Microneedle assisted delivery of the cloned components of cucurbit leaf crumple virus in yellow squash (Cucurbita pepo)

Authors: KAVALAPPARA, SARITHA - University Of Georgia; DEVENDRAN, RAGUNATHAN - University Of Georgia; Simmons, Alvin; BAG, SUDEEP - University Of Georgia

Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/25/2025
Publication Date: 9/1/2024
Citation: Kavalappara, S.R., Devendran, R., Simmons, A.M., Bag, S. 2024. Microneedle assisted delivery of the cloned components of cucurbit leaf crumple virus in yellow squash (Cucurbita pepo). Journal of Virological Methods. 329: 2024 Article 114992. https://doi.org/10.1016/j.jviromet.2024.114992.
DOI: https://doi.org/10.1016/j.jviromet.2024.114992

Interpretive Summary: The Cucurbit leaf crumple virus (CuLCrV) is an important plant virus that infects cucurbits crops such as yellow squash. Whiteflies spread this virus in fields. To screen squash cultivars and other sources for plant resistance to this virus, it is important to have an efficient procedure for plant virus inoculation. An infectious clone and a micro-needle assisted delivery system to the plant were developed. This method will be useful for scientists to rapidly screen squash germplasm lines, and potentially other crops such as beans, for resistance to CuLCrV.

Technical Abstract: Cucurbit leaf crumple virus (CuLCrV) is among the prominent viruses infecting cucurbits in the USA. Attainable procedures of virus inoculation to crops are prerequisite for screening for resistance against the virus. Because mechanical (non-vector-mediated) infection by cucurbit leaf crumple virus (CuLCrV) is inefficient in economically important crops, screening for CuLCrV resistance is currently laborious and time-consuming using transmission by viruliferous whiteflies. We constructed an infectious partial tandem repeat construct of an isolate of CuLCrV from Georgia, USA, in the plant expression binary vector pCambia2300 and transformed it into Agrobacterium tumifaciens strain EHA105. Agroinfiltration of this construct into the abaxial surface of the leaves of common bean (Phaseolus vulgaris L.) produced a systemic infection characteristic of CuLCrV, although this approach was not successful for yellow squash. However, we report a very efficient and reproducible inoculation procedure established in squash when the leaves were injured with a microneedle and rubbed with cell suspension harboring the infectious viral construct.