Cloning beta-tubulin from Fusarium fujikuroi to analyze its relationship with carbendazim resistance
水稻恶苗病菌(Fusarium fujikuroi)β-微管蛋白基因克隆及与多菌灵抗药性关系

Objective] We studied the resistance of Fusarium fujikuroi against carbendazim,to confirm whether the resistance was related to the β-tubulin gene.Methods] The isolated strains were identified based on TEF-1α gene sequence and morphological characteristics.The whole nucleotide sequence of β-tubulin gene was analyzed from different sensitive strains by the primers designed according to the nucleotide sequence of β-tubulin gene from the sequenced strain 7600 of F.verticillioides.Real-time quantitative RT-PCR(qRT-PCR) was applied to analyze the expression pattern of β-tubulin gene from different sensitive strains treated with or without carbendazim.Results] The full-length nucleotide sequence of β-tubulin gene(Accession Number: JQ026022) was cloned from different sensitive strains of F.fujikoroi,which spanned 1671 bp with 4 introns,encoding 447 amino acids.This gene showed 100% nucleotide sequence homology between sensitive and resistant strains.The expression of the β-tubulin gene in 2 sensitive strains was significantly higher than 3 resistant strains when these strains were cultured on carbendazim-free media(p=0.05).Carbendazim at the concentration of their EC50 caused the expression of the gene from the five strains increasing significantly(p=0.05) in comparison with that of strains treated without carbendazim.However,there was no significant difference between strains when treated with carbendazim.Conclusion] The resistance of F.fujikuroi against carbendazim was irrelative to β-tubulin gene.