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油菜菌核病菌β-微管蛋白基因与多菌灵抗药性相关突变的研究

Mutation in β-tubulin of Sclerotinia sclerotiorum conferring resistance to carbendazim in rapeseed field isolates

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【作者】 李红霞陆悦健周明国王晓峰

【Author】 LI Hong-xia, LU Yue-jian, ZHOU Ming-guo, WANG Xiao-feng Ministry of Agriculture P.R.China, College of Plant Protection, Nanjing Agriculture University, Nanjing 210095, China)

【机构】 南京农业大学植保学院农业部病虫监测与治理重点开放实验室南京农业大学植保学院农业部病虫监测与治理重点开放实验室 江苏南京210095江苏南京210095江苏南京210095

【摘要】 采集、分离获得江苏省通州市4乡镇油菜菌核病菌(Sclerotiniasclerotiorum)的223个菌株,对多菌灵(MBC)和乙霉威(NPC)的敏感性测定得到2种表型,其中143个菌株为MBCSNPCHR,其余80个菌株为MBCHRNPCS,后者对温度7~28℃不敏感。苯并咪唑类杀菌剂通过与抗药性相关的β-微管蛋白结合,抑制细胞的有丝分裂。克隆S.sclerotiorum6个菌株的β-微管蛋白基因,获得大小为874个碱基、编码273个氨基酸的基因片段,与典型模式菌粗糙脉孢霉(Neurosporacrassa)进行序列比较发现具高度同源性。研究证明S.sclerotiorum的β-微管蛋白198位氨基酸由谷氨酸(Glu)突变为丙氨酸(Ala),是导致油菜菌核病病原菌产生对多菌灵抗药性的主要原因;突变位点和突变类型与其他抗多菌灵真菌一致,与乙霉威之间存在明显负交互抗性。

【Abstract】 223 Scletotinia sclerotiorum strains were isolated from Tongzhou of Jiangsu Province. Two phenotypes were identified when tested for sensitivity to carbendazim (MBC) and diethofencarb (NPC). 143 isolates showed phenotype of carbendazin-sensitivity and diethofencarb-highly-resistance (MBCSNPCHR) and other 80 isolates showed carbendazim-highly-resistance and diethofencarb-sensitivity (MBCHRHPCS). The latter was not sensitive to temperature variation between 7~28℃. β-tubulin genes from 6 strains of carbenzim-resistant S.sclerotiorum were cloned and compared the gene nucleotide sequences (874bp) and their deduced amino acid sequences (273 amino acid) with those of the genes of typical carbenzim-resistant fungus, Neurospora crassa. The partial nucleotide sequence of β-tubulin and deduced amino acid of S.sclerotiorum were highly homologous to those from N.crassa. Sequence comparison between MBCS and MBCHR isolates of S.sclerotiorum revealed a single point mutation leading to change at amino acid position 198 from glutamic to alanine acid. This mutation was the same as those identified in benomyl-resistant field isolates of other phytopathogenic fungi, and there existed obvious negative cross-resistance between carbenzim and diethofencarb.

【基金】 国家"863"计划(2001AA249041);国家自然科学基金(30070510)
  • 【文献出处】 中国油料作物学报 ,Chinese Journal of Oil Crop Scieves , 编辑部邮箱 ,2003年02期
  • 【分类号】S435.654
  • 【被引频次】58
  • 【下载频次】363
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