Fig. 1.

Assessment of cross-reactivity of four species-specific primer pairs. Primer pairs viz. AG35F-AG36R, AG47F-AG48R, AG87F-AG88R, and AG79F-AG80R specific to T. palmi, S. dorsalis, T. tabaci, and F. schultzei, respectively were shortlisted for multiplex PCR assay. The cross-reactivity of the species-specific primer pairs was assessed in 25 μl PCR reactions and resolved on 0.8% agarose gel electrophoresis. Lane 1, 7: 1 kb plus DNA ladder; Lane 13, 19: 100 bp plus DNA ladder; Lane 2, 8, 14, 20: water control; Lane 3–6: PCR amplicons using T. palmi-specific primer pair with DNA templates of T. palmi (3), S. dorsalis (4), T. tabaci (5), and F. schultzei (6); Lane 9–12: PCR amplicons using S. dorsalis-specific primer pair with DNA templates of S. dorsalis (9), T. palmi (10), T. tabaci (11) and F. schultzei (12); Lane 15–18: PCR amplicons using T. tabaci-specific primer pair with DNA templates of T. tabaci (15), T. palmi (16), S. dorsalis (17), and F. schultzei (18); Lane 21–24: PCR amplicons using F. schultzei-specific primer pair with DNA templates of F. schultzei (21), T. palmi (22), S. dorsalis (23), and T. tabaci (24). PCR with primer pairs AG35F-AG36R, AG47F-AG48R, AG87F-AG88R, and AG79F-AG80R produced bands of 568 bp, 713 bp, 388 bp, and 200 bp of T. palmi, S. dorsalis, T. tabaci, and F. schultzei, respectively. No cross-reactivity was found with other thrips vectors