Gene Cloning of SCD-1a and SCD-1b in Donghai No.1 Large Yellow Croaker (Larimichthys crocea) and Their Responses to Cold Stress
LI Ming-Yun1,*, MIAO Liang1, CHEN Ying-Ying1, LI Chong1, ZHANG Li-Ning2
1 State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Ningbo University, Ningbo 315211, China; 2 Zhejiang Mariculture Research Institute, Wenzhou 325005, China
Abstract：Large yellow croaker (Larimichthys crocea) is a kind of warm-temperature fish. Low temperature in winter can cause large death of cultured L. crocea, so it is necessary to carry out researches on how to deal with low temperature stress. In order to study the sequence characteristics of stearoyl coenzyme A desaturase 1 (SCD-1) gene of Donghai No.1 large yellow croaker and their responses to cold stress, the full length of SCD-1a (GenBank No. MT325796) and SCD-1b (GenBank No. MT325797) cDNA were cloned by rapid amplification of cDNA ends (RACE) technique, and expression level under chronic and acute cold stress were analysed by qRT-PCR. The full length of SCD-1a cDNA was 3 095 bp which contained a 1 014 bp ORF and encoded 337 amino acid. The full length of SCD-1b cDNA was 2 636 bp which contained a 1 008 bp ORF and encoded 335 amino acid. SCD-1a and SCD-1b both contained 3 conserved histidine rich regions, and the sequence similarity was 72.2%. The predicted tertiary protein structure of SCD-1a and SCD-1b were highly similar and both contained 4 transmembrane helical structure. In the phylogenetic tree based on amino acids, the SCD of teleost divided into 2 clusters, the SCD-1a and SCD-1b of L. crocea located in 2 clusters separately, indicating that they were different subtypes. Under normal growth temperature (18 ℃), the expression levels of SCD-1a and SCD-1b were both high in liver and lower in other tissues. During chronic-cold stress (decreasing slowly from 12 ℃ to 6 ℃ by the method of cooling at the rate of 0.5 ℃/12 h and continuing to cool down after maintaining for 12 h), the expression of SCD-1a significantly increased in gill, skill, muscle, intestine and brain, and the expression of SCD-1b significantly increased in skill, muscle, intestine, brain and heart, nevertheless SCD-1a expression decreased significantly in liver. During acute cold stress (from 12 to 8 ℃ immediately), the expression level of SCD-1b significantly increased in all tissues, and SCD-1a expression significantly increased in all tissues except skill. The results of this study have certain reference significance for understanding the mechanism of response and adaptation of L. crocea to low temperature stress.
李明云, 苗亮, 陈莹莹, 李翀, 张立宁. 东海1号大黄鱼SCD-1a和SCD-1b基因克隆及其对低温胁迫的应答[J]. 农业生物技术学报, 2020, 28(10): 1820-1829.
LI Ming-Yun, MIAO Liang, CHEN Ying-Ying, LI Chong, ZHANG Li-Ning. Gene Cloning of SCD-1a and SCD-1b in Donghai No.1 Large Yellow Croaker (Larimichthys crocea) and Their Responses to Cold Stress. 农业生物技术学报, 2020, 28(10): 1820-1829.
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