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Bacillus subtilis and Enterobacter cloacae endophytes from healthy Theobroma cacao L. trees can systemically colonize seedlings and promote growth

  • Applied microbial and cell physiology
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Abstract

Clonal genotypes resistant to fungal diseases are an important component of the cocoa production system in southeastern Bahia state (Brazil), so that technologies for faster production of stronger and healthier plantlets are highly desirable. In this study, the effects of inoculated bacterial endophytes isolated from healthy adult cacao plants on seedlings, and aspects related to inoculation methods, colonization patterns, and photosynthesis were investigated. Sequencing of 16S rRNA, hsp-60, and rpo-B genes placed the wild-type isolates within the species Enterobacter cloacae (isolates 341 and 344) and Bacillus subtilis (isolate 629). Spontaneous rifampicin-resistant (rifR) variants for 344 were also produced and tested. Endophytic application was either by immersion of surface sterilized seeds in bacterial suspensions or direct inoculation into soil, 20 days after planting non-inoculated seeds into pots. Results from in vitro recovery of inoculated isolates showed that the wild-type endophytes and rifR variants systemically colonized the entire cacao seedlings in 15–20 days, regardless of the inoculation method. Some endophytic treatments showed significant increases in seedlings’ height, number of leaves, and dry matter. Inoculation methods affected the combined application of endophytes, which maintained the growth-promotion effects, but not in the same manner as in single applications. Interestingly, the 344-3.2 rifR variant showed improved performance in relation to both the wild type and another related variant. Photosynthetic rates and stomatal conductance increased significantly for some endophytic treatments, being partially associated with effects on growth and affected by the inoculation method. The results suggest that E. cloacae and B. subtilis endophytes from healthy adult plants (not transmitted by seeds) were able to promote vegetative growth on cacao seedlings. The development of products for large-scale use in seedlings/plantlets production systems was discussed.

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Acknowledgments

The authors are grateful to Dr. George Andrade Sodré for great technical assistance and suggestions with greenhouse experiments and soil analysis. This work was supported by grants from the International Foundation of Science (IFS), CEPLAC, FAPESB, and UESC. H.A.C. Leite and A.B. Silva were supported by Masters Fellowships from FAPESB and CNPq, respectively.

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Authors and Affiliations

  1. Departamento Ciências Biológicas (DCB), Universidade Estadual de Santa Cruz (UESC), Rod. BR 415, Km 16, Ilhéus, BA, 45662-000, Brazil

    Hianna Almeida Câmara Leite, Anderson Barbosa Silva, Fábio Pinto Gomes & Leandro Lopes Loguercio

  2. Centro de Pesquisas do Cacau (CEPEC/CEPLAC), Rod. BR 415, Km 22, Cx. Postal 07, Ilhéus, BA, 45600-970, Brazil

    Karina Peres Gramacho

  3. Departamento Ciências Exatas e Tecnológicas (DCET), Universidade Estadual de Santa Cruz (UESC), Rod. BR 415, Km 16, 45662-000, Ilhéus, BA, Brazil

    José Cláudio Faria

  4. Universidade Federal do Recôncavo da Bahia (UFRB), Centro de Ciências Agrárias, Ambientais e Biológicas (CCAAB), Cruz das Almas, BA, 44380-000, Brazil

    Jorge Teodoro de Souza

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  1. Hianna Almeida Câmara Leite
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  2. Anderson Barbosa Silva
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Correspondence to Leandro Lopes Loguercio.

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Hianna Almeida Câmara Leite and Anderson Barbosa Silva contributed equally to this work.

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Leite, H.A.C., Silva, A.B., Gomes, F.P. et al. Bacillus subtilis and Enterobacter cloacae endophytes from healthy Theobroma cacao L. trees can systemically colonize seedlings and promote growth. Appl Microbiol Biotechnol 97, 2639–2651 (2013). https://doi.org/10.1007/s00253-012-4574-2

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