Cloning And Function Of TaRIPK Gene Of Wheat Xiaoyan 6

Wheat stripe rust is an epidemic fungal disease caused by Puccinia striiformis f.sp.tritici(Pst),which occurs in major wheat producing countries all over the world.The epidemic year caused huge yield loss.Xiaoyan6(XY6)is a typical cultivar with the high-temperature seedling plant(HTSP)resistance.It has the characteristics of non-race specialization and persistent disease resistance,and can resist adverse environment.However,the molecular mechanism of HTSP resistance is not yet clear.Here,based on the transcriptome sequencing in the early stage of the laboratory,we cloned the full length of the gene of TaRIPK(RPM1-induced protein kinase).Based on sequence analysis of TaRIPK,the quantitative polymerase chain reaction analysis,gene silencing,transient expression system and yeast two-hybrid system,we explored the function of TaRIPK.The mainly results of this study are as following:1.The TaRIPK gene of Xiaoyan6 was cloned.TaRIPK,which was significantly upregulated during the HTSP resistance of Xiaoyan6 to wheat stripe rust,was screened based on the transcriptome sequencing analysis.We cloned the full length of TaRIPK,which contains an open reading frame(ORF)of 1434 bp,encoding 477 amino acids,including a serine/threonine protein kinase and a signal peptide at the N-terminal.TaRIPK has high homology with Aet RIPK(Aegilops tauschii).2.TaRIPK positively regulates the HTSP resistance of Xiaoyan6 to stripe rust.TaRIPK has the highest expression in leaves,which was 3.99 and 28.09 times higher than that in roots and stems,respectively.The transcriptional expression level of TaRIPK was induced a higher expression at 204 hpi(the inoculated plants are converted from normal temperature to high temperature for 12 h)under the infection of Pst and temperature transition.The results of virus-induced gene silencing showed that compared with unsilenced plants,the pustules of plant were significantly increased,the number of necrotic cells was significantly reduced,and the linear length of uredinia was significantly increased.3.The molecular mechanism of TaRIPK participating in the resistance of Xiaoyan 6 to stripe rust in high temperature was preliminarily clarified.The expression of TaRIPK was significantly up-regulated after treatment with exogenous hormones salicylic acid,methyl jasmonate,and ethephon for 0.5 h.The expression levels of Ta PR2 and Ta PR3 related to the defense-related genes of the salicylic acid and methyl jasmonate pathways were downregulated after knocking down of TaRIPK,and the expression level of TaRPM1 decreased.The interaction protein Ta PLCP1 of TaRIPK was screened by yeast two-hybrid,which may be related to programmed cell death and plant immunity.