Detection of each of the causal agents of groundnut rosette disease in plants and vector aphids by RT-PCR.

摘要

Detection of the three agents of groundnut rosette disease (groundnut rosette assistor virus, groundnut rosette virus and its satellite RNA) in plants and vector aphids by reverse transcription-polymerase chain reaction (RT-PCR) is reported. Three procedures for extraction of total RNA from groundnut were tested, of which two were found to be useful in giving RNA of sufficient quality for RT-PCR. Of these two, the total RNA extraction kit supplied by Qiagen was found to be the most versatile for extraction of all three agents from individual vector aphids (Aphis craccivora). Both groundnut rosette assistor virus and groundnut rosette virus could be detected from total RNA extracted from a single aphid that had been exposed to either green or chlorotic rosette-infected groundnut plants. They could be detected in aphids stored in 70% ethanol for up to 30 days at room temperature. However, satellite RNA could be amplified only when total RNA extracted from two or more aphids was used. Groundnut rosette assistor virus, groundnut rosette virus and its satellite RNA were detected by RT-PCR in aphids that had been exposed only to groundnut rosette diseased plants containing all three agents. The potential of RT-PCR in studying certain key issues of rosette disease epidemiology is discussed.