Wu Zhi, Wang Shengqiang, Yuan Yongze, Zhang Tingfu, Liu Jing, Liu Deli
Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, 152 Luoyu Road, Wuhan, 430079, Hubei, People's Republic of China.
Biotechnol Lett. 2016 Aug;38(8):1349-57. doi: 10.1007/s10529-016-2113-4. Epub 2016 May 5.
To clone a novel major facilitator superfamily (MFS, a large protein family with diverse physiological functions in all kingdoms) transporter gene, Pdmfs2, and characterize its function in Penicillium digitatum.
A novel MFS transporter gene, Pdmfs2, was isolated from P. digitatum. The full-length DNA of Pdmfs2 had a 1590 bp ORF encoding a full-size MFS transporter with 529 amino acids. In a prochloraz-resistant strain (PdHS-F6), Pdmfs2 transcript level was up-regulated compared with the prochloraz-sensitive strain (PdHS-E3) and could be induced by 7 μg prochloraz/ml. The deletion of Pdmfs2 (ΔPdmfs2) in PdHS-F6 led to increased susceptibility to prochloraz and lower EC50 value (the concentration of prochloraz producing 50 % growth inhibition) compared with the PdHS-F6 or complementation strain (COPdmfs2). The ΔPdmfs2 strain was defective in conidia yield and virulence towards citrus fruits, while the complementation of Pdmfs2 could restore the phenotypic features to a large extent.
Pdmfs2 is the second MFS transporter gene in P. digitatum and is required for prochloraz resistance, conidiation and full virulence.
克隆一个新的主要易化子超家族(MFS,一个在所有生物界具有多种生理功能的大蛋白家族)转运蛋白基因Pdmfs2,并对其在指状青霉中的功能进行表征。
从指状青霉中分离出一个新的MFS转运蛋白基因Pdmfs2。Pdmfs2的全长DNA具有一个1590 bp的开放阅读框,编码一个含529个氨基酸的完整大小的MFS转运蛋白。在对咪鲜胺耐药的菌株(PdHS-F6)中,与对咪鲜胺敏感的菌株(PdHS-E3)相比,Pdmfs2转录水平上调,并且可被7μg/ml咪鲜胺诱导。与PdHS-F6或互补菌株(COPdmfs2)相比,PdHS-F6中Pdmfs2的缺失(ΔPdmfs2)导致对咪鲜胺的敏感性增加和较低的EC50值(产生50%生长抑制的咪鲜胺浓度)。ΔPdmfs2菌株在分生孢子产量和对柑橘果实的毒力方面存在缺陷,而Pdmfs2的互补可以在很大程度上恢复表型特征。
Pdmfs2是指状青霉中的第二个MFS转运蛋白基因,是咪鲜胺抗性、分生孢子形成和完全毒力所必需的。