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β-1,3-glucanase class III promotes spread of PVYNTN and improves in planta protein production
Authors:David Dobnik  ?pela Baebler  Polona Kogov?ek  Maru?a Pompe-Novak  Dejan ?tebih  Gabriela Panter  Nikolaja Jane?  Dany Morisset  Jana ?el  Kristina Gruden
Institution:1. Department of Biotechnology and Systems Biology, National Institute of Biology, Ve?na pot 111, 1000, Ljubljana, Slovenia
2. Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000, Ljubljana, Slovenia
3. National Institute of Chemistry, Hajdrihova 19, 1000, Ljubljana, Slovenia
4. Centre of Excellence for Biosensors, Instrumentation and Process Control, Velika pot 22, 5250, Solkan, Slovenia
Abstract:Glucanases are enzymes regulating the size exclusion limit and permeability of plasmodesmata and play a role in biotic stress. In plant genomes, they are encoded as relatively large gene families divided into four classes. Most studies of plant virus interactions have focused on glucanases from classes I and II. In our study, we have evaluated the role of the β-1,3-glucanase class III (Glu-III) gene in the potato–potato virus YNTN (PVYNTN) interaction and implemented the findings to plant biotechnology application. Potato cultivars Désirée and Santé, which are tolerant and extremely resistant to PVYNTN, respectively, were stably transformed with Agrobacterium tumefaciens harbouring constructs for Glu-III overexpression. Localization of Glu-III protein in patches within the cell wall was determined by tagging the Glu-III protein with green fluorescent protein. Transgenic and non-transgenic plants were challenged with PVYNTN and its multiplication and spreading was followed. Differences in viral spread were observed between transgenic lines overexpressing Glu-III and non-transgenic lines, with stronger and faster viral spread in transgenic Désirée, and some multiplication in transgenic Santé. In addition, the ability of Glu-III to improve in planta protein production after agroinfiltration was tested. The results have shown that Glu-III overexpression enables faster spreading of vectors between cells and better protein production, which could be beneficial in improving in planta protein production system using viral vectors.
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