刺参温度相关基因在模拟不同海域冬春季水温条件下的表达研究

白雪秋，丁君，曹学顺，李成泽，常亚青

(大连海洋大学 农业部北方海水增养殖重点实验室，辽宁 大连 116023)

摘要:为探讨不同海域冬春季水温对刺参温度相关基因的表达影响，在实验室内模拟了俄罗斯沿海地区、大连沿海地区、福建沿海地区12月至翌年4月刺参Apostichopus japonicus生长的海水温度,并从已构建的刺参耐寒基因筛选cDNA文库和刺参高温胁迫正反消减cDNA文库中选取了7个差异基因:hsp70、gp96、UQCRFS1、ferritin、lysozyme、mtLSU、proeintl(2)efl，利用实时荧光定量PCR方法研究了这7个基因在3种海域冬春季模拟水温条件下的表达情况。结果表明：3个水温组刺参同一组织中hsp70基因的表达量变化不明显，但在不同组织中的表达量存在差异；模拟福建沿海地区冬春季水温组刺参各组织中gp96基因的表达量整体上均高于其他两个水温组，并且在模拟福建沿海地区冬春季水温组刺参呼吸树中的表达量显著高于该组肠和体壁(P<0.05)；模拟俄罗斯沿海地区冬春季水温组刺参各组织中UQCRFS1基因的表达量均高于其他两个水温组；3个水温组刺参各组织中ferritin基因的表达模式及表达量均不相同；3个水温组刺参各组织中lysozyme和proeintl(2)efl两个基因的表达量均在呼吸树中达到最高，且显著高于肠和体壁(P<0.05)；3个水温组刺参各组织中mtLSU基因的表达模式基本相同，其表达量均表现为体壁显著高于肠和呼吸树(P<0.05)。

关键词:刺参；温度；实时荧光定量PCR；功能基因；差异表达

DOI：10.3969/J.ISSN.2095-1388.2015.01.004

文章编号：2095-1388(2015)01-0018-07

中图分类号：S917.1

文献标志码：A

收稿日期：2014-05-17

基金项目：辽宁省教育厅优秀人才支持计划项目(LTQ2011073); 辽宁省重大科技攻关项目(2011203003); 国家海洋公益性行业科研专项(201105016-3)

作者简介：白雪秋(1988—), 女, 硕士研究生。E-mail:baixue135qiu@sina.cn

Abstract：The genes related to temperature including hsp70, gp96, UQCRFS1, ferritin, lysozyme, mtLSU, and proeintl(2)efl selected from the subtracted cDNA library were expressed in body wall, intestine and respiratory tree of sea cucumber Apostichopus japonicas using quantitative Real-time PCR under the simulated sea water temperature in Russian coastal area, Dalian coastal area, and Fujian coastal area from December to April in a laboratory. The expression of hsp70 gene was found to be not significant difference in the same tissue in the three temperature groups, but there was significant differences in different tissues at the same conditions. There was higher expression of gp96 gene in the simulation of Fujian coastal area in winter and spring group than in the other two groups, significantly higher relative expression levels in respiratory tree than in intestine and the body wall in the simulation of Fujian coastal area in winter and spring group (P<0.05). The expression of UQCRFS1 gene was higher in the simulation of Russian coastal area in winter and spring groups than in the other two groups. No same ferritin gene expression patterns and relative expression levels was observed in the three temperature groups. The maximal relative expression levels of both lysozyme and proeintl (2)efl genes were found in the respiratory tree, significantly higher than in the intestine and body wall in the three groups (P<0.05). There were essentially the same mtLSU gene expression patterns in the three groups, significantly higher relative expression levels in the body wall than that in the intestine and respiratory tree (P<0.05).

刺参Apostichopusjaponicus(Selenka)是亚洲海岸广泛分布的浅海底栖物种，栖息于潮间带至水深20~30 m的浅海，主要分布于35°N到44°N的西北太平洋沿岸，自然分布区北起俄罗斯远东地区，经朝鲜、韩国、日本,南至中国北部[1-2]。刺参可以在0~30 ℃的水温范围内生存，但刺参的生长水温为12~21 ℃，幼参生长的最适水温为15~18 ℃，水温是影响刺参生长和生理活动的重要因子之一,水温大于24 ℃时刺参开始进入夏眠阶段[3]。……