title: Comparative expression profiling reveals gene functions in female meiosis and gametophyte development in Arabidopsis.
abstract: Megasporogenesis is essential for female fertility, and requires the accomplishment of meiosis and the formation of functional megaspores. The inaccessibility and low abundance of female meiocytes make it particularly difficult to elucidate the molecular basis underlying megasporogenesis. We used high-throughput tag-sequencing analysis to identify genes expressed in female meiocytes (FMs) by comparing gene expression profiles from wild-type ovules undergoing megasporogenesis with those from the spl mutant ovules, which lack megasporogenesis. A total of 862 genes were identified as FMs, with levels that are consistently reduced in spl ovules in two biological replicates. Fluorescence-assisted cell sorting followed by RNA-seq analysis of DMC1:GFP-labeled female meiocytes confirmed that 90% of the FMs are indeed detected in the female meiocyte protoplast profiling. We performed reverse genetic analysis of 120 candidate genes and identified four FM genes with a function in female meiosis progression in Arabidopsis. We further revealed that KLU, a putative cytochrome P450 monooxygenase, is involved in chromosome pairing during female meiosis, most likely by affecting the normal expression pattern of DMC1 in ovules during female meiosis. Our studies provide valuable information for functional genomic analyses of plant germline development as well as insights into meiosis.
authors.affiliation: Center for Genomics and Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian Province, China; National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Zhao
authors.forename: Jiangman
authors.initials: J
authors.lastname: He
authors.forename: Hanyang
authors.initials: H
authors.lastname: Cai
authors.forename: Haiyan
authors.initials: H
authors.lastname: Lin
authors.forename: Yanqiang
authors.initials: Y
authors.lastname: Li
authors.forename: Renyi
authors.initials: R
authors.lastname: Liu
authors.forename: Zhenbiao
authors.initials: Z
authors.lastname: Yang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Qin
mesh_terms: D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000235:genetics*; D023902:Chromosome Pairing; D032461:Chromosomes, Plant; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D056130:Gametogenesis, Plant; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D008540:Meiosis; D009154:Mutation; D056771:Ovule / Q000235:genetics* / Q000254:growth & development*; D030821:Plants, Genetically Modified; D011938:Rec A Recombinases / Q000235:genetics*; D015203:Reproducibility of Results
chemical_list: D029681:Arabidopsis Proteins; D018797:Cell Cycle Proteins; D003577:Cytochrome P-450 Enzyme System; C546334:KLUH protein, Arabidopsis; C513034:ATDMC1 protein, Arabidopsis; D011938:Rec A Recombinases
keywords: Arabidopsis; female meiocyte; megasporogenesis; meiosis; transcriptome