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【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】 The aim of this study was to explore the molecular mechanism of gametophyte development during rice meiosis and explore genes involved in the development of gametophyte on a large scale in order to reform the process of gametophyte development more genetically. 【Method】 The 22KA silent cDNA microarray was used to analyze and analyze the expression of the expressed genes in rice and Msp1 wild-type during meiosis at genome-wide level. 【Result】 The results showed that 208 genes had significant difference (P≤0.01, lgRatio≥0.2) in the wild type and Msp1 mutants. After analyzing the nucleic acid sequences of these genes, annotating GO (Gene Ontology) and predicting their function, They are grouped into 18 categories, which include GTP, DNABinding, cell wall, kinase activity, chloroplast, meiosis, ovule, chromosome, pollen capsule, intima, cell division, ubiquitin, transport, metabolism, Ca2 + binding, RNA binding, and unknown function; statistical analysis showed that there are 99 genes associated with chromosomes, accounting for 47.60% of the total; 39 genes associated with the endomembrane system, accounting for 18.75% of the total; a gene and subtraction The results showed that the three genes were closely related to the Ca2 + binding process. The other three genes were highly expressed in the Msp1 mutant compared with the wild type, indicating that these three genes may be involved in the development and regulation of the cashmere layer. The results of microarray and RT-PCR showed that AK070642 was specific to vellusmus, indicating that it is involved in the developmental regulation of vellusmus. 【Conclusion】 A large-scale comparison of the expression of wild-type rice with its Msp1 mutant during meiosis by gene chip showed that there were 208 genes from cell metabolism, development of tapetum, Regulation of meiosis of rice in the aspects of transport, nucleic acid metabolism and kinase activity, which provided experimental support for understanding the molecular mechanism of rice meiosis and gametophyte development.