贝莱斯芽孢杆菌防治甜樱桃采后软腐病的效果和机理

doi:10.7506/spkx1002-6630-20220422-291

摘要/Abstract

摘要： 为探索贝莱斯芽孢杆菌（Bacillus velezensis）抑制甜樱桃软腐病的作用机理，实验分析贝莱斯芽孢杆菌KT对匍枝根霉（Rhizopus stolonifer）LC-7的体外和甜樱桃果实体内抑制效果，以及贝莱斯芽孢杆菌KT菌株对甜樱桃果实相关防御酶活力的影响，并通过转录组学解析在与匍枝根霉互作期间，贝莱斯芽孢杆菌KT菌株中与次级代谢物合成和生物防治相关基因的表达情况。结果表明，1×109 CFU/mL贝莱斯芽孢杆菌KT菌悬液对匍枝根霉LC-7菌丝抑制率达87.12%，此时孢子萌发率和胚芽管长度分别为21.54%和11.45 μm。在甜樱桃果实上，1×109 CFU/mL贝莱斯芽孢杆菌KT菌悬液处理48 h后，果实软腐病发病率约为20%，比对照组降低了约80%，并且贝莱斯芽孢杆菌KT菌悬液能抑制甜樱桃果实多酚氧化酶和脂氧合酶活力，诱导过氧化物酶、苯丙氨酸解氨酶、几丁质酶和β-1,3-葡聚糖酶活力增加，提高果实抗氧化和抗病能力。扫描电子显微镜观察发现，经与贝莱斯芽孢杆菌KT菌株共培养后，匍枝根霉LC-7菌丝和孢子形态扭曲，贝莱斯芽孢杆菌KT对匍枝根霉LC-7具有明显的致畸作用。转录组学分析结果表明，贝莱斯芽孢杆菌KT菌株中抗生素合成基因簇bacillaene中的pksIJ和糖醛酸磷壁酸合成基因簇中的tuaAGE基因表达上调，此外发现，表达差异显著的基因主要富集在生物学过程中的核糖核蛋白复合体组装和亚基组成及核糖体组装等一系列与核糖体活动相关的过程。贝莱斯芽孢杆菌可能通过营养和空间竞争以及分泌抑菌活性物质直接抑制匍枝根霉LC-7，还可诱导甜樱桃果实的抗病性。贝莱斯芽孢杆菌对由匍枝根霉引起的甜樱桃软腐病有良好的控制作用，具有开发成生防菌剂的潜力。

关键词: 贝莱斯芽孢杆菌；匍枝根霉；甜樱桃；软腐病；互作；抑菌机理

Abstract: To explore the mechanism of action of Bacillus velezensis against sweet cherry soft rot disease, the inhibitory effect of Bacillus velezensis KT against Rhizopus stolonifer LC-7 in vitro and in inoculated sweet cherry fruits, and the effect of B. velezensis KT on defense-related enzyme activities were investigated. Besides, the expression of genes related to secondary metabolite synthesis and biological control in B. velezensis KT during the interaction with R. stolonifer was also analyzed by transcriptomics. The results showed that the percentage inhibition of 1 × 109 CFU/mL B. velezensis KT suspension against mycelial growth of R. stolonifer LC-7 was 87.12%, and the percentage of spore germination and germ length of R. stolonifer LC-7 in the presence of B. velezensis KT were 21.54% and 11.45 μm, respectively. At 48 h after being treated with 1 × 109 CFU/mL B. velezensis KT suspension, the incidence of soft rot in cherry fruits was about 20% and was about 80% lower than that in the control group. B. velezensis KT suspension inhibited the activity of polyphenoloxidase (PPO) and lipoxidase (LOX), increased the activity of peroxidase (POD), phenylalanine ammonia lyase (PAL) chitinase (CHI) and β-1,3-glucanase (GLU), and improved disease resistance and antioxidant capacity in cherry fruits. It was observed by scanning electron microscopy that after co-cultured with B. velezensis KT, the morphology of the mycelia and spores of R. stolonifer LC-7 was distorted and malformed. Transcriptomic analysis showed that the expression of pksIJ in the biosynthetic gene cluster for antibiotic bacillaene and tuaAGE in the biosynthetic gene cluster for teichuronic acid were up-regulated, and the differentially expressed genes were mainly concentrated in ribonucleoprotein complex assembly, subunit assembly and ribosomal assembly during biological processes. The B. velezensis KT could directly inhibit R. stolonifer LC-7 probably through nutritional and spatial competition and secreting anti-bacterial substances and could also induce disease resistance in sweet cherry fruits. The results showed that B. velezensis KT had an obvious control effect on sweet cherry soft rot caused by R. stolonifer LC-7, making it a potential biocontrol agent.

Key words: Bacillus velezensis; Rhizopus stolonifer; sweet cherry; soft rot; interaction; antimicrobial mechanism

中图分类号:

TS255.36

张倩，陈雨诗，许春艳，邹曼，包汶婷，郗良卿，辛力，陈义伦. 贝莱斯芽孢杆菌防治甜樱桃采后软腐病的效果和机理[J]. 食品科学, 2023, 44(7): 229-239.

ZHANG Qian, CHEN Yushi, XU Chunyan, ZOU Man, BAO Wenting, XI Liangqing, XIN Li, CHEN Yilun. Efficacy and Action Mechanism of Bacillus velezensis on Controlling Postharvest Soft Rot of Sweet Cherry Fruits[J]. FOOD SCIENCE, 2023, 44(7): 229-239.

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贝莱斯芽孢杆菌防治甜樱桃采后软腐病的效果和机理

Efficacy and Action Mechanism of Bacillus velezensis on Controlling Postharvest Soft Rot of Sweet Cherry Fruits

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