柑橘黄龙病植株内生菌PLFAs多态性研究

Plant endophyte PLFAs polymorphism in Huanglongbing-affectedred pomelo plant

  • 摘要: 采用磷脂脂肪酸(PLFAs)生物标记法, 分析柑橘黄龙病株不同部位及健康状态叶片内生菌群落结构。结果表明, 从柑橘黄龙病植株叶片共检测到42种PLFAs, 其中完全分布的生物标记有9种, 不完全标记有33种。对PLFAs进行聚类分析表明, 柑橘黄龙病株不同部位叶片内生菌PLFAs可以分为2大类群, 类群I特点为脂肪酸生物标记均为不完全分布, 类群Ⅱ的特点为脂肪酸生物标记在被检测的柑橘黄龙病植株的大部分叶片中均有分布。柑橘黄龙病株不同部位及健康状态叶片内生菌PLFAs组成及含量存在差异。总体来看, 不同朝向中, 东面叶片内生菌PLFAs含量最大; 不同高度中, 下部叶片内生菌PLFAs含量最大; 不同健康状态中, 带有黄龙病原叶片内生菌PLFAs含量比健康植株叶片更大。此外, 不同朝向中, 南面叶片真菌/细菌值最大, 革兰氏阳性菌和革兰氏阴性菌的比值(G+/G- )较大; 不同高度叶片中, 真菌/细菌值差异不明显, G+/G- 值差异明显, 上部最大, 下部次之, 中部最小; 不同健康状态中, 健康叶片的真菌/细菌值高于黄龙病株叶片。对柑橘黄龙病株不同部位及健康状态叶片内生菌多样性研究表明, 不同朝向和不同健康状态的叶片内生菌种群多样性指数差异显著, 不同高度的叶片内生菌种群多样性指数差异不显著。主成分分析表明, 主成分1和主成分2基本上能把柑橘黄龙病株不同朝向叶片内生菌种群区分开来。对柑橘黄龙病株不同部位及健康状态叶片进行聚类, 结果表明带有黄龙病原的叶片聚为一类, 不带黄龙病原的健康叶片分聚为两类, 其中, 不同朝向的叶片分聚在不同的亚类群中, 表明柑橘黄龙病株内生菌PLFAs分布与叶片健康状态和叶片朝向均有紧密关系, 与叶片健康状态关系更为密切。

     

    Abstract: Huanglongbing (HLB, a yellow shoot disease) is the most destructive citrus disease in the world. Plant endophyte communities of red pomelo have been associated with HLB. It was therefore important to investigate the endophyte community of red pomelo plant in relation to HLB. In this paper, endophyte community structures in different spatial positions and healthy conditions of HLB-affected red pomelo plants were analyzed using phospholipid fatty acids PLFAs biomarkers based at Sherlock MIS (MIDI Inc.). Based on the results, 42 PLFAs were detected and 9 of them had wild distributions across all the samples while the other 33 were mainly distributed in different leaf orientations. Cluster analysis showed that PLFAs in different leaves in space existed into two community groups. While PLFAs in the group I belonged to an incomplete distribution, PLFAs in group Ⅱ were distributed almost in all samples. There existed some differences in PLFAs content and composition in leaves of HLB-affected red pomelo plants in different spatial positions and health conditions. When compared among each other, it was noted that PLFAs content in east-oriented leaves were maximum among different orientations. The same was true for lower parts leaf in PLFAs content. Healthy leaves contain more PLFAs than HLB-affected leaves. Furthermore, south-oriented leaves had the highest fungi/bacteria PLFAs ratio and also higher G+/G- (gram-positive bacteria/gram-negative bacteria) PLFAs ratio than north/west-oriented leaves. There was no significant difference in fungi-to-bacteria PLFAs ratio at different leaf levels. However, significant differences existed in PLFAs G+/G- ratio in different height leaves. While the highest ratio occurred in the leaves at the upper parts of plant, the lowest ratio was in leaves at the middle parts of plant. Fungi-to-bacteria PLFAs ratio was higher in healthy leaves than that in HLB-affected leaves. Diversity indexes of Shannon, Simpson and Pielou of endophyte communities among leaves with different orientations and healthy conditions were significantly different. However, no apparent differences were noted among different heights leaves. Principal component analysis showed that the first and second principal components accounted for a total of 82.99% of the variation, which may distinguish endophyte communities in leaves with different orientations. Cluster analysis divided healthy leaves in two groups, and HLB-affected leaves in the other group. The results indicated that the distribution of PLFAs was related with health conditions and orientations of leaves, and was more closely in the former than the latter.

     

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