Toxicity Analysis And Subcellular Localization Of Five Genes And Effector Proteins Of Tilletia Contraversa Kühn

Tilletia contraversa Kühn(TCK)caused by wheat dwarf smut is an important international quarantine disease and also a kind of quarantine disease in China.Its incidence rate is usually equivalent to the yield loss rate,which can reduce wheat yield by more than 80%,or even no grain harvest.In this study,the effector protein genes g10613,g13620,g6495,g10555 and g11335 were cloned from TCK genome data,and bioinformatics analysis and toxicity verification were conducted.Subcellular localization of proteins was analyzed by constructing subcellular localization vector.The main results are as follows:1.A total of 55 candidate effector protein genes were obtained through genomic screening of ANabaena glutinosa,and five genes that could be used to construct effector protein vectors were obtained after the check,which were g10613,g13620,g6495,g10555 and g11335,respectively.The results of tobacco toxicity verification showed that effector proteins g6495 and g10555 could inhibit the tobacco necrosis symptoms caused by Bax,but effector proteins g10613,g13620 and g11335 could not.The results showed that the genes g6495 and g10555 and their homonym effector proteins were toxic.The genes g10613,g13620,g10555 and their eponymous effector proteins were not toxic.2.Subcellular localization of the effector proteins encoded by the two genes with toxicity was carried out,and fluorescence signal detection was carried out by confocal microscope.It was found that the fluorescence signal of g6495 and g10555 effector proteins appeared on the cell membrane,while the negative control showed fluorescence signal on the nuclear membrane without loading.The results showed that effector proteins g6495 and g10555 were mainly expressed on cell membrane.