A Research On Interaction Between RSV NS2 And NbFib2 And The Construction Of OsSGS3 RNAi Transgenic Rice

Rice stripe virus(RSV)the causal agent of rice stripe disease is the typical member of the genus Tenuivirus.Rice stripe disease is widely distributed in temperate regions and subtropical regions in East Asia,including China's Jiangsu,Henan,Yunnan,and Shandong provinces,causing a great vield loss to rice production.NS2 encoded by RSV is a nuclear-cytoplasmic shuttling protein that interacts with the nucleolar protein(Fibrillarin2,Fib2)of Nicotiana that affects the infection process of RSV.We further studied the interacting domains of NS2 and NbFib2 proteins in this study.In addition,previous studies showed that NS2 could interact with rice gene silencing suppressor(SGS3 protein).In Arabidopsis thaliana,sgs3 mutant plants exhibit a higher susceptibility to cucumber mosaic virus(CMV).On the basis of this research,we studied the subcellular localization of two OsSGS3 encoding proteins in rice,and constructed their RNAi transgenic plants,which lays the foundation for further research on the role of OsSGS3 in RSV-rice interaction.This paper is divided into two parts:the first part studies the NbFib2 domains responsiable for its interaction with NS2.In the second part,we study the subcellular localization of OsSGS3-1 and OsSGS3-2 and construct the RNAi transgenic plants.The results obtained are as follows:1 Co-localization and BiFC results show that NS2 can interact with the three functional domains of NbFib2:NbFib2-1,NbFib2-2 and NbFib2-3.2 Subcellular localization of OsSGS3-1 and OsSGS3-2,show that OsSGS3-1 is mainly localized in the nucleus and cytoplasm,forming small bright spots in the cytoplasm,while OsSGS3-2 is localized in the cytoplasm without small point formation.3 We utilized agrobacterium mediated rice genetic transformation and produced OsSGS3-1 RNAi and OsSGS3-2 RNAi T0 generation of transgenic rice,the transgenic plants were identified and analyzed by RT-qPCR.