引发中国梨树枯死病的病原Fusarium cugenangense的鉴定

doi:10.1016/j.jia.2024.02.018

摘要/Abstract

摘要： 近年来，江苏多地梨园出现了一种未知病因的梨树枯死病。田间调查发现，该病害最初侵染梨树的根部，导致根部组织出现褐变。随着病程的发展，褐变症状逐渐向地上部分延伸，树干部分树皮褐变，并沿树体呈微螺旋状向上扩展，甚至高达地上两米以上。树干褐变部位的枝条枯萎死亡，而未褐变部位的枝条仍然保持正常生长。该病害引发的这些症状与目前已知的梨树病害均不相同。此外，该病害不仅会导致病树在显症的当年或次年死亡，而且还会向邻近梨树传播，对梨园构成了严重威胁。为了明确病因，我们在本研究中开展了病原物的分离、形态和分子鉴定以及柯赫氏法则验证等实验。基于菌落形态和产孢特征的观察，以及利用翻译延伸因子1-α（tef1）、钙调素（CaM）和RNA聚合酶Ⅱ第二大亚基（rpb2）基因序列进行的多基因系统发育分析，我们将病原分离物鉴定为Fusarium cugenangense。通过人工接种实验证实了F. cugenangense对梨树的致病性。此外，我们还构建了GFP荧光标记的F. cugenangense菌株，使用该菌株接种梨苗根部后，利用荧光显微镜和透射电子显微镜均能观察到病原菌在茎和叶组织中的定殖。从接种后发病的茎和叶中均能重新分离得到接种于根部的病原物，证明了该病原菌能对梨树产生系统性侵染。本文首次报道了 F. cugenangense 能够侵染梨树引发梨树枯死病，本文的研究结果为我国梨产区在该病害的病原鉴定、症状识别和制定有针对性的防控措施方面提供了理论依据。

Abstract: In recent years, an unusual wilt disease affecting Pyrus pyrifolia has been observed in various regions of Jiangsu, China. This disease originates from the roots and progresses with distinctive browning patterns along vascular tissues, even extending over two meters above the ground. These symptoms set it apart from recognized pear diseases and typically lead to the death of affected trees within the same or the following year. Furthermore, this disease exhibits a tendency to spread to neighboring trees even after the removal of affected trees, presenting a substantial threat to pear production. To ascertain the causative agent, the present study encompassed pathogen isolation, morphological and molecular identification, as well as validation experiments adhering to Koch's postulates. The fungal isolates obtained were identified as Fusarium cugenangense based on characteristics of the colonies and conidia, in addition to a phylogenetic analysis using DNA sequences of the translation elongation factor 1-alpha (tef1), calmodulin (CaM), and RNA polymerase second largest subunit (rpb2) genes. Pathogenicity of the isolated F. cugenangense on pear was confirmed by artificial inoculation. By introducing GFP-labeled pathogens into the roots, colonization in stem and leaf tissues was observed via fluorescence microscopy and transmission electron microscopy. Furthermore, these pathogens were successfully reisolated from stems and foliage, conclusively providing evidence of systemic infection within the pear plants. To the best of our knowledge, this is the first report of F. cugenangense causing pear wilt disease in China.

Key words: Fusarium cugenangense , , pear , , , root rot , , , necrosis , , , infection

. 引发中国梨树枯死病的病原Fusarium cugenangense的鉴定[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.02.018.

Chaohui Li, Xiaogang Li, Weibo Sun, Yanan Zhao, Yifan Jia, Chenyang Han, Peijie Gong, Shutian Tao, Yancun Zhao, Fengquan Liu. Identification of Fusarium cugenangense as a causal agent of wilt disease on Pyrus pyrifolia in China[J]. Journal of Integrative Agriculture, DOI: 10.1016/j.jia.2024.02.018.

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