Expression Of TaCDPKs Genes In The Process Of Wheat Infected By Puccinia Triticina And Screening Of TaCPK2Interacting Protein

The calmodulin-depended protein kinase abbreviat CDPK have an important Ca2+signaling receptors which are involved in various physiological and biochemical processesin plants. CDPKs contain four conserved very different functional areas, from theN-terminus variable region sequence, the catalytic region, regulatory region and theconnecting region. CDPK could serve as an important necessary condition after theoriginal infection triggers a series of plant disease resistance shoubing reaction. The thestudy had important implications CDPK to reveal the mechanisms of plant resistance topathogen infection.Using the early suppression subtractive hybridization test, it found that a part ofTaCDPK gene expression levels in wheat by leaf rust infection occurred after the changed,suggesting TaCDPK family might play a role in wheat leaf rust interaction process. Usingreal-time quantitative PCR, it confirmed that TaCPK2might be involved in signaltransduction resistant wheat leaf rust infection. Using the Western Blotting and he yeasttwo-hybrid screening technology, it found one high homology sHSP sequence（small heatshock protein, the small heat shock proteins）.For further research TaCDPK family in wheat leaf rust role in the process interaction,this experiment in14TaCDPK wheat gene for the study, based on14wheat TaCDPK genesequence specific primers were designed, the wheat varieties Lovering10, respectively,and leaf rust physiological races260and165composed of incompatible combination withaffinity combined points after inoculation time points0h,4h,8h,12h,16h,24h,48hsampled and analyzed by quantitative real-time PCR technology to detect14TaCDPKgene expression profiles. The results showed that14TaCDPK gene expression patternscan be divided into four categories: Expression of TaCPK1.TaCPK2.TaCPK6.TaCPK7four genes increased; The expression of TaCPK3.TaCPK18.TaCPK19three genes reduced;TaCPK5.TaCPK8.TaCPK9.TaCPK12.TaCPK13.TaCPK15co-expression of six genes littlechange in trend. Initially confirmed that these modes of expression correspondingTaCDPK wheat resistance genes may play in the process of leaf rust infection werepositive control, negative control and no regulatory role. For further study of leaf rustresistance TaCDPK family ’s role in the infection process wheat foundation. To have aninsight into the function of TaCPK2, the TaCPK2of wheat （Triticum aestivum L.） wascloned into PGBKT7vector firstly. Then we use Matchmaker Library Construction&Screening Kits to screened the yeast two hybrid cDNA library of wheat leaf which invadedby leaf rust. Eventally7putative positive clones were obtained. One of them is Zea mays clone155847940S ribosomal protein S5mRNA, complete cds and hypothetical proteinTRIVR3₀7388[Triticum urartu],and sHSP（small heat shock protein）.Meanwhile, the use of yeast-one conversion of verification will TaCPK2and sHSPinteraction between the two was transformed into yeast cells, initially confirmed theinteraction between the two occurred, sHSP involved in the wheat leaf rust interactionprocess. For further research sHSP leaf rust resistance in wheat infested role in the processand wheat leaf rust resistance signal transduction mechanism of the foundation.All the results above made a foundation for the further study of the TaCDPK functionand the mechanism in the reaction between the wheat and leaf rust.