Low-cost water boiling method successfully employed to extract DNA from a single whitefly to detect yellow mosaic disease-causing viruses in PCR

Abstract

A cost-effective DNA extraction approach was developed and applied to PCR-based identification of yellow mosaic disease (YMD) causing viruses in individual whiteflies. Whiteflies were collected from urdbean and eggplant, representing host and non-host plants of target viruses, respectively. DNA from whiteflies was successfully extracted using the nuclease-free water boiling method and optimised template concentration for PCR assays. An average nucleic acid content of 3.844 ± 0.14 ng/µl was extracted from individual whiteflies in 50 µl nuclease-free water. PCR assays revealed Mungbean yellow mosaic India virus (MYMIV) presence in 90% of the whiteflies collected from urdbean. Conversely, YMD-causing viruses were not present in whiteflies collected from eggplant or in the eggplant plants themselves. The use of this low-cost approach for detecting YMD-causing viruses in whiteflies is very effective for the surveillance and monitoring of viruses. Early detection facilitates the deployment of appropriate management strategies, reducing both crop losses and the potential for outbreaks.