Cloning and Physiological Function Analysis of GmCBL7 Gene in Soybean (Glycine max)
CHEN Nan, LI Jing, SHEN Xiang-Juan, PENG Ya-Nan, SUN Dan-Dan, WANG Quan-Wei*
College of Life Science and Technology, Harbin Normal University/Key Laboratory of Molecular Cytogenetics and Genetic Breeding of Heilongjiang Province, Harbin 150025, China
Abstract：Soybean (Glycine max) is main grain and oil crops in China, salinity and drought have seriously affected the yield and quality of soybeans. Calcineurin B-like protein (CBL) is a type of calcium receptor, which plays an important role in the process of plant stress response. The results of the expression pattern analysis of soybean CBL gene family in the early stage of this research group showed that soybean GmCBL7 could be induced by high salt and drought.In order to further study the relationship between GmCBL7 gene and salt tolerance and drought resistance. In this study, GmCBL7 (GenBank No. XM 006578843.3) was cloned from soybean by reverse transcription-PCR (RT-PCR) technology. The CDS sequence of the gene was 672 bp in length, encoding 223 amino acids, the molecular weight of the protein was 25.8 kD, and the theoretical isoelectric point was 4.81. It was an acidic protein with a typical EF-hand domain and no signal peptide. It was a non-secreted protein. Analysis of systematic evolution showed that GmCBL7 was in the same evolutionary branch as the homologous protein in legumes, and had the closest relationship with the homologous protein in wild soybean (Clycine soja). The plant expression vector pCPB-GmCBL7 was constructed to genetically transform tobacco (Nicotiana tabacum), and analyzed the physiological functions of the GmCBL7 transgenic tobacco for stress resistance, the results showed that after high salt (200 mmol/L NaCl) and drought (20% PEG6000) stress treatments for 6 and 12 h, the activity of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) and content of proline were significantly higher than those of wild type, while the content of malondialdehyde (MDA) was significantly lower than that of wild type. It was suggested that overexpression of GmCBL7 gene could improve salt tolerance and drought resistance of transgenic tobacco.This study provides a basis for further exploring the function of GmCBL7 gene, and provides a new gene for soybean salt and drought resistance breeding.
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