Enhancing RNA Isolation Efficiency for Tomato Brown Rugose Fruit Virus (ToBRFV) Detection in Non-primary Host Weeds and Determination of Host Potential

Abstract

Tomato brown rugose fruit virus (ToBRFV) is considered an emerging disease and a viral epidemic affecting tomato consumers. The aim of this research is to optimize the isolation of ToBRFV from the weeds Solanum nigrum and Chenopodi album and to determine its potential for use as a test plant. The detection and study of ToBRFV in these non-primary hosts requires optimized RNA isolation protocols due to challenges such as low viral load, plant inhibitors, and variations in tissue properties. This paper explores innovative approaches to improve the efficiency of RNA isolation from S. nigrum and C. album, offering a comparative analysis with standard isolation methods and proposing novel solutions to increase RNA yield and purity. By addressing the unique biochemical challenges posed by these plants, our work provides an essential foundation for accurate ToBRFV diagnosis and further virological research. The leaves taken from the samples showing symptoms were applied using SDS-LiCl method with and without phenol, CTAB-LiCI methods and the RNA isolation kit was used. Among the RNA isolation techniques, used in this study, the most suitable one was found as modifying SDS-LiCl from which good quality RNA was able to be isolated. When the two plants were evaluated in terms of their potential as indicator plants, S. nigrum plant was found to be superior to C. album plant in terms of longevity and resistance to different climatic conditions, not only showing local symptoms but also having the potential to carry the virus systemically.