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    生防细菌对油茶炭疽病病原菌的抑制作用

    Inhibition effects of biocontrol bacteria strains on the pathogen of Camellia oleifera anthracnose

    • 摘要:
        目的  从酸浆果实内分离筛选出对油茶炭疽病病原果生刺盘孢具有拮抗作用的内生细菌,并了解该菌对油茶炭疽病病原菌的抑菌机制,以期为油茶炭疽病的田间防治提供理论依据。
        方法  本研究通过组织分离法从果实中分离出细菌14株, 通过平板对峙法筛选得到对油茶炭疽病病原菌具有显著抑制作用的细菌3株,菌株依次命名为DLSB-1、DLSB-4和DLSB-13。结合菌株形态学、生理生化特征以及16S rDNA序列比对分析,对菌株进行了鉴定。在上述试验基础上,判定生防菌株能否产生蛋白酶、纤维素酶和β-1,3-葡聚糖酶,通过显微镜观察3株细菌的无菌上清发酵液对病原菌丝的生长情况,并在室内测定了油茶盆栽苗的防治效果。
        结果  3株细菌无菌发酵上清液均能抑制油茶炭疽病病原菌丝的正常生长,使其尖端膨大,畸变,扭曲。可产生蛋白酶和纤维素酶,不能产生β-1,3-葡聚糖酶;5个梯度的无菌发酵上清液(0.6 × 108、1.0 × 108、1.4 × 108、1.8 × 108、2.2 × 108 cfu/mL)与对照相比,对油茶炭疽病病原菌抑菌效果显著(P < 0.05),抑菌率最高达到81.14%。其中,菌株(DLSB-4)5个梯度的无菌发酵上清液对油茶盆栽苗的防治效果分别达到40.2%、43.9%、57.7%、68.7%和71.1%,差异显著(P < 0.05),且无菌发酵上清液的含量与防治效果呈正相关。
        结论  3株细菌通过自身产生拮抗作用酶来抑制果生刺盘孢病原菌菌丝的生长,降低油茶炭疽病的发病率和病情指数。

       

      Abstract:
        Objective  Endophytic bacteria with antagonistic action on oil tea anthracnose pathogen Colletotrichum fructicola were isolated from the fruit of Physali alkekengi, followed by the study on the antibacterial mechanism of the strain on the pathogen of Camellia oleifera anthracnose. It is expected to provide a theoretical basis for the field control of Camellia oleifera anthracnose.
        Method  In this study, fourteen strains of bacteria were isolated from fruits by tissue separation isolation, and then three strains of bacteria with significant inhibitory effects on the pathogen of Camellia oleifera anthracnose were obtained with plate-screening method, including the strains of DLSB-1, DLSB-4 and DLSB-13. Subsequently, combining strain morphology, physiological and biochemical characteristics and 16S rDNA sequence alignment analysis, the strain was identified. Furthermore, based on the above test, whether the biocontrol strain can produce protease, cellulase and β-1,3-glucanase was determined, along with the observation of the mycelial growth of pathogenic hyphae fungus by antagonistic bacteria sterile fermentative liquid with microscope, and the indoor control effect of Camellia oleifera seedlings was measured indoors.
        Result  It was found that all three strains of Bacillus sterile fermentative liquid could inhibit the normal growth of the pathogenic hyphae of Camellia oleifera anthracnose, thus making it tip-expanding, deformity and distorted. Besides, protease and cellulase could be produced, while β-1,3-glucanase could not be produced; when compared 5 concentrations (0.6 × 108, 1.0 × 108, 1.4 × 108, 1.8 × 108, 2.2 × 108 cfu/mL) of sterile fermentative liquid with control, it was found that the antibacterial effect on the pathogen of Camellia oleifera anthracnose was significant (P < 0.05), with the highest level of 81.14%. Moreover, the control effects of 5 concentrations of sterile fermentative liquid of strain DLSB-4 on potted Camellia oleifera seedlings were 40.2%, 43.9%, 57.7%, 68.7% and 71.1%, respectively. All these show that there is a positive correlation between good control effects as well as the concentration of sterile fermentative liquid and the control effect.
        Conclusion  The three strains of biocontrol bacteria could produce antagonistic enzymes which inhibiting the growth of pathogenic bacteria hyphae of Colletotrichum fructicola, thus reducing the incidence of Camellia oleifera anthracnose.

       

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