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甘蓝枯萎病菌生理小种传统鉴定方法费时费力,不能满足生产的要求,因此需要建立一种快速、可靠的分子检测技术。本研究在甘蓝枯萎病菌1号和2号生理小种基因组测序的基础上,通过比较基因组学方法筛选1、2号生理小种各自的特异基因片段并设计引物,并分别以10个甘蓝枯萎病菌1号生理小种菌株、2个2号生理小种菌株、7个尖孢镰刀菌其他专化型菌株及4个外围菌株DNA为模板进行常规PCR扩增,筛选出甘蓝枯萎病菌1号和2号生理小种特异性引物,同时引入尖孢镰刀菌通用引物W106R/W106S,建立起一步三重PCR检测甘蓝枯萎病菌1、2号生理小种的分子检测技术。结果表明,该分子检测技术实现了在一次PCR反应中快速、准确地同步检测出甘蓝枯萎病菌DNA、罹病甘蓝组织和土壤中的甘蓝枯萎病菌1号和2号生理小种,对检测甘蓝植株是否感染枯萎菌及甘蓝种植区土壤是否受到枯萎菌的污染有实用价值。
The traditional identification method of cabbage Fusarium wilt races is time-consuming and laborious, and can not meet the requirements of production. Therefore, a rapid and reliable molecular detection technique needs to be established. In this study, we sequenced the genomic sequences of F1 progeny of cabbage Fusarium oxysporum f. Sp. No. 1 and 2 and screened the specific gene segments of race 1 and race 2 by comparative genomics and designed primers. Fusarium oxysporum f. 1 strain of physiological races, 2 2 strains of physiological races, 7 strains of Fusarium oxysporum other special strains and 4 strains of peripheral DNA as a template for routine PCR amplification, screening cabbage Fusarium wilt No. 1 and 2 , And the common primers F109R and W106S of Fusarium oxysporum were introduced to establish a molecular detection technique for the detection of cabbage F1 and Fusarium oxysporum F1. The results showed that this molecular detection technique can rapidly and accurately detect the DNA of cabbage Fusarium wilt, the cabbage tissue and soil cabbage Fusarium wilt pathogen No. 1 and No. 2 in one PCR reaction simultaneously, It is of practical value to detect whether the soil infected with Fusarium oxysporum and cabbage is contaminated by Fusarium oxysporum.