The Plant Cell, Vol. 13, 1477, July 2001, www.plantcell.

org 2001 American Society of Plant Biologists

IN THIS ISSUE

Move It on Out with MATEs

Multidrug transporters form a large
class of membrane proteins present in
the cells of most organisms. These proteins bind to a variety of potentially cytotoxic compounds and remove them
from the cell in an ATP- or protondependent process (Zhelenova et al.,
2000). Traditionally, multidrug transporters have been divided into four superfamilies: the ATP binding cassette
(ABC) superfamily, the major facilitator
superfamily, the small multidrug resistance family, and the resistance-nodulation-cell division family. Brown et al.
(1999) defined a fifth family, called the
multidrug and toxic compound extrusion (MATE) family of transporters. The
MATE family is characterized by the
presence of 12 putative transmembrane
segments and by the absence of signature sequences specific to the other
multidrug transporter superfamilies.
MATE proteins are believed to function
as proton-dependent efflux transporters, based on the genetic characterization of two family members, NorM from
Vibrio parahaemolyticus and its homolog YdeH from Escherichia coli. Expression of these proteins in E. coli
confers resistance to various antibiotics
and antimicrobial agents that is dependent on the maintenance of a proton
gradient across the plasma membrane.
MATE genes are abundant in bacteria
and plantsthe Arabidopsis genome
contains at least 54 MATE family membersbut have not been found in mammals. Aside from NorM and YdeH, very
little functional information is available
on these proteins.
In this issue of The Plant Cell, Diener
et al. (pages 16251637) describe the
functional analysis of the MATE gene
ALF5 in Arabidopsis. The alf5 mutant exhibits greatly inhibited formation of lateral
roots when grown in commercial Bacto
agar (Figure 1). Interestingly, it was found

that the mutant produced roots that were

indistinguishable from wild-type roots
when grown in soil, in a different brand of
agar, or in extensively washed Bacto
agar, suggesting that the alf5 mutation
caused sensitivity to a soluble contaminant present in the Bacto agar. The alf5
locus was cloned and found to contain a
29-bp deletion in a gene, ALF5, that encodes a MATE family integral membrane
protein. Gene expression analysis using
AFL5 fused to the -glucuronidase reporter gene in transgenic plants indicated
that the ALF5 gene is highly expressed in
the root epidermis and cortex. In addition, expression of ALF5 in yeast conferred resistance to the toxic cation
tetramethylammonium, supporting the
conclusion that ALF5 is a functional
MATE efflux transporter.

PLANTS AS GREEN LIVERS

Plant cells, like the cells of most organisms, are capable of removing a large
number of potentially toxic compounds
from the cytoplasm. In plants, these
compounds are either sequestered in
vacuoles or transported to the cell wall.
Toxic compounds may be of xenobiotic
origin or produced endogenously (e.g.,
phenolics, flavonoids, and phytoalexins). The bronze-colored phenotype of
the Bronze2 mutation in maize, for example, is caused by the inability of the
mutant to transport anthocyanin from
the cytosol to the vacuoles. Bronze2 encodes a glutathione transferase (Marrs et
al., 1995), and the mutant is unable to
carry out conjugation of anthocyanin
with glutathione, a necessary step before transport of conjugated glutathione
to the vacuole.
Sandermann (1992) likened plant metabolism of toxic compounds to that of

the mammalian liver because of the

presence and activity of cytochrome
P450 monooxygenases and glutathione
transferases, which resemble the two
major enzyme systems of the liver. Plant
cytochrome P450s and glutathione
transferases are involved in the first two
phases of detoxification of a number of
polychlorinated and polycyclic hydrocarbons and related xenobiotic compounds
as well as endogenous toxins. In phase I,
cytochrome P450s prepare a substrate
for phase II via hydroxylation, and phase
II glutathione transferases carry out the
conjugation of the hydroxylated compound to reduced glutathione. Subsequently, phase III involves the transport
of the glutathione conjugate out of the
cytoplasm to the vacuole or cell wall.
There is good evidence that multidrug
transporters of the ABC superfamily are
involved in the transport of glutathione
conjugates in plant cells (Rea et al.,
1998; Theodoulou, 2000). Sandermann
(1992) described plants as green livers
that might act as a global sink for environmental pollutants of this nature. The
presence of MATE efflux proteins in
plants, which are presumed to carry out
transport of lipophilic cations and related
compounds that are not glutathione conjugates, broadens the scope of this concept and opens up more possibilities for
plant biotechnology.

EVOLUTIONARY PLANT TRICKS

Despite the wide range of chemically

and structurally distinct substrates for
multidrug transporters, transporters of
all five superfamilies show a preference
for hydrophobic (lipophilic) cations, such
as quaternary ammonium antiseptics
(Stermitz et al., 2000). Lipophilic cations, such as berberine alkaloids, are

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because of its efflux from bacterial cells

by multidrug transporters. 5-Methoxyhydnocarpin had no antimicrobial activity
alone, but it strongly potentiated the action of berberine and other toxins against
the growth of Staphylococcus aureus.
This finding suggests that the main
function of some microbial multidrug
transporters is resistance against plantproduced antimicrobial compounds.
Aside from plantpathogen interactions, one habitat in which bacteria are
likely to encounter toxic plant compounds is in root nodules. In Rhizobium
etli, multidrug transporters have been
found to play an important role in nodulation of bean (Phaseolus vulgaris).
RmrA encodes an R. etli multidrug efflux pump gene that is induced by flavonoids released from the roots of P.
vulgaris, and mutations in this gene
were found to reduce nodulation in the
bean by an average of 40% (GonzalezPasayo and Martinez-Romero, 2000).
Another habitat in which bacteria
might commonly encounter plant toxins
is in the stomachs of herbivores. In E.
coli, the transcription repressor MarR
binds various phenolic compounds
such as salicylate and regulates the expression of two multidrug transporters
to produce a more effective efflux
pump system. Sulavik et al. (1995) suggested that drug resistance in E. coli is
thus enhanced when the bacteria reside in an omnivore gut rich in plant antimicrobial compounds.

Figure 1. Roots of Wild-Type (Top) and alf5 Mutant Plants Grown in a Commercial Agar.
As a result of the disruption of the ALF5 gene, which encodes a MATE family efflux transporter,
the roots of the mutant are sensitive to a contaminant in the agar. The figure was provided by
Gerald Fink.

commonly produced by plants. Lewis

(1999) proposed that berberine alkaloids represent a larger group of cationic toxins that fueled the evolution of
microbial multidrug transporters. Interestingly, Stermitz et al. (2000) found

that several plant species in the genus

Berberis, which produce berberine, also
produce an inhibitor of multidrug transporters, identified as 5-methoxyhydnocarpin. Berberine exhibited relatively
weak antimicrobial action, presumably

DIVERSITY OF FUNCTIONS FOR

MULTIDRUG TRANSPORTERS

It is apparent that multidrug transporters constitute large superfamilies in

plants, as in other organisms. The Arabidopsis genome contains at least 60
open reading frames for ABC transporters (Davies and Coleman, 2000). As
with the less well known MATE family,
the function of the majority of these
genes is unknown, but characteriza-

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tion of a few family members suggests

a multiplicity of functions in plant
growth and development within the superfamily, in addition to their role in the
transport of xenobiotic compounds.
Sidler et al. (1998) showed that an Arabidopsis ABC transporter, AtPGP1, is
involved in the regulation of hypocotyl
elongation during photomorphogenesis. Under certain light conditions, plants
overexpressing PGP1 developed longer
hypocotyls, whereas plants with inhibited expression of PGP1 produced
shorter hypocotyls compared with the
wild type. Hypocotyl elongation in the
dark was unaffected by alterations in
PGP1 expression. In wild-type plants, the
AtPGP1 gene was found to be expressed
in the plasma membrane of root and
shoot apices, and the authors proposed
that AtPGP1 is involved in the transport
of a signal molecule, such as a peptide
hormone, from the shoot apical region.
Some mammalian ABC transporters,
such as the cystic fibrosis transmembrane conductance regulator (CFTR)
and the sulfonylurea receptor (SUR),
have been shown to act as ion channels and/or channel regulators. CFTR
functions as an outwardly rectifying Cl 
channel that also regulates other ion
channels, and the SUR acts as an ATPdependent K  channel (Theodoulou,
2000). Some researchers have begun
to look for such activity among ABC
transporters in plants and have found
evidence that ABC proteins may function as ion channel regulators in
guard cells. Gaedeke et al. (2001) and
Leonhardt et al. (1999) have investigated a slow ion channel in Arabidopsis
guard cells that shows CFTR-like characteristics and that may coordinate the
efflux of K and other ions during stomatal closure.
The MATE transporter superfamily
also may cover a diverse range of functions in plant growth and development.
Debeaujon et al. (2001) recently reported
that the TRANSPARENT TESTA12
(TT12) gene encodes another MATE
family member in Arabidopsis. The

function of TT12 appears to be in

controlling the vacuolar sequestration
of flavonoids in the seed coat (testa)
endothelium. Because of their high
chemical reactivity, flavonoids are toxic
endogenous compounds that must be
removed from the cytoplasm after their
synthesis and sequestered in the vacuole or cell wall. There is evidence that
they function as protectants against UV
light damage, oxidative stress, and
pathogen attack. The mutant seeds,
lacking the function of the TT12 MATE
protein, appear to be unable to transport and accumulate flavonoids in the
vacuoles of the seed coat endothelium.
The seeds are pale in color and also
show reduced seed dormancy, supporting the idea that flavonoids play an
important role in seed biology (WinkelShirley, 1998). Thus, it appears that we
can expect a multiplicity of functions in
growth and development for the many
other plant MATE family members. Surprisingly, Diener et al. found a second
open reading frame at the alf5 locus,
LAL5, which lies immediately downstream of ALF5 and encodes a polypeptide with 83% identity to ALF5. It is not
known if LAL5 is expressed, and this
needs to be determined, but the authors reported that the gene appeared
to be intact in the alf5 mutant. If the
gene were expressed, it would appear
to be functionally distinct from ALF5.
The discovery of multidrug sensors
that regulate the expression of some
microbial multidrug transporters suggests that the main function of these
transporters is the efflux of xenobiotic
toxins (Lewis, 1999). At least three multidrug sensors have been identified.
BmrR is a transcription factor in Bacillus subtilis that activates the expression
of the multidrug transporter gene Bmr
in response to binding a number of hydrophobic cations, many of which are
also substrates of the Bmr protein. In S.
aureus, the QacA multidrug transporter
gene is repressed by QacR, and binding of QacR to various cations induces
QacA expression. And in E. coli, re-

pression of the EmrAB transporter

gene is relieved by binding of the EmrR
repressor to various neutral compounds.
It is interesting to speculate that multidrug sensors also will be found to control the expression of plant transporter
genes such as ALF5, whose principal
function appears to be the efflux of
toxic compounds.
Nancy A. Eckardt
News and Reviews Editor
neckardt@aspp.org

REFERENCES

Brown, M.H., Paulsen, I.T., and Skurray,

R.A. (1999). The multidrug efflux protein
NorM is a prototype of a new family of
transporters. Mol. Microbiol. 31, 393395.
Davies, T.G.E., and Coleman, J.O.D. (2000).
The Arabidopsis thaliana ATP-binding cassette proteins: An emerging superfamily.
Plant Cell Environ. 23, 431443.
Debeaujon, I., Peeters, A.J.M., LonKloosterziel, K.M., and Koornneef, M.
(2001). The TRANSPARENT TESTA12
gene of Arabidopsis encodes a multidrug
secondary transporter-like protein required
for flavonoid sequestration in vacuoles of
the seed coat endothelium. Plant Cell 13,
853871.
Diener, A.C., Gaxiola, R.A., and Fink, G.R.
(2001). Arabidopsis ALF5, a multidrug
efflux transporter gene family member,
confers resistance to toxins. Plant Cell 13,
16251637.
Gaedeke, N., Klein, M., Kolukisaoglu, U.,
Forestier, C., Muller, A., Ansorge, M.,
Becker, D., Mamnun, Y., Kuchler, K.,
Schulz, B., Mueller-Roeber, B., and
Martinoia, E. (2001). The Arabidopsis
thaliana ABC transporter AtMRP5 controls
root development and stomata movement.
EMBO J. 20, 18751887.
Gonzalez-Pasayo, R., and MartinezRomero, E. (2000). Multiresistance genes
of Rhizobium etli CFN42. Mol. PlantMicrobe Interact. 13, 572577.
Leonhardt, N., Vavasseur, A., and Forestier,
C. (1999). ATP binding cassette modulators control abscisic acidregulated slow

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anion channels in guard cells. Plant Cell

11, 11411151.

xenobiotics. Trends Biochem. Sci. 17,

8284.

Lewis, K. (1999). In search of natural substrates and inhibitors of MDR pumps. J.

Mol. Microbiol. Biotechnol. 3, 247254.

Sidler, M., Hassa, P., Hasan, S., Ringli, C.,

and Dudler, R. (1998). Involvement of an
ABC transporter in a developmental pathway regulating hypocotyl cell elongation in
the light. Plant Cell 10, 16231636.

Marrs, K.A., Alfenito, M.R., Lloyd, A.M.,

and Walbot, V. (1995). A glutathione S-conjugate transferase involved in vacuolar
transfer encoded by the maize Bronze-2.
Nature 375, 397400.
Rea, P.A., Li, Z.-S., Lu, Y.-P., and
Drozdowicz, Y.M. (1998). From vacuolar
GS-X pumps to multispecific ABC transporters. Annu. Rev. Plant Physiol. Plant
Mol. Biol. 49, 727760.
Sandermann, H. (1992). Plant metabolism of

Stermitz, F.R., Lorenz, P., Tawara, J.N.,

Zenewicz, L.A., and Lewis, K. (2000).
Synergy in a medicinal plant: Antimicrobial
action of berberine potentiated by 5methoxyhydnocarpin, a multidrug pump
inhibitor. Proc. Natl. Acad. Sci. USA 97,
14331437.
Sulavik, M.C., Gambino, L.F., and Miller,
P.F. (1995). The MarR repressor of the

multiple antibiotic resistance (mar) operon

in Escherichia coli: A prototypic member
of a family of bacterial regulatory proteins
involved in sensing phenolic compounds.
Mol. Med. 1, 436446.
Theodoulou, F.L. (2000). Plant ABC transporters. Biochim. Biophys. Acta 1465, 79103.
Winkel-Shirley, B. (1998). Flavonoids in
seeds and grains: Physiological function,
agronomic importance and the genetics of
biosynthesis. Seed Sci. Res. 8, 415422.
Zhelenova, E.E., Markham, P., Edgar, R.,
Bibi, E., Neyfakh, A.A., and Brennan,
R.G. (2000). A structure-based mechanism for drug binding by multidrug transporters. Trends Biochem. Sci. 25, 3943.

APO2001: A Sexy Apomixer in Como

Long before promiscuity was discovered to bear significant risk, many flowering plants had partially abandoned
the pleasures of their sexual life to
evolve one of the most intriguing reproductive alternatives found in nature.
Apomixis is an asexual method of reproduction through seed that circumvents meiosis and fertilization to
culminate in the autonomous development of an embryo. Thus, unlike sexual
reproduction, which yields genetically
diverse progeny, apomixis produces
clonal offspring. The production of
clonal, genetically identical, seed bears
great potential for applications in plant
breeding and seed production (Hanna
and Bashaw, 1987; Savidan, 1992;
Koltunow et al., 1995). Apomixis has
evolved several times independently
from sexual ancestors and can be
viewed as a modification of the sexual
reproductive program. In angiosperms,
sexual reproduction entails complex interactions between a variety of tissues.
Female reproductive development oc-

curs in a specialized organ, the ovule,

where usually a single cell becomes
committed to the reproductive pathway (the megaspore mother cell, or
MMC). After meiosis, a single reduced
product, the functional megaspore, divides mitotically to form the mature female gametophyte or embryo sac. The
usually seven-celled embryo sac contains the egg cell and the binucleate
central cell, both of which get fertilized.
In the male reproductive organs, meiosis produces a tetrad of reduced
spores, all of which divide mitotically to
form the male gametophytes (pollen).
The male gametophyte consists of two
sperm cells, which are contained in a
large vegetative cell that delivers the
sperm cells to the female gametophyte.
During double fertilization, one sperm
fuses with the egg to form the zygote
and the second sperm fuses with the
central cell to form the endosperm.
In apomictic plants, this sexual developmental program is bypassed or
deregulated at various steps (Koltunow,

1993; Grossniklaus, 2001): (1) meiosis

is altered or absent to produce an unreduced female gametophyte with the full
complement of maternal chromosomes
(apomeiosis); (2) fertilization is avoided,
producing an autonomous embryo (parthenogenesis); and (3) endosperm development is initiated autonomously or
sexually; in the latter case, embryo sac
development or fertilization is often modified to adjust to a different genomic
context (Savidan, 2000; Grossniklaus
et al., 2001). In contrast to the modified
female reproductive program, pollen
formation usually is unaffected in
apomicts.
During the last two decades, the introduction of apomixis into sexual crops
has been perceived as one of the most
promising challenges faced by agricultural biotechnology. Apomixis could allow the fixation of any genotype, however
complex, including that of high yielding
F1 hybrids. The enormous potential of
this trait was realized as early as the
1930s by Navashin and Karpenko

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(cited by Asker, 1971). Simplified apomixis breeding programs would allow

an immediate fixation of any genotype
and the production of self-perpetuating
improved hybrids and could promise
social and economic benefits that
would challenge those of the Green
Revolution (Vielle-Calzada et al., 1996a;
Grossniklaus et al., 1998a).
The 2nd International Conference on
Apomixis took place in Como, Italy,
from April 24 to 28, 2001. The scientific
program was organized by Lucia Colombo, Thomas Dresselhaus, Yves
Savidan, and Rod Scott and was sponsored by the European Union, the Food
and Agriculture Organization of the
United Nations, the Institut de Recherche
pour de Dveloppement (IRD), the Italian National Research Center, and
many private sponsors. Following on
the success of the 1st International
Apomixis Meeting held at College Station, Texas, in 1995, the Como conference attracted 170 participants from 27
countries. All meeting abstracts of invited speakers and poster sessions are
available at http://www.apomixis.de.
Leo Beukeboom (University of Leiden, The Netherlands) gave the opening lecture on Origin and Genetics of
Parthenogenesis in Animals. Asexual
reproduction is a widespread phenomenon across the animal and plant kingdoms. There are differences in both the
terminology and the mechanisms of
asexual reproduction in plants and animals. Parthenogenesis (virgin birth),
first observed by Bonnett in 1745, is
the development of an egg without fertilization. Asexual reproduction in animals can occur either by mitotic
parthenogenesis (apomixis) or by meiotic parthenogenesis (automixis). In
contrast to the phenomenon in plants,
apomixis in animals is rarely facultative,
and most forms of automixis occur exclusively in animals. A wide range of
cytological mechanisms underlie mitotic or meiotic parthenogenesis in animals. Beukeboom presented examples
of asexual reproduction in freshwater

flatworms (Polycelis nigra), in which B

chromosomes may be associated with
parthenogenetic lineages. Although
much is known of the molecular mechanisms of fertilization in animals, remarkably little is known about the
mechanisms of parthenogenesis.

MECHANISMS AND EVOLUTION

OF APOMIXIS

Early studies demonstrated that apomixis is controlled genetically. Typically, a single dominant mendelian trait
is associated with apomixis, although
more complex modes of inheritance
have been reported (see accompanying
Insight article). Cytological descriptions
emphasized distinctions between different apomictic mechanisms and have
led to a simplified classification that is
based on the origin and the location of
cells initiating apomictic development
(reviewed in Koltunow, 1993). In
diplospory, the MMC undergoes an aberrant meiosis or divides mitotically,
producing unreduced spores that eventually form the embryo sac containing
an egg cell with the full genetic complement of the mother. In apospory, a cell
in the ovule other than the MMC produces the unreduced embryo sac.
Anna Koltunow (Commonwealth Scientific and Industrial Research Organization, Adelaide, Australia) described the
enormous variability existing in apomictic processes. In Hieracium, apomictically derived embryo sacs usually are
produced through apospory. However,
several loci modify the timing of apomictic initiation, the frequency at which
apomictic embryo sacs are formed,
and the mode of progression of
apomictic development (Koltunow et
al., 1998, 2000; Bicknell et al., 2000).
These findings indicate that the major
locus associated with apomixis might
create a competence for a variety
of reproductive developmental processes in the ovule. Koltunow hypoth-

esized that sporophytic cells in the

ovule play an active role in modulating
and controlling reproductive development. Transformation-induced alterations in ovule development that result
in significant changes of the mode of
apomixis in Hieracium add support to
this hypothesis.
For Yves Savidan (IRD, Montpellier,
France), close examination of developmental processes in the ovule indicates
that apomictic embryo sacs develop
more precociously than their sexually
derived counterparts. However, embryo sac development is always normal, no matter how strongly meiotic
processes are disturbed. This suggests
a crucial role for the timing of developmental events during reproduction and
implies that all types of gametophytic
apomixis may relate to the ectopic expression of the same master regulatory
gene(s). In this view, distinct mechanisms would result solely from differences in the timing of apomictic
induction within the ovule. Daniel
Grimanelli (IRD and Centro Internacional
de Mejoramiento de Maz y Trig
[CIMMYT], Mxico) has cytologically
analyzed the apomictic mechanism of
Tripsacum dactyloides , focusing on
chromosome and chromatin dynamics
and the characteristics of the cytoskeleton during apomeiosis. A wide
phenotypic variability occurs during
apomictic initiation, even within a single
genotype. Many of the developmental
abnormalities characteristic of apomeiotic processes in Tripsacum have striking similarities to defects found in
meiotic mutants of maize. Although
these meiotic mutants have a strong
impact on fertility, similar abnormalities
do not compromise apomictic seed formation. This suggests that developmental events occurring after apomictic
initiation can compensate for, and rescue, meiotic defects.
Whereas in Tripsacum the MMC proceeds directly to divide mitotically, in
dandelions (Taraxacum sp) the MMC
enters prophase of meiosis I without

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subsequent chromosome pairing. This

attempted first division results in a restitution nucleus enclosing a complete
set of chromosomes that undergoes the
second meiotic division (diplospory).
Hans de Jong (Wageningen University, The Netherlands) presented a detailed genetic analysis of apomixis
based on interploidy crosses between
sexual diploids and apomictic triploids
of Taraxacum. The occurrence of recombinants in which the initiation of
apomixis and the autonomous development of embryo and endosperm were
uncoupled indicates that apomixis is
controlled by at least three loci. A dominant trait linked to a microsatellite
marker and located on one of the nucleolar organizer chromosomes appears
to control diplospory. Interestingly,
this marker is absent in sexual diploid
progeny, suggesting that haploid pollen
grains cannot transmit this trait. Emidio
Albertini (University of Perugia, Italy)
confirmed that apospory in Poa pratensis can be uncoupled from the mechanisms controlling the parthenogenetic
development of embryos. Although parthenogenesis is seemingly contingent
on apomeiosis, the reverse is not the
case. The variability of the degree of
parthenogenesis suggests that it is
likely not controlled by a discrete locus
and may be under complex control.
Apomixis is reported in more than
400 species belonging to 40 families.
However, it is particularly prevalent
within the Asteraceae, Poaceae, and
Rosaceae. Focusing on the distribution
and evolutionary history of apomixis,
John Carman (Apomyx, Inc., Logan,
UT) emphasized that only 127 of more
than 14,000 genera of flowering plants
contain apomictic species, most of
which have appeared during or after
the Pleistocene. Carman suggests that
apomicts may have arisen by wide hybridization of ancestral sexual parents
having distinct phenotypic traits related
to reproduction (Carman, 2001). To test
this hybridization-derived floral asynchrony hypothesis, Carmans group

has documented variation of reproductive traits among sexual relatives of

well-known apomicts (Tripsacum and
Antennaria), finding that they are heterozygous and polygenic. Many forms
of reproductive variation, including apomixis, may have arisen after hybridization of sexual ancestors with divergent
reproductive traits. In many agamic
complexes which are composed of individuals of varying ploidy levels, diploid genotypes usually are sexual and
polyploid genotypes usually are apomictic. Tim Sharbel (Max Planck Institute
for Chemical Ecology, Jena, Germany)
studied the evolution of apomixis and
polyploidy in the Arabis holboellii agamic complex. Using chloroplast haplotypes identified in diploid, aneuploid,
and triploid individuals, he concluded
that polyploidy arose repeatedly and
independently within this complex
(Sharbel and Mitchell-Olds, 2001). The
variation in reproductive mode and
population structure suggests that apomixis might have a single evolutionary
origin, followed by multiple instances of
phenotypic expression of this trait.

BREEDING APOMIXIS INTO

SEXUAL CROPS

Among the grasses, apomixis occurs in

several economically important forage
genera (e.g., Pennisetum, Brachiaria,
Paspalum, and Poa). At least three
groups have attempted the introgression of apomixis into sexual crops via
wide hybridization using backcrossing (BC) strategies combined with
embryological or cytogenetic studies.
For close to 20 years, Wayne Hanna
(United States Department of AgricultureAgricultural Research Service,
Tifton, GA) and his colleagues have
attempted the transfer of apomixis
from Pennisetum squamulatum to
pearl millet (P. glaucum) (Hanna et al.,
1998). By screening large populations to identify partially male fertile

apomictic plants, backcrossing has

progressed to the BC 7 generation.
Apomictic tetraploid BC 7 plants have
28 or 29 chromosomes and closely
resemble pearl millet. However, they
form very little viable seed, a problem
possibly related to the dosage sensitivity of endosperm development
(Morgan et al., 1998). Seed sterility
may be overcome by exploiting the
genetic diversity found within the tetraploid germplasm pool.
Following the pioneering work of D.F.
Petrov, who realized the potential of introgressing apomixis into maize by
wide hybridization more than 50 years
ago, Victor Sokolovs group (Institute of
Cytology and Genetics, Novosibirsk,
Russia) attempted to transfer apomixis
from T. dactyloides to maize using tetraploid female parents. F1 hybrids having 20 chromosomes from maize and
36 from Tripsacum were apomictic but
male sterile. Recurrent backcrossing
to male diploid or tetraploid maize
resulted in apomictic genotypes invariably containing the same nine Tripsacum chromosomes. The absence
of any additional Tripsacum chromosomes resulted in the loss of apomixis,
suggesting polygenic control. Moreover,
apomeiosis and parthenogenesis segregate and are controlled by different loci
(Sokolov and Khatypova, 2001). Inspired by the work conducted in Russia, Yves Savidan launched an initiative
to transfer apomixis from Tripsacum to
maize in the early 1990s. Large population screening, flow cytometry, and genomic in situ hybridization were used to
obtain BC3 plants that have 20 chromosomes from maize and 18 from Tripsacum. Because little sexuality was
present in BC3 plants, the acquisition of
subsequent BC populations was difficult (Savidan, 2000). A plant with a chromosome responsible for apomixis has
yet to be found among the BC 4 generation. Olivier Leblanc (IRD-CIMMYT,
Mexico) indicated that the maize genome severely alters the expression of
apomixis in members of these BC pop-

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ulations. Specific attributes necessary

for the proper expression of apomixis in
maize could be related to gene dosage
effects of specific modifiers or to parent-of-origindependent expression (i.e.,
genomic imprinting) of key regulatory
genes that control embryo sac development and/or early seed formation.

ISOLATION OF GENES
CONTROLLING APOMIXIS

Basic knowledge of the genetic and

molecular regulation of female reproductive development in apomictic species remains poor. Although the inheritance of the trait has been studied
in several species, efforts to isolate the
genes that control apomixis are at an
early stage. The development of the
genetic and molecular tools necessary
to establish an apomictic model system
is well under way (Bicknell, 1994,
2001), and the first mutants altered in
apomictic developmental pathways have
been isolated. Ross Bicknell and his
team (Institute for Food and Crop Research, Lincoln, New Zealand) have
implemented -ray and insertional mutagenesis strategies in Hieracium spp.
At least two mutants have been isolated that have lost the ability to form
apomictic seed but still reproduce sexually, indicating that apomixis and sexuality can be uncoupled in Hieracium.
In both mutants, the differentiation of
aposporous initials early during ovule
development is affected. Further characterization promises insights into the
molecular nature of the genes involved
in apomictic initiation.
Peggy Ozias-Akins and co-workers
(University of Georgia, Tifton) have
mapped apomixis to a single locus in
both Pennisetum ciliare and Pennisetum squamulatum (Ozias-Akins et al.,
1998; Roche et al., 1999), two species related to pearl millet. Detailed analysis of
P. squamulatum revealed no recombination between 12 markers and the

apomixis locus. Interestingly, no sequences that cross-hybridize to four of

these markers are present in sexual individuals of a segregating population,
suggesting that the region is hemizygous or highly divergent in the
apomicts. On the basis of marker conservation between the two species, bacterial artificial chromosome clones
linked to apomixis were identified, revealing a series of duplications that
might explain the absence of meiotic recombination in this chromosomal region. Informative bacterial artificial
chromosome clones have been physically mapped using fluorescence in situ
hybridization. Fulvio Pupillis group
(Consiglio Nazionale delle Richerche,
Perugia, Italy) has mapped the apomixis
locus in Paspalum simplex using selected
rice probes as restriction fragment length
polymorphism anchor markers and amplified fragment length polymorphism
(AFLP) markers. They confirmed that
apomixis segregates as a single dominant trait and identified five rice markers
tightly linked to apomixis (Pupilli et al.,
2001). As in Pennisetum, the genomic region linked to apomixis shows no signs
of meiotic recombination. The authors
concluded that the apomixis locus is
contained in a chromosomal area that is
syntenic to a 15-centimorgan region on
rice chromosome 12.
Genes specific to apomictic development may be identified by comparing
gene expression in sexual and apomictic ovules among closely related genotypes of the same species (VielleCalzada et al., 1996b). Sexual and
apomictic genotypes have been characterized in the genus Brachiaria. Julio
Rodrigues from Vera Carneiross group
(Embrapa, Brasilia, Brazil) reported on a
differential display strategy to isolate
differentially expressed transcripts during specific stages of ovary development in Brachiaria brizantha. Several
dozen fragments were specific to either
sexuals or apomicts and are being analyzed further. Similarly, Gianni Barcaccia
(University of Padova, Italy) is investi-

gating gene expression during flowering of mutants of alfalfa that frequently form unreduced 2n egg cells. A
collection of 40 polymorphic cDNAAFLP clones was isolated by differential display.
In a special lecture on Approaches
to Capturing Wild Apomictic Genes
Combining Genetics and Genomics,
Michael Freeling (University of California, Berkeley) presented a strategy to
identify regulatory regions. Nick Kaplinski,
a graduate student with Freeling,
developed a sliding windowtype algorithm that identifies regions of conserved noncoding sequences both
within and between species. Such an
inverse genetics approach can be used
to identify regions that are under functional selective pressure.

OVULE AND FEMALE

GAMETOPHYTE DEVELOPMENT

The initiation of apomictic development

occurs at early stages of ovule development, during the differentiation of
meiotic or apomeiotic products. What
leads to the meiotic or apomeiotic
commitment of cells within the ovule?
Do megaspores sense their position
and communicate with other sporophytic cells? It is becoming apparent
that meiosis and female gametophyte
development are controlled by regulatory genes that may be deregulated in
space and time in apomicts (Koltunow,
1993; Grossniklaus, 2001). In fact,
many events relevant to apomixis, such
as the initiation of embryo sac development, autonomous activation of the
egg cell, and modified fertilization
mechanisms, are functions of the developing female gametophyte. Yet, we
know little about the genetic control of
these events, even in sexual model
species. A better understanding of the
molecular mechanisms underlying embryo sac development and its interactions with sporophytic cells in the ovule

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MEETING REPORT

could enhance our understanding of

apomixis dramatically.
Ueli Grossniklaus and his team (University of Zrich, Switzerland) use Arabidopsis as a model system to identify
genes involved in megasporogenesis
and embryo sac development. Using an
enhancer detection strategy (Sundaresan
et al., 1995), they identified mutants affected in female gametophyte development and double fertilization. Some of
them may be relevant to the developmental alterations that ensure the normal
endosperm formation observed in many
apomicts. For example, a new signaling
pathway between male and female gametophytes was identified by the feronia mutant: a wild-type pollen tube was
unable to release its sperm cells into a
mutant feronia embryo sac, suggesting
an active role of the female gametophyte in this process. Grossniklaus and
colleagues also identified genes expressed in specific cell types of the ovule
and embryo sac, many of which encode regulatory proteins. Promoters controlling expression in the ovule at the
site of apomictic initiation, the MMC, or
the egg could be used to induce elements of apomixis by expressing candidate genes in specific cell types.
In Arabidopsis, female meiosis results
in three cells that die, whereas a single
megaspore produces the embryo sac.
Wei-Cai Yang (Institute of Molecular
Agrobiology, Singapore) used transposon mutagenesis (Sundaresan et al.,
1995) to investigate cell fate determination among the megaspores. He identified a tagged mutant with embryo sacs
consisting of large multinucleated cells.
These cells may be derived from several surviving megaspores, indicating a
defect in cell specification, or may result from aberrant cellularization events
in the embryo sac. The isolation of regulatory genes implicated in cell fate determination may be related directly to the
developmental alternatives associated
with apomictic initiation. In that regard,
the role of transcription factors implicated
in the regulation of ovule development

in sexual species is particularly relevant. The work of Rebecca Favaro and

colleagues in Lucia Colomboss group
(University of Milan, Italy) demonstrated
that the expression of a MADS box
gene (AGL11) during ovule and seed
formation in Arabidopsis is controlled
by an intron that is crucial for determining the temporal and spatial context in
which AGL11 acts.

CELL CYCLE AND MEIOSIS

Critical aspects of cell cycle regulation

and meiosis differ between apomicts
and sexuals. Unlike sexually reproducing plants, apomicts do not undergo recombination during meiosis I, and they
produce unreduced female gametes.
Greater understanding of the (mis)regulation of the cell cycle and meiosis will
facilitate the development of systems to
induce parthogenesis and apomeiosis.
To maintain genetic stability and fertility, polyploid crops (e.g., wheat) must
both pair and segregate the closely
related chromosomes correctly during
meiosis. In polyploids, homologous chromosomes must accurately distinguish
each other from homeologous chromosomes. Peter Shaw (John Innes Centre,
Norwich, UK) described how the Ph1 locus controls the specificity of somatic
and meiotic chromosome association in
wheat. The Ph1 locus restricts chromosome pairing and recombination to true
homologs. Thus, in hexaploid wheat with
Ph1 deletions, pairing and recombination
can occur between homeologs and homologs. In the absence of Ph1, nonhomologously associated centromeres fail
to separate at the beginning of meiosis.
Shaw discussed how the Ph1 locus promotes the specificity rather than the
induction of centromere association
(Martinez-Perez et al., 2001).
Screens for male sterility in Arabidopsis by Hong Mas group (Pennsylvania State University, University Park)
identified genes regulating chromo-

some segregation during male meiosis. Two male sterile mutants (ask1 and
sds) were shown to have abnormal
chromosome segregation during meiosis I. Premature homolog dissociation
occurs in the sds mutant near the end
of prophase I, whereas the homologs
seem to remain attached at anaphase I
in the ask1 mutant. The ASK1 gene is
most similar to yeast SKP1, which is a
subunit of the SKp1-Cullin-1-F-box
(SCF) ubiquitin ligase complex. Although the yeast and human SKP1
genes regulate the mitotic cell cycle, it
was not known until recently that these
proteins also could be required for meiosis (Yang et al., 1999). Molecular
determination of the developmental
mechanisms underlying male and female meiosis is under way by Tom
Gerats team (University of Gent, Belgium), which is undertaking cDNAAFLP transcript profiling in Petunia.
Transcript profiling techniques were
applied to male meiosis, focusing on
the identification of genes involved in
synapsis and recombination. Reproducible cDNA-AFLP patterns were obtained using RNA from only three
developing anthers. In a pilot study,
100 (5%) differentially expressed
transcripts were identified, including
previously characterized meiotic genes
(e.g., DMC1-like protein).
Fifty years ago, Bcher reported unreduced pollen development in the apomict A. holboellii (Bcher, 1951). Alexei
Kravtchenko (Russian Academy of Sciences, Novosibirsk, Russia) conducted
a cytological reexamination of unreduced pollen formation in diploid and
triploid A. holboellii accessions. The
first meiotic division was equational,
and omission of the second division
produced unreduced microspores. For
both triploid and diploid plants, pollen
viability was highly variable. Yet, some
triploids exhibited greater than 90%
pollen fertility, an advantage for apomixis research on A. holboellii.
Although many meiotic mutants have
been described in plants, only six genes

July 2001

1485

MEETING REPORT

involved in meiosis have been cloned.

Christine Horlow (Institut National de la
Recherche Agronomique, Versailles,
France) showed that the Arabidopsis
SWITCH1 (SWI1) protein is required for
both sister chromatid cohesion and
bivalent formation (Mercier et al., 2001).
Horlow has identified a second swi1 allele (swi1-2) that affects both male and
female meiosis, whereas swi1-1 was reported to be female specific (Montamayor
et al., 2000). The cytological behavior
of swi1-2 during metaphase in male
meiocytes is intriguing: instead of the
normal 5 bivalents, 20 chromatids are
observed that segregate aberrantly.
Cell cycle control is a complex process that is mediated largely by protein
kinases, which activate proteins with
specific functions during the cell cycle.
Danny Geelen (University of Gent, Belgium) reviewed current research on cell
cycle progression that is under way in
Dirk Inzs laboratory (Joubes et al.,
2000; Stals et al., 2000; de Veylder et
al., 2001). Transgenic studies in which
the function of key cell cycle regulators
is disrupted demonstrated that the cell
cycle is integrated with plant development. The Inz group has conducted
an AFLP-based screening for cell cycle
genes in tobacco BY-2 cells using an
aphidicoline blocker. Approximately 500
cell cyclemodulated expressed sequence tags (ESTs) have been identified, and 50% of the clones isolated
exhibited no significant homology with
known proteins. Jim Murray (University
of Cambridge, UK) focused on the roles
of D-type cyclins (cycD) in modulating
cell division rates that affect growth
and development (Meijer and Murray,
2000). The CycD genes play an important role in deciding whether a cell enters
a division cycle. Cytokinin activates cell
division through the induction of CycD3
at the G1-S cell cycle transition (RiouKhamlichi et al., 1999). Overexpression of
CycD2 reduces the length of the G1
phase, causing faster cell cycling and accelerated plant development (Cockcroft
et al., 2000). The constitutive overexpres-

sion of the CycD2 cyclin in the shootmeristemless Arabidopsis mutant led to a

restoration of vegetative growth and
long-lived plants.

FERTILIZATION
AND PARTHENOGENESIS

Parthenogenesis is a critical element of

apomixis whereby an (unreduced) egg
cell initiates embryogenesis without fertilization. The molecular mechanisms that
trigger parthenogenesis in apomictic
plants remain largely unknown. Helmut
Bumlein (Institut fr Pflanzengenetik
und Kulturpflanzenforschung, Gatersleben, Germany) discussed embryological
and molecular studies on apomeiosis in
Poa pratensis and parthenogenesis in
wheat that his group conducted in
close cooperation with Fritz Matzk. Genetic and ploidy analysis (Matzk et al.,
2000) of crosses between obligate sexual and apomictic Poa lines demonstrated that apomixis was dominant
and that apomeiosis and parthenogenesis are not linked. A molecular
approach based on SMART subtractive suppression hybridization identified
cDNAs from (apo)meiotic stages specific to obligate sexual and apomictic
lines. The Salmon system of wheat
produces a high fraction of parthenogenetic offspring, and isogenic sexual lines
are available (Matzk, 1996). Bumlein
and co-workers isolated candidate egg
cellspecific genes from an egg cell
cDNA library of these sexual and parthenogenetic lines.
In apomicts, embryogenesis occurs
completely without the contribution
of the paternal genome. Thomas
Dresselhaus (University of Hamburg,
Germany) presented data suggesting
that maize differs from Arabidopsis with
regard to the time of activation of the
paternal genome during early embryogenesis (Vielle-Calzada et al., 2000). In
maize, the switch from maternal to
zygotic control of gene expression

appears to occur 18 to 24 hr after fertilization. Using reproductive cells isolated in vitro, Dresselhaus and coworkers developed a procedure that allows for the isolation of mRNAs present
in individual cell types of the female gametophyte or the zygote. This approach led to the identification of
differentially expressed genes (Cordts
et al., 2001) and is being implemented
in Tripsacum, allowing comparative
studies of gene expression between
sexual and apomictic cells.
Jean-Emmanuel Faure (Ecole Normale
Superieure, Lyon, France) presented a
cytological study of Arabidopsis fertilization using confocal laser scanning microscopy (Christensen et al., 1997).
Characterization of the time course of
fertilization indicates that (1) synergids
degenerate at 7 hr after pollination
(HAP), (2) there is a rapid change in egg
cell polarity, (3) karyogamy occurs 8 to 9
HAP, and (4) the first division of the primary endosperm nucleus occurs 9 to 12
HAP. Faures group identified mutants
that affect fertilization among 4000
-raymutagenized M1 plants. They
confirmed phenotypes for a collection of
200 families containing putative early fertilization mutants. Maura Cardarelli (University La Sapienza, Rome, Italy)
discussed the effects of the Agrobacterium rhizogenes rolB gene on gamete
development and embryogenesis. rolB
expression driven by AtDMC1 caused
sterility that was likely attributable to a
delay in anther dehiscence, whereas
rolB expression under the control of
the FBP7 promoter caused floral defects and a slight delay in anther dehiscence.

EMBRYOGENESIS

Despite the existence of large collections of mutants that affect plant embryogenesis, the molecular basis
underlying the developmental steps
leading to egg cell activation and early

1486

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MEETING REPORT

embryo development remains poorly

understood. Bob Goldberg (The Seed
Institute and University of California,
Los Angeles) discussed a genomics approach toward the understanding of
plant embryogenesis. The giant embryos of scarlet runner bean allowed
the microdissection and isolation of
cDNAs from either the embryo proper
or the suspensor. EST sequencing was
used to identify differentially expressed
genes in the embryo proper (2863
ESTs, 50% unique) and the suspensor
(3138 ESTs, 59% unique). Approximately 10 to 12% of the ESTs exhibited
no homology with genes currently in databases. On the basis of these expression data, the suspensor appears to be
the major source of gibberellic acid in the
developing embryo. The Seed Institute
also uses Affymetrix chips for transcript
profiling in Arabidopsis wild-type and
mutant seed. The profiles show highly
complex changes during the early steps
of seed development.
Sacco de Vries (Wageningen University, The Netherlands) discussed the role
of the Arabidopsis somatic embryogenesis receptorlike kinase (AtSERK1) in
ovule and embryo development. SERK
encodes a leucine-rich repeat transmembrane receptor kinase (Schmidt et
al., 1997) that is similar to animal receptors. The use of fluorescence spectral
imaging microscopy to study physical
interactions in plant cells showed that
AtSERK1 can oligomerize in vivo (Shah
et al., 2001) and interact directly with the
kinase-associated protein phosphatase
(KAPP) phosphatase. Plants overexpressing AtSERK1 have an increased
potential for somatic embryogenesis in
culture. To investigate the potential to
induce embryogenesis in planta, the
de Vries team developed a multiplex
marker system for testing AtSERK1
overexpressing F2 plants. Fixed heterozygosity is indicative of apomixis. Indeed, lines expressing AtSERK1 in the
ovule produced progeny without a recombination event among 14 markers
tested, whereas control crosses did not

yield such progeny among more than

1000 F2 plants. This could be attributable to apomictic embryo initiation or
suppressed recombination in these
plants. Ed Schmidt (Genetwister Technologies, Wageningen, The Netherlands) focused on the family of
transmembrane Receptor Kinaseslike
SERK (RKS) genes. There are 16 RKS
members in Arabidopsis, which can be
grouped into three classes. Genetwister Technologies characterizes the
developmental function of the RKS
family using transgenic plants, which
either overexpress or cosuppress the
different RKS genes.
Kim Boutiliers presentation (Plant
Research International, Wageningen,
The Netherlands) focused on the engineering of adventitious embryony. Using subtractive hybridization, Boutilier
isolated an embryo-expressed gene
called BABY BOOM (BBM) from microspore embryo cultures of Brassica
napus. BBM encodes an AP2 domain
transcription factor. When expressed
constitutively, it induces the spontaneous formation of somatic embryos in
young seedlings. Because the BBM
overexpression phenotype is restricted
to seedlings, Boutilier uses tissue-specific promoters with the aim of inducing somatic embryos in ovules.
Although L1 expression of BBM extends the tissue range and penetrance
of somatic embryo production, it is still
restricted to seedlings. BBM-expressing lines show a cytokinin overproduction phenotype, and mutants such as
amp1 (Chin-Atkins et al., 1996) and
other backgrounds affecting cytokinin
levels enhance the BBM somatic embryo phenotype.
Gabriella Consonni (Universita degli
Studi di Milano, Italy) presented a characterization of the maize mutant fused
leaves (fdl). The fdl mutant causes organ fusion and affects both embryo
organization and seedling growth: epidermal cells of the coleoptile and first
leaf, and the first and second leaves are
joined by a single cell wall.

ENDOSPERM DEVELOPMENT

For agricultural applications, it is essential that endosperm development in

engineered apomicts is normal. Most
apomictic seed formation requires fertilization of the central cell (pseudogamy). This poses a problem for the
transfer of apomixis to sexual crops because maize and likely most cereals require a ratio of maternal to paternal
genomes of 2m:1p for normal endosperm development (Lin, 1984). Fertilization of an unreduced central cell with
a reduced sperm will yield a 4m:1p ratio,
which is expected to cause seed abortion. In contrast, natural apomicts are
either insensitive to unbalanced endosperm or circumvent the problem by altering embryo sac development or the
mechanism of fertilization (Grossniklaus
et al., 2001).
Abed Chaudhury (Commonwealth
Scientific and Industrial Research Organization, Canberra, Australia) provided an overview of ongoing research
on the fis (fertilization-independent seed)
class of Arabidopsis mutants. The FIS
class includes MEDEA (MEA), FIS2,
and FERTILIZATION-INDEPENDENT ENDOSPERM (FIE) (Grossniklaus et al.,
1998b; Luo et al., 1999; Ohad et al.,
1999). Mutations in these genes lead to
maternal effect seed abortion, and mutant central cells are capable of endosperm development in the absence
of fertilization, a component of apomixis (reviewed in Grossniklaus et al.,
2001). Chaudhurys group made promoter::GUS fusions for all three FIS
class genes, showing that the expression of MEA and FIS2 is similar but differs significantly from the FIE pattern
(Luo et al., 2000). Only maternally inherited copies are expressed early in seed
development, suggesting regulation by
genomic imprinting. Seed abortion can
be prevented if a demethylated paternal genome is introduced. This effect is
independent of FIS gene activity (Luo et
al., 2000). Interestingly, a demethylated

July 2001

1487

MEETING REPORT

paternal genome appears to have an

effect on gene expression from the maternal genome.
Robert Fischer (The Seed Institute and
University of California, Berkeley) focused on the suppression of endosperm
development by the FIS genes. FIE and
MEA encode WD40 and suvar3-9, enhancer-of-zeste, Trithorax (SET) domain
proteins of the Polycomb group. FIE
and MEA proteins interact directly, like
their mammalian and Drosophila homologs, which form a multiprotein
complex (Luo et al., 2000; Spillane et
al., 2000; Yadegari et al., 2000). Fischer
reviewed data showing silencing of the
paternal mea locus in the endosperm
(Kinoshita et al., 1999; Vielle-Calzada et
al., 1999). He discussed how these
findings fit with the parental conflict
theory for the evolution of genomic
imprinting (Haig and Westoby, 1991)
and with the observation that many
apomicts require fertilization of the
central cell for endosperm formation.
Problems associated with the engineering of autonomous endosperm in
apomicts also were reviewed by Rod
Scott (University of Bath, UK). Endosperm abortion may be caused by
the nonequivalence of paternal and
maternal genomes at imprinted loci.
Autonomous endosperm development
observed in the Arabidopsis fie mutant
can be enhanced by combining it with
hypomethylation (Vinkenoog et al.,
2000). On the basis of the parental conflict theory (Haig and Westoby, 1991),
Scott proposes that endosperm size in
plants is an assay for gametic gender
as exemplified by interploidy crosses
(Scott et al., 1998). In Arabidopsis,
modifications of gametic gender may
be achieved through changes in the
methylation profile, which either maternalize or paternalize the gametic
genome, or as a result of mutants such
as fie, which paternalize maternal gametes (Adams et al., 2000; Vinkenoog
et al., 2000).
Fred Berger (Institut National de la
Recherche Agronomique, Lyon, France)

discussed the recent work of BoisnardLorig et al. (2001), in which a HISTONE::YFP fusion protein was used to
demonstrate that syncytial endosperm
is divided into three distinct mitotic domains. Enhancer detection screens
(Haseloff, 1999) were used to isolate
GFP-based endosperm markers, such
as KS117, which is expressed initially
in the entire endosperm but then is restricted to the chalazal region. Using this
marker, it was found that in fis class
mutants endosperm polarization is disturbed (Sorensen et al., 2001). The
KS117 line has been used in a -ray
mutagenesis screen for mutants that alter the expression of KS117. Among 4000
M1 plants, 60 lines with disturbed
KS117 GFP expression were identified.
Jos Gutierrez (University of Oxford,
UK) presented the isolation of candidate imprinted genes that are
expressed differentially in maize endosperm depending on parental origin.
Allelic display polymerase chain reaction was used to screen for imprinted
maize genes in reciprocal crosses between different inbred lines of maize.
Loci that exhibit either maternal-specific or paternal-specific expression
were identified at early endosperm
stages (10 days after pollination; 15
candidates) and at later endosperm
stages (30 days after pollination; 31
candidates). The group of Angelo Viotti
(Consiglio Nazionale delle Richerche,
Milan, Italy) investigates epigenetic
phenomena in maize endosperm. Some
alleles of the -zein and -tubulin
genes derived from specific inbred
lines are subject to genomic imprinting
(Lund et al., 1995a, 1995b). Two of the
six -tubulin genes displayed a correlation between DNA demethylation at the
locus and increased RNA accumulation
in the endosperm. -zein and -tubulin
genes are hypomethylated only when
transmitted through the female. Viottis
results confirm that the imprinting status of certain zein and tubulin alleles in
maize endosperm can be influenced by
interactions of parental factors and are

cross and genotype dependent (Ciceri

et al., 2000).
Hilde-Gunn Opsahl-Ferstad (Agricultural University of Norway, Aas) focused on the defective kernel1 (dek1)
and crinkly4 (cr4) genes that regulate
cell identity in the cereal endosperm
(Becraft et al., 2001; Olsen, 2001). The
Olsen group is studying several of the
Pioneer Hi-Bred Trait Utility System for
Maize Mutator-induced mutants to isolate genes involved in the control of
aleurone cell identity. Promoter studies
of the LIPID TRANSFER PROTEIN1
(LTP1) and LTP2 upstream sequences
have shown LTP2 to direct expression in
cereals similar to the patterns described
in dicots, whereas the LTP1 directs a
different expression pattern.

ECONOMIC AND ECOLOGICAL

IMPLICATIONS OF APOMIXIS

The long gestation period for the development of apomixis technology

(Savidan, 2000) has allowed reflection
on both economic (Jefferson, 1994;
Bicknell and Bicknell, 1999) and ecological issues (van Dijk and van
Damme, 2000) regarding the deployment of apomictic crops in agriculture.
Apomictic varieties are being developed for a few (mainly forage grass)
species. Jorge Gonzalez (Universidad
Autnoma Agraria Antonio Narro, Saltillo, Mxico) presented such a breeding program for new disease-tolerant
buffelgrass (Penn. ciliare) cultivars.
Contiguous planting of a single cultivar
of this obligate apomict over large geographic areas in the United States and
Mexico since 1949 led to genetic vulnerability to blight disease. To develop new
disease-resistant cultivars, Gonzalez and
colleagues crossed sexual lines with
the apomictic clone Zaragoza-115.
Progeny testing and multilocation evaluation trials over several years led to
the release of the hybrid AN-17-PS for
commercial seed production last year.

1488

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MEETING REPORT

Given current controversies regarding transgenic crop plants in agriculture, Peter van Dijk (Netherlands
Institute of Ecology, Heteren) suggested that biosafety issues, which
will undoubtedly arise (Ellstrand, 2001),
should be considered well in advance
of future apomixis technology deployment. He looked at the deployment of
apomixis technology from an evolutionary and population genetics viewpoint
(van Dijk and van Damme, 2000). Three
hypothetical problems were identified
with apomictic crops: (1) invasive
weeds, (2) novel weeds, and (3) infectious apomixis, which might reduce genetic diversity by freezing the gene pool.
Overall, van Dijk concluded that if pollen
production is necessary in engineered
apomictic crops, pollen flow of a dominant apomixis transgene should be
prevented. The use of inducible or
conditional systems to control apomixis could be a means to prevent
such gene flow.
In recent years, there has been an increased industrial interest in the development of apomixis technology. Marc
Albertsen (Pioneer Hi-Bred, IA) provided a well-balanced perspective on
the factors that will determine how apomixis is deployed in commercial or subsistence farming. Albertsen raised the
question of who will have access to
proprietary apomixis technology (and
the necessary supporting technologies)
and under what terms. Both intellectual
property management and freedomto-operate issues will have a major
bearing on which beneficiaries will have
access to apomixis technology. From a
strictly commercial perspective, approaches are needed that allow financial returns on technology investment
and to ensure that donated technology is not used competitively against
the original developers and their customers (G. Graaf, A. Bennett, B. Wright,
and D. Zilberman, unpublished data;
see http://www.cnr.berkeley.edu/csrd/
technology/ipcmech/). From a less
commercial perspective, there are hu-

manitarian arguments for the provision

of proprietary technologies to improve
the well-being of poorer clients such as
subsistence farmers in developing
countries. Albertsen concluded that no
single approach may encompass both
requirements and that new strategies
to deal with intellectual property rights
and licensing practices will have to be
developed.
Richard Jefferson (Center for the Application of Molecular Biology to International Agriculture, Canberra, Australia)
continued the theme of how apomixis
technology will be applied in agriculture
by focusing on the challenge to deliver
apomixis technology as a public good.
This issue arose at a meeting funded by
the Rockefeller Foundation in 1998 when
the Bellagio Declaration was issued by
many of the leading apomixis researchers (http://billie.btny.purdue.edu/apomixis).
Jefferson focused on how current intellectual property management limits
the application of enabling technologies
such as apomixis to solely commercial
objectives. He expressed concern that,
unless the research community paid
greater attention to the terms of research agreements and promoted nonexclusive licensing, a small number of
multinational companies could exercise
monopolistic control over apomixis
tech-nology worldwide. The current exclusionary intellectual property management of both the private and public
sector are nonsustainable business
strategies. Many publicly funded bodies are adopting exclusive licensing
models that are more suited to commercial than to public good objectives.
Access to proprietary enabling technologies could be achieved by a consortium approach (enabling technology
cooperative) that would develop proprietary technologies but adopt a broad
nonexclusive licensing policy, allowing
it to act as a clearinghouse for innovative proprietary technologies (G. Graaf,
A. Bennett, B. Wright, and D. Zilberman,
unpublished data; see http://www.cnr.
berkeley.edu/csrd/technology/ipcmech/).

CONCLUSION

Research presented at the 2nd International Apomixis Conference coalesced

around several themes, some of which
are new and some of which have a long
but unfinished history in apomixis research. A wide range of topicsthe
importance of existing apomictic
mechanisms and their evolutionary implications, the isolation of genes controlling apomixis, the molecular and
genetic basis of ovule and female gametophyte development, and the economic and ecological implications of
apomixiswere discussed during plenary talks. It is increasingly apparent
that sexuality and apomixis are interrelated and that they need to be investigated simultaneously to obtain a
complete understanding of plant reproduction at the developmental, ecological, and evolutionary levels. Indeed, a
number of changes were evident in the
content and focus of apomixis research
since the 1st International Apomixis
Meeting. There is now a greater acceptance that genetic and molecular approaches to study sexuality can yield
insights into the regulation and components of apomixis. This was particularly
evident in the number of new groups
entering this field. In addition to the impact of sexual model species such as
Arabidopsis and maize, it is evident
that a number of apomicts (Hieracium,
Taraxacum, Tripsacum, Paspalum, and
A. holboellii) are emerging as models
that are amenable to molecular and genetic analyses. A number of new techniques (e.g., flow cytometry, transcript
profiling, differential screening methods, and enhancer detection) will increase our knowledge of apomixis in
the coming years and bring us a step
closer to the engineering of apomixis in
sexual crops.
Charles Spillane
University of Zrich,
CH-8008 Zrich, Switzerland

July 2001

1489

MEETING REPORT

Jean-Philippe Vielle-Calzada
CINVESTAV-Plant Biotechnology Unit
36500 Irapuato, Mexico
Ueli Grossniklaus
University of Zrich,
CH-8008 Zrich, Switzerland
grossnik@botinst.unizh.ch

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How to Avoid Sex: The Genetic Control of

Gametophytic Apomixis
Apomixis is the natural ability of more
than 400 plant species to reproduce
asexually through seed (Nogler, 1984a).
Sexual embryos result from the union
of male and female gametes, which produces genetically varied offspring. In
contrast, apomictic embryos are formed
without paternal contribution. Therefore,
apomictic offspring carry the full genetic
constitution of the mother and form a
stable clone, a feature of great value for
plant breeding and seed production.
For many years, apomixis was studied only by a small group of interested
botanists (Nogler, 1984a; Asker and
Jerling, 1992) and visionary plant
breeders (Petrov et al., 1979; Hanna
and Bashaw, 1987; Savidan, 1992).
However, because of its tremendous
potential for agriculture, apomixis research has attracted much more atten-

tion during the last few years (Koltunow

et al., 1995; Vielle-Calzada et al., 1996;
Grossniklaus et al., 1998). If apomixis
could be introduced into sexual crops,
it would greatly simplify breeding
schemes and allow the fixation of any
genotype (however complex), including that of F1 hybrids. Apomixis technology could play a major role in
feeding the growing population of our
planet (Jefferson, 1994; Thoenissen,
2001) provided that it will be freely
accessible to all users, especially resource-poor farmers in developing countries, requiring innovative approaches for
technology generation, patenting, and
licensing (http://billie.btny.purdue.edu/
apomixis).
Current apomixis research focuses
on elucidating the genetic basis and
molecular mechanisms that control

apomictic reproduction (see accompanying Meeting Report). Two major

complementary approaches are being
pursued: (1) to identify genes controlling individual elements of apomixis in
well-defined sexual model species (reviewed in Grossniklaus, 2001), and (2)
to unravel the genetic control of apomixis in natural apomicts (reviewed in
Savidan, 2000). For nearly two decades, the genetic control of apomixis
had been elucidated in very few species. Recently, however, inheritance
studies for several natural apomicts
have been published that shed new
light on the genetic control of this important developmental process (van
Dijk et al., 1999; Bicknell et al., 2000;
Matzk et al., 2000; Noyes and Rieseberg,
2000; Pupilli et al., 2001; Quarin et al.,
2001).

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DEVELOPMENTAL ASPECTS
OF APOMIXIS

Apomixis occurs in many species from

more than 40 plant families and is
thought to have evolved multiple times
from sexual ancestors. Therefore, it is
likely that the controls of sexual and
apomictic reproduction are closely interrelated. Sexual reproduction and
apomixis are not mutually exclusive,
and both processes can occur in parallel, as it is typical of facultative
apomicts, which produce a mixture of
apomictic and sexual progeny. Although the mechanisms leading to
apomictic reproduction are diverse
(Koltunow, 1993; Crane, 2001), they
share the common feature that ancestral sexual processes are deregulated
in space and time (Grossniklaus, 2001).
Two distinct types of apomixis have
been described (Gustafsson, 1947a,
1947b): (1) sporophytic apomixis (adventitious embryony), in which an embryo forms directly from an unreduced
sporophytic cell, and (2) gametophytic
apomixis, which involves the formation
of an unreduced embryo sac (female
gametophyte). Although no less interesting, the genetics of sporophytic
apomixis has not been investigated in
great detail, which is why we focus on
gametophytic apomixis in this article.
Sexual reproduction involves the
generation and fusion of reduced gametes (Figure 1). Female gametogenesis and double fertilization occur within
the ovule, a specialized reproductive
organ (reviewed in Drews et al., 1998;
Grossniklaus and Schneitz, 1998; Yang
and Sundaresan, 2000). Usually a single cell within the ovule, the megaspore
mother cell (MMC) becomes committed
to the sexual pathway, undergoes meiosis, and forms a tetrad of four reduced
spores. Only one of these will divide
and ultimately form the mature embryo
sac containing the female gametes.
Double fertilization involves two pairs of
gametic cells: the egg cell fuses with

one sperm to form the embryo and give

rise to the next generation, and the
central cell fuses with a second sperm
to form the endosperm, a nutritive tissue important for seed development
and/or germination.
During gametophytic apomixis, several
of these developmental steps are bypassed or altered (Figure 1; Koltunow,
1993; Vielle-Calzada et al., 1996). (1)
Chromosome reduction is circumvented (apomeiosis) such that unreduced cells initiate embryo sac
development. These unreduced cells
can originate from an aberrant or missing meiosis of the MMC (diplospory).
Alternately, the unreduced embryo sac
forms directly from a cell within the
ovule other than the MMC (apospory)
and the sexual products degenerate.
(2) The unreduced egg cell initiates embryogenesis in the absence of fertilization (parthenogenesis). (3) The central
cell either develops autonomously or, in
the majority of apomicts, requires fertilization to initiate development (pseudogamy). The development of normal
endosperm in apomicts is important for
seed viability and often requires special
adaptations of embryo sac development
or fertilization (reviewed in Grossniklaus
et al., 2001). Recent studies suggest
that some of these developmental
steps are under independent genetic
control, at least in some apomicts (van
Dijk et al., 1999; Matzk et al., 2000;
Noyes and Rieseberg, 2000).

THE INHERITANCE OF APOMIXIS

Genetic studies largely depend on

crosses and recombination events neither of which is easily obtained in
apomicts. However, because most
apomicts produce normal, reduced
pollen, the inheritance of apomixis can
be investigated by analyzing the segregation ratios in crosses with related
sexuals. Such analyses are difficult because gametophytic apomicts are al-

most without exception polyploids,

causing complex modes of inheritance.
An assessment of the breeding system
in the hybrids requires cytological observations or, at least, time-consuming
progeny tests. Variation in the expressivity of apomixis may create an additional complication. Moreover, much of
the earlier work was done on the Rosaceae, which are extremely difficult to
analyze because the multiple MMCs
formed in sexuals make the distinction
between reduced and unreduced embryo sacs difficult. As a consequence,
for many years the genetics of apomixis seemed unclear, complex, and idiosyncratic. However, since the end of
the 1970s, a clear general pattern in the
inheritance of various types of gametophytic apomixis has emerged, first in the
Ranunculaceae and Poaceae and later
in the Compositae.
Using more suitable apomictic species and focusing on one element of
apomixis, apomeiosis, pioneer studies
by Nogler in the buttercup species Ranunculus auricomus and by Savidan in
the grass Panicum maximum indicated
that apospory in these two species
segregated as a single dominant mendelian factor (Savidan, 1982; Nogler,
1984b, and references therein). Subsequent investigations showed that both
apospory and diplospory in other species also fitted this segregation model
(Table 1). Although a dominant mendelian factor can represent any genetic
constitution from a single gene to an
entire chromosome (e.g., mammalian
sex determination), these observations
often were taken as evidence for monogenic inheritance. According to this
model, apomictic plants possess the
simplex genotype Aaaa, carrying in addition to the dominant apomeiosis allele A several recessive alleles for
sexual reproduction. Apomictic plants
thus carry the potential for sexual reproduction, but in a more or less repressed state, because of the presence
of the dominant apomixis factor. Limited penetrance of the apomixis factor

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INSIGHT

Figure 1. Scheme of Sexual and Apomictic Reproduction.

Reduced stages of the life cycle are shown in shaded ovals, and unreduced stages are shown in rectangular boxes. The key developmental
events that are affected or altered in apomictic species are highlighted in hexagons.

explains the occurrence of facultative

apomixis. The presence of recessive
sexual alleles explains how a cross between two facultative apomicts can
generate abundant purely sexual offspring. Although the occurrence of
apomixis fits this model, the degree of
apomixis often is dependent on environmental conditions (Nogler, 1984a)
and/or on modifier genes (Bicknell et al.,
2000). These as yet unspecified factors
need further investigation in the future.
Moreover, it remains to be determined

whether this general model also applies

to the many apomicts in the Rosaceae.

THE APOMEIOSIS LOCUS IS

LOCATED IN A RECOMBINATIONALLY
SUPPRESSED REGION

The segregation model described above

has been supported and refined by the
isolation of molecular markers that are
linked to the presumed apomixis loci in

several species (Table 1). In all cases in

which it has been critically tested to date,
a strong suppression of recombination
around the apomeiosis locus has been
found. For instance, strict cosegregation
with apomeiosis of many more molecular
markers than expected was found in
aposporous Pennisetum squamulatum
(Ozias-Akins et al., 1998) and diplosporous Erigeron annuus (Noyes and
Rieseberg, 2000). In Brachiaria decumbens (Pessino et al., 1998), Tripsacum
dactyloides (Grimanelli et al., 1998a), and

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Paspalum simplex (Pupilli et al., 2001),

comparative mapping with maize or rice
markers showed a lack of recombination
in the region associated with the
apospory locus. Markers that were
spread over a region ranging from 15 to
40 centimorgans in the sexual relatives
strictly cosegregated in these apomicts.
Repression of recombination could frustrate map-based cloning efforts because
closely linked markers may be at great
physical distances from the apomixis loci.
Because suppressed recombination
occurs in both dicot and monocot species, it may be a general characteristic
of apomeiosis loci. This could be related to their function as observed in
other complex loci containing several
genes involved in a common process
(coadapted gene complexes), such as
the heterostyly supergene in Primula
(Ernst, 1936), the self-incompatibility
(S) loci in Brassica (Lewis, 1962;
Awadalla and Charlesworth, 1999),
the mating-type locus in Chlamydomonas (Ferris and Goodenough,
1994), and the major histocompatibility

locus in humans (OhUigin et al., 2000).

Alternately, it could be an evolutionary
by-product of long term asexual reproduction (Judson and Normark, 1996;
Welch and Meselson, 2000). In Pennisetum species, markers that are linked
to apospory in the apomicts could not
be detected by hybridization in sexual
relatives (Ozias-Akins et al., 1998;
Roche et al., 1999), indicating that the
apomicts were either hemizygous for
the apomixis locus (A---) or that the alleles were highly divergent (A a a a),
as was observed for the Brassica S locus
(Boyes et al., 1997; Suzuki et al., 1999).

ONE MASTER APOMIXIS GENE OR

SEVERAL INDEPENDENT
APOMIXIS GENES?

Apomictic development deviates from

the sexual pathway in apomeiosis, parthenogenesis, and often endosperm
development (autonomy, altered embryo sac development, or altered fertili-

zation). Are these elements of apomixis

all controlled by a single gene or by
several genes? In the pioneering studies on R. auricomus and P. maximum,
parthenogenesis was strictly associated with apospory. Hence, apomixis
as a whole was inherited as a single
mendelian trait (Savidan, 1982; Nogler,
1984b). Similarly, in Hieracium piloselloides, all three elements are inherited
as a single genetic trait (Bicknell et al.,
2000). In these species, apomixis could
be regulated by a single master regulatory gene controlling all elements or by
a gene complex of several tightly linked
genes that are recombinationally locked.
In other species, however, crosses between sexuals and apomicts have
yielded progeny combining elements of
both the sexual and the apomictic developmental pathways. In Taraxacum
officinale, hybrids were recovered that
displayed diplospory and autonomous
endosperm development but that lacked
parthenogenesis (van Dijk et al., 1999).
Such apomixis recombinants also
have been reported in Poa pratensis

Table 1. Inheritance of Elements of Gametophytic Apomixis (Apomeiosis and Parthenogenesis) in Members of the Ranunculaceae, Poaceae,
and Compositae

Species
Apomeiosis
Ranunculus auricomus
Panicum maximum
Pennisetum squamulatum
Brachiaria decumbens
Paspalum simplex
Hieracium piloselloides
Hieracium aurantiacum
Tripsacum dactyloides
Erigeron annuus
Taraxacum officinale
Parthenogenesis
Poa pratensis
Erigeron annuus

Apomeiosis
Type
Family

Most Closely
Evidence for
Inferred
Linked Molecular Suppression
Genotype Marker
Recombination Reference

Apospory
Apospory
Apospory
Apospory
Apospory
Apospory
Apospory
Diplospory
Diplospory
Diplospory

Ranunculaceae
Poaceae
Poaceae
Poaceae
Poaceae
Compositae
Compositae
Poaceae
Compositae
Compositae

Aaaa
Aaaa
Aaaa
Aaaa
Aaaa
Aaa
Aaa
Aaaa
Aaa
Aaa

0 cM
1.2 cM
0 cM

0 cM
0 cM
4.4 cM

Yes
?
Yes

Yes
Yes
?

Nogler, 1984b
Savidan, 1982
Ozias-Akins et al.,1998
Pessino et al., 1998
Pupilli et al., 2001
Bicknell et al., 2000
Bicknell et al., 2000
Grimanelli et al., 1998a, 1998b
Noyes and Rieseberg, 2000
van Dijk et al., unpublished

Apospory
Diplospory

Poaceae
Compositae

Pppp
Ppp

6.6 cM
7.3 cM

?
No

Barcaccia et al., 1998

Noyes and Rieseberg, 2000

The genetic models are based on segregation analyses of sexual  apomictic crosses and in most cases supported by cosegregation of closely
linked molecular markers. No distinction is made between disomic (Aa/aa) and tetrasomic (Aaaa) inheritance. The distance between the apomixis
locus and the most closely linked marker is indicated in centimorgans (cM). (), not investigated; (?), no conclusive data.

July 2001

1495

INSIGHT

(Matzk et al., 2000), which suggests

that the elements of apomixis in these
species are regulated by different
genes. In Erigeron annuus, separate
genes for diplospory (A) and parthenogenesis (P) have been mapped genetically (Noyes and Rieseberg, 2000).
Apomicts in this species carry the simplex genotype for both genes (Aaa and
Ppp). In contrast to the diplospory A locus, no suppression of recombination
was observed around the parthenogenesis P locus (Table 1). In light of these
new findings, it is likely that coadapted
gene complexes are present in those
species in which all elements of apomixis cosegregate.

SEGREGATION DISTORTION OF
APOMIXIS LOCI

As mentioned above, gametophytic

apomicts are usually polyploid, whereas
related sexuals are diploid. Is gametophytic apomixis incompatible with diploidy? Again, the pioneering work on R.
auricomus by Nogler appears to have
general relevance. Nogler showed that
diploid offspring that developed parthenogenetically from reduced diploid
egg cells of tetraploid apomicts (dihaploids) or diploids produced through anther culture were able to reproduce
apomictically (Nogler, 1982). This shows
that apomixis and diploidy are not incompatible, a finding that has been
confirmed in several other species
(Bicknell, 1997; Kojima and Nagato,
1997). However, what matters is the origin of the diploid offspring, because
zygotic diploids derived from the fusion
of haploid egg cells and haploid sperm
never reproduced apomictically in Ranunculus. Nogler hypothesized that the
apospory (A) locus was recessive lethal
in the gametes. Consequently, the A locus could be transmitted via diploid gametes to generate polyploid apomicts
but not via haploid gametes to generate diploid apomicts. It is also possible

that mutations closely linked to the A

locus cause haploid gamete nonfunctionality. The net result is that haploid
gametes carrying the A locus do not
contribute to offspring production, resulting in segregation distortion of the
A locus.
More recently, additional evidence
has been obtained for segregation distortion of apomixis loci in other plant
species, such as Tripsacum dactyloides, Pennisetum squamulatum, and
E. annuus (Grimanelli et al., 1998b;
Ozias-Akins et al., 1998; Noyes and
Rieseberg, 2000). Transmission studies
of markers linked to apomixis loci in E.
annuus indicate different causes of
nontransmission: the parthenogenesis
locus P in E. annuus was not transmitted because of selection against haploid gametes, as was observed for the
A locus in R. auricomus. The diplospory
locus A, in contrast, was not transmitted because of meiotic drive. In these
triploid apomicts, the nondiplospory
chromosomes seem to pair preferentially, leaving the diplospory chromosome as a univalent that always ends
up in a diploid pollen grain (Noyes and
Rieseberg, 2000). In Hieracium piloselloides, different crossing schemes indicate that apomixis can be transmitted
via both haploid and diploid gametes,
but post-zygotic lethality rather than
segregation distortion causes the absence of apomixis in diploids (Bicknell
et al., 2000).

CONCLUSIONS

Apomixis is a complex trait involving

the modification of several steps of normal sexual development. Recent reports on the inheritance of apomixis
have revealed several common features. (1) The genetic control of apomixis is dominant; this is true for all
elements of apomixis studied so far.
This observation is often taken as evidence for apomixis being caused by a

mutated gene, but it is also compatible

with the misexpression of wild-type
genes playing key regulatory roles in
sexual development (Koltunow, 1993;
Grossniklaus, 2001). The latter view is
supported by the fact that both sexual
and apomictic reproduction coexist in
facultative apomicts and that apomictic
plants can be obtained from sexual
progenitors through hybridization (reviewed in Carman, 2001) or simple
chromosome doubling (Quarin et al.,
2001). (2) The apomeiosis locus resides
in a recombinationally suppressed region, suggesting that even in species in
which the elements of apomixis are under common control, a complex of several coadapted genes may be present.
(3) The apomixis locus is usually associated with gametic or zygotic lethality
or its transmission is reduced by some
other mechanism, with the result that
haploid gametes do not produce progeny, thereby maintaining or driving the
polyploidization associated with apomictic species.
As many reproductive mutants in
sexual species indicate, disturbances
of sexual development toward elements of apomixis can easily cause
abortion and sterility. Selection has
perfected natural apomicts, refining the
developmental modifications and integrating them into a functional developmental system with high fertility that is
likely controlled by coadapted gene
complexes. Although many of the features discussed in this article pose obstacles to understanding the molecular
basis of natural apomixis, a complementary approach using both sexual
and apomictic model systems bears
great promise that apomixis can eventually be harnessed to contribute to
sustained agricultural production.

ACKNOWLEDGMENTS

We thank Margaret Collinge, Christoph

Ringli, and three anonymous reviewers for

1496

The Plant Cell

INSIGHT

comments on the manuscript, and Peter van

Baarlen and Daniel Grimanelli for helpful
discussions.

Ueli Grossniklaus
Gian A. Nogler
Institute of Plant Biology
University of Zrich
Zollikerstrasse 107
CH-8008 Zrich, Switzerland
grossnik@botinst.unizh.ch
Peter J. van Dijk
Netherlands Institute of Ecology
P.O. Box 40
6666 ZG Heteren, The Netherlands
pjvandijk@cto.nioo.knaw.nl

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How to Avoid Sex: The Genetic Control of Gametophytic Apomixis

Ueli Grossniklaus, Gian A. Nogler and Peter J. van Dijk
PLANT CELL 2001;13;1491-1498
DOI: 10.1105/tpc.13.7.1491
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