Molecular characterization of Begomoviruses isolated from areas around Azad Kashmir

Abstract

Begomovirus of family Geminiviridae has been a considerable hindrance in producing high agricultural yields and causes severe economic losses in the majority of agro-based countries, including Pakistan. Among them only the cotton leaf curl disease (CLCuD) caused over US$ 5 billion loss to the Pakistan economy during the mid to late 1990s. Begomovirus also affects about 80 to 85 percent ornamental plants. This group of plant viruses is transmitted by whitefly (Bemisia tabaci) and infects dicotyledonous plants only. This group is composed of single stranded, circular DNA genome. A lot of data with reference to begomovirus diversity is available from Pakistan but scientific reports related to begomovirus from Azad Kashmir area are scanty. The project was designed: (I) to collect begomovirus suspected symptomatic and asymptomatic leaves samples from areas around Azad Kashmir; (II) to perform diagnostic PCR (Polymerase Chain Reaction) for the presence or absence of begomovirus; (III) molecular characterization of begomoviruses. About twenty five begomovirus suspected plant samples were collected. Total DNA was extracted, from both symptomatic and asymptomatic samples, using CTAB (Cetyl Tri methyl Ammonium Bromide) method. PCR was performed for diagnosis and amplification of begomoviruses. Only two samples i.e., Turmeric and Bottle gourd showed the amplification with universal core protein (CP) primers. The amplified partial products, DNA A and DNA betasatellite, of Turmeric and Bottle gourd respectively, were sequenced. This confirms the begomoviruses presence in these samples. Since Pakistan is an agricultural country, the diverse studies of begomoviruses, especially from far flung regions of the country, will contribute significantly for the development of scientific strategy for the problem