Agronomically important thrips: development of species-specific primers in multiplex PCR and microarray assay using internal transcribed spacer 1 (ITS1) sequences for identification.
Bull Entomol Res
; 105(1): 52-9, 2015 Feb.
Article
em En
| MEDLINE
| ID: mdl-25335450
ABSTRACT
Thrips, the sole vector of plant Tospovirus, are major pests of many agricultural crops throughout the world. Molecular approaches have been applied in recent decades to identify these minute and morphologically difficult to distinguish insects. In this study, sequences of internal transcribed spacer 1 (ITS1) region of 15 agronomically important thrips, including several virus transmission species, have been analyzed in order to design species-specific primers for multiplex PCR and probes for microarray assay. That the ITS1 sequence distances within species were smaller than those among species suggests that the ITS1 fragment can be used for thrips species identification. The specificity and stability of these primers, combined with universal paired primers, were tested and verified in multiplex PCR. Using these specific primers as probes, microarray assay showed that PCR products of all thrips species hybridized consistently to their corresponding probes, though some signals were weak. We have demonstrated that multiplex PCR using specific primers based on ITS1 sequences is a simple, reliable, and cost-effective diagnostic tool for thrips species identification. Moreover, the DNA microarray assay is expected to extend into a reliable high-throughput screening tool for the vast numbers of thrips.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Controle de Insetos
/
Análise de Sequência com Séries de Oligonucleotídeos
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Tisanópteros
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Reação em Cadeia da Polimerase Multiplex
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
País como assunto:
Asia
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article