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基于定量聚合酶链反应的定量分析揭示了叶片内生定殖水平具有高度的植物宿主特异性。

qPCR-based quantification reveals high plant host-specificity of endophytic colonization levels in leaves.

作者信息

de Paula Caio César Pires, Bárta Jiří, Borovec Jakub, Frouz Jan, Rychtecký Pavel, Sirová Dagmara

机构信息

Biology Centre CAS, Institute of Hydrobiology, Na Sádkách 702/7, České Budějovice, 37005, Czech Republic.

University of South Bohemia, Faculty of Science, Branišovská 1645/31a, České Budějovice, 37005, Czech Republic.

出版信息

Am J Bot. 2025 Jan;112(1):e16448. doi: 10.1002/ajb2.16448. Epub 2024 Dec 16.

Abstract

PREMISE

Despite the high functional importance of endophytes, we still have limited understanding of the biotic and abiotic factors that influence colonization of plant hosts along major ecological gradients and lack quantitative estimates of their colonization extent. In this study, we hypothesized that the developmental stage of the ecosystem will affect the levels of bacterial and fungal endophytic assemblages in the foliar endosphere.

METHODS

We quantified levels of bacterial and fungal endophytes in leaves of four plant hosts at four stages of vegetation succession using an optimized qPCR protocol with bacteria-specific 16S and fungi-targeting primers.

RESULTS

(1) The ecosystem developmental stage did not have a significant effect on the colonization levels of bacterial or fungal endophytes. (2) Colonization levels by bacterial and fungal endophytes were governed by different mechanisms. (3) Endophytic colonization levels and their relationship to foliar tissue stoichiometry were highly host specific.

CONCLUSIONS

Quantifying colonization levels is important in the study of endophytic ecology, and the fast, relatively low-cost qPCR-based method can supply useful ecological information, which can significantly enhance the interpretation potential of descriptive data generated, for example, by next-generation sequencing.

摘要

前提

尽管内生菌具有高度的功能重要性,但我们对影响植物宿主在主要生态梯度上定殖的生物和非生物因素的了解仍然有限,并且缺乏对其定殖程度的定量估计。在本研究中,我们假设生态系统的发育阶段会影响叶内圈中细菌和真菌内生菌组合的水平。

方法

我们使用优化的qPCR方案,采用细菌特异性16S引物和真菌靶向引物,对四个植物宿主在植被演替四个阶段的叶片中的细菌和真菌内生菌水平进行了定量。

结果

(1)生态系统发育阶段对细菌或真菌内生菌的定殖水平没有显著影响。(2)细菌和真菌内生菌的定殖水平受不同机制控制。(3)内生菌定殖水平及其与叶组织化学计量的关系具有高度宿主特异性。

结论

在内生菌生态学研究中,定量定殖水平很重要,基于qPCR的快速、相对低成本的方法可以提供有用的生态信息,这可以显著提高例如由下一代测序产生的描述性数据的解释潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ff8/11744438/23987e5da595/AJB2-112-e16448-g004.jpg

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