【目的】获得与油菜温敏雄性不育系K121S育性转换相关的功能基因。【方法】以油菜温敏雄性不育系K121S为材料,分别取可育温度下和不育温度下的花粉母细胞期、四分体期、三核期和成熟期的花蕾提取RNA,利用DDRT—PCR方法进行基因表达差异显示,共获得了44条差异表达的cDNA片段。【结果】经测序和BLAST序列比对分析,筛选出7个候选基因的差异表达与K121S的育性转换有关,推测胼胝质合成酶基因、乙醛脱氢酶基因、RNA聚合酶I转录因子RRN3基因在转录水平差异表达,而H+-ATP酶基因、I类果糖二磷酸醛缩酶基因、富含亮氨酸重复的丝氨酸,苏氨酸类受体激酶基因、碱性,中性转化酶基因在转录后水平RNA剪接时差异表达。【结论】该研究结果对揭示油菜温敏雄性不育的分子机理提供了依据。
[Objective] This study was performed to screen functional genes related to the fertility conversion of thermo-sensitive genic male sterile (TGMS) lines of Brassica juncea L. [Method] A B. juncea TGMS line K121S was selected as the experimental material. The total RNAs were isolated from fertile and sterile pollens at different development stages, including mother cell stage, tetrad stage, tricellular pollen stage and maturity stage. DDRT-PCR was carried out to identify differentially expressed genes. [Result] A total of 44 differentially expressed cDNA fragments were identified with Dot blot. And seven candidate genes related to fertility conversion of K121S were screened out by BLASTN, including callose synthase gene, aldehyde dehydrogenase gene and RNA polymerase I transcription factor RRN3 gene which were differentially expressed at the transcriptional level, H'-ATPase gene, fructose diphosphate aldolase -class I gene, teucine-rich repeat receptor-Jike serine/threonine- protein kinase gene and alkaline/neutral invertase gene, which were differentially expressed at the post-transcriptional level. [Conclusion] The results of this study will help to explain the molecular mechanism of thermo-sensitive genic male sterility of B. juncea.