摘要： 为揭示油菜黑胫病菌的致病分子机理，基于已建立的Leptosphaeria biglobosa突变体库，随机挑选16个突变体，对其致病力进行检测，筛选到12个致病力下降的突变体。并对其中6个致病力显著减弱突变体的菌落生长特性、生长速率、产孢量进行测定，结果显示，T1、T3、T4、T7突变体与野生型nm-1菌落形态相比没有明显差异，T9、T11与野生型nm-1相比，菌丝形态致密，没有产孢；T1、T3、T4、T7、T9突变体生长速率比野生型nm-1快，而T11显著低于野生型nm-1；6个突变体的产孢能力与野生型nm-1相比都显著下降；Southern Blot分析其T-DNA插入的拷贝数为单拷贝插入。并通过Hi TAIL-PCR技术扩增了6个突变体的右侧翼序列，经Blast比对分析，初步确定了T-DNA插入Leptosphaeria biglobosa基因组的位置，T-DNA插入的右侧翼序列可能为致病基因的部分序列，为进一步确定油菜黑胫病菌致病基因及其功能提供参考依据。

关键词: 油菜, 黑胫病菌, T-DNA, 致病力, 侧翼序列

Abstract: In order to reveal the pathogenic molecular mechanism of Leptosphaeria biglobosa, 16 mutants were randomly selected based on the established Leptosphaeria biglobosa mutant library and their pathogenicity was detected, and 12 mutants with reduced pathogenicity were screened.Six of these mutants with significantly reduced pathogenicity were carried out the characteristics of colony growth, growth rate and spore production quantity.The results showed that T1, T3, T4 and T7 mutants had no obvious difference in colony morphology compared with wild type nm-1, while T9 and T11 had the characteristics of the dense hyphae, but no sporulation compared with wild type nm-1;The growth rate of T1, T3, T4, T7 and T9 mutants were faster than nm-1, while T11 was significantly lower than wild type nm-1;The sporulation ability of six mutants significantly decreased compared with wild type nm-1.The Southern Blot assay revealed T-DNA insert was single copy of six mutants.Finally, the right flank sequences of six mutants were amplified by Hi TAIL-PCR, the position of T-DNA inserted into L.biglobosa genome was preliminarily identified by Blast analysis.The right flanking sequence of the six mutants might be part of the pathogenic gene sequence.This study provides the references for further screening the pathogenic genes and functions of L.biglobosa, and provides a theoretical basis for the pathogenesis of L. biglobosa.

Key words: Brassica napus, Leptosphaeria biglobosa, T-DNA, Pathogenicity, Flanking sequence

中图分类号: 

• S432.4