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Identification of spider-mite species and their endosymbionts using multiplex PCR

Flore Zélé1, Mylène Weill2, Sara Magalhães3

  • 1Centre for Ecology, Evolution and Environmental Changes (cE3c), Faculdade de Ciências da Universidade de Lisboa, Edificio C2, Piso-3 Campo Grande, 1749016, Lisbon, Portugal. fezele@fc.ul.pt.

Experimental & Applied Acarology
|February 14, 2018
PubMed

Insights

New multiplex PCR methods quickly identify common spider mite species (Tetranychidae) and their endosymbiotic bacteria. This cost-effective technique works on entire mites without DNA extraction, aiding pest management and research.

Area of Science:

  • Agricultural Entomology
  • Molecular Diagnostics
  • Microbial Ecology

Background:

  • Spider mites (Tetranychidae) are significant agricultural pests, with several species coexisting in regions like the Mediterranean.
  • Morphological identification of these mite species is challenging.
  • Endosymbiotic bacteria harbored by spider mites can influence their host's biology, necessitating their identification.

Purpose of the Study:

  • To develop cost-effective, multiplex diagnostic methods for rapid identification of key spider mite species and their endosymbionts.
  • To enable simultaneous detection of multiple mite species and symbionts in field populations and laboratory samples.
  • To provide a reliable tool for pest management and research involving Tetranychidae.

Main Methods:

  • Development and multiplexing of PCR primers for identifying three common spider mite species: *Tetranychus urticae*, *T. evansi*, and *T. ludeni*.
  • Development of a second set of multiplex PCR primers to detect common spider mite endosymbionts (*Wolbachia*, *Cardinium*, *Rickettsia*), including a generalist mite primer for quality control.
  • Application of methods to entire mites without prior DNA extraction, allowing detection of mixed species populations at low frequencies.

Main Results:

  • Successful multiplex PCR assay for simultaneous identification of *T. urticae*, *T. evansi*, and *T. ludeni*.
  • Demonstrated ability to detect multiple mite species in pooled samples, even when one species is present at a low frequency (1/100).
  • Validated multiplex PCR for detecting *Wolbachia*, *Cardinium*, and *Rickettsia* in spider mites, alongside a control primer.

Conclusions:

  • The developed multiplex PCR methods offer a simple, cost-effective, and reliable approach for identifying spider mite species and their endosymbionts.
  • These methods are suitable for analyzing natural field populations and detecting contaminations in laboratory cultures.
  • The diagnostic strategy can be readily extended to identify other spider mite species and their associated microbial communities.
Keywords:
CardiniumMultiplex diagnosis toolRickettsiaSpecies identificationTetranychidaeWolbachia

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