Abstract
BackgroundTo date, four thrips vectors have been reported to transmit five different tospoviruses in India. Their identification at an early stage is crucial in formulating appropriate pest management strategies. Since morphometric key-based thrips identification based on the adult stage is time-consuming, there is a need to develop diagnostic tools which are rapid, accurate, and independent of developmental stages. Here, we report a multiplex PCR assay to identify four major thrips vectors viz. Thrips palmi, T. tabaci, Scirtothrips dorsalis, and Frankliniella schultzei present in India.ResultsCytochrome oxidase subunit III and internal transcribed spacer region 2 were utilized to design species-specific primers. Of 38 pairs of primers tested, primer pairs AG35F-AG36R, AG47F-AG48R, AG87F-AG88R, and AG79F-AG80R amplified 568 bp, 713 bp, 388 bp, and 200 bp products from the DNA templates of T. palmi, S. dorsalis, T. tabaci, and F. schultzei, respectively at same PCR conditions. The specificity of the primer pairs was validated with a large number of known specimens and no cross-reactivity was observed with other thrips species. The multiplex PCR assay with a cocktail of all the four primer pairs detected four thrips vectors efficiently and could discriminate all of them concurrently in a single reaction.ConclusionThe multiplex PCR reported in this study could identify the major thrips vectors reported in India. The assay will be useful in ascertaining distribution profile of major thrips vectors, disease epidemiology, screening large samples, and quarantine.
Highlights
To date, four thrips vectors have been reported to transmit five different tospoviruses in India
Morphometric and cytochrome oxidase subunit I (COI)-based identification Based on the morphometric keys, four thrips vector species viz. T. palmi, S. dorsalis, T. tabaci, and F. schultzei were identified (Additional file 1: Figure S1) to establish the initial population
T. tabaci adults found on onion were whitish-yellow in color and ocelli were grey with ocellar setae within the ocellar triangle
Summary
Four thrips vectors have been reported to transmit five different tospoviruses in India. Their identification at an early stage is crucial in formulating appropriate pest management strategies. We report a multiplex PCR assay to identify four major thrips vectors viz. Thrips palmi, T. tabaci, Scirtothrips dorsalis, and Frankliniella schultzei present in India. 16 thrips vectors transmitting more than 29 tospoviruses have been reported [1,2,3,4]. Four thrips vectors viz. Thrips palmi Karny [5, 6], Scirtothrips dorsalis Hood [7], T. tabaci Lindeman [8], and Frankliniella schultzei Trybom [7] are reported to transmit five tospoviruses in India [4]. F. occidentalis (Pergande) has been recently reported in India [9, 10], its distribution and role in spreading tospoviruses under Indian conditions are yet to be studied
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