A Photo-Based Key of Thrips (Thysanoptera) Associated with Horticultural Crops in Florida

Cluever, Jeffrey D.; Smith, Hugh A.

doi:10.1653/024.100.0208

Cited by 23 publications

(16 citation statements)

References 54 publications

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“…The initial population of T. palmi was collected from a stock population maintained at Advanced Centre for Plant Virology, Indian Agricultural Research Institute (IARI), New Delhi. The identity was confirmed by morphometric keys (Bhatti 1980 ; Cluever and Smith 2017 ) and sequencing mitochondrial cytochrome oxidase subunit I (mtCoI). To develop a homogenous population, a single adult female of T. palmi was released on healthy brinjal ( Solanum melongena var.…”

Section: Methodsmentioning

confidence: 96%

A rapid field-based assay using recombinase polymerase amplification for identification of Thrips palmi, a vector of tospoviruses

et al. 2020

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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 ornamentals transmitting several tospoviruses • Morpho-taxonomic identification stage-specific, timeconsuming, 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

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Section: Methodsmentioning

confidence: 96%

A rapid field-based assay using recombinase polymerase amplification for identification of Thrips palmi, a vector of tospoviruses

et al. 2020

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“…A homogeneous population of T. palmi was developed from a single adult female collected from the stock culture maintained at Advanced Centre for Plant Virology, Indian Agricultural Research Institute (IARI), New Delhi. Identification of the thrips was based on established morphological keys [ 13 , 24 ] and on sequencing mitochondrial oxidase subunit I (mtCOI) [ 25 ]. The population was maintained on healthy eggplants ( Solanum melongena var.…”

Section: Methodsmentioning

confidence: 99%

Progression of Watermelon Bud Necrosis Virus Infection in Its Vector, Thrips palmi

Ghosh

Priti

Mandal

et al. 2021

Cells

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Thrips are important pests of agricultural, horticultural, and forest crops worldwide. In addition to direct damages caused by feeding, several thrips species can transmit diverse tospoviruses. The present understanding of thrips–tospovirus relationships is largely based on studies of tomato spotted wilt virus (TSWV) and Western flower thrips (Frankliniella occidentalis). Little is known about other predominant tospoviruses and their thrips vectors. In this study, we report the progression of watermelon bud necrosis virus (WBNV) infection in its vector, melon thrips (Thrips palmi). Virus infection was visualized in different life stages of thrips using WBNV-nucleocapsid protein antibodies detected with FITC-conjugated secondary antibodies. The anterior midgut was the first to be infected with WBNV in the first instar larvae. The midgut of T. palmi was connected to the principal salivary glands (PSG) via ligaments and the tubular salivary glands (TSG). The infection progressed to the PSG primarily through the connecting ligaments during early larval instars. The TSG may also have an ancillary role in disseminating WBNV from the midgut to PSG in older instars of T. palmi. Infection of WBNV was also spread to the Malpighian tubules, hindgut, and posterior portion of the foregut during the adult stage. Maximum virus-specific fluorescence in the anterior midgut and PSG indicated the primary sites for WBNV replication. These findings will help to better understand the thrips–tospovirus molecular relationships and identify novel potential targets for their management. To our knowledge, this is the first report of the WBNV dissemination path in its vector, T. palmi.

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“…The initial population of adult thrips was collected in plastic bags from different host plants such as brinjal, chilli, onion, and tomato from the experimental fields of Indian Agricultural Research Institute (IARI), New Delhi (28.6377°N, 77.1571°E, and 228.61 m above MSL) and taken to the laboratory. Microscopic slides were then prepared following the protocol of Silveria and Haro [43] and the thrips vectors were at first identified based on the standard morphometric keys [44,45]. Morphological characters like antennal segments, color, body shape and size, and position of ocellar setae were taken into consideration.…”

Section: Sample Collection and Identificationmentioning

confidence: 99%

“…The microscopic slides were prepared following Silveria and Haro [43]. The thrips vectors were at first identified based on the standard morphometric keys following Bhatti [44], and Cluever and Smith [45] and further confirmed by cytochrome oxidase subunit I sequences.…”

Section: Supplementary Informationmentioning

confidence: 99%

A multiplex PCR assay for rapid identification of major tospovirus vectors reported in India

et al. 2020

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Background: To 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. Results: Cytochrome oxidase subunit III and internal transcribed spacer region 2 were utilized to design speciesspecific 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. Conclusion: The 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.

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A Photo-Based Key of Thrips (Thysanoptera) Associated with Horticultural Crops in Florida

Cited by 23 publications

References 54 publications

A rapid field-based assay using recombinase polymerase amplification for identification of Thrips palmi, a vector of tospoviruses

A rapid field-based assay using recombinase polymerase amplification for identification of Thrips palmi, a vector of tospoviruses

Progression of Watermelon Bud Necrosis Virus Infection in Its Vector, Thrips palmi

A multiplex PCR assay for rapid identification of major tospovirus vectors reported in India

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