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Ultrasonic extraction of plant-parasitic nematodes from plant roots

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Abstract

Extraction of nematodes from plant roots is a very time- and labour-intensive process, so new methods offering advantages for either speeding or lessening workloads are highly desired, but must be tested before adoption. This paper reports a novel method for extraction of nematodes using ultrasonic waves that is fast and simple. It is also non-destructive to roots, and whole root systems can be sampled for migratory endoparasitic nematodes. The method was tested and optimized for extraction of Burrowing Nematodes (Radopholus similis) from the roots of the aquatic plant Anubias sp. in water. Different durations of treatment and ultrasonic frequencies were tested, and the number of nematodes and plant damage recorded. The highest extraction efficiencies per sample were obtained after 40 min or longer sonication at a frequency of 40 KHz, but the condition of the extracted nematodes was poor for identification at these long durations. Considering efficiency as number of nematodes extracted per unit time, optimal extraction was obtained with a duration of 20 min at a frequency of 40 KHz. With this duration there was little observable damage to nematodes (allowing use in experiments or inoculation trials), or plants (so that successfully replanting is possible). The ultrasonic method was compared with other extraction techniques, and was much more efficient than root incubation, shaking or mist chamber techniques, but a little less efficient per sample than the maceration-centrifugal-flotation method. The ultrasonic method is suitable for extracting migratory endoparasites and semi-endoparasites from plant roots. If present, ectoparasites are also extracted. Ultrasonic extraction requires less time and effort for root preparation than other methods, so it can be a lower cost method for routine use. Furthermore, it can be incorporated into a simple, cheap, mass-manufactured, portable and automated kit, which can be made widely available, so that the technique can be widely used.

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Acknowledgments

Most of the experimental work was supported by the Thailand Department of Agriculture. Analysis and MS preparation was carried out during laboratory visits and exchanges funded by the ASEAN Regional Diagnostic Network Project of the ASEAN-Australia-New Zealand Free-Trade Agreement (AANZFTA) Economic Cooperation Work Programme and the Crawford Fund. From these organizations we thank Dr Ian Naumann, Ms Wendy Lee and Dr Tony Fischer for their assistance in organizing funding for collaboration visits. We thank Sunil Singh and Natalie Banks for helpful comments on drafts of the MS.

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Authors and Affiliations

  1. Plant Protection Research and Development Office, Department of Agriculture, Bangkok, Thailand

    N. Tangchitsomkid

  2. Kamphaeng Saen Campus, Kasetsart University, Nathom Pathong, Thailand

    T. Chanmalee

  3. CSIRO Ecosystem Sciences, GPO Box 1700, Canberra, ACT 2601, Australia

    M. Hodda

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  1. N. Tangchitsomkid
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  2. T. Chanmalee
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  3. M. Hodda
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Correspondence to M. Hodda.

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Tangchitsomkid, N., Chanmalee, T. & Hodda, M. Ultrasonic extraction of plant-parasitic nematodes from plant roots. Australasian Plant Pathol. 44, 87–96 (2015). https://doi.org/10.1007/s13313-014-0325-4

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  • DOI: https://doi.org/10.1007/s13313-014-0325-4

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