贵州多花黄精根腐病病原菌鉴定及生防菌筛选

    Identification of Pathogen Causing Root Rot in Polygonatum cyrtonema Hua and Screening of Biocontrol Bacterias

    • 摘要:
      目的 明确贵州多花黄精根腐病病原菌,并筛选出对该病原菌有生防活性的生防菌,为多花黄精根腐病的诊断及绿色防控提供依据。
      方法 对贵州省台江县的感病多花黄精块茎进行分离纯化、致病性验证,并结合形态学特征和分子生物学技术进行病原菌鉴定。利用平板对峙试验筛选出抑菌活性高的生防菌株,综合培养形状、形态学特征和系统发育树分析,明确其分类学地位,并测定其发酵液对病原菌菌丝生长的抑制作用。
      结果 从感病多花黄精样品中分离出28株真菌,其中12株可导致明显的根腐病症状,2GF1导致的根腐病病状最为明显。形态学特征和系统发育树分析结果显示,鉴定出的12株病原菌为尖孢镰刀菌(Fusarium oxysporum)。平板对峙试验结果表明,真菌中生防效果较好的为菌株WHD01、抑制率95.21%;细菌中生防效果较好的为菌株WHD04、抑制率78.99%。结合形态学和分子生物学将WHD01、WHD02、WHD03、WHD04分别鉴定为绿木霉(Trichoderma virens)、大伏革菌(Phlebiopsis gigantea)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、贝莱斯芽孢杆菌(Bacillus velezensis)。发酵液浓度为200 mL/L时,WHD01、WHD02、WHD03、WHD04对病原菌菌丝生长的抑制效果分别为32.70%、26.72%、33.05%和33.52%。
      结论 明确了贵州省台江县多花黄精根腐病的病原菌为尖孢镰刀菌,并筛选到具有生防作用的4株生防菌株,可为多花黄精根腐病的绿色防治提供理论依据。

       

      Abstract:
      Objective The objective of this study was to identify the pathogen causing for root rot in Polygonatum cyrtonema Hua in Guizhou Province and to screen for biocontrol bacterias capable of effectively antagonizing this pathogen. Ultimately, this research aims to establish a foundation for environmentally friendly control strategies against P. cyrtonema Hua root rot.
      Method Diseased tubers of P. cyrtonema Hua were collected from Taijiang County, Guizhou Province. These samples underwent a sreices of procedures including isolation, purification, pathogenicity verification and identification. Both morphological and molecular biology techniques were employed for the identification process. Strains exhibiting significate inhibitory activity were singled out through dual-culture tests. Based on the analysis of culture shape, morphological traits and phylogenetic tree, its taxonomic status was clarified, and the inhibitory effect of its fermentation broth on the mycelial growth of pathogenic bacterias was determined.
      Result Out of 28 fungi isolated from the diseased samples, 12 were found to induce evident symptoms of root rot. Among these, strain 2GF1 exhibited the most severe disease symptoms. Both morphological traits and phylogenetic tree analysis confirmed that the all 12 pathogens belonged to Fusarium oxysporum. In the dual-culture tests, the biocontrol strain WHD01 demonstrated the most potent antagonistic effect with an inhibition rate of 95.21%. Additionally, the bacterial strain WHD04 also displayed a significant antagonistic effect with an inhibition rate of 78.99%. Based on morphological and molecular identification, the strains WHD01, WHD02, WHD03, and WHD04 were identified as Trichoderma virens, Phlebiopsis gigantea, Bacillus amyloliquefaciens, and Bacillus velezensis, respectively. The inhibitory effects on the growth of pathogenic mycelia at a fermentation broth concentration of 200 mL/L were as follows: 32.70% (Trichoderma viridans), 26.72% (Phlebiopsis gigantea), 33.05% (Bacillus amyloliquefaciens) and 33.52% (Bacillus Velez), respectively.
      Conclusion This study successfully identified F. oxysporum as the causative pathogen of root rot in P. cyrtonema Hua in Taijiang County, Guizhou Province. Moreover, four biocontrol bacterias with effective antagonistic effects against this pathogen were identified. These findings lay a the groundwork for the development of eco-friendly strategies for managing of root rot in P. cyrtonema Hua.

       

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