The Interaction Between Turnip Mosaic Virus Encoded Proteins And AtSWEET1 Protein In Arabidopsis Thaliana

Turnip mosaic virus(TuMV)is a member of the genus Potyvirus and possesses an exceptionally broad host range in terms of plant genera and families of any potyvirus.The genome of TuMV is a single-stranded,positive-sense RNA molecule of about 10,000 nt,which can be encoded 11 different structures and functions of the protein through two coding strategies,the interaction between the virus protein and the host protein is an important way for virus-host interaction.SWEET(sugers will eventually be exported transporters)protein family is a new class of sugar transporters that play an important role in the interaction of host-pathogens.In addition to participating in the functions of sugar transport,growth and development,stress,etc.It has been reported that plants infected with fungi or bacteria will induce partial expression of the SWEET gene involved in pathogen-host interaction.However,the SWEET protein family has not been found to be involved in the interaction of viral and host.Previous studies have shown that P3 protein interact with At SWEET1 protein in Arabidopsis thaliana.On the basis,the interaction between TuMV encoded other proteins and At SWEET1 in Arabidopsis thaliana.The molecular mechanism of the interaction between virus and host protein can be revealed in theory,and comprehensively understand the mechanism of interaction between Potyviruses and host.At first,in order to identity whether there is interaction between TuMV encoded protein and At SWEET1 in Arabidopsis thaliana.The bait vector of the other 10 proteins except the P3 protein were constructed using the corresponing gene sequence of the TuMV CCLB isolate(Gen Bank: KR153038).The recombinant bait vectors were verified no toxic and self-activating,then they were verified to interact with At SWEET1 by yeast two-hybrid methods.The results showed that TuMV-encoded HC-Pro,VPg and NIa-Pro interacted with At SWEET1.In addition,these intreraction was further validated on Nicotiana benthamiana and Allium cepa using Bi FC,the interaction sites were located on the cell membrane.In order to study the interaction of TuMV encoded proteins themselves,the prey vector of HC-Pro,VPg and NIa-Pro protein were constructed,and the recombinant bait vectors were verified no toxic and self-activating.The recombinant prey vector interaction with TuMV-encoded proteins was verified by yeast two-hybrid methods.The results showed that only the TuMV-encoded HC-Pro protein interacted with itself,the protein was not found to interact with VPg and NIa-Pro.In order to study which genes were induced Arabidopsis thaliana 17 At SWEET genes by virus infection.The expression of At SWEET gene was detected to infect Arabidopsis thaliana by infectious c DNA clone of TuMV.The result showed that most of the At SWEET genes induced expression,in which At SWEET7 expression was the highest,At SWEET6,At SWEET9,At SWEET10 and At SWEET14 genes were up-regulated about 100 times in Arabidopsis leaves.At SWEET3,At SWEET4,At SWEET8 and At SWEET15 genes were up-regulated about 10 times,and At SWEET1,At SWEET5 and At SWEET12 genes were up-regulated about 4 times.Therefore,we conclude that the At SWEET gene family is involved in the interaction of TuMV and Arabidopsis.This is to fill the gaps in the SWEET protein family in the interaction between the virus and the host,but also to provide a theoretical basis for further study of the virus pathogenesis.