苏云金芽胞杆菌4BM1菌株对油菜菌核病的防治潜力

doi:10.13560/j.cnki.biotech.bull.1985.2024-0279

摘要/Abstract

摘要：

【目的】前期研究显示苏云金芽胞杆菌（Bacillus thuringiensis，Bt）4BM1菌株可诱导油菜产生对核盘菌（Sclerotinia sclerotiorum）的系统抗病性。探究其诱导系统抗性机制并分析其生防潜力，为油菜菌核病的生物防治提供菌株资源。【方法】采用实时荧光定量PCR和转录组测序技术分析根部施用4BM1菌株后，油菜叶片中防御相关基因的转录水平；结合4BM1菌株全基因组序列和antiSMASH 2.0软件预测其抗病相关次级代谢产物生物合成的基因簇；采用发酵液灌根的方式将4BM1菌株接种至油菜盆栽幼苗，分析其定殖能力和促生效果。【结果】4BM1菌株可激发油菜叶片产生过敏反应；油菜根部施用4BM1菌株，叶片中参与水杨酸、茉莉酸、乙烯信号和油菜素内酯生物合成途径等基因转录水平显著上调；4BM1菌株的染色体上预测到胞外多糖和其他13个抗病相关次级代谢产物合成的基因簇；4BM1菌株在油菜根际定殖的同时可促进油菜苗期植株的生长。【结论】4BM1菌株可作为防控油菜菌核病、促进油菜幼苗生长有潜力的生物防治资源。

关键词: 苏云金芽胞杆菌, 油菜菌核病, 诱导系统抗性, 生物防治

Abstract:

【Objective】In the previous study, Bacillus thuringiensis 4BM1 strains effectively suppressed Sclerotinia sclerotiorum growth by inducing systemic resistance in Brassica campestris. The induced systemic resistance mechanism and biocontrol potential were studied, which may provide strain resources for the biological control of sclerotiniose.【Method】Quantitative real-time PCR and transcriptome sequencing were used to analyze the transcription levels of defense-related genes in B. campestris leaves after roots were exposed to the Bt 4BM1 strain. The disease-resistance related secondary metabolites biosynthesis gene clusters were predicted by using the complete genome sequence of Bt 4BM1 strain and the antiSMASH 2.0 software. The 4BM1 strain was inoculated into the potted B. campestris seedlings by fermentation liquid irrigation method, and its colonization ability and growth promotion effect were analyzed. 【Result】4BM1 strain triggered hypersensitive response in B. campestris leaves. Genes involved in salicylic acid, jasmonic acid, ethylene signaling and brassinosteroid synthesis pathways of B. campestris leaves were upregulated after the roots were exposed to the Bt 4BM1 strain. The candidate gene clusters involved in exopolysaccharide and 13 other disease-resistance related secondary metabolites biosynthesis were predicted in 4BM1 strain chromosome. The 4BM1 strain colonized in the rhizosphere of B. campestris and promoted the growth of B. campestris. 【Conclusion】4BM1 strain can be used as a potential biological control resource to control sclerotiniose and promote the growth of B. campestris seedlings.

Key words: Bacillus thuringiensis, sclerotiniose, induced systemic resistance, biocontrol

王美玲, 耿丽丽, 房瑜, 束长龙, 张杰. 苏云金芽胞杆菌4BM1菌株对油菜菌核病的防治潜力[J]. 生物技术通报, 2024, 40(9): 260-269.

WANG Mei-ling, GENG Li-li, FANG Yu, SHU Chang-long, ZHANG Jie. Control Potential of Bacillus thuringiensis 4BM1 Strain to Sclerotiniose in Brassica campestris L.[J]. Biotechnology Bulletin, 2024, 40(9): 260-269.

图/表 7

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功能 Function	基因编号 Gene ID	差异倍数的log值 log₂Fold-change	调控方式 Regulated	描述 Description
Steroid-related genes	Bra029392*	8.51283041	Up	Squalene epoxidase
Steroid-related genes	Bra031393	1.572668503	Up	Sugar phosphate/phosphate translocator
Ethylene-related genes	Bra004522	1.13850342	Up	26S protease regulatory subunit 8
	Bra002499*	2.858810471	Up	Concanavalin A-like lectin kinase-like protein
	Bra017656	2.011958293	Up	Ethylene-responsive transcription factor
Salicylic acid-related genes	Bra024318*	1.115278771	Up	U-box domain-containing protein 27
	Bra037520	1.415677575	Up	Inorganic ion transport and metabolism
	Bra013123	2.021981527	Up	Pathogenesis-related protein
	Bra013732	1.576452095	Up	WRKY transcription factor
	Bra033158	1.751241545	Up	WRKY transcription factor
	Bra008435	1.190167975	Up	WRKY transcription factor
Secondary metabolite biosynthesis-related genes	Bra040582	5.867390439	Up	Sesquiterpenoid biosynthetic
Secondary metabolite biosynthesis-related genes	Bra021965	1.653287924	Up	Abscisic acid-hydroxylase 3
Chloroplast-related genes	Bra003004	-6.601409988	Down	Protochlorophyllide reductase A

功能 Function	基因编号 Gene ID	差异倍数的log值 log₂Fold-change	调控方式 Regulated	描述 Description
Steroid-related genes	Bra029392*	8.51283041	Up	Squalene epoxidase
Steroid-related genes	Bra031393	1.572668503	Up	Sugar phosphate/phosphate translocator
Ethylene-related genes	Bra004522	1.13850342	Up	26S protease regulatory subunit 8
	Bra002499*	2.858810471	Up	Concanavalin A-like lectin kinase-like protein
	Bra017656	2.011958293	Up	Ethylene-responsive transcription factor
Salicylic acid-related genes	Bra024318*	1.115278771	Up	U-box domain-containing protein 27
	Bra037520	1.415677575	Up	Inorganic ion transport and metabolism
	Bra013123	2.021981527	Up	Pathogenesis-related protein
	Bra013732	1.576452095	Up	WRKY transcription factor
	Bra033158	1.751241545	Up	WRKY transcription factor
	Bra008435	1.190167975	Up	WRKY transcription factor
Secondary metabolite biosynthesis-related genes	Bra040582	5.867390439	Up	Sesquiterpenoid biosynthetic
Secondary metabolite biosynthesis-related genes	Bra021965	1.653287924	Up	Abscisic acid-hydroxylase 3
Chloroplast-related genes	Bra003004	-6.601409988	Down	Protochlorophyllide reductase A

4BM1	CDS	Bases	Gene	misc_RNA	rRNA	tRNA	Intergenetic region/%	GC/%	Gene GC/%	Intergenetic GC/%
染色体Chromosome	5529	5498518	5811	131	42	108	15.22	35.53	36.38	30.77
质粒1 Plasmid1	313	333074	318	5	0	0	21.78	33.37	34.29	30.06
质粒2 Plasmid2	205	218923	207	2	0	0	26.69	33.2	34.28	30.24

4BM1	CDS	Bases	Gene	misc_RNA	rRNA	tRNA	Intergenetic region/%	GC/%	Gene GC/%	Intergenetic GC/%
染色体Chromosome	5529	5498518	5811	131	42	108	15.22	35.53	36.38	30.77
质粒1 Plasmid1	313	333074	318	5	0	0	21.78	33.37	34.29	30.06
质粒2 Plasmid2	205	218923	207	2	0	0	26.69	33.2	34.28	30.24

基因编号 Gene ID	功能预测 Predicted function	4F5菌株中预测的胞外多糖基因簇 Predicted eps gene cluster in Bt 4F5 strain	4BM1与4F5之间的氨基酸序列相似性Amino acid identity between 4BM1 and 4F5/%
4BM1_Chr1_00581	EpsC	4F5_Chr1_00600	N/A
4BM1_Chr1_00582	UTP-glucose-1-phosphate uridylyltransferase（EC:2.7.7.9）	4F5_Chr1_00601	94.5
4BM1_Chr1_00583	Lipopolysaccharide synthesis sugar transferase	4F5_Chr1_00602	94.9
4BM1_Chr1_00584	Nucleotide sugar dehydrogenase	4F5_Chr1_00603	23
4BM1_Chr1_00585	Asparagine synthase	4F5_Chr1_00604	N/A
4BM1_Chr1_00586	Polysaccharide biosynthesis protein	4F5_Chr1_00605	N/A
4BM1_Chr1_00587	Glycosyltransferase	4F5_Chr1_00606	N/A
4BM1_Chr1_00588	Hypothetical protein	4F5_Chr1_00607	N/A
4BM1_Chr1_00589	Hypothetical protein	4F5_Chr1_00608	N/A
4BM1_Chr1_00590	Glycosyltransferases group 1	4F5_Chr1_00609	N/A
4BM1_Chr1_00591	Glycosyltransferase, group 1 family protein	4F5_Chr1_00610	N/A
4BM1_Chr1_00592	Glycosyltransferase	4F5_Chr1_00611	N/A
4BM1_Chr1_00593	Glucose-1-phosphate thymidylyltransferase（EC:2.7.7.24）	4F5_Chr1_00612	N/A
4BM1_Chr1_00594	dTDP-4-dehydrorhamnose 3,5-epimerase（EC:5.1.3.13）	None
4BM1_Chr1_00595	dTDP-glucose 4,6-dehydratase	None
4BM1_Chr1_00596	dTDP-4-dehydrorhamnose reductase（EC:1.1.1.133）	None
4BM1_Chr1_00597	Membrane-bound transcriptional regulator LytR	4F5_Chr1_00613	88.5
4BM1_Chr1_00598	EPSX protein	4F5_Chr1_00614	89.9
4BM1_Chr1_00599	UDP-glucose 4-epimerase GalE	4F5_Chr1_00616	96.1
4BM1_Chr1_00600	Two component transcriptional regulator	4F5_Chr1_00617	93.3
4BM1_Chr1_00601	Integral membrane sensor signal transduction histidine kinase	4F5_Chr1_00618	93
4BM1_Chr1_00602	Hypothetical protein	4F5_Chr1_00619	88.9
4BM1_Chr1_00603	ABC transporter substrate-binding protein	4F5_Chr1_00620	93.6
4BM1_Chr1_00604	glpT; glycerol-3-phosphate transporter	4F5_Chr1_00621	89.6