OVERVIEW OF PLANT DEVELOPMENT 6 3 7 (A) (B) (C) Signals from Signals from embryo

(B)

AN OVERVIEW OF PLANT DEVELOPMENT 6 3 7 (A) (B) (C) Signals from Signals from embryo
(C) Signals from Signals from embryo suspensor suppress cells promote embryonic embryonic development
(C)
Signals from
Signals from
embryo
suspensor
suppress
cells promote
embryonic
embryonic
development
development
in suspensor
cells

The globular shape of the embryo is lost as cotyledons (“first leaves”) begin to form. Dicots have two cotyledons, which give the embryo a heart-shaped appearance as they form. In monocots, such as maize, only a single cotyledon emerges.The axial body plan is evident by this heart stage of development (Figure 20.15B). Hormones (specifically, auxins) may mediate the transition from radial to bilateral symmetry (Liu et al. 1993).

The hormone auxin plays

a key role in establishing the apical-basal axis of the

embryo. Very early in embryogenesis, the family of pin- formed (PIN) auxin efflux carriers are asymmetrically dis- tributed (Figure 20.16A; Friml et al. 2003, 2004). The pinoid (PID) protein kinase aids in the asymmetric localization of PIN proteins. At the 4-cell stage, auxin moves apically, but

AUXIN AND THE APICAL-BASAL AXIS

by the globular stage, the efflux carriers direct the flow of auxin to the hypophysis, which organizes as the root meris- tem (Figure 20.16B). Disruption of the auxin morphogenet-

ic gradient, either through mutation or overexpression of

(A) Radial patterning

layer layer)
layer
layer)

Globular stage

Protoderm

Ground layer

Procambium

(vascular

(B) Axial patterning

Ground layer Procambium (vascular (B) Axial patterning Heart stage Cotyledon Shoot meristem Procambium Root

Heart stage

Cotyledon

Shoot

meristem

Procambium

Root meristem

Root–shoot

axis

FIGURE 20.14 The SUS gene (a DCL1 allele) suppresses embryonic development in the suspensor. (A) Wild-type embryo and suspensor. (B) The suspensor of a sus mutant devel- ops like an embryo (arrow). (C) Model showing how the embryo proper suppresses embryonic development in the suspensor and the suspensor provides feedback information to the embryo. (Photographs courtesy of D. Meinke.)

Radial and axial patterns

develop as cell division and differentiation continue (Fig- ure 20.15). The cells of the embryo proper divide in trans- verse and longitudinal planes to form a globular stage embryo with several tiers of cells. Superficially, this stage bears some resemblance to cleavage in animals, but the nucleus/cytoplasm ratio does not necessarily increase. The emerging shape of the embryo depends on regulation of the planes of cell division and expansion, since the cells are not able to move and reshape the embryo. Cell division planes in the outer layer of cells become restricted, and this layer, called the protoderm, becomes distinct. Radial pat- terning emerges at the globular stage as the three tissue systems (dermal, ground, and vascular) of the plant are initiated (Figure 20.15A). The dermal tissue (epidermis) will form from the protoderm and contribute to the outer protective layers of the plant. Ground tissue (cortex and pith) forms from the ground meristem, which lies beneath the protoderm. The procambium, which forms at the core of the embryo, gives rise to the vascular tissue (xylem and phloem), which will function in support and transport. The differentiation of each tissue system is at least partially independent. For example, in the keule mutant of Arabidop- sis, the dermal system is defective while the inner tissue systems develop normally (Mayer et al. 1991).

RADIAL

AND

AXIAL

PATTERNING

FIGURE 20.15 Radial and axial patterning. (A) Radial patterning in angiosperms begins in the globular stage and results in the establishment of three tissue systems. (B) The axial pat- tern (shoot-root axis) is established by the heart stage.

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638 CHAPTER 20

638 CHAPTER 20 (A) Embryo Suspensor Embryo Suspensor Auxin transport via PIN family auxin efflux carriers
(A) Embryo Suspensor
(A)
Embryo
Suspensor

Embryo

Suspensor

Auxin transport via PIN family auxin efflux carriers Cells accumulating auxin Cells producing auxin

Auxin transport via PIN family auxin efflux carriers

Cells accumulating auxin Cells producing auxin

FIGURE 20.16 An auxin gradient specifies the shoot-root axis. (A) A family of PIN auxin efflux carriers are responsible for the early apical flow of auxin in the embryo and the shift to basal auxin flow as the apical end of the globular stage embryo begins to produce auxin. (B,C) The PID protein kinase localizes the PIN auxin efflux carriers. (B) In wild-type globular embryos, PID expression leads to the accumulation of PIN1 at the basal end of the embryo. When an auxin-response gene is fused with GFP and expressed, basal accumulation and normal embryonic development occur. (C) Ectopic overexpression of the PID gene (induced by fusion to a viral promoter) eliminates basal accumu- lation of PIN1. The GFP-labeled auxin response gene is then expressed throughout the embryo proper rather than in the basal portion of the embryo.

PID allows auxin to accumulate more apically and root development is abnormal or inhibited (Figure 20.16C). Although auxin accumulation is necessary for apical-basal polarity, an extensive study of the PIN proteins reveals that auxin alone may not be sufficient to establish polarity in the embryo (Weijers and Jürgens 2005). The discovery of the auxin receptor provided a link between the asymmetric distribution of auxin and the expression of auxin-induced genes (Dharmasiri et al. 2005; Kipinski and Leyser 2005). In the absence of auxin, the auxin response factors (ARF) are in a repressed state (Fig- ure 20.17A). These genes form a heterodimer with Aux/IAA protein, which inhibits the transcription of auxin-induced genes. In the presence of auxin, the enzyme SCF binds Aux/IAA, catalyzing Aux/IAA ubiquitination (Figure 20.17B). Ubiquitination targets Aux/IAA for degra- dation by the 26S proteosome. Freed from the Aux/IAA repressor, ARF can act on its own to stimulate transcrip- tion or form a homo/heterodimer with another ARF to fur- ther modulate gene expression.

COTYLEDONS In many plants, the cotyledons aid in nour- ishing the plant by becoming photosynthetic after germina-

(B) Wild type Concentration of PIN1 auxin efflux carriers
(B) Wild type
Concentration
of PIN1 auxin
efflux carriers
Auxin-induced gene expression
Auxin-induced
gene expression

(C) PID overexpression

Auxin-induced gene expression (C) PID overexpression No PIN 1 auxin efflux carriers Auxin-induced gene

No PIN 1 auxin efflux carriers

Auxin-induced gene expression
Auxin-induced
gene expression

tion (although those of some species never emerge from the ground). In some cases—peas, for example—the food reserve in the endosperm is used up before germination, and the cotyledons serve as the nutrient source for the ger- minating seedling.* Even in the presence of a persistent endosperm (as in maize), the cotyledons store food reserves such as starch, lipids, and proteins. In many monocots, the cotyledon grows into a large organ pressed against the endosperm and aids in nutrient transfer to the seedling. Upright cotyledons can give the embryo a torpe- do shape. In some plants, the cotyledons grow sufficient- ly long that they must bend to fit within the confines of the seed coat. The embryo then looks like a walking stick. By this point, the suspensor is degenerating. The Arabidopsis LEAFY COTYLEDON1 (LEC1) gene, first identified by a mutant with leaflike cotyledons (Meinke 1994), is necessary to maintain the suspensor early in devel- opment, to specify cotyledeon identity, to initiate matura- tion, and to prevent early germination of the seed. LEC1 belongs to a gene class that is unique among the embryo- genesis genes in acting throughout the course of embryo development (Harada 2001; Kwong et al. 2003).

MERISTEM ESTABLISHMENT The shoot apical meristem and root apical meristem are clusters of stem cells that will per-

*Mendel’s famous wrinkled-seed mutant (the rugosus or r allele) has a defect in a starch branching enzyme that affects starch, lipid, and protein biosynthesis in the seed and leads to defective cotyle- dons (Bhattacharyya et al. 1990).

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sist in the postembryonic plant and give rise to most of the sporophyte body (see Jurgens 2001 for a review of apical- basal pattern formation). The root meristem is partially derived from the hypophysis in some species (see Figure 20.15B). All other parts of the sporophyte body are derived from the embryo proper. Genetic evidence indicates that the formation of the shoot and root meristems is regulated independently. From an evolutionary perspective, this is not surprising. One of the major adaptations to terrestrial life involved the evo- lution of a root system.* This independence is demonstrat- ed by the dek23 maize mutant and the shootmeristemless (STM) mutant of Arabidopsis, both of which form a root meristem but fail to initiate a shoot meristem (Clark and Sheridan 1986; Barton and Poethig 1993). The STM gene, which has a homeodomain, is expressed in the late globu- lar stage, before cotyledons form. Genes have also been identified that specifically affect the development of the root axis during embryogenesis. Mutations of the HOBBIT gene in Arabidopsis, for example, affect the hypophysis derivatives and eliminate root meristem function (Willem- son et al. 1998). While it is clear that root and shoot devel- opmental programs are different, what triggers root or

*It should be noted, however, that the nonvascular land plants, including the mosses, did not develop root systems.

(A) Auxin (Indole-3-acetic acid)

N
N

H

CH 2 COOH

(B) No Auxin present

SCF (enzyme) Aux/IAA (inhibitor) No transcription ARF (transcription factor) Promoter Auxin-induced gene
SCF (enzyme)
Aux/IAA (inhibitor)
No transcription
ARF (transcription
factor)
Promoter
Auxin-induced gene
(C) Auxin present Auxin SCF 26S proteosome Aux/IAA + Ubiquitin Protein
(C) Auxin present
Auxin
SCF
26S proteosome
Aux/IAA
+
Ubiquitin
Protein

degradation

AN OVERVIEW OF PLANT DEVELOPMENT

639

shoot development is less tractable. The TOPLESS gene in Arabidopsis may provide some clues. A single gene muta- tion has been identified that converts a shoot into a root, but how the wild-type gene functions is still a puzzle (Long et al. 2002). The shoot apical meristem will initiate leaves after ger- mination and, ultimately, the transition to reproductive development. In Arabidopsis, the cotyledons are produced from general embryonic tissue, not from the shoot meris- tem (Barton and Poethig 1993). In many angiosperms, a few leaves are initiated during embryogenesis. In the case of Arabidopsis, clonal analysis points to the presence of leaves in the mature embryo, even though they are not morphologically well developed (Irish and Sussex 1992). Clonal analysis has demonstrated that the cotyledons and the first two true leaves of cotton plants are derived from embryonic tissue rather than an organized meristem (Christianson 1986). Clonal analysis experiments provide information on cell fates, but do not necessarily indicate whether or not cells are determined for a particular fate. Clonal analysis has demonstrated that cells that divide in the wrong plane and “move” to a different tissue layer often differentiate accord- ing to their new position. Position, rather than clonal ori- gin, appears to be the critical factor in embryo pattern for- mation, suggesting some type of cell-cell communication (Laux and Jürgens 1994).

Dormancy

From the earliest stages of embryogenesis, there is a high level of zygotic gene expression. As the embryo reaches maturity, there is a shift from constructing the basic body plan to creating a food reserve by accumulating storage carbohydrates, proteins, and lipids. Genes coding for seed storage proteins were among the first to be characterized by plant molecular biologists because of the high levels of specific storage protein mRNAs that are present at differ- ent times in embryonic development. The high level of

FIGURE 20.17 Mechanism of auxin action. (A) In the absence of auxin, auxin response factors (ARF) are in a repressed state. These genes form a heterodimer with Aux/IAA protein, which inhibits the transcription of auxin-induced genes. (B) In the pres- ence of auxin, the enzyme SCF binds Aux/IAA, catalyzing Aux/IAA ubiquitination. Ubiquitination targets Aux/IAA for degradation by the 26S proteosome. Freed from the Aux/IAA repressor, ARF can act on its own to stimulate transcription or form a homo or heterodimer with another ARF to further modu- late gene expression.

Transcription ARF Promoter Auxin-induced gene
Transcription
ARF
Promoter
Auxin-induced gene

Auxin-induced

mRNA

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640 CHAPTER 20

FIGURE 20.18 Viviparous maize mutant. Each kernel on this maize ear contains an embryo. Viviparous embryos do not go through a dormant phase, but begin germinating while still on the ear.

metabolic activity in the developing embryo is fueled by continuous input from the parent plant into the ovule. Eventually metabolism slows, and the connection of the seed to the ovary is severed by the degeneration of the adjacent supporting sporophyte cells. The seed desiccates (loses water), and the integuments harden to form a tough seed coat. The seed has entered dormancy, officially ending

embryogenesis. The embryo can persist in a dormant state for weeks or years, a phenomenon that affords tremendous survival value. There are even cases where seeds found stored in ancient archaeological sites have germinated after thousands of years of dormancy. Maturation leading to dormancy is the result of a pre- cisely regulated program. The viviparous mutation in maize, for example, produces genetic lesions that block dorman- cy (Steeves and Sussex 1989). The apical meristems of vivip- arous mutants behave like those of ferns, with no pause before producing postembryonic structures. The embryo continues to develop, and seedlings emerge from the ker- nels on the ear attached to the parent plant (Figure 20.18).

A group of plant genes have been identified that belong to

the Polycomb group, which regulates early development

in mammals, nematodes, and insects (Preuss 1999). These

genes encode chromatin silencing factors, which may play an important role in seed formation. Plant hormones are critical in dormancy, and linking them to genetic mechanisms is an active area of research. The hormone abscisic acid is important in maintaining dormancy in many species. Gibberellins, another class of

hormones, are important in breaking dormancy.

Germination

The postembryonic phase of plant development begins

with germination. Some dormant seeds require a period

of after-ripening, during which low-level metabolic activ-

ities continue to prepare the embryo for germination. High-

ly evolved interactions between the seed and its environ-

ment increase the odds that the germinating seedling will survive to produce another generation. Temperature, water, light, and oxygen are all key in determining the success of germination. Stratification is the requirement for chilling (5°C) to break dormancy in some seeds. In temperate climates, this adaptation ensures that germination takes place only after the winter months have passed. In addition, seeds have maximum germina- tion rates at moderate temperatures of 25–30°C and often will not germinate at extreme temperatures. Seeds such as lettuce require light (specifically, the red light wavelengths) for germination; thus seeds will not germinate so far below

for germination; thus seeds will not germinate so far below ground that they use up their

ground that they use up their food reserves before photo- synthesis is possible. Desiccated seeds may be only 5–20 percent water. Imbi- bition is the process by which the seed rehydrates, soak- ing up large volumes of water and swelling to many times its original size. The radicle (primary embryonic root) emerges from the seed first to enhance water uptake; it is protected by a root cap produced by the root apical meris- tem. Water is essential for metabolic activity, but so is oxy- gen; a seed sitting in a glass of water will not survive. Some species have such hard protective seed coats that they must be scarified (scratched or etched) before water and oxy- gen can cross the barrier. Scarification can occur when the seed is exposed to the weather and other natural elements over time, or by its exposure to acid as the seed passes through the gut of a frugivore (fruit eater). The frugivore thus prepares the seed for germination, as well as dispers- ing it to a site where germination can take place. During germination, the plant draws on the nutrient reserves in the endosperm or cotyledons. Interactions between the embryo and endosperm in monocots use gib- berellin as a signal to trigger the breakdown of starch into sugar. As the shoot reaches the surface, the differentiation of chloroplasts is triggered by light. Seedlings that germi- nate in the dark have long, spindly stems and do not pro- duce chlorophyll. This environmental response allows plants to use their limited resources to reach the soil sur- face, where photosynthesis will be productive.

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Terminal flower Axillary Inflorescence flower Vegetative axillary bud Internode Node Leaf blade Epicotyl
Terminal
flower
Axillary
Inflorescence
flower
Vegetative
axillary bud
Internode
Node
Leaf
blade
Epicotyl
Cotyledons
Hypocotyl
Root apical
meristem

FIGURE 20.19 Morphology of a generalized angiosperm sporophyte.

Vegetative Growth

When the shoot emerges from the soil, most of the sporo- phyte body plan remains to be elaborated. Figure 20.19 shows the basic parts of the mature sporophyte plant, which will emerge from meristems.

Meristems

As has been mentioned, meristems are clusters of cells that allow the basic body pattern established during embryoge- nesis to be reiterated and extended after germination. Meristematic cells are similar to stem cells in animals.* They divide to give rise to one daughter cell that continues to be meristematic and another that differentiates. Meristems fall into three categories: apical, lateral, and intercalary. Apical meristems occur at the growing shoot and root tips. Root apical meristems produce the root cap, which consists of lubricated cells that are sloughed off as the meristem is pushed through the soil by cell division and

*The similarities between plant meristem cells and animal stem cells may extend to the molecular level, indicating that stem cells existed before plants and animals pursued separate phylogenetic pathways. Homology has been found between genes required for plant meristems to persist and genes expressed in Drosophila germ line stem cells (Cox et al. 1998).

AN OVERVIEW OF PLANT DEVELOPMENT

641

elongation in more proximal cells. The root apical meris- tem also gives rise to daughter cells that produce the three tissue systems of the root (Figure 20.20A). New root api- cal meristems are initiated from tissue within the core of the root and emerge through the ground tissue and der- mal tissue. Root meristems can also be derived secondari- ly from the stem of the plant; in the case of maize, this is the major source of root mass. The shoot apical meristem produces stems, leaves, and reproductive structures. In addition to the shoot apical meristem initiated during embryogenesis, axillary shoot apical meristems (axillary buds; Figure 20.20B) derived from the original one form in the axils (the angles between leaf and stem). Unlike new root meristems, these arise from the surface layers of the meristem.

(A) Root meristems Lateral root Pericycle Root hairs Protoderm Ground tissue Procambium Apical meristem Root
(A) Root meristems
Lateral root
Pericycle
Root hairs
Protoderm
Ground
tissue
Procambium
Apical
meristem
Root cap

(B) Shoot meristems

Axillary Apical meristem meristem Leaf Protoderm primordium Leaf primordium Ground tissue Procambium
Axillary
Apical meristem
meristem
Leaf
Protoderm
primordium
Leaf
primordium
Ground
tissue
Procambium

FIGURE 20.20 Both shoots and roots develop from apical meristems, with undifferentiated cells clustered at their tips. (A) The root meristem is protected by the root cap as it pushes through the soil. Lateral roots are derived from pericycle cells deep within the root. (B) The lateral organs of the shoot (leaves and axillary branches) have a superficial origin in the shoot api- cal meristem.

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642 CHAPTER 20

(A) Apical meristem

(B)
(B)

tity in the cells positioned above, and the CLV signaling pathway is proposed to negatively feed back on WUS expression to control the size of the meristem. It is possible that POL expression is a target in CLV1 signal transduction. STM, like POL and WUS, plays an important role in maintaining an undifferenti- ated population of meristematic cells, and STM may positively reg- ulate WUS (Clark and Schiefelbein 1997). The interactions of these gene products balance the rate of cell division (which enlarges the meri- stem) and the rate of cell differenti- ation in the periphery of the meri- stem (which decreases meristem size) (Meyerowitz 1997). Lateral meristems are cylindri- cal meristems found in shoots and roots that result in secondary growth (an increase in stem and

root girth by the production of vas- cular tissues). Monocots do not have lateral meristems, but often have intercalary meristems insert- ed in their stems between mature tissues. The popping sound you can hear in a cornfield on a summer night is caused by the rapid increase in stem length due to inter- calary meristems.

increase in stem length due to inter- calary meristems. L1 L2 L3 FIGURE 20.21 Organization of
L1 L2 L3
L1
L2
L3

FIGURE 20.21 Organization of the shoot apical meristem. (A) Angiosperm meristems

have two or three outer layers of cells that are histologically distinct (L1, L2, and L3). While cells in certain layers tend to have certain fates, they are not necessarily committed to those fates. If a cell is shifted to a new layer, it generally develops like the other cells in that layer. (B) The fates of the cell layers can be seen in a chimeric tobacco plant. One por- tion of the meristem contains three layers of wild-type cells, while the other portion has an L2 that lacks chlorophyll. This section of the meristem has given rise to the variegated leaves. In wild-type plants, the L1 layer always lacks chlorophyll (except in guard cells), but in this plant the L2, too, is genetically unable to produce chlorophyll; the L3 remains green. The L3 does not contribute to the outer edges of leaves, which is why they appear

white in this plant. (Photograph courtesy of M. Marcotrigiano.)

Angiosperm shoot apical meristems are composed of up to three layers of cells (labeled L1, L2, and L3) on the plant surface (Figure 20.21A). One way of investigating the contributions of different layers to plant structure is by con- structing chimeras (Figure 20.21B). Plant chimeras are com- posed of layers having distinct genotypes with discernible markers. When L2, for example, has a different genotype than L1 or L3, all pollen will have the L2 genotype, indi- cating that pollen is derived from L2. Chimeras have also been used to demonstrate classical induction in plants, in which (as in animal development) one layer influences the developmental pathway of an adjacent layer. The size of the shoot apical meristem is precisely con- trolled by intercellular signals, most likely between layers of the meristem (see Bäurle and Laux 2003). Mutations in the Arabidopsis CLAVATA (CLV) genes, for example, lead to increased meristem size and the production of extra organs. CLV1, CLV2, and CLV3 all limit the number of undifferentiated, stem cells in both vegetative and floral meristems (Figure 20.22). CLV1 is a serenine-threonine kinase that, along with the receptor-like transmembrane CLV2 protein, form a receptor for CLV3, which is localized in the extracellular space between cell layers of the meri- stem (Clark et al. 1997; Jeong et al. 1999; Rojo et al. 2002). Reddy and Meyerowitz (2005) have demonstrated that CLV3 restricts its own domain of expression to the central zone by preventing the differentiation of surrounding cells. POLTERGEIST (POL) and WUSCHEL (WUS) are redun- dant in function and appear to keep genes in an undiffer- entiatied state (Yu et al. 2000). WUS specifies stem cell iden-

Stem cell population CLV3 CLV3 STM STM Cell Cell WUS differentiation differentiation CLV1/2 L1 L2
Stem cell
population
CLV3
CLV3
STM
STM
Cell
Cell
WUS
differentiation
differentiation
CLV1/2
L1
L2
L3

FIGURE 20.22 The WUS and STM proteins act to keep meris- tem cells in an undifferentiated state, while the products of the CLAVATA genes CLV1, CLV2, and CLV3 all limit the number of undifferentiated meristem cells. The presence of WUS indirectly induces expression (upward arrow) of CLV3 in L1 and L2. CLV3 is a ligand that binds to the CLV1/CLV2 receptor in L3 cells. This binding triggers a negative feedback signal cascade that inhibits WUS expression and regulates the number of undiffer- entiated cells in the meristem, thus counterbalancing the roles of STM and WUS in keeping cells undifferentiated and dividing.

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Root development

Radial and axial patterning in roots begins during embryo- genesis and continues throughout development as the pri- mary root grows and lateral roots emerge from the pericy- cle cells deep within the root. Both clonal analyses and laser ablation experiments that elilminate single cells have demonstrated that cells are plastic, and that position is the primary determinant of fate in early root development. Analyses of mutations in root radial organization have revealed genes with layer-specific activity (Scheres et al. 1995; Scheres and Heidstra 1999). We will illustrate these findings by looking at two Arabidopsis genes that regulate ground tissue fate. In wild-type Arabidopsis, there are two layers of root ground tissue. The outer layer becomes the cortex while the inner layer becomes the endodermis, which forms a tube around the vascular tissue core. The SCARECROW (SCR) and SHORT-ROOT (SHR) genes have mutant phenotypes with only a single layer of root ground tissue (Benfey et al. 1993). The SCR gene is necessary for an asymmetrical cell division in the initial layer of cells, yielding a smaller endo- dermal cell and a larger cortex cell (Figure 20.23). The scr mutant expresses markers for both cortex and endodermal cells, indicating that differentiation progresses in the absence of cell division (Di Laurenzio et al. 1996). SHR is responsible for endodermal cell specification. Cells in the shr mutant do not develop endodermal features. Axial patterning in roots may be morphogen-depend- ent, paralleling some aspects of animal development. A variety of experiments have established that the distribu- tion of the plant hormone auxin organizes the axial pat- tern. A peak in auxin concentration at the root tip must be perceived for normal axial patterning (Sabatini et al. 1999; Ueda et al. 2005). As discussed earlier, distinct genes specifying root and shoot meristem formation have been identified; however, root and shoot development may share common groups of genes that regulate cell fate and patterning (Benfey 1999). This appears to be the case for the SCR and SHR genes. In the shoot, these genes are necessary for the normal gravit- ropic response, which is dependent on normal endodermis formation (a defect in mutants of both genes; see Figure 20.29C). It is important to keep in mind that there are a number of steps between establishment of the basic pattern and elaboration of that pattern into anatomical and mor- phological structure. Uncovering the underlying control mechanisms is likely to be the most productive strategy in understanding how roots and shoots develop.

Shoot development

The unique aboveground architectures of different plant species have their origins in shoot meristems. Shoot archi- tecture is affected by the amount of axillary bud outgrowth. Branching patterns are regulated by the shoot tip—a phe- nomenon called apical dominance—and plant hormones

AN OVERVIEW OF PLANT DEVELOPMENT 643 Positional information (A) Endo- Daughter Cortex dermal cell cell
AN OVERVIEW OF PLANT DEVELOPMENT
643
Positional information
(A)
Endo-
Daughter
Cortex
dermal
cell
cell
cell
Cortex/
Cortex/
Cortex/
endodermal
endodermal
endodermal
initial
initial
initial
(B) Root
(C) Shoot
endodermal initial initial initial (B) Root (C) Shoot (D) Wild type Ep EpEp C C En

(D) Wild type

Ep EpEp C C En EnEn V V P P
Ep EpEp
C C
En
EnEn
V V
P P
(C) Shoot (D) Wild type Ep EpEp C C En EnEn V V P P (E)

(E) scr mutant

P P M M V V Ep EpEp
P P
M M
V V
Ep EpEp

(F) shr mutant

EpEp Ep V V M M P P
EpEp Ep
V V
M M
P P

FIGURE 20.23 SCR and SHR

regulate endodermal differ- entiation in root radial devel- opment. (A) Diagram of nor- mal cell division yielding cortical and endodermal cells.

SCR regulates this asymmetri- cal cell division. (B,C) SCR expression in root and shoot. The SCR promoter is linked to the gene for GFP (green fluo- rescent protein). (D–F) Cross sections of primary roots of (D) wild-type Arabidopsis, (E) scr mutant, and (F) shr mutant. Ep, epi- dermis; C, cortex; En, endodermis; M, mutant layer; P, pericycle; V, vascular tissue; St, stele. (A after Scheres and Heidstra 1999; B–F photographs courtesy of P. Benfey.)

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644 CHAPTER 20

appear to be the regulating factors. Auxin is produced by young leaves and transported toward the base of the leaf. It can suppress the outgrowth of axillary buds. Grazing and flowering often release buds from apical dominance, at which time branching occurs. Cytokinins can also release buds from apical dominance. Axillary buds can ini- tiate their own axillary buds, so branching patterns can get quite complex. Branching patterns can be regulated by environmental signals so that an expansive canopy in an open area maximizes light capture. Asymmetrical tree crowns form when two trees grow very close to each other. In addition to its environmental plasticity, shoot architec- ture is genetically regulated. In several species, genes have now been identified that regulate branching patterns. Leaf primordia (clusters of cells that will form leaves) are initiated at the periphery of the shoot meristem (see Figure 20.20). The union of a leaf and the stem is called a node, and stem tissue between nodes is called an intern- ode (see Figure 20.19). In a simplistic sense, the mature sporophyte is created by stacking node/internode units together. Phyllotaxy, the positioning of leaves on the stem, involves communication among existing and newly form- ing leaf primordia. Leaves may be arranged in various pat- terns, including a spiral, 180º alternation of single leaves, pairs, and whorls of three or more leaves at a node (Jean and Barabé 1998). Experimentation has revealed a number of mechanisms for maintaining geometrically regular spac- ing of leaves on a plant, including chemical and physical interactions of new leaf primordia with the shoot apex and with existing primordia (Steeves and Sussex 1989). It is not clear how a specific pattern of phyllotaxy gets started. Descriptive mathematical models can replicate the observed patterns, but reveal nothing about the mecha- nism. Biophysical models (e.g., of the effects of stress or strain on deposition of cell wall material, which affects cell division and elongation) attempt to bridge this gap. Devel- opmental genetics approaches are promising, but few phyl- lotactic mutants have been identified.* Currently there is much interest in the role of local auxin maxima and mini- ma in determining where the next leaf primordium will form on a meristem. The PIN gene family that plays an important role in embryo axis formation has also been implicated in phyllotaxy because of the correlation between localization of PIN auxin efflux carriers and primordia sit- ing (Fleming 2005).

Leaf development

Leaf development includes the cells’ commitment to become a leaf; establishment of the leaf axes; and morpho- genesis, which gives rise to a tremendous diversity of leaf

*One candidate phyllotactic mutant is the terminal ear mutant in maize, which has irregular phyllotaxy. The wild-type gene is expressed in a horseshoe-shaped region, with a gap where the leaf will be initiated (Veit et al. 1998). The plane of the horseshoe is per- pendicular to the axis of the stem.

shapes. Culture experiments have assessed when leaf pri- mordia become determined for leaf development. Research on ferns and angiosperms indicates that the youngest vis- ible leaf primordia are not determined to make a leaf; rather, these young primordia can develop as shoots in cul- ture (Steeves 1966; Smith 1984). The programming for leaf development occurs later. The radial symmetry of the leaf primordium becomes dorsal-ventral, or flattened, in all leaves. Two other axes, the proximal-distal and lateral axes, are also established. The unique shapes of leaves result from regulation of cell division and cell expansion as the leaf blade develops. There are some cases in which selective cell death (apop- tosis) is involved in the shaping of a leaf, but differential cell growth appears to be a more common mechanism (Gif- ford and Foster 1989).

Plant biologists refer

to the surface of the leaf that is closest to the stem as the adaxial side and the more distant surface is the abaxial side. As the leaf begins to form, the Arabidopsis genes PHABU- LOSA (PHB) and PHAVOLUTA (PHV) initially have uni- formly expressed RNA throughout the primordium (Fig- ure 20.24A). The PHB and PHV proteins are postulated to be receptors for an adaxial signal, which leads to the accu- mulation of PHB and PHV on the adaxial leaf surface (McConnell et al. 2001; Byrne 2005). Exclusion of PHB and PHV from the abaxial side is caused by microRNA binding to the transcripts of the two genes, which leads to their degradation (Kidner and Martienssen 2005). In addition, the PHB- and PHV-specific microRNAs appear to increase methylation of their complementary PHB and PHV DNA sequences, suppressing transcription (Bao et al. 2004). Dom- inant gain-of-function mutants have disrupted microRNA binding sites such that these genes are expressed on both sides of the primordium, leading to a leaf with two adaxial sides. The sequences of PHB and PHV make it likely that they are activated by a lipid ligand and that this activation is followed by the development of adaxial cells. KANADI (KAN) genes initiate abaxial cell differentia- tion in Arabidopsis (Kerstetter et al. 2001). In situ hybridiza- tion shows that KAN is transiently expressed on the abax- ial side of cotyledons, leaves, and initiating floral organ primordia (Figure 20.24B). KAN contains a GARP domain found in transcription factors. KAN and PHB/PHV mutual- ly suppress each other to maintain abaxial and adaxial fates, respectively. Abaxial fate also appears to be specified by three mem- bers of the YABBY gene family (Siegfried et al. 1999). The exact mechanism of YABBY genes in leaf polarity is unknown, although promoter deletion experiments reveal that FILAMENTOUS, a gene in the YABBY family, is active- ly excluded from the adaxial side of the primordium. KAN and YABBY likely have redundant functions. KAN gene expression is restricted to the abaxial side by PHB and PHV, while KAN restricts PHB and PHV expression to the adaxial side (Figure 20.24C).

DORSAL-VENTRAL PATTERNING IN LEAVES

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AN OVERVIEW OF PLANT DEVELOPMENT 645 Adaxial surface Adaxial development Abaxial surface
AN OVERVIEW OF PLANT DEVELOPMENT
645
Adaxial
surface
Adaxial
development
Abaxial
surface
(A) Leaf primordium
(A)
Leaf
primordium
(B)
(B)
development Abaxial surface (A) Leaf primordium (B) Abaxial development (C) ac Ab ia f a x
development Abaxial surface (A) Leaf primordium (B) Abaxial development (C) ac Ab ia f a x

Abaxial

development

(C) ac Ab ia f a x x A l e i a r d
(C)
ac
Ab
ia
f
a
x
x
A
l
e
i
a
r
d
a
l
s
s
u
u
r
f
a
c
e

Normal

Adaxial signalA l e i a r d a l s s u u r f a

PHB, PHVr d a l s s u u r f a c e Normal Adaxial signal

Micro RNAs bind PHB and PHV RNAl s s u u r f a c e Normal Adaxial signal PHB, PHV KAN

KANAdaxial signal PHB, PHV Micro RNAs bind PHB and PHV RNA YABBY development FIGURE 20.24 Patterning

YABBYAdaxial signal PHB, PHV Micro RNAs bind PHB and PHV RNA KAN development FIGURE 20.24 Patterning

development

FIGURE 20.24 Patterning of adaxial and abaxial leaf surfaces of Arabidopsis. (A) PHB and PHV proteins are uniformly distrib- uted throughout the leaf primordium. MicroRNAs expressed on the abaxial side specifically degrade PHB and PHV, leading to degradation by DICER and blocking translation. In addition, microRNAs increase methylation of the complementary regions in PHB and PHV DNA, suppressing transcription. (B) Transient expression of KAN on the abaxial side of the leaf leads to the expression of YABBY genes, which in turn lead to the transcrip- tion of other abaxial genes. (C) PHB and PHV block KAN adaxial activity. Expression of PHB, PHV, and KAN is transient and occurs early in leaf development.

Leaves fall into two categories, simple and compound (Figure 20.25; see review by Sinha 1999). There is much variety in simple leaf shape, from smooth-edged leaves to deeply lobed oak leaves. Compound leaves are composed

of individual leaflets (and sometimes tendrils) rather than

a single leaf blade. Whether simple and compound leaves

develop by the same mechanism is an open question. One perspective is that compound leaves are highly lobed sim-

ple leaves. An alternative perspective is that compound leaves are modified shoots. The ancestral state for seed

plants is believed to be compound, but for angiosperms it

is simple. Compound leaves have arisen multiple times in

the angiosperms, and it is not clear if these are reversions to the ancestral state. Developmental genetic approaches are being applied to

leaf morphogenesis. The Class I KNOX genes are home- obox genes that include STM and the KNOTTED 1 (KN1)

gene in maize. Gain-of-function mutations of KN1 cause meristem-like bumps to form on maize leaves. In wild-type plants, this gene is expressed in meristems. KNOX genes stimulate meristem initiation and growth. Although some YABBY genes specify abaxial leaf identity (see Figure 20.24), others function by downregulating KNOX genes,

SIMPLE LEAF Midrib Petiole Blade COMPOUND LEAF Vein Stipule Tendril Petiole Rachis Leaflet
SIMPLE LEAF
Midrib
Petiole
Blade
COMPOUND LEAF
Vein
Stipule
Tendril
Petiole
Rachis
Leaflet

FIGURE 20.25 Simple and compound leaves.

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646 CHAPTER 20

(A) (B) (C)
(A)
(B)
(C)

FIGURE 20.26 Overexpression of Class 1 KNOX genes in tomato. The photograph shows the single leaves of (A) a wild- type plant, (B) a mouse ears mutant, with increased leaf com- plexity, and (C) a transgenic plant that uses a viral promoter to overexpress the tomato homologue (LeT6) of the KN1 gene from maize. (Photographs courtesy of N. Sinha.)

thereby restricting where meristems form (Kumaran et al. 2002). If too much KN1 is present, YABBY is insufficient to control KN1 expression. When KN1, or the tomato homologue LeT6, has its pro- moter replaced with a promoter from cauliflower mosaic virus and is inserted into the genome of tomato, the gene is expressed at high levels throughout the plant, and the leaves become “super compound” (Figure 20.26; Hareven et al. 1996; Janssen et al. 1998). Simple leaves become more lobed (but not compound) in response to overexpression of KN1, consistent with the hypothesis that compound leaves may be an extreme case of lobing in simple leaves (Jackson 1996). The role of KN1 in shoot meristem and leaf

development, however, is consistent with the hypothesis that compound leaves are modified shoots. Looking at pat- terns of KN1 expression over a broad range of angiosperm taxa, Bharathan et al. (2002) demonstrated that KN1 expres- sion is associated with compound leaf primordia, but is not present in the developing leaf primordia of simple leaves (Figure 20.27). These data support the conclusion that com- pound leaves maintain shootlike activity, at least for some time. How KN1 works in meristems and in leaf primordia is an area of active research. In tomato leaf, KN1 causes a

(A) (B) (C) (C)(C) (D) (E) (F) (F)(F) LP LPLPLP MM LPLPLP (G) (H) (I)
(A)
(B)
(C) (C)(C)
(D)
(E)
(F) (F)(F)
LP
LPLPLP
MM LPLPLP
(G)
(H)
(I) (I)(I)

FIGURE 20.27 KNOX is known to occur in leaf primordia in species making complex leaves. Here we show that species with simple leaves can also express KNOX in leaf primordia that start out complex but undergo secondary alteration to become sim- ple. (A) Coffee simple leaf. (B) SEM of primordia (arrow) showing early complexity. (C) Expression of KNOX genes in these early leaves. (D) Anise simple leaf. (E). SEM showing early complexity. (F) KNOX expression in these primordia. (G) Amborella showing a simple leaf that is simple throughout development (H) and has no KNOX expression (I). LP, leaf primordia; M, meristem. (From Bharathan et al. 2002; photographs courtesy of N. Sinha.)

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AN OVERVIEW OF PLANT DEVELOPMENT

647

(A)
(A)
(C)
(C)
(B)
(B)
(D)
(D)

FIGURE 20.28 Leaf morphology mutants in peas. (A) Wild-type pea plant. (B) The tl mutant, in which tendrils are converted to leaflets. (C) The af mutant, in which leaflets are converted to tendrils. (D) An af tl double mutant, which results in a “parsley leaf” phe- notype. (Photographs courtesy of S. Singer.)

decrease in the expression of a gene needed for the produc- tion of the plant hormone gibberellic acid (Hay et al. 2002). A second gene, LEAFY, is essential for the transition from vegetative to reproductive development and also appears to play a role in compound leaf development. LEAFY was identified in Arabidopsis and snapdragons (in which it is called FLORICAULA) and has homologues in other angiosperms. The pea homologue, UNIFOLIATA, has

a mutant phenotype in which compound leaves are

reduced to simple leaves (Hofer and Ellis 1998). This find-

ing is also indicative of a regulatory relationship between shoots and compound leaves. In an intriguing evolution- ary twist, UNI appears to have taken on the role of KN1 genes in leaf development in peas (Sinha 2002). It appears

to have been co-opted from its role in flowering in this

highly derived legume. In some compound leaves, developmental decisions about leaf versus tendril formation are also made. Muta- tions of two leaf-shape genes can individually and in sum dramatically alter the morphology of the compound pea leaf. The acacia (tl) mutant converts tendrils to leaflets; afil- ia (af) converts leaflet to tendrils (Marx 1987). The af tl dou- ble mutant has a complex architecture and resembles a parsley leaf (Figure 20.28). At a more microscopic level, the patterning of stomata (openings for gas and water exchange) and trichomes (hairs) across the leaf is also being investigated. In mono- cots, the stomata form in parallel files, while in dicots the distribution appears more random. In both cases, the pat- terns appear to maximize the evenness of stomata distri- bution. Genetic analysis is providing insight into the mech- anisms regulating this distribution. A common gene group appears to be working in both shoots and roots, affecting the distribution pattern of both trichomes and root hairs (Benfey 1999).

The Vegetative-to-Reproductive Transition

Unlike most animal systems, in which the germ line is set

aside during early embryogenesis, the germ line in plants

is established only after the transition from vegetative to

reproductive development (flowering). The vegetative and

reproductive structures of the shoot are all derived from the shoot meristem formed during embryogenesis. Clon-

al analysis indicates that no cells are set aside in the shoot

meristem of the embryo to be used solely in the creation of reproductive structures (McDaniel and Poethig 1988). In maize, irradiating seeds causes changes in the pigmen- tation of some cells. These seeds give rise to plants that have visually distinguishable sectors descended from the mutant cells. Such sectors may extend from the vegetative portion of the plant into the reproductive regions (Figure 20.29), indicating that maize embryos do not have distinct reproductive compartments. Maximal reproductive success in angiosperms depends on the timing of flowering and on balancing the number of seeds produced with the resources allocated to individ-

ual seeds. As in animals, different strategies work best for different organisms in different environments. There is a great diversity of flowering patterns among the over 300,000 angiosperm species, yet there appears to be an underlying evolutionary conservation of flowering genes and common patterns of flowering regulation. A simplistic explanation of the flowering process is that

a signal from the leaves moves to the shoot apex and induces flowering. In some species, this flowering signal

is a response to environmental conditions. The develop-

mental pathways leading to flowering are regulated at numerous control points in different plant organs (roots, cotyledons, leaves, and shoot apices) in various species,

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648 CHAPTER 20

FIGURE 20.29 Clonal analysis can be used to construct a fate map of a shoot
FIGURE 20.29 Clonal analysis can
be used to construct a fate map of a
shoot apical meristem in maize.
Seeds that are heterozygous for cer-
tain pigment genes (anthocyanins)
are irradiated so that the dominant
allele is lost in a few cells (a chance
occurrence). All cells derived from
the somatic mutant will be visually
distinct from the nonmutant cells.
Plants A and B have mutant sectors
that reveal the fate of cells in the
Tassel
(male flowers)
SECTOR A
Length =
4 internodes
Width =
1/8 stem
circumference
shoot meristem of the seed. The
mutant sector in A includes both
vegetative and reproductive (tassel) internodes.
Thus there is no distinct developmental compart-
ment that forms the tassel. The mutant sector in A
is longer and wider than the mutant sector in B.
This indicates that more cells were set aside to
contribute to the lower than to the upper intern-
odes in the shoot meristem in the seed. The actual
number of cells can be calculated by taking the
reciprocal of the fraction of the stem circumfer-
ence the sector occupies. Sector A contributes to
1/8 of the circumference of the stem; thus 8 cells
were fated to contribute to these internodes in the
seed meristem. Sector B is only 1/24 of the stem
circumference; thus 24 cells were fated to con-
tribute to these internodes. In this example, only
cells derived from the L1 are being analyzed. It is
also important to consider the possible contribu-
tions of the L2 and L3 cell layers of the shoot
meristem. (Data from McDaniel and Poethig 1988;
photographs courtesy of C. McDaniel.)
Plant A
Plant B

resulting in a diversity of flowering times and reproduc- tive architectures. The nature of the flowering signal, how- ever, remains unknown. There is now evidence that RNA with developmental functions can move through the phloem, and the role of very small pieces of RNA in signaling developmental mechanisms has recently been recognized.

Juvenility

Some plants, especially woody perennials, go through a juvenile phase during which the plant cannot produce reproductive structures even if all the appropriate environ- mental signals are present (Lawson and Poethig 1995). The transition from the juvenile to the adult stage may require the acquisition of competence by the leaves or meristem to

SECTOR B

Length =

2 internodes

Width =

1/24 stem

circumference

respond to an internal or external signal (McDaniel et al. 1992; Singer et al. 1992; Huala and Sussex 1993). Even in herbaceous plants, there is a phase change from juvenile to adult vegetative growth. For example, the EARLY PHASE CHANGE (EPC) gene in maize is required to maintain the juvenile state (Vega et al. 2002). Mutant epc plants flower early because they have fewer juvenile leaves which are dis- tinguished from adult leaves by the presence of wax and the absence of hairs. These epc mutants, however, have the same number of adult leaves as wild-type plants. Juvenile traits are not expressed in the absence of EPC. The mechanisms of phase change genes are just beginning to be understood. The Arabidopsis juvenility gene HASTY (HST), is necessary for microRNA processing and export from the nucleus. Loss- of-function mutants undergo early phase change because microRNAs are trapped in the nucleus (Park et al. 2005).

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AN OVERVIEW OF PLANT DEVELOPMENT

Floral meristem

identity genes

649

(A)

TFL1 Flowers FT + FD
TFL1
Flowers
FT
+
FD
TFL1 Flowers FT LFY/ + API FD
TFL1
Flowers
FT
LFY/
+ API
FD

Phase change

Floral organ identity genes (ABC genes)

(HST, EPC)

Floral organ identity genes ( ABC genes) ( HST , EPC ) FT transcript CO FT
Floral organ identity genes ( ABC genes) ( HST , EPC ) FT transcript CO FT
FT transcript CO FT Possible florigen?
FT
transcript
CO
FT
Possible florigen?

Light

(B)

FIGURE 20.30 The vegetative-to-reproductive transition. (A) Signals promoting or inhibiting flowering can move from the roots, cotyledons, or leaves to the shoot apex, where meristem competence determines whether or not the plant will respond to the signals. Leaves may also need to develop competence to respond to envi- ronmental signals before they can produce floral promoters. Leaves and meristems that are com- petent to respond to flowering signals have undergone a juvenile-to-adult phase change. (B) Internal and external factors regulate whether a meristem produces vegetative or reproductive structures. Not all of the regulatory mechanisms shown are used in all species, and some species

flower independently of external environmental signals.

Photoperiodic Autonomous Vernalization Giberellin pathway pathway pathway pathway Phase change Floral meristem
Photoperiodic
Autonomous
Vernalization
Giberellin
pathway
pathway
pathway
pathway
Phase change
Floral meristem
Floral organ
genes
identity genes
identity genes

Adult

vegetative

growth

identity genes identity genes Adult vegetative growth Inflorescence growth Juvenile vegetative growth Floral

Inflorescence

growth

genes Adult vegetative growth Inflorescence growth Juvenile vegetative growth Floral signals Flower
Juvenile vegetative growth Floral signals
Juvenile
vegetative
growth
Floral signals

Flower

formation

1999). Leaves, cotyledons, and roots have been identified as sources of floral inhibitors in some species (McDaniel et al. 1992; Reid et al. 1996). A critical balance between inhibitor and promoter is needed for the reproductive tran- sition. In addition, vernalization, a period of chilling, can enhance the competence of shoots and leaves to perceive or produce a flowering signal. The “black box” between environmental signals and the production of a flower is vanishing rapidly, especially in the model plant Arabidopsis (Searle and Coupland 2004; Achard et al. 2006). The signaling pathways from light via different phytochromes to key flowering genes are being elucidated (Figure 20.30A). CONSTANS (CO) responds to day length, promoting flowering under long-day condi- tions. CO activates transcription of a gene called FLOW- ERING LOCUS T (FT). FT is transcribed only in the leaves, but the transcript travels through the phloem (transport tissue) from the leaf to the shoot (see Blázquez 2005). FT is likely translated when it arrives at the shoot meristem. The FT protein interacts with the transcription factor FD, which is expressed only in the shoot tip. Together these two pro- teins are responsible for the expression of regulatory genes that lead to the production of flowers.

Grafting and organ culture experiments, mutant analyses, and molecular analyses give us a framework for describ- ing the reproductive transition in plants. Grafting experi- ments have identified the sources of signals that promote or inhibit flowering and have provided information on the developmental acquisition of meristem competence to respond to these signals (Lang et al. 1977; Singer et al. 1992; Reid et al. 1996). Analyses of mutants and molecular char- acterization of genes are yielding information on the mechanics of these signal-response mechanisms (Levy and Dean 1998; Hempel et al. 2000; Hecht et al. 2005). Leaves produce a graft-transmissible substance that induces flowering. In some species, this signal is produced only under specific photoperiods (day lengths), while other species are day-neutral and will flower under any photoperiod (Zeevaart 1984). Not all leaves may be com- petent to perceive or pass on photoperiodic signals. The phytochrome pigments transduce these signals from the external environment. The structure of phytochrome is modified by red and far-red light, and these changes can initiate a cascade of events leading to the production of either floral promoter or floral inhibitor (Deng and Quail

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650 CHAPTER 20

Is FT the long-sought florigen? Possibly, but we don’t know if other proteins aid in the movement of FT, or if other transcripts must also move through the phloem before flowering can occur. Some of the genes that CONSTANS protein activates in addition to FT are also activated by transcription factors in other, non-light-dependent, flowering pathways. Mol- ecular explanations are revealing redundant pathways that ensure that flowering will occur. In Arabidopsis, four sepa- rable pathways leading to flowering are being elucidated. Light-dependent, vernalization-dependent, gibberellin, and autonomous pathways that regulate the floral transi- tion have been genetically dissected (Figure 20.30B). The availability of the Arabidopsis genome sequence now makes it possible to search on a broader scale for classes of genes that regulate critical events such as time of flowering (Rat- cliffe and Riechmann 2002). Schmid and colleagues (2005) created a developmental gene expression profile of almost all the Arabidopsis genes using microarrays. The current challenge is to integrate the vast amount of expression data with functional analysis. One promising approach is the search for quantitative trait loci (QTL) that control responses to environmental and hormonal factors. In Arabidopsis, two lines, including a wild accession with natural variation in light and hormone responses, were used to identify new genes (Borevitz et al. 2002). Once QTL are mapped, the Arabidopsis genome map makes it more likely that researchers will be able to asso- ciate function with a specific gene.

Inflorescence development

The ancestral angiosperm is believed to have formed a terminal flower directly from the terminal shoot apex (Stebbins 1974). In modern angiosperms, a vari- ety of flowering patterns exist in which the terminal shoot apex is indetermi- nate, but axillary buds produce flowers. This observation introduces an interme- diate step into the reproductive process:

the transition of a vegetative meristem to an inflorescence meristem, which initiates axillary meristems that can pro- duce floral organs, but does not directly produce floral parts itself. The inflores- cence is the reproductive backbone (stem) that displays the flowers (see Fig- ure 20.19).

(A)

The inflorescence meristem probably arises through the action of a gene that suppresses terminal flower formation. The CENTRORADIALUS (CEN) gene in snapdragons sup- presses terminal flower formation by suppressing expres- sion of FLORICAULA (FLO), which specifies floral meris- tem identity (Bradley et al. 1996). Curiously, the expression of FLO is necessary for CEN to be turned on. The Arabidop- sis homolog of CEN (TERMINAL FLOWER 1 or TFL1) is expressed during the vegetative phase of development as well, and has the additional function of delaying the com- mitment to inflorescence development (Bradley et al. 1997). Overexpression of TFL1 in transgenic Arabidopsis extends the time before a terminal flower forms (Ratcliffe et al. 1998). TFL1 must delay the reproductive transition.

Floral meristem identity

The next step in the reproductive process is the specifica- tion of floral meristems—those meristems that will actu- ally produce flowers (Weigel 1995). In Arabidopsis, LEAFY (LFY), APETALA 1 (AP1), and CAULIFLOWER (CAL) are floral meristem identity genes (Figure 20.31). LFY is the homologue of FLO in snapdragons, and its upregulation during development is key to the transition to reproduc- tive development (Blázquez et al. 1997; Blázquez and Weigel 2000). The LEAFY transcription factor can actually move between cell layers in the meristem before activat- ing other flowering genes (Sessions et al. 2000). Analysis of the promoter of LFY reveals that there are separate sites for activation by the photoperiod pathway and the autonomous pathway. The autonomous pathway works through the binding of giberellin to the LFY promoter.

(B)

through the binding of giberellin to the LFY promoter. (B) (D) (C) FIGURE 20.31 Floral meristem

(D)

the binding of giberellin to the LFY promoter. (B) (D) (C) FIGURE 20.31 Floral meristem identity
the binding of giberellin to the LFY promoter. (B) (D) (C) FIGURE 20.31 Floral meristem identity

(C)

the binding of giberellin to the LFY promoter. (B) (D) (C) FIGURE 20.31 Floral meristem identity

FIGURE 20.31 Floral meristem identity mutants. (A) Wild-type Arabidopsis. (B) The leafy mutant. (C) The apetala1 mutant. (D) The leafy apetala1 double mutant. (Pho- tographs courtesy of J. Bowman.)

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AN OVERVIEW OF PLANT DEVELOPMENT 6 5 1 AP1 gene family and most likely arose

AN OVERVIEW OF PLANT DEVELOPMENT

651

AP1 gene family and most likely arose as a result of gene duplication and divergence. The FUL gene is partially redundant to AP1 and CAL (Ferrandiz et al. 2000). CAL, however, arose only within the brassicas through an AP1 gene duplication event, and appears to have accompanied the domestication of cauliflower and broccoli (Purugganan et al. 2000). Floral meristem identity genes initiate a cascade of gene expression that turns on region-specifying (cadastral) genes, which further specify pattern by initiating transcrip- tion of floral organ identity genes (Weigel 1995). SUPER- MAN (SUP) is an example of a cadastral gene in Arabidop- sis that plays a role in specifying boundaries for the expression of organ identity genes. Three classes (A, B, and C) of organ identity genes are necessary to specify the four whorls of floral organs (Figure 20.33; Coen and Meyerowitz 1991). They are homeotic genes (but not Hox genes; rather, most are members of the MADS box gene family that had its origins before the divergence of animals and plants) and include AP2, AGAMOUS (AG), AP3, and PISTILLATA (PI) in Arabidopsis. Class A genes (AP2) alone specify sepal development. Class A genes and class B genes (AP3 and PI) together specify petals. Class B and class C (AG) genes are necessary for stamen formation; class C

Se Se Ca
Se
Se
Ca

FIGURE 20.32 Arabidopsis double mutant of ap1 and cal. Since cal alone gives a wild-type phenotype, the double mutant demonstrates the redundancy of these two genes in the flower- ing pathway. (Photograph courtesy of J. Bowman.)

Expression of floral meristem identity genes is necessary for the transition from an inflorescence meristem to a floral meristem. Strong lfy mutants tend to form leafy shoots in the axils where flowers form in wild-type plants; they are unable to make the transition to floral development. If LFY is overexpressed, flowering occurs early. For example, when aspen was transformed with an LFY gene that was expressed through- out the plant, the time to flowering

was dramatically shortened from years to months (Weigel and Nilsson 1995). AP1 and CAL are closely related and redundant genes. The cal mutant looks like the wild-type plant, but ap1 cal double mutants produce inflores- cences that look like cauliflower heads (Figure 20.32). More recently another gene, FRUITFUL (FUL), with close sequence similarity to AP1 and CAL, has been characterized. The FRUITFUL gene family is closely related to the

FIGURE 20.33 The ABC model for floral organ specification. Three classes of genes—A, B, and C—regulate organ iden- tity in flowers. The central diagram repre- sents the wild-type flower; surrounding diagrams represent mutants that are miss- ing one or more of these gene functions (indicated by the lowercase a, b, or c). Se, sepal; Pe, petal; St, stamen; Ca, carpel; Pe/St, a hybrid petal/stamen; Lf, leaf; Se*, a modified sepal indicating that other genes (possibly ovule genes) also regulate floral organ specification. (Model of Coen and Meyerowitz 1991.)

Ca Ca Se St Pe St Pe Ca Se* b a A c A +
Ca
Ca
Se
St
Pe
St
Pe
Ca
Se*
b
a
A
c
A + B
B + C
C
Wild-type
Ca St Pe Se
abc
ac
ab
Ca
bc Pe/St
Ca
Pe/St
Ca
Pe/St
Ca
Se
Pe/St
Se
Se
Se
bc Pe/St Ca Pe/St Ca Pe/St Ca Se Pe/St Se Se Se Lf Lf Lf Lf
Lf Lf Lf Lf
Lf
Lf
Lf
Lf

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652 CHAPTER 20

(A)

Genes

Flower

structure

(B)

Genes

Flower

structure

(C)

Genes

Flower

structure

(D)

Genes

Flower

structure

Wild-type

 

B B

 

A

A C

C

Sepal

Petal

Stamen

Carpel

ap2 (A )

 

B B

   

C

 

C C

C

Carpel

Petal

Stamen

Stamen

Carpel

ap3 and pi (B )

A

A

C

C

Sepal

Sepal

Carpel

Carpel

ag (C )

 

B B

 

A

A A

A

Sepal

Petal

Petal

Sepal

  A A A A Sepal Petal Petal Sepal FIGURE 20.34 Wild-type and mutant pheno- types
  A A A A Sepal Petal Petal Sepal FIGURE 20.34 Wild-type and mutant pheno- types
  A A A A Sepal Petal Petal Sepal FIGURE 20.34 Wild-type and mutant pheno- types
  A A A A Sepal Petal Petal Sepal FIGURE 20.34 Wild-type and mutant pheno- types

FIGURE 20.34 Wild-type and mutant pheno- types of the Arabidopsis class A (ap2), class B (ap3, pi), and class C (ag) floral organ identity

genes. (Photographs courtesy of J. Bowman.)

organ identity genes. (Photographs courtesy of J. Bowman.) dinated with that of the carpel, one would

dinated with that of the carpel, one would

expect more ancient, independent pathways to exist. Transcription of floral organ identity genes is actually the beginning rather than the end of flower development. One of the amazing attributes of angiosperms is the tremendous diversity of flower phenotypes, many of which attract specific pollinators. Adaxial/abaxial asymmetry in flowers is one contributing factor to diverse floral morphologies that attract pollinators. Phylogenetic evidence indicates that this trait arose independently many times, as well as being lost many times. The cloning of the CYCLOIDEA (CYC) gene in snapdragons has led to extensive discussion of the molecular mechanisms involved in the evolution of asym- metry (Donoghue et al. 1998; Cubas et al. 2001). In snap- dragons, CYC expression is observed on the adaxial side of the floral primordium early in development. In cyc mutants, flowers have a more radial symmetry. Did CYC evolve independently numerous times, or are there many ways to make an asymmetrical flower? Putative CYC orthologues have been identified in other species, and one plausible explanation for the multiple origins of asymme- try is that the CYC gene was recruited for the same func- tion multiple times. The combination of developmental and phylogenetic approaches to the study of patterning in plants promises to provide insight into the origins of the myriad morphological novelties that are found among the angiosperms.

genes alone specify carpel formation. When all of these homeotic genes are not expressed in a developing flower, floral parts become leaflike (Figure 20.34). The ABC genes code for transcription factors that initiate a cascade of events leading to the actual production of floral parts. While the ABC model is compelling, it is not sufficient to support the hypothesis that flowers evolved from leaves. Overexpressing the ABC genes in leaves does not produce petals or other flower parts. About a decade after the ABC model was proposed, a fourth class of floral organ identi- ty genes, SEPALLATA (SEP), was identified (see Jack 2001). These MADS box genes can convert a leaf into a petal when ectopically expressed in the leaf. In the absence of SEP func- tion, flowers become whorls of sepals (Figure 20.35). SEP transcription factors form dimers with ABC or other SEP transcription factors to initiate floral development. In addition to the ABC and SEP genes, class D genes are now being investigated that specifically regulate ovule development. The ovule evolved long before the other angiosperm floral parts, and while its development is coor-

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(A)

Wild-type

 

B B

   

A

A C

 

C

 

SEP1

 

SEP2

 
     

SEP3

 
  SEP2         SEP3   Sepal Petal Stamen Carpel   Wild-type
  SEP2         SEP3   Sepal Petal Stamen Carpel   Wild-type
  SEP2         SEP3   Sepal Petal Stamen Carpel   Wild-type
  SEP2         SEP3   Sepal Petal Stamen Carpel   Wild-type

Sepal

Petal

Stamen

Carpel

 

Wild-type

 
 

B B

   

A

A C

 

C

 

SEP1

 

SEP1

 
     

SEP1

 
     
     
 
     
 
     
 

Sepal

Sepal

Sepal

Sepal

      Sepal Sepal Sepal Sepal Ectopic expression of SEP in Arabidopsis leaf Genes

Ectopic expression of SEP in Arabidopsis leaf

Genes

Flower

structure

(B)

Genes

Flower

structure

(C)

leaf Genes Flower structure (B) Genes Flower structure (C) Normal Arabidopsis leaf FIGURE 20.35 SEPALLATA (

Normal Arabidopsis

leaf

FIGURE 20.35 SEPALLATA (SEP) genes are a fourth class of organ identity genes. Arabidopsis plants with the triple mutation sep1 sep 2 sep 3 produce whorls of sepals and lack the other three floral organs. Ectopic expression of all three SEP genes in leaves causes them to convert to petals.

Snapshot Summary
Snapshot
Summary

Plant Development

1. Plants are characterized by alternation of genera- tions; that is, their life cycle includes both diploid and haploid multicellular generations.

2. Land plants have evolved mechanisms to protect embryos. Angiosperm embryos develop deeply embedded in parent tissue. The parent tissue pro- vides nutrients and some patterning information. This evolutionary theme of an increasingly protected embryo is shared by both plants and animals.

3. A multicellular diploid sporophyte produces hap- loid spores via meiosis. These spores divide mitoti- cally to produce a haploid gametophyte. Mitotic divisions within the gametophyte produce the gametes. The diploid sporophyte results from the fusion of two gametes.

4. In angiosperms, the male gamete, pollen, arrives at the style of the female gametophyte and effects fer- tilization through the pollen tube. Two sperm cells move through the pollen tube: one joins with the ovum to form the zygote, and the other is involved in the formation of the endosperm.

AN OVERVIEW OF PLANT DEVELOPMENT

653

Senescence

Senescence, a developmental program leading to death,

is closely linked to flowering in many angiosperms. In some species, individual flower petals senesce following pollination; orchids, which can stay fresh for long periods of time unless they are pollinated, are a good example.

Fruit ripening (and ultimately overripening) is an exam- ple of organ senescence. Whole-plant senescence leads to

the death of the entire sporophyte generation. Monocarpic plants flower once and then senesce. Polycarpic plants, such as the bristlecone pine, can live thousands of years (4,900 years is the current record) and flower repeatedly.

In polycarpic plants, death is accidental; in monocarpic plants, it appears to be genetically programmed. Flowers and fruits play a key role in senescence, and their removal can sometimes delay the process. In some legumes, senescence can be delayed by removing the developing seed—in other words, the embryo may trigger senescence in the parent plant. During flowering and fruit development, nutrients are reallocated from other parts of the plant to support the development of the next genera- tion. The reproductive structures become a nutrient sink, and this can lead to whole-plant senescence.

5.

Early embryogenesis is characterized by the estab- lishment of the shoot-root axis and by radial pattern- ing yielding three tissue systems. Pattern emerges by regulation of planes of cell division and the direc- tions of cell expansion, since plant cells do not move during development.

6.

As the angiosperm embryo matures, a food reserve is established. Only the rudiments of the basic body plan are established by the time embryogenesis ceas- es and the seed enters dormancy.

7.

Pattern is elaborated during postembryonic devel- opment, when meristems construct the reiterative structures of the plant.

8.

Unlike most animals, the germ line is not set aside early in plant development. Coordination of signal- ing among leaves, roots, and shoot meristems regu- lates the transition to the reproductive state.

9.

Floral meristem identity genes and organ identity genes enable the angiosperm flower to display a tremendous amount of morphological diversity.

10.

Reproduction may be followed by genetically pro- grammed senescence of the parent plant.

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654 CHAPTER 20

For Further Reading

Bao, N., K.-W. Lye and M. K. Barton. 2004. MicroRNA binding sites in Ara- bidopsis class III HD-ZIP mRNAs are required for methylation of the tem- plate chromosome. Dev. Cell 7: 653–662. Blázquez, M. A. 2005. The right time and place for making flowers. Science 309: 1024–1025. Brownlee, C. 2004. From polarity to pat- tern: Early development of in fucoid algae. Ann. Plant Rev. 12: 138–156.

Byrne, M. E. 2005. Networks in leaf development. Curr. Opin. Plant Biol. 8: 59–66. Dharmasiri, N., S. Dharmasiri, and M. Estelle. 2005. The F-box protein TIR1 is an auxin receptor. Nature 435: 441–445. Fleming, A. J. 2005. Formation of pri- mordia and phyllotaxy. Curr. Opin. Plant Biol. 8: 53–58.

Friml, J., X. Yang, M. Michniewicz, D. Weijers, A. Quint, O. Tietz, R. Ben- jamins, et al. 2004. A PINOID-depend- ent binary switch in apical-basal PIN polar targeting directs auxin efflux. Science 306: 862–865. Jack, T. 2001. Relearning our ABCs:

New twists on an old model. Trends Plant Sci. 6: 310–316.

© 2010 Sinauer Associates, Inc. This material cannot be copied, reproduced, manufactured, or disseminated in any form without the express written permission of the publisher.

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