湖北省生姜白星病病原菌鉴定与室内药剂筛选

Pathogen identification and indoor fungicides screening for white scab disease of ginger(Zingiber officinale)in Hubei Province

  • 摘要: 【目的】 明确湖北省生姜白星病病原菌种类和生物学特性,室内筛选对病原菌菌丝生长有抑制效果的药剂,为生姜白星病的田间防治提供理论依据。【方法】 生姜白星病病叶采集自湖北省恩施州来凤县生姜种植基地,利用组织分离法对生姜白星病病原菌进行分离纯化,根据柯赫氏法则进行致病性测定,利用特异性引物对病原菌的ITS、EF-1αACTCALTUB2GAPDH序列进行扩增,结合形态特征和分子鉴定结果,确定病原菌的种类;分析病原菌的生物学特性,并采用菌丝生长速率法测定8种常用药剂对病原菌的抑制效果。【结果】 从生姜白星病病叶中分离纯化的真菌经致病性测定后,获得形态一致的病原菌5株。选取代表性菌株23-5进行形态学和分子生物学鉴定,其形态特征与Neoscirrhia matteucciicola一致;经比对分析后选取覆盖率和相似性均较高的参考菌株序列,按照ITS-TUB2拼接序列后构建系统发育树,结果显示菌株23-5与Neoscirrhia matteucciicola聚于同一分支,确定生姜白星病病原菌为Neo‐ scirrhia matteucciicola。病原菌的生物学特性测定结果表明,病原菌致死温度为54℃;菌丝生长较适宜pH为6~8,pH为7时,菌落直径最大,达6.45 cm;病原菌最适碳源为可溶性淀粉,适宜氮源为蛋白胨、酵母提取物和牛肉浸粉。8种供试药剂中,恶醚唑对菌株23-5菌丝生长的抑制效果最佳,抑制中浓度(EC50)为0.05 mg/L。【结论】 确定湖北省生姜白星病病原菌为Neoscirrhia matteucciicola,恶醚唑对该病原菌具有较好的抑制效果,可作为生姜白星病田间防治的候选药剂。

     

    Abstract: 【Objective】 To study the species and biological characteristics of the pathogen causing ginger(Zingiber officinale)white scab disease,and to screen effective fungicides for inhibiting mycelium growth of the pathogen in the laboratory,which could provide basis for field control of ginger white scab disease. 【Method】 Diseased ginger leaves were col‐lected from a ginger plantation in Laifeng County,Enshi Prefecture,Hubei Province. The pathogen was isolated and purified by tissue separation method,and pathogenicity was assessed based on Koch’s rule. Sequences of ITS,EF-1αACTCALTUB2,and GAPDH of the pathogen were amplified using specific primers. The pathogen was identified by combining morphological characteristics and molecular identification. The biological characteristics of the pathogen and the inhibitory effects of 8 commonly used fungicides on mycelium growth were evaluated using the mycelium growth rate method. 【Result】 Five strains of pathogens with consistent morphology were isolated and purified from the diseased ginger leaves. The representative strain 23-5 was selected for morphological and molecular biological identification. Its morphological characteristics were consistent with those of Neoscirrhia matteucciicola;after comparative analysis,the reference strain sequences with high coverage and similarity were selected. The phylogenetic tree was constructed after splicing the ITS-TUB2 sequence. The results showed that strain 23-5 and Neoscirrhia matteucciicola were clustered in the same branch. The pathogen of ginger white scab disease was determined to be Neoscirrhia matteucciicola. The determination results of the biological characteristics of the pathogen indicated that the lethal temperature of the pathogen was 54 ℃;the mycelium growth was more suitable at pH of 6 to 8. When the pH was 7,the colony diameter was the largest,reaching 6.45 cm. The optimal carbon source for pathogen was soluble starch,and the suitable nitrogen sources were peptone, yeast extract and beef extract powder. Among the 8 tested fungicides,difenoconazole had the best inhibitory effect on the mycelium growth of strain 23-5,with an inhibitory medium concentration(EC50)of 0.05 mg/L. 【Conclusion】 This study is the first to identify Neoscirrhia matteucciicola as the pathogen of ginger white scab disease in Hubei Province. Difenoconazole demonstrates a good inhibitory effect on the mycelium growth of this pathogen and can be recommended as a candidate fungicide for field control of ginger white scab disease.

     

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