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Screening and Identification of NsylCBL Family Members Interacting with Protein Kinase NsylCIPK24a in Nicotiana Sylvestris

The CIPK (CBL-interacting protein kinase) kinase family in plants is a type of serine/threonine protein kinase family. The members of this family interact with the upstream CBL protein (Calcineurin B-like protein) to form the CBL-CIPK signal system, involved in regulating plant growth and development as well as stress response processes . Early research found that AtCIPK24 interacts with AtCBL4 and AtCBL10, respectively, to activate downstream target proteins to respond to high salt stress. Earlier in this study, NsylCIPK24a homologous to AtCIPK24 from Arabidopsis thaliana was obtained from Nicotiana sylvestris , but CBL family members interacting with NsylCIPK24a and its specific function are unclear. Therefore, a genome-wide prediction of CBL family members of N . sylvestris was carried out ; P redicted NsylCBL genes were cloned by RT-PCR, and the analyses of gene structure, protein conserved domain and expression pattern were then conducted. The NsylCBL members interacting with NsylCIPK24a were screened by the yeast two-hybrid system. Results showed that there are 12 potential NsylCBL genes in N . sylvestris , and all of them were cloned successfully. Four members including NsylCBL4, NsylCBL5, NsylCBL9 and NsylCBL10 could interact with NsylCIPK24a in yeast. There might be some similar CBL-CIPK pathways in tobacco plants, compared with Arabidopsis . The study provides experimental data for the functional identification of NsylCIPK24a , and increases understanding of CBL-CIPK network in N . sylvestris .