Comparative expression profiling reveals gene functions in female meiosis and gametophyte development in Arabidopsis

• Published in: The Plant journal : for cell and molecular biology Vol. 80; no. 4; pp. 615 - 628
• Main Authors: Zhao, Lihua, He, Jiangman, Cai, Hanyang, Lin, Haiyan, Li, Yanqiang, Liu, Renyi, Yang, Zhenbiao, Qin, Yuan
• Format: Journal Article
• Language: English
• Published: England Blackwell Scientific Publishers and BIOS Scientific Publishers in association with the Society for Experimental Biology 01.11.2014; Blackwell Publishing Ltd
• Subjects: Arabidopsis; Arabidopsis - genetics; Arabidopsis Proteins - genetics; Cell Cycle Proteins - genetics; Cell division; Chromosome Pairing; Chromosomes, Plant; cytochrome P-450; Cytochrome P-450 Enzyme System - genetics; female fertility; female meiocyte; Gametogenesis, Plant; gametophytes; Gene expression; Gene Expression Profiling; Gene Expression Regulation, Plant; genes; genetic techniques and protocols; Genomics; megaspores; megasporogenesis; Meiosis; mutants; Mutation; Ovule - genetics; Ovule - growth & development; ovules; Plant biology; Plants, Genetically Modified; Rec A Recombinases - genetics; Reproducibility of Results; sorting; transcriptome; meiosis; Arabidopsis; megasporogenesis; transcriptome; female meiocyte
• ISSN: 0960-7412; 1365-313X
• DOI: 10.1111/tpj.12657

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.