Research On The Resistance Of Rice GLK Mutant To Rice Black-streaked Dwarf Disease

Rice black-streaked dwarf virus(RBSDV)belongs to Fijivirus of the Family Reoviridae and is mainly transmitted by grey planthopper without egg transmission.The virus can infect rice,corn and wheat.After infecting rice,it causes severe plant shrinkage,tillering,low seed setting rate and other symptoms,causing serious economic loss.In recent years,with the enhancement of the effect of warm winter,the density of grey planthopper,the main source of virus transmission,and the rate of carrying virus,the incidence of rice black stripe dwarf disease has been on the rise,and it has been spreading in the rice planting areas nationwide,which has caused great threat and economic loss to China's rice production.Our previous studies showed that rice varieties(GLK mutants)had certain resistance to RBSDV.But how GLK becomes resistant to RBSDV is unclear.This paper mainly focused on the effect of rice GLK mutants on RBSDV resistance in the field,and the expression of genes related to the interaction resistance between GLK and RBSDV was studied and discussed.The main results are as follows:1)Field resistance detection of GLK and RBSDVThe effect of RICE GLK mutant on RBSDV resistance was studied by artificial inoculation.The results showed that RBSDV resistance of rice was significantly improved in GLK overexpressed strains.In GLK deletion mutant strain,RBSDV resistance of rice decreased significantly.Through the above field results,it was confirmed again that GLK could participate in the rice resistance mechanism.2)Interaction between GLK and RBSDVThe interaction between the candidate resistance gene GLK and RBSDV was studied by yeast double hybridization and luciferase validation in vivo.The results showed that GLK did not interact directly with the 13 gene fragments of RBSDV,that is,GLK did not interact with RBSDV,since there was no direct interaction,but whether there was an indirect interaction.In this paper,the relationship between GLK,RX1 and rice RBSDV was proved through the interaction verification of rice homologous genes RX1 and GLK.By applying the above verification methods,it was also found that GLK,RX1 and RBSDV had no direct or indirect interaction.3)Expression and detection of resistance genesIn order to further clarify the internal mechanism of GLK induced resistance,the transcription factor GLK in rice could regulate the expression of WEKY40 to respond to the mechanism of shedding pathway.Resistance genes expressed in JA pathway and SA pathway and WRKY40 in plants of different varieties were tested to confirm whether GLK was involved in disease resistance.By fluorescence quantitative PCR method,tested in five kinds of plants,wild type,expression(GLK1),east japonica varieties,single mutant varieties(glk2)double mutant varieties(glk1,2),JA pathway in SA approach the expression of resistance genes and WRKY40,verified under the participation of GLK,JA and SA way of expression of resistance genes and WRKY40 situation.The results showed that in GLK overexpressed plants,the related resistance genes in JA pathway and SA pathway were significantly increased,improving disease resistance.But WRKY40 expression is almost the same.In GLK single mutant and double mutant plants,the expression of related resistance genes was reduced.Thus,it can be determined that GLK participates in the process of disease resistance of plants and plays a key role in disease resistance of plants.However,the mechanism of intrinsic resistance in the system remains to be further explored.