P.49. A simple method for isolating DNA from single specimen of Thrips tabaci Lindman and T. palmi Karry for PCR

Isolation of DNA from individual insects of minute size such as thrips is often difficult for further applications like PCR and sequencing. The various factors that effect isolation of DNA are, proper crushing, toughness of the integument, extraction buffer, boiling time etc. Thrips are important pests of wide range of agricultural and horticultural crops worldwide. Thrips show a high degree of similarity particularly at immature stages, which make identification extremely difficult. Further lack of conventional taxonomic tools for identification of immature stages, availability of specialized taxonomists when and where needed could hamper correct and timely identification. One of the main difficulties in the management of the thrips vector lies in the correct identification of the vector species. Identification of thrips vectors at the larval stage is important since larva only can acquire the virus while adults can transmit. In this situation molecular tools can effectively be employed for the correct identification of the thrips vectors at any developmental stages. Previously various authors employed different methods of DNA isolation with varying degrees of difficulty from various Thrips species. In this communication we report a simple method of DNA isolation from single larva and adult specimen of T. tabaci and T. palmi for the amplification of mitochondrial cytochrome oxidase I gene. The PCR products were cloned and sequenced to corroborate with morphological identification.