PCR BASED AMPLIFICATION AND DETECTION OF PAPAYA LEAF CURL VIRUS (PALCUV)

Detection of Papaya leaf curl virus in twelve papaya samples displaying leaf curl symptoms was attempted using PCR based amplification of partial and full length coat protein gene.
PCR was performed on total genomic DNA using two sets of primers from the coat protein regions of PaLCuV. Twelve infected papaya samples and Ageratum spp. (positive control) with the primer set PaLCuV1/ PaLCuV2 amplified fragments of around 1 kb (which includes the conserved nonanucleotide, precoat protein gene and partial fragment of coat protein gene) was obtained as expected.
Similarly PaLCuV1/ PaLCuV3 primer set designed from PaLCuV DNA genome, amplified fragments of around 1.14 kb which cover the entire coat protein gene (including the conserved nonanucleotide, precoat protein gene and coat protein gene), while nothing could be amplified from healthy papaya plants.
We conclude that the PCR amplified fragment of expected size indicate DNA to be of geminiviral nature and also confirm the specificity of the primers used to detect geminivirus.
Information on genomic variability of PaLCuV isolates is very important, as this study could have an impact on the control strategy to be developed for disease management against PaLCuV in India with respect to PaLCuV resistance transgenic papaya.
Investigation of the sequence homology levels of variability among PaLCuV isolates, is the first step towards selecting appropriate sequence(s) for use as transgene and secondly to assist in the prediction of stability of resistance.
This concept is gaining importance as more and more viral genomes are being sequenced.

II International Symposium on Papaya

N. Srivastava, R. Chandra, S. Saxena, A. Bajpai

papaya, PaLCuV, geminivirus

https://doi.org/10.17660/ActaHortic.2010.851.36