FIG. 5.

Immunocapture-PCR assay for detection of TYLCV CP-GroEL interactions. (A) Extracts from B-biotype strains B-HRc, B-HRs, and B-H Neg03E and from Q-biotype strains Q-A, Q-WA, and Q-RA AV04E (see the legend to Fig. 1) were subjected to SDS-PAGE, Western blotting, and immunodetection with anti-Buchnera GroEL antibody to confirm reactivity with all possible symbionts in whiteflies. The positions of molecular mass markers (lane M) (in kilodaltons) are indicated to the left of the gel. (B) PCR tubes coated with the GroEL-specific antibody and virus-specific primers were used to detect the interaction. Lane M contains 100-bp ladder marker. Lanes 1, 3, and 5 contain extracts from BHRc, BHRs, and B-H Neg03E populations, and lanes 2, 4, and 6 are no-antibody controls for each respective population. Lanes 7, 9, and 11 contain extracts from Q-A, Q-WA, and Q-RA AV04E populations, and lanes 8, 10, and 12 contain PCR mixtures from the three Q-biotype populations to verify TYLCV acquisition. Lane 13 is a negative no-template control, lane 14 is a positive PCR control plasmid (bearing the full TYLCV sequence), and lane 15 is a positive PCR control from a TYLCV-infected plant.