Breeding of antagonistic bacteria against Penicillium expansum and study on its inhibition mechanism
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摘要: 为选育有效抑制扩展青霉（Penicillium expansum）的拮抗菌,并初步探讨其抑菌机制。从苹果表面分离到拮抗扩展青霉的菌株BA-16,经形态学、生理生化及16S rRNA基因序列分析,对该菌进行鉴定,并采用低能N+注入技术对其进行诱变选育。采用双酶反应体系检测野生株和突变株对扩展青霉分泌磷脂酶的抑制效果以检测突变效果并探究其抑菌机制。经鉴定,BA-16-8该菌被鉴定为解淀粉芽孢杆菌（Bacillus amyloliquefaciens）。低能N+注入技术诱变选育出的突变株BA-16-8抑菌性能显著提高且遗传性能稳定。磷脂酶活性结果显示,相对于野生株,突变株代谢产物可显著抑制病原菌所分泌的磷脂酶A的活性,且其抑制效果随浓度的增高而增强,故推测拮抗菌可能通过该机制起到抑制扩展青霉的作用。本研究对于苹果采后青霉病的生物防治具有良好应用开发前景。Abstract: To breed the antagonistic strain which can inhibit Penicillium expansum effectively,and discuss its antibacterial mechanism preliminarily.An antagonistic strain BA-16,which could inhibit Penicillium expansum,was isolated from apple surface and identified based on phenotypic,physiological,biochemical and phylogenetic( 16 S r DNA) studies.In order to enhance its antagonistic capability,the mutation of BA-16 was carried out by using low energy N+implantation.Double enzyme reaction system was used to detect the phospholipase A( PLA) activity of Penicillium expansum under the action of wild- type antagonistic strain and its mutant. The strain BA- 16 was identified as Bacillus amyloliquefaciens.After the low energy N + implantation,the mutant BA-16-8,which showed the strongest antagonist capability and stable hereditary stability was selected out. In phospholipase activity detection,mutant showed stronger inhibition against PLA activity secreted by Penicillium expansum than its wild-type strain. Moreover,as the concentration of fermentation broth increased,the inhibition effect enhanced,which indicated that it was possibly the inhibitory mechanism of antagonistic strain against Penicillium expansum. The mutant BA-16-8 will be a potential biological control agent against apple blue mold decay.
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