Abstract
The native plant microbiome is composed of diverse microbial communities that influence overall plant health, with some species known to promote plant growth and pathogen resistance. Here, we show the antibacterial and growth promoting activities of autoclaved culture metabolites (ACM) from native endophytic bacteria (NEB). These NEB were isolated from a papaya cultivar (var. Cariflora) that is tolerant to bacterial crown rot (BCR) caused by Erwinia mallotivora. In this cultivar, bacterial colonization in tissues recovering from the disease was observed before onset of tissue regeneration or ‘regrowth’. We further isolated and characterized these bacteria and were able to identify two culturable stem NEB related to plant endophytic genera Kosakonia sp. (ex. Enterobacter sp., isolate EBW), and to Sphingomonas sp. (isolate EBY). We also identified root NEB under genus Bacillus (isolates BN, BS, and BT). Inhibition assays indicated that ACM from these NEB promptly (within 18-30 h) and efficiently inhibited (60–65% reduction) E. mallotivora proliferation in vitro. When surface-sterilized papaya seeds were soaked in ACM from isolates EBY and EBW, germination was variably retarded (20–60% reduction) depending on plant genotype, but plant biomass accumulation was significantly stimulated, at around two-fold increase. Moreover, greenhouse experiments show that ACM from all isolates, especially isolate EBW, significantly reduced BCR incidence and severity in a susceptible genotype (var. Solo), at around two-fold. In general, our observations of pathogen antagonism and plant growth promotion leading to disease reduction, suggested the influence of native endophytic bacteria to increased fitness in plants, and tolerance against the re-emerging crown rot disease of papaya.
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Acknowledgements
The authors would like to thank Valeriana Justo, Lorele Trinidad, Ireneo Pangga, Teresita Dalisay, Fe dela Cueva, Demetrio Alvarez, Ramon Cortez, Gil Cueto, Besseluz Abayon and Marcelino Gregorio of the University of the Philippines Los Baños in Laguna, Philippines, and Eka Mirnia of the Indonesian Agency of Agricultural Research and Development in West Sumatra, Indonesia, for their technical support during the conduct of the experiments. The authors would also like to thank Karla Manigbas, and the staff of Diamed Enterprises in Laguna, Philippines for their DNA sequencing logistics services.
Availability of data and Supplementary materials
Sequences of the 16S rRNA gene for each bacterial isolate are deposited in NCBI GenBank, with accession numbers MW405488, MW405489, MW405490, MW405491 and MW405492. All supplementary materials from this study can be downloaded through this link: https://doi.org/10.6084/m9.figshare.13499409.v1
Funding
The authors are very grateful for the support received from two funding institutions: the University of the Philippines Los Baños (UPLB) Basic Research Program (Project number 9116004), and the Australian Centre for International Agricultural Research (ACIAR, Project code HORT/2012/113).
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MPSR and PMM conceptualized the study, and acquired funding. PMM supervised all experiments. Materials preparation, experimental setup and data collection were performed by MPSR, with substantial assistance from EPP and SFMD. MPSR did the analysis of raw data, and wrote the first draft of the manuscript, with all the tables and figures. All authors contributed revisions and proofreading, and approved the final manuscript.
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Rivarez, M.P.S., Parac, E.P., Dimasingkil, S.F.M. et al. Influence of native endophytic bacteria on the growth and bacterial crown rot tolerance of papaya (Carica papaya). Eur J Plant Pathol 161, 593–606 (2021). https://doi.org/10.1007/s10658-021-02345-1
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DOI: https://doi.org/10.1007/s10658-021-02345-1