禾谷镰孢菌α-微管蛋白基因克隆及其与多菌灵抗药性关系分析

Cloning of α-tubulin gene from Fusarium raminearum and analyzing its relationship with carbendazim-resistance

根据禾谷镰孢菌参考菌株NRRL310 84 (PH 1)的α- 微管蛋白基因核苷酸序列设计 4对引物 ,采用PCR方法克隆并测序了禾谷镰孢菌 (Fusariumgraminearum)对多菌灵 (MBC)不同敏感性表型的 6个中国菌株的α 微管蛋白基因全序列。DNA序列对照表明中国的 3个敏感菌株和 3个抗药菌株的α- 微管蛋白基因核苷酸序列同源性没有差异 ,多菌灵抗药性与α- 微管蛋白无关。该基因全长 1718bp ,含有 6个内元 ,编码 4 4 9aa ;与NRRL310 84的α- 微管蛋白基因核苷酸序列同源性为 99% ,存在 5个差异核苷酸 ,与其所编码的氨基酸序列同源性为 99 78% ;与其他 6种真菌α- 微管蛋白基因所编码的氨基酸序列同源性为 37%～ 86 %。

The full-length nucleotide sequence of α-tubulin gene from each of 6 Fusarium graminearum strains from China which had different carbendazim (MBC) sensitivity phenotypes were separated using PCR with 4 primer sets designed in accordance with nucleotide sequence of the gene from the reference isolate, NRRL 31084 (PH-1). The DNA sequence comparison showed that there was no difference in the nucleotide sequence of α-tubulin gene amongst 3 sensitive and 3 resistant strains from China. This result demonstrates that there is no relationship between MBC-resistance and α-tubulin gene. The full-length of the gene spans 1718 bp, including 6 introns, encoding 449 amino acids. With 99 homology, there is 5 nucleotide differences inα-tubulin gene between PH-1 isolate and the 6 strains from China. The homology of the deduced amino acid sequence of the gene is 99.78 between the 6 strains and PH-1 isolate, and 37～86 between the 6 strains and other 6 species of fungi.