张溪香芋软腐病病原菌的分离鉴定及其拮抗菌筛选

    Isolation and Identification of Pathogens Causing Soft Rot in Zhangxi Taro and Screening of Its Antagonistic Bacteria

    • 摘要:
      目的 香芋〔Colocasia esculenta (L.) Schott〕是广东地区的重要经济作物之一,近年来韶关张溪地区香芋软腐病发生频繁,严重影响香芋品质与种植效益。为此,本研究旨在探索香芋软腐病绿色防控途径。
      方法 采用组织分离法从香芋病株球茎中分离具有优势生长的菌落,并进行致病性测定。通过菌落形态观察、生理生化特性测定(包括氧化酶、吲哚测定、明胶液化、硝酸盐还原等)及16S rDNA基因序列比对,确定病原菌类型。此外,采用递减稀释法和平板对峙法从健康香芋根际土壤中筛选获得拮抗菌株,对具有明显抑菌活性的菌株进行形态特征、生理生化测定及16S rDNA基因序列分析测定,明确拮抗菌分类地位,并测定其最适生长温度和pH。
      结果 从患软腐病的香芋球茎组织中分离病原菌株,经致病性测定明确菌株B1为香芋软腐病病原菌。结合形态观察、理化测定和分子鉴定,确定该病原菌为方中达迪基氏菌(Dickeya fangzhongdai),最适生长温度为28℃,最适生长pH为5.0。从健康香芋根际土壤中筛选出一株对香芋软腐病菌具有显著抑制效果的生防菌株J1,其在平板对峙试验中形成明显抑菌圈,平均直径约为18.3(± 0.6)mm,经鉴定该菌株为枯草芽孢杆菌(Bacillus subtilis),最适生长温度为37℃,最适生长pH为7.0。
      结论 韶关张溪香芋软腐病的病原菌为方中达迪基氏菌,枯草芽孢杆菌对该菌具有拮抗作用。

       

      Abstract:
      Objective Taro〔Colocasia esculenta (L.) Schott〕is one of the important economic crops in Guangdong region. In recent years, taro soft rot disease has occurred frequently in Zhangxi area, Shaoguan, seriously affecting the quality and planting benefits of taro. Therefore, this study aims to explore the green prevention and control methods of taro soft rot.
      Method Colonies with dominant growth were isolated from the bulbs of diseased plants by tissue isolation method, and pathogenicity was determined. The type of pathogenic bacteria was determined through colony morphology observation, determination of physiological and biochemical characteristics (including oxidase, indole determination, gelatin liquefaction, nitrate reduction, etc.) and 16S rDNA gene sequence alignment. In addition, antagonistic bacteria were screened from the rhizosphere soil of healthy taro by the decreasing dilution method and the plate confrontation method. Morphological characteristics, physiological and biochemical determinations, and 16S rDNA gene sequence analysis were conducted on the strains with obvious antibacterial activity to clarify the taxonomic status of antagonistic bacteria, and their optimal growth temperature and pH were determined.
      Result Pathogenic strains were isolated from the bulb tissues of taro affected by soft rot disease. Pathogenicity determination confirmed that strain B1 was the pathogen of taro soft rot disease. Combined with morphological observation, physicochemical determination and molecular identification, it was determined that the pathogen was Dickeya fangzhongdai, with the optimal growth temperature being 28℃ and the optimal growth pH being 5.0. A biocontrol strain J1 with a significant inhibitory effect on taro soft rot pathogen was screened out from the rhizosphere soil of healthy taro. It formed a distinct inhibition zone, with an average diameter of approximately 18.3 (±0.6) mm in the plate confrontation test. It was identified as Bacillus subtilis, and the optimal growth temperature was 37℃. The optimal pH for growth is 7.0.
      Conclusion The pathogen of soft rot disease of Zhanxi taro in Shaoguan is D. fangzhongdai. B. subtilis has antagonistic effect on this bacteria.