Comparison of methods for isolating DNA of viral infections from Bemisia tabaci (Genn.) using the case of tomato yellow leaf curl virus (TYLCV)

Abstract

Silverleaf whitefly (Bemisia tabaci Genn.) is one of the key vectors of phytopathogenic viruses, primarily begomoviruses (family Geminiviridae), which include tomato yellow leaf curl virus (TYLCV).
Since TYLCV has phytosanitary status and is regulated in accordance with Decision No. 158 of the Council of the Eurasian Economic Commission dated November 30, 2016, it is particularly important to ensure its early detection and identification. This study aimed to determine the feasibility of isolating TYLCV DNA from silverleaf whitefly individuals stored in different ways — in a 70% ethanol solution and frozen. These individuals fed on TYLCV-infected tomato plants. DNA extraction using different commercial kits was compared, after which DNA concentrations were measured. Real-time PCR was then performed.
Additionally, to analyze the genetic sequence of the B. tabaci cytochrome oxidase gene fragment after total NC isolation, classical PCR was applied to the target fragment. The study demonstrated that the DNA product was detected using both isolation methods, and these methods are applicable in practice. Since inhibition of the reaction was observed with the ProbaNK kit for silverleaf whitefly samples stored in 70% ethanol, the FitoSorb kit is recommended for use.