贝莱斯芽孢杆菌J28的分离鉴定及其对番木瓜炭疽病的防治效果

    Isolation, Identification, and Biocontrol Efficacy of Bacillus velezensis J28 Against Anthracnose Disease of Carica papaya L.

    • 摘要:
      目的 针对采后番木瓜易受炭疽病为害,导致番木瓜果实品质和产量大幅度下降问题,分离筛选对暹罗炭疽菌(Colletotrichum siamense)有抑菌作用的优质生防菌,并探究其对炭疽病的防治效果。
      方法 从广东省湛江市感病番木瓜根际土壤中分离筛选对暹罗炭疽菌具有强拮抗活性的菌株,通过形态观察、理化特性分析及系统发育树分析完成菌株分类鉴定。采用平板对峙法探究菌株对暹罗炭疽菌T3及其他7种病原菌的抑菌效果。进一步对番木瓜果实和幼苗进行接种,验证菌株对炭疽病的防治效果。
      结果 成功筛选到1株拮抗炭疽菌的菌株J28,根据形态特征、理化特性及系统发育树结果,鉴定该菌株为贝莱斯芽孢杆菌(Bacillus velezensis)。菌株J28能水解淀粉和石蕊牛奶、液化明胶、还原硝酸盐,还能利用柠檬酸盐、葡萄糖、蔗糖、麦芽糖,并表现出接触酶活性、参与甲基红和V-P反应的能力。菌株J28具有高度耐盐性,在125 g/L NaCl的LB平板上表现出良好生长能力。菌株J28能抑制暹罗炭疽菌T3菌丝生长,抑制率为61.81%;还能抑制番木瓜胶孢炭疽菌、葡萄座腔菌以及其他5种病原真菌的生长,抑制率在60.28%~70.94%。100%浓度的J28发酵液对暹罗炭疽菌T3萌发抑制率达65.89%。活体接种试验表明,菌株J28在番木瓜果实与叶片中均能高效抑制暹罗炭疽菌T3,防治效果分别为72.35%和93.33%。
      结论 菌株J28具有广谱抑菌能力,对番木瓜炭疽病有良好的防治效果,可为后续耐盐菌种资源挖掘以及开发植物广谱性生物抗菌剂提供理论基础和技术支持。

       

      Abstract:
      Objective To address the significant decline in papaya fruit quality and yield caused by postharvest anthracnose, this study aimed to isolate and screen high-quality antagonistic bacteria, evaluate their antifungal activity against Colletotrichum siamense, and preliminarily investigate their mechanisms of action against anthracnose.
      Method Rhizosphere soil samples were collected from diseased papaya plants in Zhanjiang City, Guangdong Province, to isolate and screen bacterial strains exhibiting strong antagonistic activity against C. siamense. Taxonomic identification of the target strain was performed through morphological observation, physiological and biochemical characterization, and phylogenetic analysis. The antagonistic activity of bacteria against C. siamense T3 and seven pathogens was assayed using the dual-culture plate method. Furthermore, the biocontrol efficacy of the antagonistic bacteria against anthracnose was validated through in vivo inoculation of papaya fruits and leaves.
      Result A biocontrol strain J28 with antagonistic activity against C. siamense T3 was successfully isolated. Morphological characteristics, physiological and biochemical properties, and phylogenetic analysis identified the strain J28 as Bacillus velezensis. The strain J28 was capable of hydrolyzing starch and litmus milk, liquefying gelatin and reducing nitrate. It could also utilize citrate, glucose, sucrose, and maltose, and exhibited catalase activity, as well as the ability to participate in the Methyl Red and Voges-Proskauer reactions. Strain J28 exhibited high salt tolerance, maintaining robust growth capability on LB plates containing 125 g/L NaCl. B. velezensis J28 significantly inhibited the mycelial growth of C. siamense T3 isolated from papaya, with an inhibition rate of 61.81%. Concurrently, strain J28 exhibited broad-spectrum antifungal activity, inhibiting the growth of C. gloeosporioides, Botryosphaeria dothidea, and five other pathogenic fungal species, with inhibition rates ranging from 60.28% to 70.94%. In addition, the fermentation broth of strain J28 inhibited the germination of C. siamense T3 by 65.89% at a concentration of 100%. In vivo inoculation experiments confirmed that B. velezensis J28 effectively inhibited C. siamense in both papaya fruits and leaves, achieving biocontrol efficacies of 72.35% and 93.33%, respectively.
      Conclusion In conclusion, strain J28 possesses broad-spectrum antifungal activity and exhibits significant biocontrol potential against papaya anthracnose. This research provides a theoretical basis and technical support for future exploration of salt-tolerant microbial resources and the development of broad-spectrum biological antifungal agents for plants.