野生型和Msp1突变体水稻减数分裂期转录水平上的基因表达分析(英文)

Transcript level analyses of genes expressed during rice meiosis in wild type and Msp1 mutant

【目的】深入探索水稻减数分裂期间配子体发育的分子机理,大规模发掘与水稻配子体发育相关的基因,以便更有把握地利用基因工程手段改造水稻配子体发育的进程。【方法】利用22KAilentcDNA芯片,在全基因组水平上对野生型水稻和Msp1突变体在减数分裂期间表达基因的表达情况进行研究和分析。【结果】有208个基因在野生型和Msp1突变体中的表达差异极显著(P≤0.01,lgRatio≥0.2);对这些基因的核酸序列进行分析、GO(GeneOntology)注释以及功能预测后,将其划归为18个大类,分别涉及到GTP、DNABinding、细胞壁、激酶活性、叶绿体、减数分裂、胚珠、染色体、花粉囊、内膜、细胞分裂、泛素、转运、代谢、蛋白解、Ca2+结合、RNA结合、以及未知功能等;统计分析表明,有99个基因与染色体有关,占总数的47.60%;39个基因与内膜系统有关,占总数的18.75%;有1个基因与减数分裂紧密相关,3个基因与Ca2+结合过程有关;另外有3个基因在Msp1突变体中的表达极显著高于野生型,表明这3个基因很可能参与了绒粘层的发育调控。基因芯片和RT-PCR检测结果表明,水稻基因AK070642特异于绒粘层,说明其参与了绒粘层的发育调控。【结论】研究通过基因芯片技术,将野生型水稻与其Msp1突变体在减数分裂期间表达基因的表达情况进行大规模对比后发现,有208个基因分别从细胞代谢、绒毡层的发育、离子转运、核酸代谢、激酶活性等方面调控水稻的减数分裂过程,为深入理解水稻减数分裂和配子体发育的分子机理提供了实验支撑。

[Objective] This study aimed to find some candidate genes for modifying rice development process by biotechnology,and further understand the molecular mechanism of rice gametogenesis during meiosis. [Method] The 22 K Ailent cDNA microarray was used to study the gene expression difference between wild type and Msp1 mutant during meiosis stage. [Result] After comparing the genes expressed during meiosis between wild type rice and Msp1 mutant by microarray,208 genes whose expressions changed significantly （P≤0.01 and 1g Ratio≥0.2） when rice meiosis were explored. The potential functions of these genes were predicted by ontology analysis and clustered in 18 groups based on GO classification, ranging from GTP, DNA Binding, cell wall, kinase activity, chloroplast, meiosis, ovule, chromosome, anther, endomembrane, cell division, metabolism, ubiquitin, transport, proteolysis,Ca^2＋ binding,RNA binding,and unknown. It showed that 99 genes,47.60% of 208 genes, were involved in chromosome activity;39 genes, 18.75 % of 208 genes, were involved in endomembrane and transport process respectively. Among these 208 genes,one gene was related to meiosis intimately;three of them were related to Ca^2＋ binding; the expression of three genes was significantly higher in Msp1 mutant compared with wild type. It implies they might control the development of tapetum. Moreover,microarray and RT-PCR results demonstrated that rice OSC6 （oxidosqualene cyclase） homolouges gene Os11g0582500 （AK064672） was tapetum specific and involved in tapetum development. The relationship between meiosis and ubiquitin, and the role of three Ca^2＋ binding genes in rice meiosis were discussed. [Conclusion] After comparing the gene expression between wild type and Mspl mutant by 22 K chips,208 genes were found involved in rice meiosis control at various aspects such as cell metabolism, tapetum development, ion transport, nucleic acid metabolism,kinase activity. This study provided an experimental support for the further understanding of the molecular mechanism of rice meiosis and gametogenesis.