Research Projects

Tulip (Tulipa spp.) belongs to the family of Liliaceae, and is one of important ornamental flower crops worldwide. Tulips are being cultivated in different agro-climatic zones due to their huge diversity in cultivars. For successful cultivation and maintenance of vast diversity of tulips ensuring high phytosanitary state is essential. However, tulips have been found susceptible to several viral species which significantly affect both quality and quantity of crops. Tulips being propagated vegetatively via bulbs, the viral pathogens can survive in dormant bulbs due to their systemic nature, thereby aggravating this problem further. Moreover these virus infected dormant bulbs can be transported to far distant places through international trade. In India, tulip cultivation is largely dependent on bulb imports, primarily from the Netherlands. Each year, India imports about 20–25 lakh tulip bulbs and cut flowers. The Kashmir Valley holds a significant share in this import, with approximately 15 lakh bulbs brought annually to curate Asia’s largest tulip garden—the Indira Gandhi Memorial Tulip Garden. So, there is a considerable risk of introduction of viral pathogens into India via dormant bulbs. Recently it was reported that tulips in Kashmir are infected by three viruses namely, tulip breaking virus (TBV), tulip virus X (TVX) and potato virus Y (PVY). This is a concerning indication of introduction of virus-infected material through imported bulbs. So, there is a need to prevent the further introduction and spread of tulip viruses in India, especially in high-risk areas. The existing diagnostics such as ELISA, RT-PCR and LAMP assays have a variety of limitations. Therefore, there is an urgent need to develop a rapid, highly sensitive and specific onsite detection and diagnosis system for tulip viruses. Recently CRISPR-Cas systems have been used for detection of pathogens which provide onsite viral detection with high sensitivity and specificity. Considering the significant damage caused by identified viruses (TBV, TVX and PVY) in tulips, and the need to prevent their introduction and spread in India, the present study aims to develop a multiplex RT-RPA based CRISPR-Cas12a diagnostic assay capable of simultaneously detecting three target viruses. The assay will be designed to offer high sensitivity and specificity, with field deployability. Moreover assay will be paired with methods that will enable direct detection from crude extract with visual readouts.