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FgLEU1在禾谷镰刀菌亮氨酸生物合成及产毒致病中起关键作用

引用本文：方欣,王菲菲,范中玥,汪爽,刘馨,徐剑宏,史建荣.FgLEU1在禾谷镰刀菌亮氨酸生物合成及产毒致病中起关键作用.植物保护学报,2022,49(2):497-507

DOI：10.13802/j.cnki.zwbhxb.2022.2021141

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作者	单位	E-mail
方欣	江苏省农业科学院农产品质量安全与营养研究所, 南京 210014 江苏大学食品与生物工程学院, 镇江 212013
王菲菲	新乡医学院三全学院, 生命科学技术学院, 河南新乡 453002
范中玥	新乡医学院三全学院, 生命科学技术学院, 河南新乡 453002
汪爽	江苏省农业科学院农产品质量安全与营养研究所, 南京 210014 江苏大学食品与生物工程学院, 镇江 212013
刘馨	江苏省农业科学院农产品质量安全与营养研究所, 南京 210014 江苏大学食品与生物工程学院, 镇江 212013	xinliu@jaas.ac.cn
徐剑宏	江苏省农业科学院农产品质量安全与营养研究所, 南京 210014 江苏大学食品与生物工程学院, 镇江 212013
史建荣	江苏省农业科学院农产品质量安全与营养研究所, 南京 210014 江苏大学食品与生物工程学院, 镇江 212013

中文摘要:为挖掘新型药剂的潜在靶标，利用靶向基因敲除和互补技术研究赤霉病病原菌禾谷镰刀菌Fusarium graminearum中必需氨基酸亮氨酸合成酶编码基因FgLEU1的功能，并测定禾谷镰刀菌的生物学表型。结果表明，FgLEU1编码亮氨酸合成途径中的3-异丙基苹果酸脱水酶，其敲除突变体表现亮氨酸营养缺陷。生物学表型测定结果显示，与野生型菌株相比，FgLEU1敲除突变体的产孢量和孢子萌发率显著下降，产孢量仅为野生型菌株的20.96%，培养4 h后孢子萌发率下降了49.45%，且合成脱氧雪腐镰刀烯醇（呕吐毒素）能力丧失，在麦穗上的致病力下降，仅能侵染接种小穗，赤霉病症状不能扩展。外源添加一定量的亮氨酸、FgLeu1催化产物或导入含启动子的全长FgLEU1基因可以恢复敲除突变体表型缺陷。表明FgLEU1基因在禾谷镰刀菌亮氨酸合成、菌丝孢子形成及产毒致病过程中发挥着重要作用，可作为新型安全杀菌剂的潜在研发靶标，用于持续有效控制麦类赤霉病和镰刀菌毒素。

中文关键词:禾谷镰刀菌支链氨基酸 FgLEU1基因生物学功能产毒致病

FgLEU1 is crucial for leucine biosynthesis, virulence and DON production in fungal pathogen Fusarium graminearum

Author Name	Affiliation	E-mail
FANG Xin	Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Jiangsu Province, China
WANG Fei-fei	School of Life Science and Technology, Sanquan College of Xinxiang Medical University, Xinxiang 453002, Henan Province, China
FAN Zhong-yue	School of Life Science and Technology, Sanquan College of Xinxiang Medical University, Xinxiang 453002, Henan Province, China
WANG Shuang	Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Jiangsu Province, China
LIU Xin	Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Jiangsu Province, China	xinliu@jaas.ac.cn
XU Jian-hong	Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Jiangsu Province, China
SHI Jian-rong	Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu Province, China School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, Jiangsu Province, China

Abstract:To explore the potential usage of essential amino acid biosynthetic enzymes in novel drug design, targeted gene deletion and complementation strategy were used to characterize the Fusarium graminearum FgLEU1 gene encoding a leucine(Leu)-biosynthetic enzyme, 3-isopropylmalate dehydratase(3-IPDH), which is homologous to the yeast Leu1. The deletion mutant of FgLEU1 gene was Leu autotrophic and showed reduced mycelial growth rate and aerial hyphae formation. Compared with the wildtype strain, the deletion mutant of FgLEU1 gene showed reduced conidial formation and germination as well as deoxynivalenol(DON) production. The conidial formation of the FgLEU1 deletion mutant was only 20.96% of that of the wild-type strain and the mutant showed decreased conidial germination rates(reduced by 49.45%), and lost ability to synthesize deoxynivalenol. Plant infection assay on the flowering wheat heads indicated that the mutant could only infect the inoculated spikelet but could not spread to other spikelets. Exogenous addition of Leu or the FgLeu1-catalyzing intermediate 3-isopropylmalate(3-IPA), or re-introduction of the full-length FgLEU1 gene with its native promoter could restore the phenotypic defects of the FgLEU1 deletion mutant. Taken together, the above results indicated that FgLEU1 plays crucial roles in Leu biosynthesis, vegetative growth, DON production and virulence in F.graminearum, and is potentially a good target in designing safe fungicides for the substantial control of Fusarium head blight and toxins in wheat.

keywords:Fusarium graminearum branched-chain amino acids FgLEU1 gene biological functions deoxynivalenol production and virulence

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