瓜类作物枯萎病病原菌的分子鉴定及其ITS序列差异性分析.docVIP

瓜类作物枯萎病病原菌的分子鉴定及其ITS序列差异性分析 郑雪芳 蓝江林 曹宜 肖荣凤 葛慈斌 刘波* (福建省农业科学院生物技术研究所 福州 350003) 摘 要:比较和分析了福建省几种重要瓜类作物枯萎病病原菌的ITS区序列，结果表明，供试的26个菌株属于两种不同类型的镰孢菌：尖孢镰孢和串珠镰孢，其中尖孢镰孢所占比例较大（54%），为优势种。对26株供试菌进行亲缘关系分析表明，在同源系数为92%的条件下，26株镰刀菌被划分为2个类群：第1类为尖孢镰孢，第2类为串珠镰孢。其中，第2类群中的串珠镰孢在同源系数为99%的条件下又被细分为2个类群。另外，对尖孢镰孢和串珠镰孢的ITS区序列进行比对分析，结果显示，这两种类型菌株的差异主要表现在ITS2区间，差异达20.4%，而ITS1区间的序列完全相同或只有1个碱基的差异；对尖孢镰孢各菌株的ITS序列进行多重比较发现，14株尖孢镰孢存在3种类型，类型I菌株与类型Ⅱ菌株在376bp处有1个碱基（C/T）的差异，类型Ⅱ菌株比类型Ⅲ菌株少1个碱基（A）；而对串珠镰孢各菌株的ITS序列进行多重比较发现，12株串珠镰孢也存在3种类型，类型Ⅰ菌株与类型Ⅱ菌株在320bp处有1个碱基差异（T/C），类型Ⅰ菌株与类型Ⅲ菌株有3个碱基的差异，分别是在38bp处有1个碱基差异（A/C），在395bp处有1个碱基差异（T/C），在429bp处有1个碱基的差异（T/A）。 关键词： 镰孢菌；鉴定；ITS序列分析 Identification and ITS sequence analysis of Fusarium wilt from cucumber, melon，watermelon and bottle gourd ZHENG Xue-Fang LAN Jiang-Lin CAO Yi XIAO Rong-Feng ＧE Ci-bin LIU Bo (Biotechnology Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China) ABSTRACT: An experiment was conducted to compare and analyze sequence of ITS-rDNA lied? in Fusarium wilt from cucumber, melon，watermelon and bottle gourd in Fujian. Result showed that 26 test strains were belonged to two different types: Fusarium oxysporum and F. moniliforme.The percent of F. oxysporum, which was dominant species, was 54% in the total tested strains. The phylogenetic tree of 26 tested strains indicated that 26 tested strains was divided into two major groups while the homology coefficient was 92%.The first group was F. oxysporum and the second group was F. moniliforme, which was subdivided into two groups while the homology coefficient was 99%.ITS sequence BLAST analysis was applied in F. oxysporum and F. moniliforme, result showed that main difference of two type Fusarium strains lied in ITS2, the difference of them was significant at 20.4% level, however, ITS1 sequences of them were absolutely identical or one base difference. Multiple sequence alignment of F. oxysporum showed that 14 strains had 3 types, type I and type Ⅱhad one base difference at the positio