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Isolation and molecular characterization of five entomopathogenic nematode species and their bacterial symbionts from eastern Australia

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Abstract

Entomopathogenic nematodes (EPNs) are used in biological control of pest insects but their potential may be limited by strain availability from different bioregions and effectiveness against specific pests. Here, we isolated and characterized EPNs and their symbiotic bacteria from Australia where their diversity is scarcely known. We collected 198 soil samples from citrus orchards, grasslands and forests across temperate, subtropical and tropical eastern Australia. EPNs were isolated by baiting with mealworm, greater wax moth and Queensland fruit fly, The Australia’s most significant horticultural pest. We obtained 36 isolates which, according to DNA sequence analyses, represented five species, Heterorhabditis bacteriophora, Heterorhabditis indica, Heterorhabditis marelatus, Heterorhabditis zealandica and Steinernema feltiae, including the first report of H. marelatus from Australia, and H. indica and H. zealandica from New South Wales. Thirty-five isolates were baited with mealworm, one with fruit fly, and none with wax moth. Heterorhabditis marelatus was recovered from forests, H. bacteriophora from citrus orchards, S. feltiae from citrus orchards and grasslands, H. indica and H. zealandica from all three habitats. According to bacterial DNA analyses, Photorhabdus heterorhabditis occurred in H. zealandica and a reference strain of H. bacteriophora, Photorhabdus laumondii in H. bacteriophora and H. marelatus, Photorhabdus tasmaniensis in H. indica and H. bacteriophora, and Photorhabdus namnaonensis in H. zealandica. Unexpectedly, Pseudomonas protegens and Delftia acidovorans were found in S. feltiae while its expected symbiont Xenorhabdus remained undetected, possibly due to our approach. The newly isolated EPNs should be tested as biological control agents against pest insects.

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Data availability

All sequence data have been deposited in GenBank (NCBI). All other data are contained within the manuscript and supplementary material.

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Acknowledgements

We thank Geraldine Tilden for technical support with fruit fly rearing, Alexander Robertson, Giles Ross, Alihan Katlav and Kylie Baker for help with field sampling, Michael Duncan for supply of wax moths and bees wax, and Roy Akhurst and Ian Broughton for advice.

Funding

This research was supported by the Australian Research Council Industrial Transformation Training Centre (ARC-ITTC) Fruit Fly Biosecurity Innovation (IC150100026), with a PhD scholarship to SA, and the Department of Agriculture, Water and the Environment’s Strengthening Australia’s Fruit Fly System Research Program; project: A national biocontrol program to manage pest fruit flies in Australia (4-EKSH327).

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Authors and Affiliations

  1. Hawkesbury Institute of the Environment, Western Sydney University, Penrith, NSW, 2751, Australia

    Sitaram Aryal, Uffe N. Nielsen & Markus Riegler

  2. Department of Biological Sciences, College of Science and Mathematics, Mindanao State University-Iligan Institute of Technology, Iligan City, Philippines

    Nanette H. Sumaya

  3. Department of Agriculture and Fisheries (DAF), Mareeba, QLD, Australia

    Stefano De Faveri

  4. Ecogrow Environment Pty Ltd., P.O. Box 241, Westgate, NSW, 2048, Australia

    Craig Wilson

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  1. Sitaram Aryal
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  2. Uffe N. Nielsen
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  3. Nanette H. Sumaya
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  4. Stefano De Faveri
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  5. Craig Wilson
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  6. Markus Riegler
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Contributions

SA, MR and UNN conceptualized and designed the experimental work. SA and MR collected material with support of SDF. CW provided additional material. SA performed the experiments, collected and analyzed the data, under guidance of MR and UNN, and advice of NHS and CW. MR was responsible for research funding. SA wrote the manuscript together with MR and UNN and with input of all other authors. All authors agree with the submission of the manuscript.

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Correspondence to Sitaram Aryal or Markus Riegler.

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Conflict of interest

The authors do not have a conflict of interest, except that CW works at Ecogrow, a company that supplied three EPN isolates which were characterised as part of this study.

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Handling Editor: Ralf-Udo Ehlers

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Aryal, S., Nielsen, U.N., Sumaya, N.H. et al. Isolation and molecular characterization of five entomopathogenic nematode species and their bacterial symbionts from eastern Australia. BioControl 67, 63–74 (2022). https://doi.org/10.1007/s10526-021-10105-7

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