The Arabidopsis MutS homolog AtMSH4 functions at an early step in recombination: evidence for two classes of recombination in Arabidopsis.
MSH4, a meiosis-specific member of the MutS-homolog family of genes, is required for normal levels of recombination and fertility in budding yeast, mouse, and Caenorhabditis elegans. In this paper, we report the identification and characterization of the Arabidopsis homolog of MSH4 (AtMSH4). We demonstrate that AtMSH4 expression can only be detected in floral tissues, consistent with a role in reproduction. Immunofluorescence studies indicate that its expression is limited to early meiotic prophase I, preceding the synapsis of homologous chromosomes. A T-DNA insertional mutant (Atmsh4) exhibited normal vegetative growth but a severe reduction in fertility, consistent with a meiotic defect; this was confirmed by cytological analysis of meiosis. RNAi-induced down-regulation of the MSH4 gene resulted in a similar fertility and meiotic phenotype. We demonstrate that prophase I chromosome synapsis is delayed and may be incomplete in Atmsh4, and metaphase I chiasma frequency is greatly reduced to approximately 15% of wild type, leading to univalence and nondisjunction. We show that these residual chiasmata are randomly distributed among cells and chromosomes. These features of chiasma frequency and distribution in Atmsh4 show close parallels to MSH4-independent crossovers in budding yeast that have been proposed to originate by a separate pathway. Furthermore, the characteristics of the MSH4-independent chiasmata in the Atmsh4 mutant closely parallel those of second-pathway crossovers that have been postulated from Arabidopsis crossover analysis and mathematical modeling. Taken together, this evidence strongly indicates that Arabidopsis possesses two crossover pathways.
Higgins
James D
JD
School of Biosciences, The University of Birmingham, Birmingham B15 2TT, United Kingdom.
Armstrong
Susan J
SJ
Franklin
F Christopher H
FC
Jones
Gareth H
GH
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D032461:Chromosomes, Plant / Q000235:genetics / Q000502:physiology*; D003434:Crossing Over, Genetic / Q000235:genetics / Q000502:physiology*; D003584:Cytological Techniques; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D035264:Flowers / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D040461:Gene Components; D017404:In Situ Hybridization, Fluorescence; D049469:Meiotic Prophase I / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference; D012098:Reproduction / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D029681:Arabidopsis Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; C494377:MSH4 protein, Arabidopsis; C036483:T-DNA