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期 刊 ：植物保护学报 2010年

关键词：油菜籽；油菜茎基溃疡病菌；油菜黑胫病菌；检测；

Keywords:Brassica napus seed, Leptosphaeria maculans, Leptosphaeria biglobosa, detection,

摘 要 ：为准确鉴定从进境澳大利亚油菜籽样品中分离的真菌分离物,利用形态学特征、PCR检测、序列分析以及致病性测试等方法对分离物6382-43和6382-51进行了鉴定试验。结果表明,分离物6382-43的形态特征和油菜茎基溃疡病菌Leptosphaeria maculans相似,菌丝生长较慢,菌落边缘不规则,不产生色素。油菜茎基溃疡病菌特异性引物LmacF/LmacR检测为PCR阳性;ITS区序列和油菜茎基溃疡病菌的序列相似性为99.8%;接种幼嫩油菜子叶产生油菜茎基溃疡病的典型症状。分离物6382-51的形态特征和油菜黑胫病菌L.biglobosa相似,菌丝生长较快,菌落边缘规则,产生色素;油菜黑胫病菌特异性引物LbigF/LmacR检测为PCR阳性;ITS区序列和油菜黑胫病菌的序列相似性为100%;接种幼嫩油菜子叶产生油菜黑胫病的典型症状。根据分离物的形态特征、PCR检测结果、序列分析以及致病性测试结果,将进境澳大利亚油菜籽样品中的真菌分离物6382-43和6382-51分别鉴定为油菜茎基溃疡病菌Leptosphaeria maculans和油菜黑胫病菌L.biglobosa。

Abstract:The isolates 6382-43 and 6382-51 were intercepted from oilseed rape samples imported from Australia. The isolate 6382-43 with similar morphological characteristics to Leptosphaeria maculans showed a slower growth rate of 3.0mm/d with irregular margin of colony and no pigment production. The isolate 6382-51 with a faster growth rate of 4.8mm/d and regular margin of colony produced diffusible pigment. The result of PCR detection showed that the isolate 6382-43 could be amplified by L.maculans-specific primer LmacF/LmacR and the isolate 6382-51 got PCR product of 444bp by L.biglobosa-specific primer LbigF/LmacR. The sequence analysis revealed that the isolate 6382-43 had 99.8% sequence identity with L.maculans, and isolate 6382-51 showed 100% sequence similarity with L.biglobosa. Pathogenicity was confirmed on cotyledons of rape seed by artificial inoculation with typical symptoms of L.maculans and L.biglobosa. Based on the morphological characteristics, PCR detection, sequence analysis and pathogenicity test, the isolates 6382-43 and 6382-51 from Brassica napus seed sample were identified as Leptosphaeria maculans and L.biglobosa, respectively.

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