Pollen tetrad-based visual assay for meiotic recombination in Arabidopsis

Recombination, in the form of cross-overs (COs) and gene conversion (GC), is a highly conserved feature of meiosis from fungi to mammals. Recombination helps ensure chromosome segregation and promotes allelic diversity. Lesions in the recombination machinery are often catastrophic for meiosis, resul...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2007-03, Vol.104 (10), p.3913-3918
Main Authors: Francis, Kirk E, Lam, Sandy Y, Harrison, Benjamin D, Bey, Alexandra L, Berchowitz, Luke E, Copenhaver, Gregory P
Format: Article
Language:English
Subjects:
Alleles
Arabidopsis - genetics
Arabidopsis Proteins - genetics
Arabidopsis thaliana
Biological Sciences
Cell division
chromatids
Chromosomes
crossing over
crossover interference
cytogenetic analysis
DNA
Flowers
Fluorescence
Fluorescent Dyes - pharmacology
fluorescent proteins
Gametes
gametogenesis
Gene Conversion
Genes, Plant
Genetic Linkage
Genetic loci
Genetic recombination
Genomics
Heterozygote
Meiosis
Models, Genetic
Mutagenesis
Nonnative species
Plants
Pollen
Pollen - genetics
Pollen - metabolism
pollen tetrads
Proteins
Recombination, Genetic
reporter genes
Temperature
Transgenes
Weeds
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Summary:Recombination, in the form of cross-overs (COs) and gene conversion (GC), is a highly conserved feature of meiosis from fungi to mammals. Recombination helps ensure chromosome segregation and promotes allelic diversity. Lesions in the recombination machinery are often catastrophic for meiosis, resulting in sterility. We have developed a visual assay capable of detecting Cos and GCs and measuring CO interference in Arabidopsis thaliana. This flexible assay utilizes transgene constructs encoding pollen-expressed fluorescent proteins of three different colors in the qrt1 mutant background. By observing the segregation of the fluorescent alleles in 92,489 pollen tetrads, we demonstrate (i) a correlation between developmental position and CO frequency, (ii) a temperature dependence for CO frequency, (iii) the ability to detect meiotic GC events, and (iv) the ability to rapidly assess CO interference.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0608936104