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    伦莹莹, 王慧敏, 吕全, 刘会香, 姚洪锡, 姜文忠, 张星耀. 松材线虫病木内天牛蛀道蛹室的优势长喙壳真菌分析[J]. 北京林业大学学报, 2019, 41(6): 102-110. DOI: 10.13332/j.1000-1522.20180157
    引用本文: 伦莹莹, 王慧敏, 吕全, 刘会香, 姚洪锡, 姜文忠, 张星耀. 松材线虫病木内天牛蛀道蛹室的优势长喙壳真菌分析[J]. 北京林业大学学报, 2019, 41(6): 102-110. DOI: 10.13332/j.1000-1522.20180157
    Lun Yingying, Wang Huimin, Lü Quan, Liu Huixiang, Yao Hongxi, Jiang Wenzhong, Zhang Xingyao. Dominantly ophiostomatoid fungus inhabiting tunnels and pupal chambers of Monochamus alternatus in pines infected by Bursaphelenchus xylophilus[J]. Journal of Beijing Forestry University, 2019, 41(6): 102-110. DOI: 10.13332/j.1000-1522.20180157
    Citation: Lun Yingying, Wang Huimin, Lü Quan, Liu Huixiang, Yao Hongxi, Jiang Wenzhong, Zhang Xingyao. Dominantly ophiostomatoid fungus inhabiting tunnels and pupal chambers of Monochamus alternatus in pines infected by Bursaphelenchus xylophilus[J]. Journal of Beijing Forestry University, 2019, 41(6): 102-110. DOI: 10.13332/j.1000-1522.20180157

    松材线虫病木内天牛蛀道蛹室的优势长喙壳真菌分析

    Dominantly ophiostomatoid fungus inhabiting tunnels and pupal chambers of Monochamus alternatus in pines infected by Bursaphelenchus xylophilus

    • 摘要:
      目的松材线虫病与传媒昆虫天牛、伴生真菌的关系密切,研究与松材线虫病关系密切的长喙壳真菌区系,利用生理特征、形态特征和基于多基因序列的系统发育关系对优势种进行准确鉴定,为揭示病害、传媒天牛与真菌的互作关系提供理论基础。
      方法以华东地区松材线虫病老疫区浙江省和新疫区山东省染病的马尾松、黑松树体内松墨天牛幼虫坑道和蛹室样本为对象,分离和鉴定与松材线虫病伴生的可培养真菌区系组成,重点分析长喙壳类真菌。采用组织分离法对伴生菌区系进行分离培养,在培养特征和形态学鉴定的基础上,依据地域、寄主的不同选取代表性菌株,进行基于多位点DNA(ITS和β-tubulin)序列的系统发育分析。
      结果从98份样本内分离获得254株长喙壳类真菌,占所有可培养真菌区系组成的20.6%,为优势类群;培养特性和显微形态学特征支持其中235株属于齿小蠹长喙壳,占92.5%,同时该种下存在明显的菌落和显微形态特征的分化,代表性菌株的生长速率和系统发育分析进一步支持了该种基于表型的鉴定结果。
      结论本研究首次明确了齿小蠹长喙壳是我国松材线虫病伴生真菌区系中的绝对优势种,今后针对该种真菌与传媒昆虫天牛及松材线虫病的关系、病害真菌区系对松材线虫种群扩散流行提供生态缓冲适应性的研究十分必要,也可为病害的科学防控提供新策略。

       

      Abstract:
      ObjectiveCerambycid vectors and fungi are important components in system of pine wilt disease. Illustrating the fungal flora, especially the ophiostomatoid fungi associated with pine wilt disease, to accurately identify the dominant species with combination of their physiological and morphological characteristics and further phylogenetic analysis based on multiple DNA sequences, could provide a solid basis for understanding relationship among the disease, cerambycid vectors and associated fungi.
      MethodThe tunnels and pupal chambers of sawyer beetle Monochamus alternatus (the vector of the nematode pathogen, Bursaphelenchus xylophilus) inside the infected Pinus massoniana and P. thunbergii from newly epidemic areas of Shandong Provinces and historical epidemic areas of Zhejiang Provinces were collected to investigate the ophiostomatoid fungal flora. The tissue isolation method was used to obtain the associated fungi. Based on cultural characteristics and morphological identification, the representative strains were selected according to their geographical and host distribution to conduct a phylogenetic analysis based on the rDNA-ITS and β-tubulin sequences.
      ResultTwo hundred and fifty-four ophiostomatoid fungal strains were isolated from 98 samples, accounting for 20.6% of the fungal flora, indicating the dominant status of the group. According to the cultural and morphological characteristics, 235 strains among them were identified as Ophiostoma ips, which accounted for 92.5% of the ophiostomatoid fungi, giving rise to its predominant status. The species exhibited obvious internal differentiations on colony and micro-morphological characteristics. The temperature dependent growths and phylogenetic analyses based on rDNA-ITS and β-tubulin sequences of representative strains further support the phenotype status of the species.
      ConclusionThis is the first report confirmed by the comprehensive identification that O. ips as the predominantly species extensively and intensively associated with the pine wilt disease involved by both the sawyer beetle and the pine wood nematode in China. The studies on the association mechanism between O. ips and the pine wilt disease, and on the fungus roles reacting on the nematode population dynamics are further necessary, which can lead to new strategies for the effective prevention and control of the disease.

       

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