Cloning β-tubulin from Fusarium fujikuroi to analyze its relationship with carbendazim resistance

Abstract:

Abstract:［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.