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Diversity of endophytic bacteria from the cuprophytes Haumaniastrum katangense and Crepidorhopalon tenuis

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Abstract

Haumaniastrum katangense and Crepidorhopalon tenuis are two cuprophytes characteristic of the Katangan Copper Belt flora. We have studied the endophytic bacteria of H. katangense and C. tenuis as a first step to evaluate their potential contribution to plant adaptation to copper excess. Although their number varied considerably from sample to sample, culturable bacteria were found in roots and shoots of most plants. More than 800 isolates were screened for each plant species. Identification of isolates based on the sequence of the 16S rRNA gene, allocated them to 31 taxonomic units, belonging to 17 genera, mainly Proteobacteria. A great proportion of the bacteria were cupro-resistant and often resistant to other metals, especially zinc and cobalt, as well as nickel for the Methylobacterium isolates. Direct PCR amplification of the polymorphic bacterial internal transcribed spacer (ITS) from the plants’ organs DNA revealed a more diverse endophytic community, with more Gram+, among which a Rubrobacteridae that was never found associated with plants before. This work represents the first study of endophytes in Katangan cuprophytes.

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Acknowledgments

A.C.K was supported by a PhD fellowship from the Coopération Technique Belge. C.-L. M. is supported by a grant on the Interuniversity Attraction Pole Programme VI/33 (Belgian Science Policy). The project is supported by a grant from the Fonds de la Recherche Scientifique (FRFC n°2.4558.08). Special thanks are due to M. G. Colinet and Professors P. Meerts and M. Ngongo in charge of the PIC REMEDLU project, for their assistance and advices.

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Authors and Affiliations

  1. Laboratoire de Physiologie et de Génétique Moléculaire des Plantes, Université Libre de Bruxelles, Campus Plaine-CP242, Bd du Triomphe, 1050, Brussels, Belgium

    Alfred Cubaka Kabagale, Claire-Lise Meyer & Nathalie Verbruggen

  2. Institut de Recherches Microbiologiques JMW, Av. E. Gryson, 1, 1070, Brussels, Belgium

    Bertrand Cornu, Françoise van Vliet & Corinne Vander Wauven

  3. Laboratoire de Microbiologie, Université Libre de Bruxelles, Av. E. Gryson, 1, 1070, Brussels, Belgium

    Max Mergeay & Louis Droogmans

  4. Molecular and Cellular Biology, Unit for Microbiology, Belgian Center for Nuclear Energy (SCK*CEN), Boeretang, 200, 2400, Mol, Belgium

    Max Mergeay

  5. Laboratoire de Chimie Organique, Université de Lubumbashi, Katanga, DRC

    Alfred Cubaka Kabagale & Jean-Baptiste Lumbu Simbi

  6. Laboratoire de Physiologie Végétale et de Microbiologie Appliquée, Université Officielle de Bukavu, BP 570 Bukavu, Sud-Kivu, DRC

    Alfred Cubaka Kabagale

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  1. Alfred Cubaka Kabagale
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Correspondence to Corinne Vander Wauven.

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Table S1

Number of plant specimens containing endophytic isolates from the same taxonomic group as the mentioned isolate. +, presence of isolate; −, absence of isolate. n = 7 (H. katangense) and 6 (C. tenuis). Results from Pearson’s chi square exact tests are indicated for each taxon. Isolates were considered for the statistical analysis when they were detected more than once (DOC 34 kb).

Table S2

Distribution between plant species and among organs of the ITS amplicons obtained from more than one plant-extracted DNA sample. +, number of DNA samples from which the sequence was obtained; −, number of samples from which it was not obtained. n = 8 for H. katangense; n = 6 for C. tenuis leaves and stems and n = 4 for C. tenuis roots. No significant differences were found between species. Significant differences among organs are in bold and indicated as follows: **, P < 0.001; *, P < 0.05. Ampl. for amplicon (DOC 47 kb).

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Cubaka Kabagale, A., Cornu, B., van Vliet, F. et al. Diversity of endophytic bacteria from the cuprophytes Haumaniastrum katangense and Crepidorhopalon tenuis . Plant Soil 334, 461–474 (2010). https://doi.org/10.1007/s11104-010-0396-0

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