Cloning of LbCER1 Gene and Expression Analysis under Osmotic Stress in Lycium barbarum ssp. Bianguo

doi:10.7668/hbnxb.201751628

Abstract

Abstract: In order to identify the molecular characterization of fatty-aldehyde decarboxylase gene (ECERIFERUM1, CER1) involved in osmotic stress of xerophyte, CER1 gene was cloned from the cDNA of Lycium barbarum ssp. Bianguo by reverse transcription PCR (RT-PCR) and RACE method, which was a kind of salt-tolerant and drought-tolerant plant. Then the sequence was analyzed by bioinformatics software. Furthermore, the gene expression level of CER1 in L. barbarum ssp. Bianguo was detected in leaf, stem and root under osmotic stress treatment by quantitative Real-time PCR (qRT-PCR). The results showed that the gene sequence of CER1 in L.barbarum ssp. Bianguo contained 2 168 bp in full length and an open reading frame of 1 881 bp, which was named LbCER1, encoding 626 amino acids. Based on bioinformatics analysis, the protein contained four transmembrane regions, a fatty acid hydroxylase domain and a Wax2 domain of C-terminal. The calculated molecular weight, isoelectric point, aliphatic index, instability index and average hydropathicity (GRAV) of LbCER1 was 72.47 ku, 7.40, 91.28, 27.88 and -0.070, respectively. LbCER1 is a stable hydropathic protein possessing alkalinity and high-temperature tolerance, which is located in the membrane of plastic reticulum. Based on the secondary structure prediction of LbCER1, the majority of its amino acid compose alpha-helix (44.09%) and loop (45.21%), which was confirmed by tertiary structure prediction. In addition, LbCER1 was predicted to have 100% homologous with Solanaceae CER1 by phylogenetic analysis. Expression pattern of LbCER1 showed that it express in roots, stems, and leaves. The LbCER1 was also induced by 160 mmol/L sorbitol for 6 h in leaf, stem, and root to 1.8, 1.2, and 1.1 fold, respectively. On contrary, LbCER1 was only sensitive to 80 mmol/L sorbitol in leaves after 6 h-treatment. These results indicate that LbCER1 may play a role in osmotic stress response in L. barbarum ssp. Bianguo.

Key words: Lycium barbarum ssp. Bianguo, LbCER1 gene, Cloning, Gene expression

CLC Number:

S567.03
Q78

YUAN Huijun, MA Qianguo, GAO Ze, LI Xueyong, BAO Jingting, WANG Chunmei, LI Hujun. Cloning of LbCER1 Gene and Expression Analysis under Osmotic Stress in Lycium barbarum ssp. Bianguo[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.7668/hbnxb.201751628.

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