Development of PCR-based diagnostic probe to detect begomoviruses infecting chilli in the hot arid region of Rajasthan

Abstract

A survey was conducted in major chilli-growing hot arid regions of Rajasthan, namely, Bikaner, Nagur, Jodhpur and Jalore districts, during November 2009. Among the four districts surveyed for chilli leaf curl disease (ChLCD), maximum disease incidence was recorded in Jodhpur (98%) followed by Jalore district (88%). The number of whiteflies was also counted in top, middle and bottom leaf of chilli plants grown in these areas. The average number of whiteflies per plant ranged from 0.0 to 4.0. Higher number of whiteflies (4.0) was recorded in Jodhpur and lowest (1.8) in Jalore district. On the basis of conserved region in the genome of begomoviruses infecting chilli, a set of primers was designed to amplify all begomoviruses infecting chilli by PCR; ChCPF 5'-ATTAGGGCTAAGAATTATGTC-3' and ChCPR 5'-AAATTCCAATCTTTATTAATT-3'. These primers were validated by cloning and sequencing (HM004433) of PCR-amplified products and detection from infected chilli leaf samples. These primers were utilised while screening chilli cultivars against begomovirus infection in the asymptomatic plants. Our investigation suggests that the leaf curl disease of chilli is widespread in the hot arid regions of Rajasthan and is caused by a begomovirus associated with a satellite DNA β. The PCR primers designed in this study could be highly useful in chilli breeding programmes.