The production of the sperm cells in angiosperms requires coordination of cell division and cell differentiation. In Arabidopsis thaliana, the germline-specific MYB protein DUO1 integrates these processes, but the regulatory hierarchy in which DUO1 functions is unknown. Here, we identify an essential role for two germline-specific DUO1 target genes, DAZ1 and DAZ2, which encode EAR motif-containing C 2 H 2 -type zinc finger proteins. We show that DAZ1/DAZ2 are required for germ cell division and for the proper accumulation of mitotic cyclins. Importantly, DAZ1/DAZ2 are sufficient to promote G2-to M-phase transition and germ cell division in the absence of DUO1. DAZ1/DAZ2 are also required for DUO1-dependent cell differentiation and are essential for gamete fusion at fertilization. We demonstrate that the two EAR motifs in DAZ1/DAZ2 mediate their function in the male germline and are required for transcriptional repression and for physical interaction with the corepressor TOPLESS. Our findings uncover an essential module in a regulatory hierarchy that drives mitotic transition in male germ cells and implicates gene repression pathways in sperm cell formation and fertility.

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The objectives of this study were to determine drought tolerant novel mutant of alfalfa (Medicago sativa L.) genotypes by screening EMS mutagenized 340675 M3 seeds at germination stages in the presence of osmotic stress of 35% PEG6000. Root growth assay provided several drought tolerant candidate mutants. Of those, 4 mutants were further evaluated at water deficit conditions applied for 24 days after the first cutting at flowering bud stage. The results revealed that mutants determined as drought tolerant at germination stage were also tolerant to water deficit conditions. Protein content and superoxide dismutase values were found to be higher in all mutants than controls. Ascorbate peroxides, glutton reductase and lipid peroxidase values varied based on the mutant genotype and duration of drought stress. Drought stress significantly changed transcriptional levels of MtP5CS, MtDehyd, MseIF-2, MtRD2 and MsNAC genes. These results indicated that in vitro screening of alfalfa mutant seeds for osmatic tolerance at germination and early seedling growth stages was successfully able to determine the drought tolerant alfalfa mutants which were also tolerant to water deficit conditions after the first cutting at flowering bud stage.