Deciphering the multiplication behaviour of Rice tungro bacilliform virus by absolute quantitation through real-time PCR

Abstract

Rice tungro virus disease is one of the most destructive diseases that cause extensive damage to the rice crop. To elucidate the multiplication behaviour of Rice tungro bacilliform virus (RTBV), real-time Polymerase chain reaction (PCR) experiments were performed on rice and insect vector green leafhopper (GLH). SYBR green chemistry-based real-time PCR assay for the quantification of RTBV was developed. A standard curve using plasmid DNA was constructed to determine the absolute quantity of RTBV genome copies in different plant tissues and GLH vector. Here, 6.309 × 104, 7.943 × 105, 3.162 × 106 and 3.162 × 103 RTBV genome copies per ng of total DNA were estimated in root, shoot, leaf and panicles, respectively, on virus-infected rice cultivar TN1. In addition, 5.011 × 103 copies of virus in an individual GLH were quantified. Also, RTBV was quantified at different time interval after inoculation. The real-time assay was performed with five different RTBV isolates that showed differential accumulation pattern of virus isolates in a same host. These results provide new insight into the biology of the economically important interaction between rice, GLH and RTBV.