论文部分内容阅读
本研究利用cDNA-AFLP技术和生物信息学技术分析乌塌菜核不育系败蕾过程中的基因表达差异。筛选到64个与花蕾败育相关的差异表达片段(TDF)。选取42个进行测序,获得的序列信息在BRAD和NCBI数据库上进行同源性比对,其中同源性大于90%的TDF有16条,功能涉及能量代谢、逆境响应、转录和翻译、信号转导、抗病与衰老、氨基酸的合成与加工、跨膜运输等方面。通过QRT-PCR技术对表达模式进行验证,结果与cDNA-AFLP一致,无假阳性条带。选取3个同源性较高的TDF,采用QRT-PCR分析其组织表达模式。结果表明,H49在不育系败蕾期表达受到抑制,而在开花期大花蕾、叶片、萼片、花瓣等组织中高水平表达;与之相反,H41和H59在不育系开花期表达受抑制,而在败蕾期大花蕾、萼片、花瓣、花药中高水平表达。
In this study, cDNA-AFLP technology and bioinformatics techniques were used to analyze the gene expression differences in the process of aborting buds of the sterile plant of Brassica campestris. A total of 64 differentially expressed fragments (TDFs) related to flower bud abortion were screened. Forty-two sequences were sequenced, and the sequence information was aligned on the BRAD and NCBI databases. There were 16 TDFs with greater than 90% homology, including energy metabolism, stress response, transcription and translation, Guide, disease resistance and aging, amino acid synthesis and processing, transmembrane transport and so on. The expression pattern was verified by QRT-PCR and the results were consistent with that of cDNA-AFLP. There was no false positive band. Three TDFs with high homology were selected and their tissue expression patterns were analyzed by QRT-PCR. The results showed that the expression of H49 was inhibited in the budding stage of the male sterile line, but was highly expressed in the buds, leaves, sepals, petals and other tissues at the flowering stage. On the contrary, the expression of H41 and H59 was inhibited in the flowering stage of the male sterile lines, In the budding buds, sepals, petals, anthers high level expression.