A rapid field-based assay using recombinase polymerase amplification for identification of Thrips palmi , a vector of to
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A rapid field‑based assay using recombinase polymerase amplification for identification of Thrips palmi, a vector of tospoviruses Priti1 · Sumit Jangra1 · V. K. Baranwal1 · Ralf G. Dietzgen2 · Amalendu Ghosh1 Received: 13 March 2020 / Revised: 11 September 2020 / Accepted: 21 September 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Thrips palmi (Thysanoptera: Thripidae) is an important pest of vegetables, ornamentals, and legumes worldwide. Besides damage caused by feeding, it transmits several tospoviruses. Identification of T. palmi at an early stage is crucial in implementing appropriate pest management strategies. Morpho-taxonomic identification of T. palmi based on the adult stage is time-consuming and needs taxonomic expertise. Here, we report a rapid, on-site, field-based assay for identification of T. palmi based on recombinase polymerase amplification (RPA), its first application in insects. RPA primers designed based on 3′ polymorphisms of the Internal Transcribed Spacer 2 region efficiently discriminated T. palmi without any cross-reactivity to other predominant thrips species. RPA was performed with crude DNA, extracted from single T. palmi simply by crushing in sterile distilled water and could be completed within 20 min by holding the reaction tubes in the hand. The assay was further simplified by using fluorescent as well as colorimetric dyes thus eliminating the gel-electrophoresis steps. The presence of T. palmi was visualized by a change in color from dark blue to sky blue. The assay was validated with known thrips specimens and found to be effective in diagnosing the presence of T. palmi in natural vegetation. This on-site, rapid assay for diagnosis of T. palmi can be used by non-expert personnel in the field of quarantine and pest management. Keywords RPA · Melon thrips · Insect vector · Tospovirus transmission · Thrips diagnostics
Key message • Thrips palmi an important pest of vegetables and orna-
mentals transmitting several tospoviruses
• Morpho-taxonomic identification stage-specific, time-
consuming, and needs taxonomic expertise
• Rapid, on-site, field-based assay for identification of T.
palmi using recombinase polymerase amplification
• Assay uses crude extract of T. palmi, completed within
20 min by holding the reaction tubes in hand without use of sophisticated laboratory instruments • The presence of T. palmi visualized by a change in reaction color • Useful for non-expert personnel in field-based identification, quarantine, and adopting suitable pest management strategies
Introduction Communicated by M. Traugott . * Amalendu Ghosh [email protected] 1
Insect Vector Laboratory, Advanced Centre for Plant Virology, ICAR—Indian Agricultural Research Institute, New Delhi 110012, India
Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St. Lucia, QLD 4072, Australia
2
Thrips (order Thysanoptera) are important agricultural pests that cause significant losses in yield and quality of a
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