菰黑粉菌转录调节因子Rbf1的克隆及表达分析

Cloning and Expression Analysis of Transcription Factor Rbf1 from Ustilago esculenta

为研究菰黑粉菌的形态转换机制及灰茭形成机理,利用菰黑粉菌的转录组数据,分别从正常茭白中的菰黑粉菌(MT型)和灰茭中的菰黑粉菌(T型)中扩增得到Rbf1基因及开放阅读框序列。结果表明,2种菰黑粉菌的Rbf1基因序列相同,开放阅读框全长为1 281 bp,无内含子,NCBI中blast及进化分析表明,菰黑粉菌Rbf1基因与玉米黑粉菌具有较高的同源性;另外,菰黑粉菌不同生长阶段的Rbf1基因的表达量检测结果发现,Rbf1基因在菌丝生长前后表达量有显著差异,在菌丝生长12 h后表达量明显升高,表明Rbf1基因在菰黑粉菌的菌丝形成及生长过程中具有重要作用。

For the study of Ustilago esculenta morphological transformation mechanism and formation mechanism of grey Jiaobai, the Rbf1 gene was cloned in two U. esculenta strains isolated from white Jiaobai（MT-type） and grey Jiaobai（Ttype）, based on analysis of the genome and transcriptome sequencing data. Sequence analysis results indicated that the Rbf1 gene sequence was conservative in the two strains, with the open reading frame（ORF） of 1 281 bp without intron.The NCBI blast and phylogenetic analysis showed that the Rbf1 had highest homology compared to that in U. maydis. In addition, microscope observation and real-time analysis results showed that Rbf1 was up-regulated after the hyphae formed and to be the highest 12 h later, indicating the importance in hyphae formation and growth.