蜜蜂患白垩病虫体内一株球囊菌拮抗细菌的分离与鉴定

doi:10.3864/j.issn.0578-1752.2012.05.019

摘要/Abstract

摘要： 【目的】寻找抑制蜜蜂球囊菌的拮抗菌，用于白垩病的生物防治。【方法】利用细菌纯化技术从患白垩病的蜜蜂幼虫体内分离纯化了一株对蜜蜂球囊菌(Ascosphaera apis)有抑制作用的细菌，并结合形态学、革兰氏染色以及16S rDNA序列分析技术进行鉴定，同时通过蜜蜂体内、体外接种试验研究其对蜜蜂球囊菌的抑制作用。【结果】该细菌被初步鉴定为蜡样芽胞杆菌(Bacillus cereus)，它在培养基中能够完全抑制蜜蜂球囊菌的生长，但其发酵液对蜜蜂球囊菌的生长没有影响。将含菌饲料喂蜜蜂幼虫，蜜蜂幼虫的死亡率和生长发育没有受到影响，说明该细菌对蜜蜂幼虫没有致病性。蜜蜂幼虫体内同时接种细菌和球囊菌孢子，发现细菌对球囊菌在蜜蜂体内的萌发和初期菌丝的生长没有影响，蜜蜂依然能够患病，但生长后期该细菌能够降解球囊菌菌丝，对球囊菌子代孢子的产生表现出一定的抑制作用。【结论】该研究结果为蜜蜂白垩病的生物防治提供了理论依据。

关键词: 白垩病, 蜜蜂球囊菌, 蜡样芽孢杆菌, 拮抗, 分离

Abstract: 【Objective】 The objective of this study is to explore Ascosphaera apis antagonistic bacteria for biological control of honeybee chalkbrood disease.【Method】An antagonistic bacterial strain was isolated from honeybee larvae suffered from chalkbrood disease on PDA medium plate. It was identified by combining of morphology, gram stain and 16S rDNA sequence analysis. Its inhibiting activity to Ascosphaera apis was determined in vitro and in vivo. 【Result】 The bacterium was identified as Bacillus cereus. It could inhibit the growth of Ascosphaera apis in PDA medium, but its fermentation solution could not. The death rate and development of the honeybee larvae was unchanged when feeding diet containing the bacteria suggesting that the strain was not pathogenic to honeybee. By inoculating together with the spore of Ascosphaera apis to honeybee larvae, the bacterium could not inhibit the growth of Ascosphaera apis at early infecting stage, but could effectively degrade the hyphae soon after. Ascosphaera apis could not form the offspring spore, thus lost the capability of spreading. 【Conclusion】This study provides a theoretical basis for biological control of honeybee chalkbrood disease.

Key words: chalkbrood disease, Ascosphaera apis, Bacillus cereus, antagonism, isolation

李江红, 郑志阳, 洪双燕, 齐香凤, 梁勤. 蜜蜂患白垩病虫体内一株球囊菌拮抗细菌的分离与鉴定[J]. 中国农业科学, 2012, 45(5): 973-980.

LI Jiang-Hong, ZHENG Zhi-Yang, HONG Shuang-Yan, QI Xiang-Feng, LIANG Qin. Isolation and Identification of an Antagonistic Bacterial Strain Against Ascosphaera apis from Honeybee Larvae Infected with Chalkbrood Disease[J]. Scientia Agricultura Sinica, 2012, 45(5): 973-980.

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