| Abstrakt: |
As one of calmodulin families, calmodulin-like (CMLs) regulate plant growth and development, hormone regulation, stresses, which have great significance to the further study of calmodulin-like protein involved in pollen germination and growth. In this study, to explore the interaction between CMLs and NPG1 related genes in pollen development of Brassica oleracea L. var. capitata, and to explore the function of CMLs in pollen development and germination, 12 CMLs genes related to pollen development and germination, and pollen germination genes NPG1, NPGR1, and NPGR2 were cloned from "ZG". Interactions between them were detected by yeast two-hybrid test. and the relative expression levels of NPG1, NPGR1, and NPGR2 after different pollination time was detected. The results showed that the open reading frames (ORFs) of 12 CMLs genes ranged from 447 bp to 1095 bp, among which CML20, CML48, CML49, and CML50 contained 4 introns, CML21 contained 2 introns, and the others contained no introns. All of them contained no signal peptide and transmembrane domain, all of them were extracellular proteins and hydrophilic proteins which contain 2--4 EF-hands, with amino acids range of 148--364 aa. Subcellular localization was mainly located in cell membrane, vacuole, and nucleus. NPG1, NPGR1, and NPGR2 were clustered on different evolutionary branches, which were the closest to Brassica rape and Brassica napus, and closer to Arabidopsis thaliana. Yeast two-hybrid showed that CML13, CML21, CML24, CML42 could not interact with NPG1, NPGR1, and NPGR2 in vitro, CML15, CML20, CML25, CML39, CML41, CML48, and CML49 could interact with NPG1, NPGR1, and NPGR2, CML50 could interact with NPGR1 and NPGR2, but not with NPG1. The relative expression level of NPG1 increased significantly after pollination, while the relative expression levels of NPGR1 and NPGR2 didn't change significantly. The relative expression levels of CMLs genes were varied at different pollination times, and CML15, CML20, CML39, CML41, and CML49 were a trend of first increasing and then decreasing. In conclusion, several CMLs proteins were screened which can interact with NPG1 and NPGRs, indicating that CMLs can participate in the process of pollen germination and pollen tube elongation, and it was expected to provide references for the research on the mechanism of Ca2+ signaling system participating in pollen germination and pollen tube elongation. [ABSTRACT FROM AUTHOR] |
