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A Multiphasic Approach for the Identification of Endophytic Bacterial in Strawberry Fruit and their Potential for Plant Growth Promotion

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Abstract

This study used a multiphasic approach, characterized by the simultaneous use of culture-dependent and culture-independent methods, to investigate endophytic bacterial communities in strawberry (Fragaria ananassa) fruit. A total of 92 bacterial endophytes were isolated and initially grouped by their repetitive extragenic palindromic (rep)-PCR banding pattern and biochemical features. Phylogenetic analysis of the 16S rRNA gene sequences of 45 representatives showed that the isolates belonged to the species Bacillus subtilis (eight isolates), Bacillus sp. (seven isolates), Enterobacter sp. (seven isolates), Enterobacter ludwigii (six isolates), Lactobacillus plantarum (six isolates), Pseudomonas sp. (five isolates), Pantoea punctata (three isolates), and Curtobacterium citreum (three isolates). Nucleic acids were extracted from the strawberry fruit and subjected to 16S rRNA gene directed polymerase chain reaction denaturing gradient gel electrophoresis (16S rRNA PCR-DGGE). The species B. subtilis, Enterobacter sp., and Pseudomonas sp. were detected both by isolation and DGGE. The DGGE fingerprints of total bacterial DNA did not exhibit bands corresponding to several of the representative species isolated in the extinction dilution (L. plantarum, C. citreum, and P. punctata). In contrast, bands in the DGGE profile that were identified as relatives of Arthrobacter sp. and one uncultivable Erythrobacter sp. were not recovered by cultivation techniques. After isolation, the nitrogen fixation ability and the in vitro production of indole-3-acetic acid (IAA) equivalents and siderophores were evaluated. A high percentage of isolates were found to possess the ability to produce siderophores and IAA equivalents; however, only a few isolates belonging to the genera Pseudomonas and Enterobacter showed the ability to fix nitrogen. Plant growth promotion was evaluated under greenhouse conditions and revealed the ability of the Bacillus strains to enhance the number of leaves, shoot length, root dry weight, and shoot dry weight. The activity of the bacterial isolate identified as B. subtilis NA-108 exerted the greatest influence on strawberry growth and showed a 42.8% increase in number of leaves, 15.26% for high shoot, 43.5% increase in root dry weight, and a 77% increase in shoot dry weight when compared with untreated controls.

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Acknowledgments

The authors acknowledge Horto Florestal e Plantas Medicinais (UFLA) for providing the strawberry samples for use in this study and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Fundação de Amparo à pesquisa do Estado de Minas Gerais (FAPEMIG) for financial support and Dr. Disney Ribeiro Dias for collaboration in manuscript preparation.

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Authors and Affiliations

  1. Department of Biology, Federal University of Lavras (UFLA), CEP 37200-000, Lavras, Minas Gerais, Brazil

    Gilberto Vinícius de Melo Pereira, Karina Teixeira Magalhães, Thiago Pereira Souza & Rosane Freitas Schwan

  2. Department of Plant Pathology, Federal University of Lavras (UFLA), CEP 37200-000, Lavras, Minas Gerais, Brazil

    Emi Rainildes Lorenzetii

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  1. Gilberto Vinícius de Melo Pereira
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Correspondence to Rosane Freitas Schwan.

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de Melo Pereira, G.V., Magalhães, K.T., Lorenzetii, E.R. et al. A Multiphasic Approach for the Identification of Endophytic Bacterial in Strawberry Fruit and their Potential for Plant Growth Promotion. Microb Ecol 63, 405–417 (2012). https://doi.org/10.1007/s00248-011-9919-3

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