Key Points
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Arabidopsis thaliana is the model system of choice for plant molecular geneticists. Forward genetic screens in Arabidopsis offer a powerful means for the functional characterization of various aspects of the plant life cycle. • Morphological and brute force biochemical screens have been highly successful in identifying the genes that are involved in many aspects of plant metabolism and development.
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The success of genetic screens depends on the appropriate genetic background, and an easy and tight procedure to identify mutants of interest.
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Unbiased modifier screens often yield fundamental new insights into complex developmental processes.
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Engineered genetic backgrounds often allow the isolation of mutants using selection rather than screening procedures.
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Use of reporter genes allows the specific tailoring of genetic backgrounds for mutagenesis. This allows the construction of an easily scorable phenotype for nearly any hidden aspect of the plant's life cycle.
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Insertional mutagenesis offers an efficient way to screen for gametophytic mutants by screening for a non-Mendelian segregation ratio distortion.
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Taking advantage of natural variation bears great potential to gain insights into plant growth and development in an evolutionary context.
Abstract
Molecular genetic studies rely on well-characterized organisms that can be easily manipulated. Arabidopsis thaliana — the model system of choice for plant biologists — allows efficient analysis of plant function, combining classical genetics with molecular biology. Although the complete sequence of the Arabidopsis genome allows the rapid discovery of the molecular basis of a characterized mutant, functional characterization of the Arabidopsis genome depends on well-designed forward genetic screens, which remain a powerful strategy to identify genes that are involved in many aspects of the plant life cycle.
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Acknowledgements
We thank M. Curtis and three anonymous reviewers for helpful comments on the manuscript, J.-P. Vielle-Calzada, J. Bowman and N. Huck for providing images, and J.-J. Pittet for help with original artwork. D.P. was supported by a Fellowship of the Roche Research Foundation and U.G. is a Searle Scholar. Support in U.G.'s lab comes from the Kanton of Zürich, the Swiss National Science Foundation, Novartis, and grants from the Bundesamt für Bildung und Wissenschaft as part of the APOTOOL and EXOTIC Projects in Framework V of the European Union.
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Glossary
- FLORAL DIPPING
-
A T-DNA transformation method in which plants are simply dipped into an Agrobacterium suspension.
- OVULE
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The organ that develops into a seed after fertilization.
- ENHANCER DETECTION
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A method originally developed in Drosophila that allows the identification of genes on the basis of their expression patterns. Engineered insertion elements carry a reporter gene construct under a minimal promoter that can respond to cis-acting regulatory elements near the insertion site.
- GENE TRAPPING
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A modification of enhancer detection first used in mice. The minimal promoter driving the reporter gene is replaced by splice acceptor sites such that expression depends on integration within a gene.
- ACCESSION
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A sample of a plant variety collected at a specific location and time. The terms ecotype, wild type and accession are not uniformly used in the Arabidopsis field and often cause confusion. The term accession is probably the most appropriate way to describe the Arabidopsis laboratory lines collected initially from the wild.
- GAMETOPHYTE
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The plant generation that produces the gametes and usually has a reduced chromosome number.
- PHYTOHORMONE
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A plant hormone that controls, or regulates, germination, growth, metabolism or other physiological functions.
- CARPEL
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A leaf-like structure that encloses the ovules and is the defining character of flowering plants. In some species, multiple carpels might be present in a compound structure, called an ovary.
- PLACENTA
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The tissue in the female reproductive organ of a plant that produces the ovules.
- NECTARY
-
The nectar-secreting gland in a flower.
- MERISTEMATIC CELL
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Totipotent, undifferentiated cells in the meristems.
- CpNpG METHYLATION
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In contrast to animals, in which 5-methylcytosine that is found in the sequence CpG is the main site of DNA methylation, both CpG and CpNpG (where N = A, T, C or G) are methylated in plants.
- STAMEN
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The male, pollen-bearing organ of the flower.
- GYNOECIUM
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The seed-bearing organ of the flower, which consists of the ovary, stigma (a sticky surface to which pollen grains attach and germinate) and style (which connects the stigma to the ovary).
- CO-SUPPRESSION
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Silencing of an endogenous gene due to the presence of a homologous transgene or virus.
- EPIALLELES
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Epigenetic variant that can confer a phenotype similar to a mutant variant of the same locus, but that has no associated changes at the DNA level.
- QUELLING
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Specifically used to describe transgene-induced silencing in Neurospora crassa.
- RNA INTERFERENCE
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(RNAi). The process by which double-stranded RNA specifically silences the expression of homologous genes through degradation of their cognate mRNA.
- SPOROPHYTE
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The multicellular diploid form in plants that undergo alternation of generations. The sporophyte results from a union of haploid gametes and meiotically produces haploid spores that grow into the gametophyte generation.
- CHALAZA
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The region of the ovule that surrounds the proximal pole of the female gametophyte and that gives rise to the integuments (protective coats) of the ovule.
- MICROPYLE
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An opening at the distal pole of the ovule through which the pollen tube enters to effect double fertilization.
- EPISTATIC
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When one gene masks the expression of another. If mutant a gives phenotype A and mutant b gives phenotype B, and if the double mutant ab gives phenotype A and not B, then gene a is epistatic to gene b.
- RECOMBINANT INBRED LINE
-
Homozygous plant line obtained after crossing two parental accessions and subsequent inbreeding over several generations.
- QUANTITATIVE TRAIT LOCUS
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A genetic locus identified through the statistical analysis of complex traits (such as plant height or body weight). These traits are typically affected by more than one gene, and also by the environment.
- NEAR-ISOGENIC LINE
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Homozygous plant lines obtained after crossing two parental accessions, then repeatedly backcrossing the progeny to one of the parents. Near-isogenic lines differ from the parental line only in a small genomic region.
- ECOTYPE
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A subdivision of a species that survives as a distinct population through environmental selection and reproductive isolation.
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Page, D., Grossniklaus, U. The art and design of genetic screens: Arabidopsis thaliana. Nat Rev Genet 3, 124–136 (2002). https://doi.org/10.1038/nrg730
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DOI: https://doi.org/10.1038/nrg730
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