Development of papaya breeding lines with transgenic resistance to Papaya ringspot virus

Papaya (Carica papaya) was transformed via Agrobacterium-mediated transformation with four constructs containing either the unmodified or modified coat protein (CP) gene of Florida isolate HIK of Papaya ringspot virus (PRSV). The CP genes were in the sense orientation (S-CP), antisense orientation (AS-CP), sense orientation with a frame-shift mutation (FS-CP), or sense orientation mutated with three-in-frame stop codons (SC-CP). In all, 256 putative transgenic lines with the CP constructs were inoculated mechanically with PRSV HIK. None of the lines was immune to PRSV; however, highly resistant lines were found in each CP transgene group. For breeding purposes, 21 PRSV-resistant lines representing the four transgene constructs were selected and crossed with six papaya genotypes. The lines from the FS-CP and SC-CP transgene groups were highly fertile, but those from the S-CP and AS-CP transgene groups were practically infertile. Plants derived from 54 crosses and representing 17 transgenic lines were planted in the field. After 1 year in the field, 293 of the 1,258 the plants (23.3%) became naturally infected with PRSV; whereas, 29 of 30 of the nontransgenic control plants (96.7%) became infected. The incidence of PRSV infection varied in the R-1 progeny depending on both the transgenic line and the nontransgenic parent.