Dobnik David, Baebler Spela, Kogovšek Polona, Pompe-Novak Maruša, Stebih Dejan, Panter Gabriela, Janež Nikolaja, Morisset Dany, Zel Jana, Gruden Kristina
Department of Biotechnology and Systems Biology, National Institute of Biology, Večna pot 111, 1000 Ljubljana, Slovenia.
Plant Biotechnol Rep. 2013;7(4):547-555. doi: 10.1007/s11816-013-0300-5. Epub 2013 Aug 27.
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 Y (PVY) interaction and implemented the findings to plant biotechnology application. Potato cultivars Désirée and Santé, which are tolerant and extremely resistant to PVY, respectively, were stably transformed with 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 PVY 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 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 protein production system using viral vectors.
葡聚糖酶是调节胞间连丝的大小排阻极限和通透性的酶,在生物胁迫中发挥作用。在植物基因组中,它们被编码为相对较大的基因家族,分为四类。大多数关于植物病毒相互作用的研究都集中在I类和II类葡聚糖酶上。在我们的研究中,我们评估了III类β-1,3-葡聚糖酶(Glu-III)基因在马铃薯-马铃薯Y病毒(PVY)相互作用中的作用,并将研究结果应用于植物生物技术。分别对PVY具有耐受性和极强抗性的马铃薯品种Désirée和Santé进行稳定转化,使其携带用于Glu-III过表达的构建体。通过用绿色荧光蛋白标记Glu-III蛋白来确定其在细胞壁内斑块中的定位。用PVY对转基因和非转基因植物进行挑战,并跟踪其增殖和传播情况。在过表达Glu-III的转基因株系和非转基因株系之间观察到病毒传播存在差异,在转基因Désirée中病毒传播更强、更快,在转基因Santé中病毒有一些增殖。此外,还测试了Glu-III在农杆菌浸润后提高蛋白质产量的能力。结果表明,Glu-III的过表达能够使载体在细胞间更快地传播,并提高蛋白质产量,这可能有助于改进使用病毒载体的蛋白质生产系统。