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Development of silverleaf assay, protein and nucleic acid-based diagnostic techniques for the quick and reliable detection and monitoring of biotype B of the whitefly, Bemisia tabaci (Gennadius)

Published online by Cambridge University Press:  04 October 2007

K.S. Shankarappa ,
K.T. Rangaswamy ,
D.S. Aswatha Narayana ,
A.R. Rekha ,
N. Raghavendra ,
C.N. Lakshminarayana Reddy ,
T.C.B. Chancellor  and
M.N. Maruthi
K.S. Shankarappa
Affiliation:
Department of Plant Pathology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India
K.T. Rangaswamy
Affiliation:
Department of Plant Pathology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India
D.S. Aswatha Narayana
Affiliation:
Department of Plant Pathology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India
A.R. Rekha
Affiliation:
Department of Plant Pathology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India
N. Raghavendra
Affiliation:
Department of Plant Pathology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India
C.N. Lakshminarayana Reddy
Affiliation:
Department of Plant Pathology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India
T.C.B. Chancellor
Affiliation:
Natural Resources Institute, University of Greenwich, Central Avenue, Chatham Maritime, Kent ME4 4TB, UK
M.N. Maruthi*
Affiliation:
Natural Resources Institute, University of Greenwich, Central Avenue, Chatham Maritime, Kent ME4 4TB, UK
*
*Author for correspondence Fax: +44 (0)1634 883379 E-mail: M.N.Maruthi@greenwich.ac.uk
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Abstract

The aim of this study was to develop and optimize silverleaf bioassay, esterase analysis and PCR-based techniques to distinguish quickly and reliably biotype B of the whitefly, Bemisia tabaci (Gennadius), from Indian indigenous biotypes. Zucchini and squash readily develop silverleaf symptoms upon feeding by the B biotype, but they are not readily available in Indian markets. A local pumpkin variety ‘Big’ was, therefore, used in silverleaf assay, which developed symptoms similar to those on zucchini and squash and can be used reliably to detect B biotype. Analysis of non-specific esterases of B and the indigenous biotypes indicated both quantitative and qualitative differences in esterase patterns. Two high molecular weight bands were unique to B biotype and they occurred in abundance. These esterases were used to develop quick and field-based novel detection methods for differentiating B from the indigenous biotypes. Development of these simple and cost-effective protocols has wider application as they can be potentially used to identify other agricultural pests. Mitochondrial cytochrome oxidase I gene sequences and randomly amplified polymorphic DNA (RAPD) polymorphisms, generated using the primer OpB11, were also found useful for detecting B. tabaci biotypes. A B biotype-specific RAPD band of 800 bp was sequenced, which was used to a develop sequence characterized amplified region (SCAR) marker. The SCAR marker involved the development of B biotype-specific primers that amplified 550 bp PCR products only from B biotype genomic DNA. Silverleaf assay, esterases, RAPDs or a SCAR marker were used in combination to analyse whitefly samples collected from selected locations in India, and it was found that any of these techniques can be used singly or in combination to detect B biotype reliably. The B biotype was found in southern parts of India but not in the north in 2004–06.

Keywords

biotypescytochrome oxidase I geneesterasemolecular variabilityRAPD-PCRwhitefly
Type
Research Article
Information
Bulletin of Entomological Research , Volume 97 , Issue 5 , October 2007 , pp. 503 - 513
Copyright
Copyright © Cambridge University Press 2007

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