炭疽病是茶树的重要病害之一,对茶叶的生长和产量造成严重的危害。目前该病的防治主要依靠化学药剂,开发生物防治产品是推广茶树绿色防控技术主要措施之一。用生理生化特征分析和分子生物学技术鉴定一株茶树根际生防菌JT68,评估其对茶炭疽病菌的抑菌效果以及菌液对菌丝生长和孢子萌发的影响。用对扣法检测该菌株的挥发性有机物对茶炭疽菌的抑制效果,并鉴定有机物的成分。用离体叶片接种的方法测试了菌液对炭疽病的防效。鉴定结果表明,菌株JT68为解淀粉芽孢杆菌。JT68菌液对茶炭疽病菌的平板抑制率为80.94%,对孢子萌发抑制率为99.18%。其发酵液富含细胞壁降解酶,处理菌丝后使病菌菌丝萎缩变形和产生厚垣孢子。JT68菌株产生的挥发性有机物对茶炭疽菌的抑菌率为50.73%,GC-MS检测发现挥发物中主要含有酮类物质。离体接种结果表明,菌液稀释100倍、稀释10倍和原液对炭疽菌相对抑制率分别为72.66%、79.70%和83.20%。抑菌谱试验表明该菌株对稻瘟病、香蕉枯萎病、辣椒炭疽病菌、希金斯炭疽菌、大丽轮枝菌和齐整小核菌等6种植物病原菌抑菌率达到70.0%~93.2%。本研究鉴定的茶树根际解淀粉芽孢杆菌,具有优越的防治病害效果,目前已经投入生产,用于制备茶树专用叶面肥并推广应用,该产品的使用可以替代或减少化学农药的使用,实现茶树的绿色防控。
Abstract
Tea anthracnose (Colletotrichum gloeosporioides) is one of the most important fungal diseases of Camellia sinensis worldwide, which causes serious damage to tea growth and production. The control of tea anthracnose is mainly dependent on chemical fungicides. To promote green prevention and control in tea plantation, development of biocontrol agents is critically important. A bacterium named JT68 isolated from tea rhizosphere was identified based on physiological and biochemical analysis and PCR. The inhibition effects of fermented broth JT68 on confrontation culture, mycelia growth and spores germination were determined. The effect of volatile organic compounds (VOCs) of JT68 was tested and the components were identified by GC-MS. The control effect of JT68 was determined using detached leaf method. The results of this study shows that the strain JT68 was identified as Bacillus amyloliquefaciens. Plate confrontation shows that the inhibition rate of fermentation broth of JT68 against C. gloeosporioides was 80.94%. Co-culture shows that the inhibition rate of spore germination of C. gloeosporioides was 99.18%, and the mycelia of pathogen shrank and formed chlamydospores. The VOCs of JT68 could inhibit 50.73% mycelia growth of C. gloeosporioides. Ketones such as 2-Nonanone and 2-Undecanone were revealed as major components in VOC through GC-MS analysis. Leaf detached inoculation shows that the relative inhibition rates of the original fermentation broth, 10-fold, and 100-fold dilutions were 83.20%, 79.70% and 72.66%, respectively. Furthermore, our study found that JT68 strongly inhibited the growth of Magnaporthe grisea, Fusarium oxysporum, C. capsici, C. higginsianum, Verticillium dahlia and Sclerotium rolfsii with the inhibition rates of 70.0%- 93.2%. Our study provided a biocontrol agent B. amyloliquefaciens from tea rhizosphere, which showed superior biocontrol effect against C. gloeosporioides. This strain had been applied to develop biofertilizer and widely used in the field, which would reduce the use of chemical fungicide and implement prevention and control in tea plantation.
关键词
茶树炭疽菌 /
解淀粉芽孢杆菌 /
生物防治 /
抑菌作用
Key words
Bacillus amyloliquefaciens /
biological control /
inhibition effect /
tea anthracnose
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基金
广东省自然科学基金（2021A1515012091）、广州市科技计划（202102020290）、广东省重点领域研发计划（2019B020218009）
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