水稻花期特异表达基因的cDNA和RNA阵列检测

Monitoring and Analyzing the Genes Specific-Expressed at Flowering Stage of Rice (Oryza sativa)

用水稻孕穗期、开花期总mRNA为探针与含有 2 2 0 0个独立基因的cDNA阵列杂交 ,以检测开花期群体基因的表达谱。结果发现 ,在所获得的 172 0个有效克隆当中 ,有 4 72条基因表达下调 (占 2 7.4 4 % ) ,892条为持家基因 (占5 1 86 % ) ,35 6条基因表达上调 (占 2 0 .70 % )。表达上调的基因中有 2 5 0个功能已知 (占 14 .5 3% ) ,10 6个是功能未知的新基因 (占 6 .16 % )。其中S 腺苷甲硫氨酸还原酶、查尔酮合酶、酰基载体蛋白Ⅱ基因、过敏反应蛋白、醇溶蛋白、3 酮脂酰载体蛋白合酶Ⅲ和β D (1→ 3) 葡聚糖外水解酶等基因参与水稻开花和胚胎发育过程。为了进一步验证它们与开花过程的相关性 ,选取查尔酮合酶、酰基载体蛋白Ⅱ、醇溶蛋白、S 腺苷基甲硫氨酸还原酶和过敏反应蛋白等 5个基因为探针 ,与 75 5个水稻RNA斑点阵列进行杂交。结果证实醇溶蛋白和过敏反应蛋白基因受发育调节在乳熟成穗中高表达 ,查尔酮合酶和S 腺苷基甲硫氨酸还原酶基因在叶片中受光诱导、在根中受缺氮胁迫时高表达。在所有表型中 ,查尔酮合酶、S 腺苷基甲硫氨酸还原酶和酰基载体蛋白Ⅱ基因有相近的表达趋势 ,过敏反应蛋白和醇溶蛋白基因的表达趋势也相近。

A 2200 uni-gene cDNA array was hybridized with the total mRNA from rice at flowering stage for monitoring the expression profile. Results showed that 356 genes were up-regulated(20.70%), 472 genes were down-regulated(27 44%) and 892 genes were housekeeping(51.86%). Many genes expressed during the flowering process and embryo development such as putative peptide methionine sulfoxide reductase, chalcone synthase, acyl carrier protein Ⅱ gene, gene for allergenic protein, prolamine, 3-ketoacyl carrier protein synthase Ⅲ, and beta-d-glucan exohydrolase-like protein. In order to confer these genes transcription on flowering, mRNA for chalcone synthase, acyl carrier protein Ⅱ gene, prolamine, methionine sulfoxide reductase and gene for allergenic protein were chosen to hybridize with a 755-RNA (bio-phenotypes) array. Results verified that prolamine and allergenic protein were high-expressed in filling grains after being regulated by development, chalcone synthase and methionine sulfoxide reductase were high-expressed in the leaves induced by light, in the root stressed for nitrogen deficiency. Acyl carrier protein Ⅱ was a house-kept gene at all the phases except for at seedling with significant down-regulated characteristics. A co-regulation existed in the expression among chalcone synthase, methionine sulfoxide reductase and acyl carrier protein Ⅱ, and so did allergenic protein and prolamine.