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炭疽病对核桃不同品种果实附生细菌群落的影响

The Effect of Anthracnose on the Epiphytic Bacterial Community of Fruits from Different Walnut Varieties

  • 摘要:
    目的 明确核桃炭疽病不同抗性品种果面附生细菌群落的差异,为利用有益微生物防控该病害提供依据。
    方法 采用组织分离法、形态学、多基因系统发育分析和致病性测定相结合的方法,分离并鉴定病原菌,验证采集样品的有效性;利用16s 扩增子测序技术分析比较核桃炭疽病不同抗性品种‘香玲’(易感品种)、‘鲁康1号’(抗性品种)、‘秋香’(抗性品种)以及感病与未感病果面附生细菌群落的组成和差异。
    结果 ‘香玲’、‘鲁康1号’、‘秋香’感病果实的病原为Colletotrichum gloeosporioides,健康果实未分离到病原菌;病原菌的入侵对易感和抗性品种果面附生细菌群落的Chao1指数影响显著;与易感品种相比,病原菌的入侵对抗性品种果面细菌群落的Shannon和Pielou’s evenness指数影响不显著;易感与抗性品种健康果实表面优势属分别为Methyloversatilis spp.和罗尔斯通菌属Ralstonia spp.;易感品种感病果面优势属为Methyloversatilis spp.和罗尔斯通菌属Ralstonia spp.,抗性品种感病果面优势属为泛菌属Pantoea spp.;抗性品种‘鲁康1号’感病果实表面富集的特有属为Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium,‘秋香’感病果实表面富集的特有属为Ochrobactrum spp.。 MetagenomeSeq分析结果显示病原菌侵染核桃炭疽病不同品种后,果面富集的高丰度ASVs数量不同,依次为‘鲁康1号’、‘秋香’、‘香玲’,富集的ASVs主要属于欧文菌科(Erwiniaceae)和根瘤菌科(Rhizobiaceae)。
    结论 核桃炭疽病易感与抗性品种果面微生物群落的组成不同,不同抗性品种感病果实表面招募的微生物不同。

     

    Abstract:
    Objective To clarify the differences in epiphytic bacterial communities on the fruit surfaces of walnut varieties with different resistances to anthracnose, and to provide a basis for the prevention and control of this disease using beneficial microorganisms.
    Method The pathogen was isolated and identified through tissue isolation, morphology, multi-gene phylogenetic analysis, and pathogenicity determination to verify the validity of the collected samples. The 16s amplicon sequencing technology was used to analyze and compare the composition and differences of epiphytic bacterial communities on the fruit surfaces of walnut varieties with different resistances to anthracnose, including "Xiangling" (susceptible variety), "Lukang No.1" (resistant variety), "Qiuxiang" (resistant variety), as well as on diseased and healthy fruits.
    Result Colletotrichum gloeosporioides was identified as the pathogen infecting diseased fruits of all three varieties, while no pathogen was isolated from healthy fruits. Pathogen invasion significantly affected the Chao1 index of epiphytic bacterial communities on the fruit surfaces of both susceptible and resistant varieties. Compared with susceptible varieties, the invasion of the pathogen had no significant impact on the Shannon and Pielou’s evenness indices of bacterial communities on the fruit surfaces of resistant varieties. The dominant genera on healthy fruits of susceptible and resistant varieties were Methyloversatilis spp. and Ralstonia spp. respectively. On diseased fruits of susceptible varieties, Methyloversatilis spp. and Ralstonia spp., remained dominant, whereas diseased fruits of resistant varieties were predominantly colonized by Pantoea spp. The unique genus enriched on the surface of diseased fruits of the resistant variety "Lukang No. 1" was Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, and the unique genus enriched on the surface of diseased fruits of "Qiuxiang" was Ochrobactrum spp. The results of MetagenomeSeq analysis showed that after the pathogen infected different walnut varieties with anthracnose, the number of high-abundance ASVs enriched on the fruit surfaces varied, in the order of "Lukang No. 1", "Qiuxiang", and "Xiangling". The enriched ASVs mainly belonged to Erwiniaceae and Rhizobiaceae.
    Conclusion The epiphytic bacterial communities on fruit surfaces differ between walnut varieties susceptible and resistant to anthracnose. Moreover, resistant varieties recruit distinct microbial taxa on diseased fruits, which may contribute to their resistance mechanism.

     

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