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OPTIMIZING CROP CULTIVATION ON THE INTERNATIONAL SPACE STATION BY IMPROVING PLANT DESICCATION-TOLERANCE WITH TARDIGRADE PROTEINS
Abstract
Despite advancements in crop growth on the International Space Station (ISS), unforeseen dry conditions have highlighted the demand for research into enhanced plant desiccation-tolerance. Tardigrades are extremotolerant microorganisms which can survive abiotic conditions such as drought, radiation, and extreme temperatures. This study successfully establishes a method to introduce tardigrade intrinsically disordered proteins (TDPs) in a plant vector, Nicotiana benthamiana, via agroinfiltration. Five TDP constructs were designed and tagged with mVenus, a yellow fluorescent protein to visualize TDP expression. The TDP expression constructs were successfully co-infiltrated with an abscisic acid (ABA) autoluminescent reporter in a transient transformation of N. benthamiana leaves via agroinfiltration. After three days, the leaves were excised from the plants and subjected to either dry conditions or floated on water. Luminescent imaging of whole leaves was used to evaluate osmotic stress and ABA-induced luminescence over time. While our initial constructs failed to express the TDP-mVenus fusion, we validated our methods for measuring luminescence reporters of ABA signaling in response to the simulated drought stress in detached leaves. In addition to agroinfiltration with corrected constructs, floral dip of Arabidopsis thaliana will be used to generate transgenic lines for further examination of the effects of TDP expression on plant desiccation-tolerance.
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