核桃炭疽病生防菌的分离鉴定及生防潜力研究

doi:10.16409/j.cnki.2095-039x.2024.02.058

摘要/Abstract

摘要： 炭疽病是一种对核桃品质和产量造成严重影响的毁灭性病害。在病害防治中，生物防治因其对环境友好、不易产生抗药性和安全性高等优点，有望替代化学防治发挥优势。本研究采用平板对峙法，从核桃叶片内生细菌中筛选出一株对核桃炭疽病菌（包括Colletotrichum gloeosporioides、C. fructicola、C. siamense、C.fioriniae和C. aenigma）具有较强抑制作用的菌株GYX7，该菌株对核桃炭疽病菌抑制率在70.56%～80.78%，镜检发现对峙培养下的炭疽菌菌丝出现畸形和裂解现象。菌株GYX7具有较广的抑菌谱，对其他13种供试植物病原菌也表现出明显的拮抗效果。通过对其形态学、生理生化特性以及16S rDNA和gyrB基因序列进行系统发育分析，鉴定该菌株为贝莱斯芽胞杆菌Bacillus velezensis。对菌株GYX7的抑菌机理研究表明，该菌株可产生对核桃炭疽病菌菌丝生长具有抑制作用的挥发性物质，抑制率最高可达51.39%；浓度为20%的菌株GYX7无菌滤液对果生刺盘孢菌丝生长和孢子萌发具有良好的抑制效果，抑制率分别为82.24%和98.17%；菌株GYX7能产生多种胞外水解酶，如蛋白酶、纤维素酶、果胶酶、β-1，3-葡聚糖酶；该菌株具有编码多种抗菌化合物的基因。此外，离体接种试验结果显示，菌株GYX7能有效降低核桃炭疽病的发病率和严重程度，防效高达86.53%。这些结果表明菌株GYX7在防治核桃炭疽病方面具有较高的潜在应用价值。

关键词: 核桃炭疽病, 贝莱斯芽胞杆菌, 生物防治, 拮抗作用

Abstract: Walnut anthracnose, caused by several Colletotrichum genus species, is recognized as one of the most destructive and economically important diseases, resulting in considerable losses. The implementation of biological control is expected to provide a favorable alternative to chemical methods for disease management, as it is environmentally friendly, less prone to resistance development, and offers high safety levels. In the present study, an endophytic bacteria strain GYX7, isolated from healthy walnut leaves, exhibited the most potent inhibitory activity against the pathogens of walnut anthracnose, including C. gloeosporioides, C. fructicola, C. siamense, C. fioriniae and C. aenigma, with inhibition rates ranging from 70.56% to 80.78%. Microscopic examination indicated that strain GYX7 induced morphological distortions and lysis of the mycelium during confrontation culture. Furthermore, strain GYX7 demonstrated a broad-spectrum antifungal, inhibiting 13 phytopathogens except Pythium aphanidermatum. Based on morphological, physiological, and biochemical characteristics and a phylogenetic tree constructed using 16S rDNA and gyrB gene sequences, strain GYX7 was identified as Bacillus velezensis. The volatile compounds produced by strain GYX7 were found to contribute to its biocontrol capabilities. The aseptic filtrate of strain GYX7 at a 20% concentration significantly inhibited the growth and spore germination of C. fructicola, with inhibition rates of 82.24% and 98.17%, respectively. Enzyme activity assays indicated that strain GYX7 produced various hydrolytic enzymes, including protease, cellulase, pectinase, and β-1,3-glucanase. Additionally, the strain was found to possess genes that encode antimicrobial substances, including surfactin, bacillomycin, fengycin, iturin, bacillaene, bacillibactin, bacilysin, macrolactin and difficidin, as identified through polymerase chain reaction (PCR). In vitro inoculation experiments showed that strain GYX7 effectively reduced the incidence and severity of walnut anthracnose. These findings suggest that strain GYX7 has the potential to be a biocontrol agent for managing walnut anthracnose.

Key words: walnut anthracnose, Bacillus velezensis, biological control, antagonism

中图分类号:

S476

谷玉锌, 王树和, 王盛豪, 刘圣明, 康业斌. 核桃炭疽病生防菌的分离鉴定及生防潜力研究[J]. 中国生物防治学报, 2024, 40(5): 1099-1112.

GU Yuxin, WANG Shuhe, WANG Shenghao, LIU Shengming, KANG Yebin. Isolation and Identification of Endophytic Bacteria from Walnut Leaves and Their Biocontrol Potential against Walnut Anthracnose[J]. Chinese Journal of Biological Control, 2024, 40(5): 1099-1112.

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