Construction of Yeast Two-hybrid cDNA Library on Tomato Spotted Wilt Tospovirus-infected Frankliniella occidentalis

  Objective  A two-hybrid eDNA library was constructed for studying the interaction between tomato spotted wilt tospovirus (TSWV) and Frankliniella occidentalis to decipher the mechanism of the disease transmission by thrip and the factors involved in the virus infection.

  Method  A highly infected F. occidentalis population was created by inoculating TSWV into the thrip nymphs. Thereby, a yeast two-hybrid three-frame cDNA library on TSWV-infected F. occidentalis was constructed using the SMART technology.

  Result  The established 3 reading frame cDNA libraries had the capacities of 3.0×10⁶, 2.0×10⁶, and 2.0×10⁶ cfu. The actual amplification base of the library was more than 1.5×10⁶ cfu or 5 ×10⁵ cfu each with an average insert fragment of 0.5–3.0 kb in length. From the library 16 clones were randomly selected, sequenced, and compared with the GenBank database for of homology verification on the insert fragments.

  Conclusion  The established cDNA library had an ample capacity and high recombination rate adequate for studies on the interactions between TSWV and F. occidentalis.