%A Xiao-Jing BAI,Xiao-Ping LIAN,Yu-Kui WANG,He-Cui ZHANG,Qian-Ying LIU,Tong- Hong ZUO,Yi-Zhong ZHANG,Qin-Qin XIE,Deng-Ke HU,Xue-Song REN,Jing ZENG,Shao-Lan LUO,Min PU,Li-Quan ZHU %T Cloning and analysis of BoCDPK14 in self-incompatibility Brasscia olerace %0 Journal Article %D 2019 %J Acta Agronomica Sinica %R 10.3724/SP.J.1006.2019.94040 %P 1773-1783 %V 45 %N 12 %U {https://zwxb.chinacrops.org/CN/abstract/article_6776.shtml} %8 2019-12-12 %X

Brasscia oleracea self-incompatibility (SI) is the reaction of rejection or inhibition of stigma to pollens from the same haplotype. Calcium-dependent protein kinase (CDPK) is an important component of plants involved in resistance to stress signals. In this study, we identified an up-regulated gene named BoCDPK14 based on the stigma transcriptome data in 0-60 min self-pollination. The gene was highly homologous to the calcium dependent protein kinase gene involved in stress signaling in Arabidopsis lyrata. BoCDPK14 gene contained an open reading frame (ORF) with the length of 1599 bp, encoding a protein with the length of 533 amino acid residues. It was a hydrophilic protein expressed in cytoplasm, and could be induced to express a 60.4 kD protein in the cytoplasm of E. coli Transetta (DE3). It was suggested that BoCDPK14 is an active cytoplasmic protein. The 2000 bp upstream of BoCDPK14 translation initial codon contained elements for responses to stress, hormone, and metabolic regulation. BoCDPK14 was expressed in stigma, pollen, flower bud, petal, and leaf, with lower expression level in stigma than in pollen. The results of qRT-PCR revealed that BoCDPK14 mRNA expression level after self- and cross-pollinations for 0 min to 60 min was consistent with that of RNA-seq data. The interaction between BoCDPK14 protein kinase domain and glutamate receptor channel protein BoGLR2.8d was identified by yeast two-hybrid, suggesting that BoCDPK14 might be a novel protein involved in SI reaction. These results suggest that BoCDPK14 may act as a Ca 2+ signal element to participate the process of response to pollen stimulation in the stigma of Brasscia oleracea, which provides a new insight into the further research and utilization of self-incompatibility of Brasscia oleracea.