枯草芽胞杆菌8-32对病原真菌的拮抗效应及对植物促生长机制

doi:10.3969/j.issn.1674-7968.2019.05.014

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Abstract Plant diseases caused by pathogenic fungi are one of the main reasons for the decline in crop quality and yield in countries around the world, which result in 20% reduction on the yields of global grain and other crops.Therefore, biological control has become one of the hotspots in plant fungal disease controls.In previous study, a strain Bacillus subtilis 8-32 was screened, which was tested to have strong antagonism to Fusarium oxysporum causing soybean root rot, and 8-32 had significant growth-promoting and biocontrol effect in greenhouse experiments.In this study, the growth-promoting and antagonistic mechanism of B.subtilis 8-32 was investigated for the further development and application of this strain.In detail, the antifungal spectrum of B.subtilis 8-32 was studied by plate confrontation method.The inhibition of B.subtilis 8-32 on spore germination, germ tube elongation and mycelium growth of main plant pathogenic fungi such as Fusarium oxysporum was testified in this study.Besides, molecular level study was carried out to reveal related antagonistic genes according to the reported literature, these genes were directly amplified by PCR using the main antagonistic gene primers of Bacillus such as iturin, surfactin, fengycin.Meanwhile, the lipopeptide antagonists were identified by high performance liquid chromatography-mass spectrometry (HPLC-MS).Plant growth-promoting substances (including indole acetic acid and siderophores) secreted by B.subtilis 8-32 were detected.The results showed that the antifungal spectrum of B.subtilis 8-32 strain was extensive.It could antagonize 7 pathogenic fungi which caused different crop diseases such as soybean root rot and wheat wilt, among which the effects were stronger for Rhizoctonia solani and Pomacea graminis, and the inhibition zone radius was 1.2 cm and 1.1 cm, respectively.The results showed that the strain 8-32 could inhibit spore germination and germ tube elongation of pathogenic fungi, and the maximum inhibition ratio reached to 100% and 95.40%, respectively.And the strain 8-32 also inhibited the growth of pathogenic fungi such as Fusarium oxysporum, Fursarium solani, Colletotrichum gloeosporivides, Alternaria brassicea through mycelial breakage.The results of PCR amplification of the antagonistic genes showed that 4 distinct single bands were amplified from 7 pairs of primers.After sequencing and NCBI Blast, the similarities between sequences and lipopeptide antibiotic genes subtilosin A (sboA), subtilisin gene (qk), fengycin gene (FenB) and hypothetical protein yndj gene (yndj) were 100%, respectively.Finally, the antagonistic substances secreted from B.subtilis 8-32 identified by HPLC-MS, including a family of lipopeptides, which were identified as homologues of iturin, that had antifungal effect.In addition, the quantitative detection on plant growth-promoting substances showed that B.subtilis 8-32 had a weak ability to produce siderophores, but strong on IAA production, which reached to 45.29 mg/L after 72 h fermentation.All the above results will lay a foundation for further study on the biological control of B.subtilis 8-32 at the molecular level, and will provide a theoretical basis for the commercial application of B.subtilis 8-32.

Key words： Bacillus subtilis Antagonistic mechanism Spore germination Germ tube growth Peptide lipid compounds

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	WANG Huan
	JIA Tian-Hui
	YANG Ke-Xin
	SHI Xiang-Zhe
	GAO Tong-Guo*
	ZHU Bao-Cheng

Cite this article:

WANG Huan,JIA Tian-Hui,YANG Ke-Xin, et al. Antagonistic Effect on Pathogenic Fungi and Plant Growth Promoting Mechanism of Bacillus subtilis 8-32[J]. 农业生物技术学报, 2019, 27(5): 908-918.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2019.05.014 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2019/V27/I5/908

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