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, 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.
Francis
Kirk E
KE
Department of Biology and Carolina Center for Genome Scientists, University of North Carolina, Chapel Hill, NC 27599, USA.
Lam
Sandy Y
SY
Harrison
Benjamin D
BD
Bey
Alexandra L
AL
Berchowitz
Luke E
LE
Copenhaver
Gregory P
GP
D000483:Alleles; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics; D005456:Fluorescent Dyes / Q000494:pharmacology; D005785:Gene Conversion; D017343:Genes, Plant; D008040:Genetic Linkage; D006579:Heterozygote; D008540:Meiosis; D008957:Models, Genetic; D016296:Mutagenesis; D011058:Pollen / Q000235:genetics* / Q000378:metabolism*; D011995:Recombination, Genetic; D013696:Temperature; D019076:Transgenes
D029681:Arabidopsis Proteins; D005456:Fluorescent Dyes