| 中华蛸糖代谢相关基因克隆及表达分析 |
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| 引用本文： | 陈小灵,孙玉龙,朱友芳,张子平,王艺磊.中华蛸糖代谢相关基因克隆及表达分析[J].中国水产科学,2022,29(12):1738-1749. |
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| 作者姓名： | 陈小灵 孙玉龙 朱友芳 张子平 王艺磊 |
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| 作者单位： | 集美大学水产学院, 福建 厦门 361021;福建农林大学海洋学院, 福建 福州 350002;莆田市水产科学研究所, 福建 莆田 351100 |
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| 基金项目： | 福建省科技计划项目(2022N0035)； 福建省科技计划项目(2022T3028). |
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| 摘 要： | 头足类的养殖主要依赖于天然饵料, 目前无人工配合饲料可用, 这一问题成为其发展大规模养殖的制约因素, 研究中华蛸(Octopus sinensis)的糖代谢模式可为其配合饲料中碳水化合物的添加比例提供参考依据。本研究克隆了中华蛸葡萄糖-6-磷酸异构酶(glucose phosphate isomerase, GPI)、磷酸甘油酸激酶(phosphoglycerate kinase, PGK) 和丙酮酸激酶(pyruvate kinase, PK) 3 个糖代谢酶的相关基因, 并对其表达进行了检测。结果显示, 中华蛸 OsGPI 的开放阅读框(open reading frame, ORF)1725 bp, 编码 574 个氨基酸; OsPGK 的 ORF1248 bp, 编码 415 个氨基酸, OsPK 的 ORF1683 bp, 编码 560 个氨基酸。qRT-PCR 结果显示, 这 3 个基因在不同组织/器官中均有表达, 但在脑、 消化腺和后唾液腺中的表达水平最高。3 个基因在胚胎不同发育时期皆有表达, 总体趋势是初孵幼体的表达量高于多细胞期。在幼体饥饿胁迫过程中, 随着饥饿时间的推移, 3 个基因在饥饿 2 d 后的表达水平显著降低, 紧接着在饥饿 3 d 和 4 d 后表达水平显著升高, 最后在饥饿 5 d 后的表达水平开始回落并降低。在投喂配合饲料(compound feeds)、锐齿蟳(Charybdis acuta)和沙丁鱼(Sardina pilchardus)的实验中, 与投喂 0 d 相比, 配合饲料组和沙丁鱼组在投喂 21 d 后, OsGPI、OsPGK 和 OsPK 在中华蛸消化腺的表达均有不同程度的升高, 在投喂 42 d 后, OsGPI、OsPGK 和 OsPK 的表达量出现不同程度的回落。锐齿蟳组在投喂 21 d 和 42 d 后, OsGPI、OsPGK 和 OsPK 的表达量相较投喂 0 d 时的均显著升高。OsGPI、OsPGK 和 OsPK 在胚胎发育、饥饿和不同饲料投喂过程中的表达模式说明其参与了中华蛸糖代谢过程的调节, 本研究可为中华蛸饲料的选择和配合饲料中碳水化合物的添加提供基础数据。
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| 关 键 词： | 中华蛸 糖代谢 饥饿 饲料 表达分析 |
Cloning and expression analysis of genes related to glucose metabolism in Octopus sinensis |
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| CHEN Xiaoling,SUN Yulong,ZHU Youfang,ZHANG Ziping,WANG Yilei.Cloning and expression analysis of genes related to glucose metabolism in Octopus sinensis[J].Journal of Fishery Sciences of China,2022,29(12):1738-1749. |
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| Authors: | CHEN Xiaoling SUN Yulong ZHU Youfang ZHANG Ziping WANG Yilei |
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| Institution: | Fisheries College, Jimei University, Xiamen 361021 , China;College of Marine Science, Fujian Agriculture and Forestry University, Fuzhou 350002 , China;Putian Municipal Institute of Fishery Science, Putian 351100 , China |
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| Abstract: | At present, the cultivation of cephalopods mainly relies on natural feeds. The absence of artificial compound feeds is a restrictive factor for the large-scale development of the cultivation of cephalopods. The genes related to glucose metabolism in Octopus sinensis, including glucose phosphate isomerase (GPI), phosphoglycerate kinase (PGK), and pyruvate kinase (PK), were cloned, and their expression levels were examined to study the glucose metabolism pattern of O. sinensis and provide a reference for optimizing the proportion of carbohydrates in its compound feed. The results showed that the open reading frame (ORF) length of OsGPI was 1725 bp encoding 574 amino acids, the ORF length of OsPGK was 1248 bp encoding 415 amino acids, and the ORF length of OsPK was 1683 bp encoding 560 amino acids. The sequence alignment and phylogenetic tree results showed that the identity of O. sinensis GPI, PGK, and PK proteins was the closest to that of Octopus bimaculoides. qRT-PCR results showed that these three genes were expressed in different tissues/organs of O. sinensis, with OsGPI having the highest expression level in the digestive gland and brain, and OsPGK and OsPK in the brain, followed by posterior salivary glands and the digestive glands. This difference in expression distribution may be related to the biological functions performed by different tissues/organs. OsGPI, OsPGK and OsPK were expressed in different developmental stages, with the highest expression at the hatchling stage. During starvation stress in larvae, the expression levels of OsGPI, OsPGK, and OsPK decreased significantly after 2 days (d) of starvation, followed by a significant increase in expression levels after 3 d or 4 d of starvation. Finally, the expression levels started to decrease after 5 d of starvation. In the experiment of feeding compound feeds, Charybdis acuta or Sardina pilchardus, the expression levels of OsGPI, OsPGK, and OsPK in the digestive gland of O. sinensis were increased to varying degrees in the compound feed group and S. pilchardus group after 21 d feeding. The expression levels of OsPGK and OsPK decreased to varying degrees. The expression levels of OsGPI, OsPGK, and OsPK were significantly increased in the C. chinensis group after feeding for 21 d and 42 d, compared to those at 0 d. After 42 d of feeding, the expression levels of OsGPI, OsPGK, and OsPK decreased to varying degrees. The expression levels of OsGPI, OsPGK, and OsPK were significantly increased in the C. acuta group after feeding for 21 d and 42 d, compared to those at 0 d. The expression patterns of OsGPI, OsPGK, and OsPK during different embryonic development stages, starvation, and feeding of different feeds indicated that they were involved in regulating glucose metabolism in O. sinensis. These results may aid in the selection of diets and the addition of carbohydrates in compound feeds. |
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