Evaluation of Rice stripe virus transmission efficiency by quantification of viral load in the saliva of insect vector.

Abstract

Persistent plant viruses transfer from insect gut to the hemolymph, and finally to the salivary glands before inoculation into the plant hosts with saliva during insect feeding. Virus accumulation in saliva is an important indicator for the transmission ability of an insect vector. In order to evaluate the transmission ability of the small brown planthopper to rice stripe virus (RSV), we successfully measured accumulation of RSV in the saliva of planthoppers via the absolute real-time quantitative polymerase chain reaction method by quantifying the copy numbers of viral genes. After feeding on an artificial diet for 24 h, the copy numbers of viral genes of capsid protein (CP) and disease-specific protein (SP) can be detected in the saliva collected from as few as ten viruliferous planthoppers and ten non-viruliferous planthoppers after infected with RSV for 7days. When the expression of planthopper G protein pathway suppressor 2 or c-Jun N-terminal kinase was knocked down, the copy numbers of CP and SP in the saliva varied accordingly. Our study provided an accurate and convenient detection system to evaluate the transmission efficiency of RSV by small brown planthoppers, and this method may also be suitable for other persistent plant viruses. © 2019 Society of Chemical Industry.