Expression Of TaCPK2 Gene In The Process Of Wheat Infected By Puccinia Triticina And Screening Of TaCPK2 Interacting Protein

Calcium-dependent protein kinases (CDPKs) are crucial sensors of calcium concentration changes in plant cells under diverse endogenous and environmental stimuli.The TaCPK2-EST was isolated from the wheat SSH cDNA library induced by leaf rust, which has a high level expression in 8h after inoculated leaf rust in incompatible combination. CDPKs(calcium-dependent protein kinases) are key elements in decoding the changes in cytosolic calcium concentrations during the plant defense responses. The western blotting analysis and RT-PCR analysis were used to further investigate relative expression of the TaCPK2 in different time points after wheat inoculated leaf rust. These results showed that the expression of TaCPK2 was obviously increased in the incompatible combination compared with the control whether at the levels of mRNA or protein at 4 h after inoculation, and the level of mRNA was declined to that in control level at 16 h which the protein level reached the maximum at the meantime. In compatible combination, TaCPK2 was detectable at 8 h after inoculation, then declined to the normal level. These results implied that TaCPK2 was involved in leaf rust resistance in wheat. To have an insight into the function of TaCPK2, the Ta CPK2K of wheat (Triticum aestivum L.) was cloned into PGBKT7 vector firstly. Then we use Matchmaker Library Construction & Screening Kits to screened the yeast two hybrid cDNA library of wheat leaf which invaded by leaf rust. Eventally 4 putative positive clones were obtained. One of them is small heat shock protein.All the results above made a foundation for the further study of the Ta CPK2’s function and the mechanism in the reaction between the wheat and leaf rust.