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Ann. Rev. Genet. 1987. 21:93-111

Copyright©1987by AnnualReviewsInc. All rights reserved

ARABIDOPS1S THALIANA
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Elliot M. Meyerowitz
Division of Biology, California Institute of Technology,Pasadena, California 91125
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CONTENTS

THE
PLANT
........................................................................................... 93
CLASSICAL ANDBIOCHEMICAL GENETICS .............................................. 94
MOLECULAR
GENETICS ......................................................................... 98
Genomic Size andOrganization ............................................................... 98
ClonedGenes...................................................................................... 100
APROPOSAL........................................................ ~ ............................... 104

Arabidopsis thaliana is a small weed in the mustard family. It has been a

convenient subject for studies in classical genetics for over forty years (35,
76). Recently, investigators have recognized that this flowering plant also has
a genome size and genomic organization that recommend it for certain
experiments in molecular genetics (41, 56, 57). As a result of the ease with
which this plant lends itself to work in both classical and molecular genetics,
Arabidopsis is coming to be widely used as a model organism in plant
molecular genetics, development, physiology, and biochemistry.

THE PLANT
WhileArabidopsisis in the samefamilyas economically importantcropssuch
as cabbage,broccoli, andhorseradish,it is of no economic value. Theexact
relation of Arabidopsisand the other members of Brassicaceaeis unclear,
with different taxonomicclassifications of the mustardsaccordingdifferent
taxonomicpositions (2, 19, 21, 84). Arabidopsisis believedto be native to
the Old World,althoughits exact geographicorigin is unknown. It has been
collected or reported in manydifferent regions and climates, rangingfrom
highelevationsin the tropics to the cold climateof northernScandinavia,
and
93
0066-4197/87/1215
-0093$02.00
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94 MEYEROWITZ

including locations in Europe, Asia, Africa, Australia, and North America

(23, 33, 74, 83).
A mature Arabidopsisthaliana plant consists of a rosette of small leaves,
with a mainstem topped by an inflorescence. Older plants have branches from
the mainstem and secondarystems arising from the rosette. At its tallest, the
plant mayreach 30 or 40 cm, although size depends on nutrition and other
factors. In depleted soil a plant maymature and produceseed whenonly a few
centimeters in height. The inflorescence is a racemecontaining a series of
flowers, with the youngest flowers at the top and successively older flowers
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below. The flowers are typical of membersof the mustard family, consisting
of four sepals surrounding and alternating with four white petals. Inside the
whorl of petals are six stamens, twoshort and four long, each consisting of a
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filament cappedby a pollen-bearing anther. In the floral center is an ovary of

two carpels. Ovules are borne along the ovary wall. Mature flowers are
approximately 3-mmlong, with a diameter near 1 nun. The flowers normally
self-fertilize, althoughthey can be easily cross-fertilized in the laboratory;
after fertilization the ovaryelongatesand developsinto a fruit called a silique.
Each silique maycontain 30-60 seeds at maturity, which is about two weeks
after fertilization at 25°C.The seeds are small, each weighingon average less
than 20/xg, and only a few hundred micrometersin their longest dimension.
Detailed descriptions of plant and floral development (62), and embryo-
genesis (53, 61, 81,103) are available. Embryogenesisin Arabidopsis resem-
bles that in its well-studied relative Capsella (85-89).
Oncemature, a seed maybe stored in dry conditions for years without loss
of germinability. If soaked in water, the seed germinates within a few days,
and in optimal growth conditions forms a rosette, bolts, and flowers within
four weeks. The rate of developmentand time to flowering dependon various
factors, including nutrition (poor nutrition can lead to very rapid flowering
and a very short generation time, but with low seed set); temperature; day
length (Arabidopsis flowers rapidly in long days or continuous light, but in
short days the time to flowering is delayed severalfold); and genetic back-
ground. Undercontinuous light, at 25°C, with good nutrition, the commonly
used ecotypes Columbiaand Landsberghave a generation time of around six
weeks. Since the plants continue to produceflowers for months,it is possible
to collect well over 10,000 seeds from an individual plant. Arabidopsisgrows
well in soil, and can also be grownin sterile nutrient agar (37) or floating
liquid medium(78).

CLASSICAL AND BIOCHEMICAL GENETICS

All of the properties of the plant described above---smallsize (and consequent

ability to growlarge numbersof plants in little space; for example,several can
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ARABIDOPSISTHALIANA 95

be grownto maturity in an area of 1 cm2), short generation time, high seed

set, ease of growth, self- or cross-fertilization at will--makeArabidopsisa
convenientsubject for studies in classical genetics. Theinitial genetic studies
were performed by Laibach, who established the haploid chromosomenum-
ber as 5 in 1907(34; see also 101), collected with various collaborators many
different ecotypes (83), and with his coworkerReinholz performedthe first
mutagenesis experiments (35, 79, 80). Since this work several laboratories
have used various mutagensto isolate visible, biochemical, and physiological
mutants (5, 32, 49, 50, 57, 82, 97).
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Mutagenesis is performed on seeds. Each seed contains only a few cells

that will give rise to the reproductive structures of the matureplant (17, 53,
64). A mutation induced in one of these cells in a seed (the M1seed)
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replicated in the progenyof this cell, and is thus present in heterozygousform

in a sector of the mature plant. Whenthe flowers that arise from this mutant
sector self-fertilize, M2seeds are produced. One-quarter of them are
homozygous for the newly induced mutation, and two-thirds of the remaining
seeds are heterozygous. Any newly induced mutation will thus occur in
homozygous form in numeroussibling seeds; if the mutation gives a recessive
sterile or lethal phenotype, heterozygousseeds can be found in the siliques
from which the homozygoteswere obtained. Mutagenesis of large numbersof
M1seed can be done in small volumesowingto the small size of Arabidopsis
seeds, and the small size of the mature plant allows manythousands of M1
seeds to be grownto maturity in the laboratory. M1plants can be harvested
individually or en masse, and the resulting collection of M2seeds screenedor
selected to obtain new mutations with desired phenotypes.
The type of mutations most easily found are those that give an embryo-
lethal phenotype (54, 63). These mutations becomeevident on the M1plants
as sectors containing siliques with about one-quarter of the seeds aborted. By
scoring siliques of M1plants for recessive embryolethals, the mutagenic
potential of manyseed treatments has been assessed; over eighty chemical
substances as well as X rays and fast neutrons are mutagenic to Arabidopsis
(74). Amongthe most potent mutagens are alkylating agents such as ethyl
methanesulfonic acid ester (EMS).Manyof the existing Arabidopsis muta-
tions have been induced by soaking seeds in EMSsolutions, or by exposing
either dry or imbibed seeds to X rays.
Morphologicalvariants comprisethe largest class of characterized mutants.
Mutationsaffecting virtually all parts of the plant are known(32, 49, 50, 57):
angustifolia (40, 71) and asymmetric leaves (77) give narrow or asymmetric
leaves whenhomozygous;glabra and distorted trichomes mutations (15, 26,
40) removeor changethe shape of leaf hairs; disposition and size of stemsare
altered by the erecta (72, 77) and compacta(25, 32) mutations. Pedicels are
reduced in brevipedicellus (32) homozygotes;petals are reduced (and some-
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96 MEYEROWITZ

times apparently replaced by additional flowers) in apetala-1 (26, 49, 50)

plants; epidermal wax morphologyis altered in homozygotesfor the various
eceriferum mutations (12).
Of particular interest for developmental biology is a set of recessive
mutations that cause homeiotic variations in floral morphogenesis (69).
Amongthemare apetala-2 (25, 69), which causes partial or complete conver-
sion of petals to stamens, and at the same time converts sepals to leaves;
pistillata (32, 69), which causes petals to develop as sepals; and apetala-3
(M. Koornneef, personal communication; E. Meyerowitz, unpublished),
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whichconverts petals to sepals and also converts stamens into carpels. Other
floral mutations, whichare not strictly homeiotic, althoughthey cause meris-
tic variation (1), include agarnousand multipetala (8, 25, 69), which cause
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the stamens and carpels to be absent and replaced by numerousadditional

petals and sepals, and the clavata mutations (32), whichhave an ovary of four
rather than two carpels.
Anothergroup of visible mutations includes color variants. Abnormalplant
color can result from absence of anthocyanins, as in the transparent testa
mutations (24, 32); from abnormalchloroplast differentiation resulting from
one of several nuclear mutations (82); from absence or reduction of chloro-
phyll, as in chlorina-1 homozygotes,which lack chlorophyll b (20); or from
chloroplast rnutator, a nuclear mutation that apparently causes mutagenesisof
the chloroplast genome(75).
Manytypes of recessive lethal mutations have been studied. These include
over forty EMS-inducedembryo lethals, which display a broad range of
phenotypes,with different mutations causing changes in color, size, morphol-
ogy, and developmentalstage of embryonicarrest (47, 51, 52, 54, 63). Other
recessive lethals include nonphotosynthetic mutants (82) and conditional
lethals such as th-1 and th-2, whichrequire thiamine(36, 38, 43), or tz, which
is rescued by either thiamine or the thiamine precursor 4-methyl-5-hy-
droxyethyl thiazole (14, 73). While the exact biochemical lesions in th-1,
th-2, and tz are not known, manyother mutants lacking specific enzyme
activities have been selected. Amongthese are plants lacking alcohol de-
hydrogenase activity due to mutations in the single Arabidopsis alcohol
dehydrogenasegene (13). These were selected by their ability to survive
treatment with allyl alcohol, whichis convertedto a toxic aldehydeby alcohol
dehydrogenaseactivity. Plants resistant to sulfonylurea herbicides have also
beenselected; this dominantresistance results from mutationsin the acetolac-
tare synthase gene (18; C. Somerville, personal communication).Selection
for chlorate resistance has resulted in plants with reduced nitrate reductase
activity (4, 68). An extensive series of experimentsin whichplants that can
only survive in nonphotorespiratory conditions were selected has led to
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ARABIDOPSISTHALIANA 97

recessive nuclear mutations that reduce or eliminate various enzymaticactivi-

ties. Theseenzymeactivities include those of phosphoglycolatephosphatase,
serine-glyoxylate aminotransferase, chloroplast glutamate synthase, and two
mitochondrial enzymes, glycine decarboxylase and serine transhydroxy-
methylase (92-96).
A further class of mutations might be characterized as physiological or
hormonal; they affect the ability of the plant to synthesize or respond to
growth regulators. Amongthese are recessive mutations that cause the ab-
sence of gibberellins. Five different complementationgroups give this pheno-
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type; when mutations in them are homozygousthe plants are dwarfed, and
depending on the allele that is homozygous,mayfail to germinate unless
exposed to exogenousgibberellins (28, 30). Anothermutant strain does not
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produce abscisic acid (22, 28, 29). This deficiency results in reduced seed
dormancy and abnormal water balance. A dominant mutation that confers
resistance to the auxin 2,4-dichlorophenoxyacetic acid at concentrations
2000-fold higher than normal has also been selected (60, 66). This mutation
produces dwarfed plants with agravitropic roots whenheterozygous, and is
homozygouslethal. Additional auxin-resistant and agravitropic recessive
mutations have been reported (46, 59, 60). There are also mutations that fail
to induce flowering in the long-day conditions that normally serve for floral
induction. Plants homozygous for these mutations maytake as long to flower
in long days as wild-type plants take in a short-day regime(32, 71,102). The
growth regulator affected in these mutations is not known.
Nearly one hundredof the visible, conditional lethal, and hormonemuta-
tions have been placed on a genetic map(32; M. Koornneef, unpublished),
which has five linkage groups corresponding to the five chromosome pairs of
the diploid plant. The order and distance of the markershave been established
by analysis of segregation of different mutations in meiosis; the correspon-
dence of linkage groups and chromosomes,and the location of the centro-
meres in three of the five linkage groups, has been determined by com-
plementation analysis using trisomic and telotrisomic stocks (31). The total
mapis currently 437 centimorgans in length, with individual chromosomes
ranging from 51 centimorgans for the second chromosometo 126 centimor-
gans for the first (32; M. Koornneef,unpublished). Multiply markedstrains
are available for use in mappingcrosses (27).
Arabidopsisis thus well characterized genetically: a wealth of useful and
interesting mutations have been induced, analyzed, and mapped. This fact
does not distinguish Arabidopsis from other experimentally useful and genet-
ically well-studied plants such as tomato and maize, however. While the
small size, short generation time, high seed set, and ease of mutagenesisin
Arabidopsismakeit easier and faster to induce, select, and characterize new
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98 MEYEROWlTZ

mutations in this species than in most other angiosperms,the properties that

truly set Arabidopsis apart from other plants are those that relate to ex-
periments in molecular genetics.

MOLECULAR GENETICS
Genomic Size and Organization
Arabidopsis has the smallest knowngenomeamongthe higher plants. Both
DNAreassociation kinetics (41) and quantitative genome blotting ex-
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periments (70) indicate a haploid nuclear genomesize of roughly 70,000

kilobase pairs (kb). This recently measuredgenomesize is consistent with
previous reports that Arabidopsis has the smallest nuclear volume among
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flowering plants (99), since nuclear volumein plants is roughly correlated

with genomesize. Arabidopsisis also far moreresistant to ionizing radiation
than are other angiosperms (98), another property having somecorrelation
with genomesize. Seventy thousand kb is only five times the size of the
Saccharomyces cerevisiae genome(39), and only fifteen times the size of the
Escherichia coli chromosome(11).The contrast of the Arabidopsis genome
with that of other plants frequently used in moleculargenetic workis striking:
tobacco, for example, has a haploid nuclear genomeof 1,600,000 kb; the pea
haploid genomeis 4,500,000 kb; and the wheat haploid genomeis 5,900,000
kb (see 58).
The significance of this small DNA content for moleculargenetics is that a
genomiclibrary of Arabidopsis chromosomalfragments is easy to make, and
simple and economical to screen. Only 16,000 random lambda clones of
20-kb average insert size must be screened to have a 99%chance of obtaining
any Arabidopsis fragment. In contrast, in tobacco 370,000 clones from a
similar library would have to be screened to have the same expectation of
success; the comparable number for pea is 1,000,000, and for wheat
1,400,000, lambdaclones (58). It is thus rapid and inexpensiveto repeatedly
screen Arabidopsis genomic libraries, which is important for the
chromosomal-walkingexperiments discussed below. A second advantage of a
genomeas small as that of Arabidopsis is the enhancementof signal relative
to noise in genomicgel blot experiments. This enhancementallows detection
of weaksignals from heterologous probe hybridizations, and consequently
allows detection of hybridization between very diverged probes and an an-
giosperm genome.
In addition to its remarkablylow content of nuclear DNA,Arabidopsis has
a genomic organization that makes it uniquely suited to certain types of
molecular cloning experiments. Analysis of DNAreassociation results (41)
and of randomly chosen genomic clones (70) indicates that the Arabidopsis
nuclear genome has a very low content of repeated sequences: perhaps
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ARABIDOPSISTHALIANA 99

10-15%of the genomeis highly repeated or foldback DNA,7.5% is com-

prised of tandemrepeats of ribosomal-RNA coding sequences, and as little as
1% may be dispersed repeats. The highly repeated DNAis as yet poorly
characterized, although someconsists of 4,000-6,000 repeats of a 180 base
pair sequence that exists in one or more long tandemarrays (48). This single
repeat class accounts for 1-1.5% of the total nuclear genomeand approx-
imately 10% of the highly repeated or foldback sequences seen in the
reassociation analysis. The ribosomal DNAis 570 copies per haploid genome
of largely or entirely tandemrepeats of a 9.9-10.7 kb unit. Anexact size for
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this unit cannot be given because there is a small variable region in it that
showslength heterogeneity, and therefore causes a range of repeat unit sizes
(70).
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Other than these two types of tandem repeats, the only other DNAse-
quencesthat have beenidentified as belongingin a middleor highly repetitive
sequence class are dispersed repeats that are found surroundedby DNA that is
single-copy by genomeblot hybridization criteria. The frequency with which
these repetitive elements are found in randomly chosen genomic clones is
low. More than 90%of lambda clones (with 13-kb average insert size)
deriving from nuclear DNA,and not containing tandem rDNArepeats, are
entirely single-copy; fewer than 10%contain one or more small repeated
elements surrounded by single-copy sequence (70). If the genomic library
from whichthese clones originated is indeed representative, this/observation
indicates that there is on average one repeat element (or cluster of elements)
every 125 kb. The length of the elements is one to several kb, and each type
so far found exists in approximately 20-50 copies per haploid genome(K.
Mossie & E. Meyerowitz, unpublished). If they are separated by 125 kb of
nonrepetitive DNA,there are around 600 elements per haploid genome;if all
of themare in 40-member families there are about 15 different families. Since
the elements have not been studied in sufficient detail to knowthe exact
numberof elements per family, it is possible that there are either more or
fewer families than indicated by this estimate. It is not knownwhether these
elements are transposable, as are manyof the dispersed repeat elements in
Drosophila (100).
The Arabidopsis genomeis thus nearly devoid of dispersed repeat ele-
ments, and those elements that do exist are usually very far from each other.
In this respect Arabidopsisis quite different from other angiospermsfor which
similar information is available. For example, DNAreassociation ex-
periments showthat the meanlength of uninterrupted single-copy DNA in the
tobacco genomeis 1.4 kb; in the pea genomeit is 0.3 kb (65, 106). The
importanceof this fact is that it is possible to performchromosome-walking
experiments conveniently in Arabidopsis: by successively isolating overlap-
ping genomic clones, DNAfrom large contiguous regions of the Arabidopsis
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100 MEYEROWITZ

genomecan be obtained for study. The interspersed repeats of other an-

giosperms makeit difficult to select single-copy probes for each step of a
chromosome walk, and their large genomes make it time-consuming to
repetitively screen complete genomiclibraries.

Cloned Genes

ManyArabidopsis genes have been cloned and characterized. This cloning

has provided genes for use in manydifferent types of experiments; in addi-
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tion, the aggregatepicture that these genes present allows further conclusions
about the organization and evolution of the Arabidopsis genome.
One of the first Arabidopsis genes to be cloned and sequenced was that
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coding for alcohol dehydrogenase(7). Genetic evidence (13) had shown

Arabidopsis has only one such gene, in contrast to other flowering plants,
whichhave two or moregenes coding this activity. The clone was obtained by
reduced-stringency cross-hybridization with a fragment of the maize Adhl
gene. That it indeed represents the homologousArabidopsis gene has been
confirmed by demonstration of identical induction of the RNAcoded by the
gene and of ADHenzymeactivity in anaerobic conditions, by DNAsequenc-
ing and comparisonof the predicted protein coded by the gene with other Adh
proteins, and by introduction of the cloned gene to the genomeof Adh-
mutant plants, which causes restoration of Adhenzymeactivity (C. Chang
E. Meyerowitz, unpublished; see below). Genomicblot analysis shows the
Arabidopsis Adh gene to be single-copy, as expected from the genetic data.
DNAsequence analysis reveals a gene interrupted by six introns, each in a
position identical to the positions of six of the nine introns present in the only
other plant Adhgenes (maize Adhl and Adh2) for which sequence information
is published. The total length of intronic sequencein the Arabidopsisgene is
567 base pairs (bp); in maizeAdhlit is 1837bp, and in the maizeAdh2geneit
is 1695bp. All three of the sequenced plant Adhgenes code for a protein of
379 amino acids. The smaller size and single copy of the Arabidopsis gene
allow about 3-fold less DNA to be used to code for Adhproteins in Arabidop-
sis than in maize. The greater than 20-fold difference in low-copyDNAin the
respective genomes(16, 41) cannot be accounted for by differences of this
small magnitude.Comparisonof the nucleic acid and aminoacid sequences of
the maize and Arabidopsis genes shows that the two maize genes are more
closely related to each other than either is to the Arabidopsis gene. The
simplest modelfor the evolution of these genes is thus that a single ancestral
gene was present at the time the maizeand Arabidopsislineages diverged, and
that this genesubsequentlyduplicated in the line of descent leading to maize.
This Adh gene work points to two preliminary conclusions about the
Arabidopsisgenome:first, that it is possible to cross-hybridize genesof other
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ARABIDOPSISTHALIANA 101

plants, even monocotssuch as maize, with their Arabidopsis homologues;and

second, that Arabidopsismayhave fewer genes in its gene families than other
plants have in homologousfamilies. Other molecular cloning experiments
support these conclusions. A second class of cloned Arabidopsis gene in-
cludes the chlorophyll aJb light-harvesting protein genes (42). The chloro-
phyll aJb binding proteins are a major componentof the chloroplast thylakoid
membrane,functioning as part of a light-harvesting complexthat includes
chlorophylls a and b, and carotenoid pigments. They have been studied in
manyplants, and are knownto be coded by multiple nuclear genes. Thesecab
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genesare identified by hybridization of cloned probes; by this criterion there

are at least 16 cab genes in Petunia, 8 in pea, 7 in wheat, and 10-12 in the
aquatic monocot Lemna(see 42). Arabidopsis cab genes were cloned by
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cross-hybridization with a Lemnaprobe; using the cloned Arabidopsis genes

as a probe of genomicblots and genomiclibraries reveals three Arabidopsis
cab genes. All three are coded within the same6.5-kb DNA region, and are so
closely related that all of the genes code for identical mature proteins. No
introns are evident in these genes. The principles of cross-hybridization and
reduced copy numberrelative to other plants are thus demonstratedby the cab
genes as well as by the Adh gene.
Another set of genes displaying these properties are the 12S-globulin
seed-storage protein genes. The 12S globulins are one of the predominant
seed-storage proteins of dicots, and are madeof subunits originally cleaved
from a single translation product. Arabidopsis genes coding for abundant
seed-specific RNAswere cloned by using seed RNAas a template for the
production of radioactively labeled cDNA,and then using this cDNAas a
hybridization probe of an Arabidopsis genomic library (R. Pruitt & E.
Meyerowitz, unpublished). Amongthe genes obtained from a library made
from DNA of the Landsbergerecta strain of Arabidopsis were four that coded
for abundant 1.7-kb embryo-specific transcripts.
DNAsequencing, which is completefor two of the genes and partial for the
other two (R. Pruitt & E. Meyerowitz, unpublished; P. Pang & E. Meyero-
witz, unpublished), showsthat all four genescode for variants of 12S storage
proteins. Twoof the genes exist as a tandemrepeat, with the 5’ end of one
separated from the 31 end of the other by approximately 2 kb. These two
genes are closely related to each other. The other two genes are so distantly
related to the tandempair and to each other that they do not cross-hybridize,
even in conditions of greatly reduced stringency. The four genes thus
represent three different gene families (as defined by hybridization), one with
two membersand the other two with only one membereach. All three gene
families code for membersof the same protein family (defined by sequence
comparison).The structure of the genescoding for this protein family has also
been analyzed in the Columbiastrain of Arabidopsis thaliana (R. Pruitt &E.
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102 MEYEROWITZ

Meyerowitz,unpublished), and found to be even simpler: in this strain, each

of the three 12S-protein gene families consists of a single gene.
In other dicots a similar situation has been found. For example, the
12S-globulin (legumin) genes of the bean Vicia faba are found in two
different gene families (defined by hybridization and cDNA clone sequence
criteria), the legA family and the legB family. There are multiple genes in
each family (105). In no other plants have 12S-globulin gene families been
found that are as simple as those in Arabidopsis. The sequencedArabidopsis
12S genes showthree introns at positions identical to those at whichthe three
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introns of a Pisumlegumin gene are found (45). The Arabidopsis gene of one
family cross-hybridizes to a similar gene from the mustard Brassica napus;
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this Brassica gene cross-hybridizes to several related genes in the Brassica

napus genome(M. Crouch, personal communication). Thus, the principle
cross-hybridization of homologousgenes between different genera is pre-
served, though it appears from the great divergence in the DNA sequences of
the legumeand Arabidopsis 12S genes that cross-hybridization between 12S
genes from the two families wouldnot occur. The evolutionary relationship of
the sequenced 12S genes in mustards and legumes resembles that of the Adh
genes in Arabidopsis and maize: the genes of the legumes are more closely
related to each other than to any of the mustard genes, and the Arabidopsis
and Brassica genes are all moreclosely related to each other than any of them
is to the bean and pea genes. It thus appears that a single ancestral gene was
inherited by each lineage, with gene duplication occurring after divergenceof
the lines leading to the legumesand the mustards (P. Pang &E. Meyerowitz,
unpublished).
Manyother Arabidopsis genes have been cloned by cross-hybridization
with genes from other plants. Nitrate reductase clones have been obtained
using the squash gene as a probe (10). The single RNApolymerase II gene
has been cloned by use of a soybean probe (H. Klee, personal communica-
tion). An ADPglucose pyrophosphorylase gene has been cloned with a rice
probe (J. Preiss &C. Somerville, personal communication).The glycollate
oxidase gene has been obtained using a spinach gene (C. Somerville, personal
communication). Three different glyceraldehyde-3-phosphate dehydrogenase
genes have been cloned with tobacco probes (90; H. Goodman,personal
communication). The phytochrome gene has been cloned using a squash
probe (P. Quail, personal communication). Finally, one gene coding for
5-enolpyruvylshikimate-3-phosphate synthase (EPSPS)has been cloned using
a Petunia sequence as a hybridization probe (H. Klee, personal communica-
tion). Genomeblots indicate that there is a second EPSPSgene in Arabi-
dopsis; it has not as yet been cloned. The cloned EPSPSgene has been
sequenced.It has six introns in positions identical to those of introns in the
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ARABIDOPSISTHALIANA 103

Petunia gene, although each of the Arabidopsis introns is smaller than its
Petunia counterpart.
Other Arabidopsis genes have been isolated using cross-hybridization
across even broader evolutionary distances. A Chlamydomonas alpha-tubulin
gene has been used in isolating clones of two (of at least four) Arabidopsis
alpha-tubulin genes. Thesegeneseach havefour introns in identical positions.
Thepositions differ from those in whicheither of the intervening sequencesis
found in the Chlamydomonas gene, despite high (90%) conservation of the
proteins coded by the higher plant and algal genes. One of the Arabidopsis
genes is expressedin leaves, roots, and flowers, while the other is expressed
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pdmarly in flowers (D. P. Snustad, personal communication).The wild-type

acetolactate synthase gene has been cloned using a Saccharomycesgene as a
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probe (B. Mazur, personal communication; H. Klee, personal communica-

tion). A mutantacetolactate-synthase gene that causes resistance to sulfonyl-
urea herbicides due to a single base changefrom wild type has been cloned as
well (C. Somerville, personal communication).Even animal genes have been
used as probes for cloning Arabidopsis genes; an exampleis the cloning of
three heat-shock protein (hsp70) homologuesobtained from an Arabidopsis
genomic library by use of a Drosophila probe. One of the three genes is
constitutively expressed. Anotheris also expressedat all times, but is induced
about fourfold after heat shock (C. Somerville, personal communication).
One example of an Arabidopsis gene family that is not smaller than its
counterpart in other flowering plants is the beta-tubulin family. There are at
least 7, and perhaps as manyas 11, individual gene or pseudogenemembers
of this family in Arabidopsis; genomicblots reveal someother species to have
more, but others to have fewer, of these genes (D. Weeks,personal com-
munication; D. P. Snustad, personal communication). Another Arabidopsis
gene family comparable in size to the homologous family in other an-
giosperms is that coding for the small subunit of ribulose bisphosphate
carboxylase. There are four genes that code for this subunit in the Columbia
strain of Arabidopsis,with three of themclosely related and tightly linked (M.
Timko,personal communication).The numberof small subunit genes in other
plants ranges from as few as two to as manyas twelve per haploid genome.As
moreArabidopsis clones are characterized, additional exceptions will likely
be found; nonetheless it seemsto be true that manyArabidopsisgenes exist in
smaller gene families, and as smaller genes, than their homologuesin other
higher plants. Theseproperties are useful in the study of multigenefamilies,
and in the DNAsequence analysis of individual genes. The wide cross-
hybridization of plant genes implies that a gene conveniently cloned from the
small Arabidopsis genomemight be used as a probe for the isolation of
homologoussequences from plants of economic importance.
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104 MEYEROWITZ

A PROPOSAL

All of the properties of Arabidopsisdiscussed so far showit to be a remark-

ably convenient organismfor experimentsin classical genetics and molecular
cloning, but the work described could have been done with other higher
plants. There is one type of experiment, though, for which Arabidopsis is
uniquely suited. Due to its small genomesize and low level of interspersed
repetitive DNA,there is a potential methodfor molecularcloning of any gene
for which only a mutant phenotype and a genetic mapposition are known.
Suchcloning requires three steps: first, finding a molecular probe that maps
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org

near the desired gene; second, isolating overlapping probes that cover the
contiguous region betweenthe starting probe and the desired genetic region;
by CORNELL UNIVERSITY on 07/14/06. For personal use only.

and third, finding which clone from the contiguous region contains the gene
of interest.
Thefirst step, the acquisition of a molecularprobenear a desired locus, can
be done by producing a restriction fragment length polymorphism(RFLP)
map(3). Manydifferent ecotypes of Arabidopsis are available, including the
Landsberg and Columbia strains mentioned above. Randomlychosen geno-
mic lambdaclones that represent single-copy sequences (as do most randomly
chosenlambdaclones; 70) frequently hybridize to different patterns of restric-
tion fragments in the isolated genomicDNAof different ecotypes (58; R.
Pruitt & E. Meyerowitz, unpublished; C. Chang & E. M. Meyerowitz,
unpublished). In fact, about 50%of randomlambda clones reveal one or more
polymorphismsbetween the strains Landsberg, Columbia, and Niederzenz,
all rapid-flowering ecotypes that originated from Europeanpopulations, when
three different restriction enzymes(EcoRI, BgllI, or XbaI) are used for
digestion of their genomicDNA.Most of the visible and biochemical muta-
tions knownin Arabidopsis were induced in either the Columbiaor Landsberg
genetic background.
Standard genetic mapping crosses between a Columbia strain multiply
markedwith visible mutations and a Niederzenz plant, followed by segrega-
tion analysis of the visible markers and the polymorphicchromosomal regions
hybridized by the lambda clones, have resulted in a genetic map. This map
includes both the visible markers (whichare already on the standard genetic
map) and, as new genetic loci, the locations of the RFLPsidentified by the
lambda clones. This genetic mapcurrently has over 40 RFLPsmapped,and is
rapidly becoming more dense (C. Chang &E. M. Meyerowitz, unpublished).
Since the entire Arabidopsis genomeis 70,000 kb, as few as 100 markers
wouldgive a mapwith lambdaclones spaced an average of only 700 kb apart.
The current genetic mapof Arabidopsis is 437 centimorgans; thus, one map
unit is on average around 160 kb. Any mutation mapping one or two map
units from an RFLPmarker could be cloned relatively easily, by successive
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ARABIDOPSISTHALIANA 105

isolation of overlapping clones; and with a mapof not manymore than i00
lambdamarkers, a polymorphism probe is likely to be within this distance Of
any mutation.
Arabidopsis is uniquely suited to cloning by this method. The genomesize
and the relation of physical to genetic distance makeit is feasible to produce
an RFLPmap with a density of markers sufficient to give probes within
chromosome-walkingdistance of any genomic region. This is not true of
plants with larger genomes,Also, Arabidopsis is singularly useful for the
chromosome-walking process itself: the near absence of dispersed repeats
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org

makesit simple to isolate single-copy probes from one clone, to use in the
isolation of overlapping clones.
Once a walk has gone far enough to include the gene whose molecular
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clones are of interest, howdoes one knowto stop? Onepossibility is to use the
cloned DNAto complement mutations in the gene, using DNA-mediated
transformation. Foreign DNAcan be introduced to the Arabidopsis genome,
and integrated into nonhomologouslocations in the Arabidopsis chromo-
somes (44). This is done by the methods that have become standard for
dicotyledonous plants (6), involving recombinantconstructions based on the
Ti-plasmidofAgrobacteriurntumefaciens, and using the ability of the bacteri-
umto cause the insertion of the T-DNA region of the plasmid into chromo-
somesof infected plant cells. The Arabidopsis cells that have been used are
those that grow from woundedleaf surfaces; once foreign DNAcontaining a
selectable markersuch as hygromycinresistance is introduced to these cells,
the transformed cells grow into callus. After selection for drug resistance,
callus is regenerated into plants, which, uponself-fertilization, give hygromy-
cin-resistant progenyin Mendelianratios (44).
Recent experiments have shownthat a cloned copy of an Arabidopsis gene
can be introduced into the genomeof a plant mutant for the endogenouscopy
of the gene, and the mutant phenotype be complemented(C. Chang & E.
Meyerowitz,unpublished). In these experiments, Adh-leaf cells were trans-
formed, and one or more copies of the cloned wild-type alcohol de-
hydrogenase gene integrated into their chromosomes.A bacterial hygromy-
cin-resistance gene fused to a constitutive plant promoterwas adjacent to the
Adhgene in the Ti-plasmid variant used in the transformation. Transformed
leaf-derived callus ceils were selected for hygromycinresistance, then re-
generatedto hygromycin-resistantplants. After self-fertilization these plants
produced progeny seeds in which alcohol dehydrogenase activity and hy-
gromycinresistance cosegregated. The inheritance of the introduced genes
indicates whether one copy, or multiple unlinked copies of the introduced
genes are present in the chromosomesof any individual transformed plant.
The progeny of the transformed plants have normal amounts of alcohol
dehydrogenaseactivity in those cells wherethe activity is foundin wild-type
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106 MEYEROWlTZ

plants; the introduced Adhgenes also restore the wild-type phenotypeof allyl
alcohol sensitivity.
These experiments imply that any Arabidopsis DNAfragment could be
tested for its ability to complementrecessive mutations, allowing the genesin
the DNAmolecule to be identified. If each step in a chromosomewalk is
tested by transformation, the end of the walk will be signaled by com-
plementationof a mutationin the genethat is the goal of the walk. All of the
steps in the proposed general cloning method have thus been tested suc-
cessfully; what remains is to apply all of them to the molecular cloning of
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org

individual genes.
An additional general method for molecular cloning of any Arabidopsis
gene is also madefeasible by the tiny genomeof the plant. This methodis the
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ordering of a library of genomicclones, by finding overlaps betweenrandom-

ly chosen clones. Whenoverlaps between enough clones are established,
clones of an entire genomeof any genetic region available will be ordered.
This method has been developed and used effectively in establishment of
partial collections of ordered genomic fragments of DNAfrom the yeast
Saccharomycescerevisiae (67) and the nematodeCaenorhabditis elegans (9).
The Caenorhabditis genomeis approximately the same size as the Arabidop-
sis genome,and the chromosomenumberof the two organisms is similar. The
methodsdeveloped for the nematodeshould be directly applicable to Arabi-
dopsis. Evenwith a fully ordered set of clones representing the Arabidopsis
genome, RFLPmap data would be necessary to knowthe relation between
sets of ordered clones and the genetic map that includes the visible or
biochemical markers whose DNAis desired.
A final possibility for cloning any gene of Arabidopsis, knowingno more
about it than its phenotype, is shotgun transformation. If transformation of
Arabidopsis can be performed at a high enough frequency, and enough
transformed plants are generated, direct selection or screening for cloned
DNAfragments that will complement any recessive mutation should be
possible (91). A cosmidlibrary with average 30-kb inserts could contain the
entire Arabidopsis genomeonce in fewer than 250~0 clones; if 10,000 in-
dividual transformed plants were generated from random cosmid clones, it
wouldbe highly likely that any individual gene wouldbe found. For this to be
true, though, the library would have to be random, with no differential
efficiency of conjugation from E. coli into Agrobacterium,and no differential
transfer of different T-DNAregions into the plant. Also, the shotgun
transformation process wouldhave to be repeated each time a new gene was
cloned. Considerable improvementover the current labor-intensive leaf disc
transformation methodand a better understanding of the dynamics of pop-
ulations of cosmidsin Agrobacteriurn will be required before a direct com-
plementation methodinvolving regenerated plants is practicable. Recent re-
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ARABIDOPSISTHALIANA 107

ports of a method for production of transformed Arabidopsis plants by

treatment of seeds with Agrobacterium-bearingrecombinant Ti-plasmids in-
dicates the possibility that transformants maybe producedrapidly and without
regeneration (K. Feldmann& D. Marks, submitted for publication). Perhaps
such methodswill eventually allow shotgun transformation procedures to be
used.
It thus seemslikely that it will soon be possible to clone any Arabidopsis
gene for which a mutant phenotype is known.Amongthe genes that might be
cloned are those whose mutations affect developmental and physiological
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org

processes whose mechanismsare unknown.Molecular clones of such genes

will lead to information on the mechanismsof these processes in several
ways. First, use of current methods for in situ hybridization to RNAin
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Arabidopsis tissues (55) will allow the developmentalstage and cell type
gene expression to be determined. For developmental mutations such as the
floral homeoticmutants, this sort of information wouldimmediatelylead to
hypotheses for the action of the wild-type gene in pattern formation. For
instance, if a gene whosemutations cause conversion of petals to stamens
(apetala-2) is expressed in the cells that will differentiate abnormally, it
wouldbe reasonable to hypothesizethat the function of the wild-type gene is
in the reception of positional informationby presumptivepetal cells, or in the
biochemical pathwayleading to petal differentiation. If the gene is only
expressed in presumptive sepal cells, one could imagine that signalling of
positional cues betweenneighboring cells is mediatedby the wild-type gene.
If the gene is expressed in roots, one wouldbe led to hormonalmodels of
positional specification in the shoot apex. Use of the cloned gene to over-
express the protein in a prokaryote, and use of this protein as an antigen,
could provide antibodies that might allow subcellular localization of the gene
product, and thus lead to hypothesesfor its function within the cell. It is even
possible that DNA sequencingand conceptual translation of the coded protein
wouldindicate likely protein activities.
Newinformation on physiological processes may also be obtained by
molecular cloning. For example, clones of genes whose mutations prevent
synthesis of abscisins, or prevent normal response to them(29), might allow
the biochemicalpath of abscisin synthesis and responseto be clarified. Clones
of genes necessary for normal photoinduction of flowering (32, 71,102) may
indicate where, when, and in response to what stimuli different products
necessary for floral induction are made.
The process of cloning any gene whose mutant phenotype is known, a
process for whichArabidopsisis uniquely suited, will open newareas of plant
physiology and plant developmental biology for biochemical and molecular
analysis. Arabidopsis thaliana thus provides a valuable and unique system in
which classical and molecular genetics can be used as a tool for studying
 Annual Reviews
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108 MEYEROWITZ

biochemical, physiological, and developmental processes. Arabidopsis is

indeed becomingthe "botanical Drosophila" that it was first called over forty
years ago (10z~).

ACKNOWLEDGMENTS

I wouldlike to thank mycolleagues workingwith Arabidopsisfor allowing

meto cite their unpublished
results; andfor.their adviceandsuggestions.I
would’alsolike to thankthe members of mylaboratory:JohnBowman, Caren
Chang,Dr. KevinMossie,Dr. PattyPang,andDr. MartyYanofsky for their
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org

commentson, andcriticisms of, the manuscript..OurArabidopsisworkis

supportedI~y NSFgrant PCM-8408504.
by CORNELL UNIVERSITY on 07/14/06. For personal use only.

LiteratureCited
1. Bateson, W. 1894. Materials for the
Study of Variation, London: Macmillan.
598 pp.
2. Berger, B. 1965. The taxonomic confu-
sion within Arabidopsis and allied
genera. In Arabidopsis Research, Report
of an International Symposium,ed. (3.
R6bbelen, pp. 19-25. G6ttingen: Arabi-
dopsis Inf. Serv.
3. Botstein, D., White; R. L., Skolnick,
M., Davis, R. W. 1980. Construction of
a genetic linkage map in 1nan using
restriction fragment length polymor-
phisms. Am. J. Hum. Genet. 32:314-31
4. Braaksma,F. J., Feenstra, W. J. 1982.
Isolation and characterization of nitrate-
deficient mutants of Arabidopsis tha-
liana. Theor. Appl. Genet. 64:83-90
5. Btirger, D. 1971. Die morphologischen
Mutanten des G6ttinger Arabidopsis-
Sortiments, einschliesslich der Mutanten
mit abweiehender Samen~arbe. Arab.
Inf. Serv. 8:36-42
6. Caplan, A., Herrera-Estrella, L., Inz6,
D., Van Hante, E., Van Montagu, M.,
et al. 1983.. Introduction of genetic
material into plant cells. Science
222:815-21
7. Chang, C., Meyerowitz, E. M. 1986.
Molecular cloning and DNAsequence of
the Arabidopsis thaliana alcohol de-
hydrogenase gene. Proc. Natl. Acad.
Sci. USA 83:1208-12
8. Conrad, D. 1971. l~lber eine ROntgen-
mutante von Arabidopsis thaliana (L.)
Heynh. mit ver’~indertem Bli~tenbau and
BlOtenstand. Biol. Zentralbl. 90:137-44
9. Coulson, A., Sulston, J., Brenner, S.,
Kbarn, J. 1986. Towarda physical map
of the genome of the nematode
Caenorhabditis elegans. Proc. Natl.
Acad. Sci. USA 83:7821-25
10. Crawford, N. M., Campbell, W. H.,
Davis, R. W. 1986. Nitrate reductase
from squash: cDNAcloning and nitrate
regulation. Proc. Natl. Acad. Sci. USA
83:8073-76
11. Daniels, D. L., Blattner, F. R. 1987.
Mappingusing gene encyclopedias. Na-
ture 325:831
12. Dellaert, L. M. W., Van Es, J. Y. P.,
Koornneef, M. 1979. Ecedferum
mutants in Arabidopsis thaliana (L.)
Heynh.: II. Phenotypicand genetic anal-
ysis. Arab. Inf. Serv. 16:10-26
13. Dolferus, R., Jacobs, M. 1984.
Polymorphismof alcohol dehydrogenase
in Arabidopsis thaliana (L.) Heynh::
genetical and biochemical characteriza-
tion. Biochem. Genet. 22:817-38
14. Feenstra, W. J. 1964. Isolation of nutri-
tional mutants in Arabidopsis thaliana.
Gene~ica 35:259-69
15. Feenstaa, W. J. 1978. Contiguity of
linkage groups 1 and 4, as revealed by
linkage relationships of two newly iso-
lated markersdis-1 and dis-2. Arab. Inf.
Serv. 15:35-38
16. Hake, S., Walbot, V. 1980. The
geuomeof Zea mays, its organization
and homologyto related grasses. Chro-
mosomu 79:251-70
17. Harle, J. R. 1972. A revision of muta-
tion breeding procedures in Arabidopsis
based on a fresh analysis of the mutant
sector problem. Can. J. Genet. Cytol.
14:559-72
18. Haughn, G. W., Somerville, C. 1986.
Sulfonylurea-resistant mutants of Arabi-
dopsis thaliana. Mol. Gen. Genet.
201:430-34
19. Hedge, I. C.~ 1976. A systematic and
geographical survey of the old world
Cruciferae. In The Biology and Chemis-
 Annual Reviews
www.annualreviews.org/aronline
try of the Cruciferae, ed. J. G. Vaughan,
A. J. Macleod, B. M. G. Jones, pp.
i-45. London: Academic
20. Hirono, Y., R6dei, G. P. 1963. Multiple
allelic control of chlorophyll b level in
Arabidopsis thaliana. Nature 197:1324-
25
21. Janchen, E. 1942. Das system der Cru-
eiferen. Oesterr. Bot. Z. 91:1-28
22. Karssen, C. M., Brinkhorst-van der
Swan, D. L. C., Breekland, A. E.,
Koomneef,M. 1983. Induction of dor-
mancy during seed development ’by
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org
endogenous abscisic acid: studies on
abscisic acid deficient genotypesof Ara-
bidopsis thaliana (L.) Heynh. Planta
157:158-65
by CORNELL UNIVERSITY on 07/14/06. For personal use only.
23. Kirchheim, B., Kranz, A. R. 1981. New
population samples of the AIS-seed
bank. Arab. Inf. Serv. 18:173-76
24. Koomneef, M. 1981. The complex syn-
dromeof ttg mutants. Arab. Inf. Serv.
18:45-51
25.’Koomneef, M., De Bruine, J. H.,
Goettsch, P. 1980. A provisional mapof
chromosome 4 of Arabidopsis. Arab.
Inf. Serv. 17:11-18
26. Koomneef,M., Dellaert, L. W. M., van
der Veen, J. H. 1982. EMS-and radia-
tion-induced mutation frequencies at~t~i-
dividual loci in Arabidopsis thaliana.
Mut. Res. 93:109-23
27. Koornneef, M., Hanhart, C. J. 1983.
Linkage marker stocks of Arabidopsis
thaliana. Arab. Inf. Serv. 20:89-92
28. Koornneef, M., Jorna, M. L., Brink-
horst-van der Swan,D. L. C., Karssen,
C. M. 1982. The isolation of abscisic
acid (ABA)deficient mutants by selec-
tion of induced revertants in non-
germinatinggibberellin sensitive lines of
Arabidopsis thaliana (L.) Heynh.
Theor. Appl. Genet, 61:385-93
29. Koomneef, M., Reuling, G., Karssen,
C. M. 1984. The isolation and
characterization of abscisic acid-
insensitive mutants of Arabidopsis tha-
liana. Physiol Plant. 61:377-83
30. Koornneef, M., van der Veen, J. H.
1980. Induction and analysis of gib-
berellin sensitive mutantsin Arabidopsis
thaliana (L.) Heynh. Theor. Appl.
Genet. 58:257-63
31. Koornneef, M., van der Veen, J. H.
1983. The trisomics of Arabidopsis and
the location of linkage groups. Genetica
61:41--46
32. Koomneef,M., van Eden, J., Hanhart,
C. J., Stam, P., Braaksma,F. J., Feen-
stra, W. J. 1983. Linkagemapof Arabi-
dopsis thaliana. J. Hered. 74:265-72
33. Kranz, A. R. 1978. Demonstration of
ARABIDOPSIS THALIANA 109
new and additional population samples
and mutant lines of the AIS-seed bank,
Arab. Inf. Serv. 15:118-39
34. Laibach, F. 1907. Zur Frage nach der
IndividualiSt der Chromosomen im
Pflanzenreich. Beih. Bot. Zentralbl. 1
Abt. 22:191-210
35. Laibach, F. 1943. Arabidopsis thaliana
(L.) Heynh. als Objekt fiir genetische
und entwicklungsphysiologische Un-
tersuchungen. Bot. Archiv. 44:439-55
36. Langridge, J. 1955. Biochemical muta-
tions in the crucifer Arabidopsisthaliana
(L.) Heynh. Nature 176:26(L61
37. Langridge, J. 1957. The aseptic culture
of Arabidopsis ~haliana (L.) Heynh.
Aust. J. Biol. Sci. 10:243-52
38. Langridge, J. 1958. Ahypothesis of de-
velopmental selection exemplified by
lethal and semi-lethal mutants of Arabt-
dopsis. Aust. J. Biol. Sci. 11:58~68
39. Lauer, G. D., Roberts, T. M., Klotz, L.
C. 1977. Determination of the nuclear
DNAcontent of Saccharomyces cere-
visiae and implications for the organiza-
tion of DNAin yeast chromosomes. J.
Mol. Biol. 114:507-26
40. Lee-Chert, S., Steinitz-Sears, L. M.
1967. The location of linkage groups in
Arabidopsis thaliana. Can. J. Genet.
Cytol. 9:381-84
41. Leutwiler, L. S., Hough-Evans,B. R.,
Meyerowitz, E. M. 1984. The DNAof
Arabidopsis thaliana. Mol. Gen. Genet.
194:15-23
42. Leutwiler, L. S., Meyerowitz, E. M.,
Tobin, E. M. 1986. Structure and ex-
pression of three light-harvesting chloro-
phyll a/b binding proteins in Arabidopsis
thaliana. Nucleic Acids Res. 14:4051-
43. Li, S. L., R6dei, G. P. 1969. Thiamine
mutants of the crucifer, Arabidopsis.
Biochem. Genet. 3:163-70
44. Lloyd, A. M., Barnason, A. R., Rogers,
S. G., Byrne, M. C., Fraley, R. T.,
Horsch, R. B. 1986. Transformation of
Arabidopsis thaliana with Agrobacter-
ium tumefaciens. Science 234:464-
66
45. Lycett, G. W., Croy, R. R. D., Shirsat,
A. H., Boulter, D. 1984. The complete
sequence of a legumin gene from pea
(Pisum sativum L.). Nucleic Acids Res.
12:4493-506
46. Maher,E. P., Martindale, S. J. B. 1980.
Mutants of Arabidopsis thaliana with
altered responses to auxins and gravity.
Biochem. Genet. 18:1041-53
47. Marsden, M. P. F., Meinke, D. W.
1985. Abnormaldevelopmentof the sus-
pensor in an embryo-lethal mutant of
 Annual Reviews
www.annualreviews.org/aronline
110 MEYEROWITZ
Arabidopsis thaliana. Am. J. Bot.
72:1801-12
48. Martinez-Zapater, J. M., Estelle, M.
A., Somerville, C. R. 1986. A highly
repeated DNAsequence in Arabidopsis
thaliana. Mol. Gen. Genet. 204:417-23
49. McKelvie,A. D. 1962. A list of mutant
genes in Arabidopsis thaliana (L.)
Heynh. Radiat. Bot. 1:233~-1
50. McKelvie, A. D. 1963. Studies in thc
induction of mutations in Arabidopsis
thaliana (L.) Heynh. Radiat. Bot.
3:105-23
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org
51. Meinke, D. W. 1985. Embryo-lethal
mutants of Arabidopsisthaliana: analysis
of mutants with a wide range of lethal
phases. Theor. Appl. Genet. 69:543-52
by CORNELL UNIVERSITY on 07/14/06. For personal use only.
52. Meinke, D. W., Franzmann, L., Baus,
A., Patton, D., Weldon,R., et al. 1985.
Embryo-lethal mutants of Arabidopsis
thaliana. In Plant Genetics, UCLA
Syrup. Mol. Cell. Biol., ed. M. Freeling,
35:129-46. NewYork: Liss
53. Meinke, D. W., Sussex, I. M. 1979a.
Embryo-lethal mutants of Arabidopsis
thaliana: A model system for genetic
analysis of plant embryo development.
Dev. Biol. 72:50-61
54. Meinke, D. W., Sussex, I. M. 1979.
Isolation and characterization of six
embryo-lethal mutants of Arabidopsis
thaliana. Dev. Biol. 72:62-72
55. Meyerowitz, E. M. 1987. In situ
hybridization to RNAin plant tissue.
Plant Mol. Biol. Rep. In press
56. Meyerowitz, E. M., Chang, C. 1987.
Molecular biology of plant growth and
development:Arabidopsis thaliana as an
experimental system. In Developmental
Biology: A Comprehensive Synthesis,
Vol. 5, ed. L. Browder. NewYork: Ple-
num. In press
57. Meyerowitz, E. M., Pruitt, R. E. 1984.
Genetic Variations of Arabidopsis tha-
liana. Pasadena:Calif. Inst. Technol.39
pp.
58. Meyerowitz, E. M., Pruitt, R. E. 1985.
Arabidopsis thaliana and plant molecu-
lar genetics. Science 229:1214-18
59. Mirza, J. I. 1987. The effects of light
and gravity on the horizontal curvature
of roots of gravitropic and agravitropic
Arabidopsis thaliana L. Plant Physiol.
83:118-20
60. Mirza, J. I., Olsen, G. M., Iversen, T.-
H., Maher, E. P. 1984. The growth and
gravitropic responses of wild-type and
auxin-resistant mutants of Arabidopsis
thaliana. Physiol. Plant. 60:516-22
61. Misra, R. C. 1962. Contribution to the
embryology of Arabidopsis thalianum
(Gay & Monn.). Agra Univ. J. Res. Sci.
11:191-99
62. M~iller, A. 1961. Zur Charakterisierung
der Bliiten und Infloreszenzen yon Ara-
bidopsis thaliana (L.) Heynh. Kultur-
pflanze 9:364-93
63. Mfiller, A. J. 1963. Embryonenstest
zumNachweis rezessiver Letalfaktoren
bei Arabidopsis thaliana. Biol. Zen-
tralbl. 83:133-63
64. M011er, A. J. 1965. The chimerical
structurc of MIplants and its bearing on
the determination of mutation frequen-
cies in Arabidopsis. In Induction of
Mutations and the Mutation Process,
pp. 46-52. Prague: Czech. Acad. Sci.
65. Murray, M. G., Cuellar, R. E., Thomp-
son, W. F. 1978. DNAsequence organi-
zation in the pea genome. Biochemistry
17:5781-90
66. Olsen, G. M., Mirza, J. I., Maher, E.
P., Iversen, T.-H. 1984. Ultrastructure
and movementsof cell organelles in the
root cap of agravitropic mutantsand nor-
mal seedlings of Arabidopsis thaliana.
Physiol. Plant. 60:523-31
67. Olsen, M. V., Dutchik, J. E,, Graham,
M. Y., Brodeur, G. M., Helms, C., et
al. Randomclone strategy for genomic
restriction mapping in yeast. 1986.
Proc. .Natl. Acad. Sci. USA83:7826-30
68. Oostindier-Braaksma, F.-J., Feenstra,
W. J. 1973. Isolation and characteriza-
tion of chlorate-resistant mutantsof Ara-
bidospsis thaliana. Mut. Res. 19:175-85
69. Pruitt, R. E., Chang, C., Pang, P. P.-
Y., Meyerowitz, E. M. 1987. Molecular
genetics and development of Arabidop-
sis. In Genetic Regulation of Develop-
ment, 45th Syrup. Soc. Dev. Biol., ed.
W. Loomis, pp. 327-38. New York:
Liss
70. Pruitt, R. E., Meyerowitz, E. M. 1986.
Characterization of the genomeof Ara-
bidopsis thaliana. J. Mol. Biol.
187:169-83
71. R6dei, G. P. 1962. Supervital mutants
of Arabidopsis. Genetics 47:443-60.
72. R6dei, G. P. 1962. Single locus heter-
osis. Z. Vererbungsl. 93:164-70
73. R6dei, G. P. 1965. Genetic blocks in the
thiamine synthesis of the angiosperm
Arabidopsis. Am. J. Bot. 52:834-41
74. R6dei, G. P. 1970. Arabidopsis thaliana
(L.) Heynh. A review of the genetics
and biology. Bibliogr. Genet. 20:1-151
75. R6dei, G. P. 1973. Extrachromosomal
mutability determined by a nuclear gene
locus in Arabidopsis. Mut. Res. 18:149-
62
76. R6dei, G. P. 1975. Arabidopsis as a
genetic tool. Ann. Rev. Genet. 9:111-27
77. R6dei, G. P., Hirono, Y. 1964. Linkage
studies. Arab. Inf. Serv. 1:9-10
78. R6dei, G. P., Perry, C. M. 1971. Sub-
 Annual Reviews
www.annualreviews.org/aronline
mergedaseptic culture of intact plants in
liquid medium.Arab. Inf. Serv. 8:34
79. Reinholz, E. 1947. Ausl6sung von
R6ntgenmutationenbei Arabidopsis tha-
liana (L.) Heynh. und ihre Bedeutung
fOr die Pfianzenz0chtung und Evolu-
tionstheorie. Field Inf. Agency Tech.
Rep. 1006:1-70
80. Reinholz, E. 1947. R6ntgenmutationen
bei Arabidopsis thaliana
Naturwissenschaften (L.) Heynh.
34:26-28
81. Reinholz, E. 1959. Beeinflussung der
Morphogenese embryonaler Organe
durch ionisierende Strahlungen. I.
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org
Keimlingsanomalien durch R6ntgenbes-
trahlung yon Arabidopsis thaliana-
Embryonenin verschiedenen Entwicko
by CORNELL UNIVERSITY on 07/14/06. For personal use only.
lungsstadien. Strahlentherapie 109:537-
53
82. RObbelen, G. 1957. Untersuchungen an
strahleninduzierten Blattforbmutanten
yon Arabidopsis thaliana (L.) Heynh.
Indukt. Abst. Vererb. 88:189-252
83. R6bbelen, G. 1965. The Laibach stan-
dard collection of natural races. Arab.
Inf. Serv. 2:36-47
84. Schulz, O. E. 1936. Crueiferae. In Das
Natiirlichen Pflanzenfamilien, ed. A.
Engler, H. Harms, 17b:227-658. Leip-
zig: Engelmann
85. Schulz, P., Jensen, W. A. 1969. Cap-
sella embryogenesis: the suspensor and
basal cell. Protoplasma67:139-63
86. Schulz, P., Jensen, W. A. 1971. Cap-
sella embryogenesis: the chalazal pro-
liferating tissue. J. Cell. Sci. 8:201-27 100. Spradling, A. C., Rubin, G. M. 1981.
87. Schulz, R., Jensen, W. A. 1968. Cap-
sella embryogenesis: the early embryo.
J. Ultrastruc. Res. 22:376-92
88. Schulz, R., Jensen, W. A. 1968. Cap-
sella embryogenesis: the synergids be-
fore and after fertilization. Am. J. Bot.
55:541-52
89. Schulz, R., Jensen, W. A. 1968. Cap-
sella embryogenesis: the egg, zygote
and young embryo. Am. J. Bot. 55:807-
19
90. Shih, M.-C., Lazar, G,, Goodman,H.
M. 1986. Evidencein favor of the sym-
biotic origin of chloroplasts: primary
structure and evolution of tobacco
glyceraldehyde-3-phosphate dehydroge-
nases. Cell 47:73-80
91. Simoens, C., Alliote, T., Mendel, R.,
MOiler, A., Schiemann,J., et al. 1986.
Abinary vector for transferring genomic
libraries to plants. Nucleic Acids Res.
14:8073-90
92. Somerville, C. R., Ogren, W. L. 1979.
A phosphoglycolate phosphatase-
deficient mutant of Arabidopsis. Nature
280:833-36
93. Somerville, C. R., Ogren, W. L. 1980a.
ARABIDOPSIS THALIANA 111
Photorespiration mutants of Arabidopsis
thaliana deficient in serine-glyoxylate
aminotransferase activity, Proc. Natl.
Acad. Sci. USA 77:2684-87
94. Somerville, C. R., Ogren, W. L. 1980.
Inhibition of photosynthesis in Arabi-
dopsis mutants lacking leaf glutamate
synthase activity, Nature 286:257-59
95. Somerville, C. R., Ogren, W. L. 1981.
Photorespiration-deficient mutants of
Arabidopsis thaliana lacking mitocbon-
drial sedne transhydroxymethylase
activity. Plant Physiol. 67:666-71
96. Somerville, C. R., Ogren, W. L. 1982.
Mutants of the cmciferous plant Arabi-
dopsis thaliana lacking glycine de-
carboxylase activity. Biochem. J.
202:373-80
97. Somerville, C. R,, Ogren, W. L. 1982b.
Isolation of photorespiration mutants in
Arabidopsis thaliana. In Methods in
Chloroplast Molecular Biology, ed. M.
Edelman, R. B. I-lallick, N. H. Chua,
pp. 129-38. NewYork: Elsevier Bio-
medical
98. Sparrow, A. H., Miksche, J. P. 1961.
Correlation of nuclear volnme and DNA
content with higher plant tolerance to
chronic radiation. Science 134:282-83
99. Sparrow, A. H., Price, H. J., Un-
derbrink, A. G. 1972. A survey of DNA
content per cell and per chromosomeof
prokatyotic and eukaryotic organisms:
some evolutionary considerations.
Brookhaven Symp. Biol. 23:451 94
Drosophila genome organization: con-
served and dynamic aspects. Ann. Rev.
Genet. 15:219-64
101. Steinitz-Sears, L. M. 1963. Chromo-
some studies in Arabidopsis thaliana.
Genetics 48:483-90
102. Van der Veen, J. H. 1965. Genes for
late flowering in Arabidopsis thaliana.
See Ref. 2, pp. 62-71
103. Vandendries, R. 1909. Contribution ~
l’histoire du d~veloppementdes cruci-
f~res. Cellule 25:412-59
104. Whyte, R. O. 1946. Crop Production
and Environment. London: Faber & Fa-
ber
105. Wobus, U., Baumlein, H., Bassiiner,
R., Helm, U., Jung, R., et al. 1986.
Characteristics of two types of legumin
genes in the field bean (Vicia faba L.
var. minor) genome as revealed by
cDNAanalysis. FEBS Lett. 201:74-
80
106. Zimmerman, J. L., Goldberg, R. B.
1977. DNAsequence organization in the
genomeof Nicotiana tabacum. Chromo-
soma 59:227-52
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org
by CORNELL UNIVERSITY on 07/14/06. For personal use only.
Annu. Rev. Genet. 1987.21:93-111. Downloaded from arjournals.annualreviews.org
by CORNELL UNIVERSITY on 07/14/06. For personal use only.