Line 63-1 is a `Sunset'-derived transgenic papaya expressing the coat protein (CP) gene from a mild mutant of a Hawaiian isolate of Papaya ringspot virus (PRSV). Previous work showed that line 63-1 R1 plants exhibited a range of resistance to severe PRSV isolates from Hawaii (HA), Jamaica (JA), Thailand (TH), and Brazil (BR). Genetic and molecular data obtained in this study confirm that line 63-1 has two CP transgene insertion sites; segregation analysis shows that the CP and the npt II genes are present at both loci. To study the potential effect of gene dosage on resistance, various populations of R1, R2, and R3 seedlings were challenged by PRSV HA, BR, and TH. A R1 population obtained by self-pollination of line 63-1 hermaphrodite R0 plant exhibited resistance to all three isolates. The percentage of plants resistant to all three PRSV isolates increased in 63-1-derived populations as a result of recurrent selection. Additional genetic studies demonstrate that the number of resistant plants in a 63-1-derived population is directly correlated with the number of plants with multiple transgene copies. We conclude that transgene dosage plays a major role in affecting the resistance of 63-1 to PRSV isolates from various geographical locations.
Manoel T. Souza Jr., Paula F. Tennant, and Dennis Gonsalves
Paula Tennant, Manoel T. Souza Jr., Dennis Gonsalves, Maureen M. Fitch, Richard M. Manshardt, and Jerry L. Slightom
The disease resistance of a transgenic line expressing the coat protein (CP) gene of the mild strain of the papaya ringspot virus (PRSV) from Hawaii was further analyzed against PRSV isolates from Hawaii and other geographical regions. Line 63-1 originated from the same transformation experiment that resulted in line 55-1 from which the transgenic commercial cultivars, `Rainbow' and `SunUp', were derived. Plants of line 63-1 used in this study consisted of a population from a self pollinated R0 bisexual plant. ELISA and PCR tests provided evidence that there are at least two segregating CP loci. To allow for comparison with reactions of the previously reported line 55-1, virus isolates from Hawaii, Brazil, Thailand, and Jamaica were used to challenge seedlings of 63-1. Unlike line 55-1, a significant percentage of inoculated transgenic plants were susceptible to isolates from Hawaii. However, a proportion of plants were resistant to the non-Hawaiian isolates. In contrast, previous work showed that all plants of the hemizygous line 55-1 were susceptible to PRSV isolates from Brazil, Thailand, and Jamaica. Line 63-1, therefore, presents Hawaii with PRSV-resistant transgenic germplasm that could be used as a source of transgenes for resistance to PRSV isolates within and outside of Hawaii.