Mutation in β-tubulin of Sclerotinia sclerotiorum conferring resistance to carbendazim in rapeseed field isolates
Mutation in β-tubulin of Sclerotinia sclerotiorum conferring resistance to carbendazim in rapeseed field isolates
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发表时间：
2003
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通讯作者：
Wang Xiao-feng
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作者：
Wang Xiao-feng
223株<s:1>菌核菌<e:1>菌株分离自江苏通州。在检测多菌灵（MBC）和嘧虫威（NPC）敏感性时，鉴定出两种表型。143株菌株表现为多菌灵敏感型和乙硫虫威高耐型（MBC SNPC HR）， 80株菌株表现为多菌灵高耐型和乙硫虫威敏感型（MBC HR HPC S）。后者对7~28℃的温度变化不敏感。6株耐卡苯齐姆菌株的β-微管蛋白基因。克隆了菌核菌（sclerotiorum）的基因序列（874bp）及其推导出的氨基酸序列（273个氨基酸），并与典型的耐卡本辛真菌——Neurospora进行了比较。得到了β-微管蛋白的部分核苷酸序列和推断出的氨基酸序列。sclerotiorum与高度同源。MBC与MBC HR分离株的序列比较。菌核苗（Sclerotiorum）显示了一个单点突变，导致198号氨基酸位置从谷氨酸变为丙氨酸。该突变与其他植物病原真菌抗苯甲酰田间分离株的突变一致，且卡苄星与嘧虫威之间存在明显的负交叉抗性。
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 (MBCSNPCHR) and other 80 isolates showed carbendazim-highly-resistance and diethofencarb-sensitivity (MBCHRHPCS). 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 MBCS and MBCHR 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.