伯克霍尔德氏菌JP2-270抗水稻纹枯病菌机理的初步研究

doi:10.16409/j.cnki.2095-039x.2022.02.003

摘要/Abstract

摘要： 水稻纹枯病是由立枯丝核菌Rhizoctonia solani引起的，是水稻的三大病害之一。本研究采用基因敲除、荧光定量PCR和靶向代谢物差异分析等方法，初步探索了伯克霍尔德氏菌Burkholderia sp.JP2-270抑制水稻纹枯病菌的作用机制。首先，选取了水稻稻瘟病菌Magnaporthe oryza、香蕉枯萎病菌Fusarium oxysporum、番茄斑枯病菌Septoria lycopersici Speg.、小麦赤霉病菌Fusarium graminearum和烟草枯萎病菌Fusarium oxysporum等病原真菌进行抗真菌活性测定，结果表明，菌株JP2-270具有广谱抗真菌活性。其次，通过靶向基因敲除明确了菌株JP2-270所产生的硝吡咯菌素pyrrolnitrin是抑制立枯丝核菌GD118的主要次生代谢物。最后，qRT-PCR分析表明硝吡咯菌素的合成在转录水平上受到菌株JP2-270中LysR家族转录调控蛋白BysR的正向调控；并且，差异代谢物谱分析也证明BysR能够正调控硝吡咯菌素的合成。在伯克霍尔德氏菌中LysR类型的转录调控蛋白BysR作为硝吡咯菌素合成调控的研究属于首次报道，BysR蛋白可作为基因工程的靶点，通过遗传改良提高硝吡咯菌素的生物合成量。因此，菌株JP2-270具有开发成生物防治剂防治植物真菌病害的潜能。

关键词: 伯克霍尔德, 水稻纹枯病, 硝吡咯菌素, 转录调控蛋白BysR

Abstract: Sheath blight, caused by Rhizoctonia solani, is one of the three major diseases of rice. In this study, we performed gene knockout, real time quantitative PCR and differential analysis of targeted metabolites to preliminarily uncover the inhibitory mechanism of Burkholderia sp. JP2-270 against rice sheath blight. Magnaporthe oryza, Fusarium oxysporum, Fusarium graminearum and Septoria lycopersici Speg were selected as the target fungi for inhibition activity analysis, and the results showed that JP2-270 has broad spectrum inhibitory activities against these phytopathogenic fungi. Targeted gene knockout analysis indicated that pyrrolnitrin produced by strain JP2-270 was the main secondary metabolite inhibiting R. solani GD118. qRT-PCR analysis showed that the expression of gene cluster (prnA-D) responsible for the production of pyrrolnitrin was positively regulated by a LysR transcriptional regulator BysR in strain JP2-270, and the differential metabolite spectrum analysis further confirmed that BysR could positively regulate the synthesis of pyrrolnitrin. LysR type transcriptional regulatory protein BysR was reported for the first time as the regulator of pyrrolnitrin production in Burkholderia. The transcriptional regulator BysR could be used as a target of genetic engineering to improve the production of pyrrolnitrin. Thus, Burkholderia sp. JP2-270 has the potential to be developed as a biocontrol agent against plant fungal disease.

Key words: Burkholderia, rice sheath blight disease, pyrrolnitrin, transcriptional regulator BysR

中图分类号:

S476

吴丽娟, 韩聪, 王惠梅, 王磊, 鄂志国. 伯克霍尔德氏菌JP2-270抗水稻纹枯病菌机理的初步研究[J]. 中国生物防治学报, 2022, 38(1): 230-241.

WU Lijuan, HAN Cong, WANG Huimei, WANG Lei, E Zhiguo. Study on the Resistance Mechanism of Burkholderia sp. JP2-270 against Rice Sheath Blight[J]. Chinese Journal of Biological Control, 2022, 38(1): 230-241.

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