刺参Apostichopus japonicus生态免疫与夏眠的基础研究

IOCAS-IR > 海洋环流与波动重点实验室

	刺参Apostichopus japonicus生态免疫与夏眠的基础研究
其他题名	Mechanism of Ecoogical Immunity and Aestivation of Apostichopus japonicus
	王方雨
学位类型	博士
	2008-06-07
学位授予单位	中国科学院海洋研究所
学位授予地点	海洋研究所
关键词	刺参生态免疫环境胁迫儿茶酚胺类激素夏眠抑制消减杂交
摘要	针对目前刺参养殖过程中亟需解决的种质、环境和产品品质之间日益突出的矛盾，本论文从刺参的自身防御和内分泌机制入手，比较系统地研究了环境胁迫与细胞免疫和应激激素之间的作用机理；并从生理和分子的角度对刺参的夏眠机理进行了初步研究，主要研究成果如下： 1. 查阅国内外大量文献资料，较为系统地综述了海参的免疫体系、生态免疫学研究进展、当前夏眠机理的研究状况和刺参夏眠的基础研究。对刺参生态免疫及其夏眠机理研究提出了新的见解和思路。 2. 研究了温度(0、8、16、24、32℃)和盐度(20、25、35)的急性胁迫(72h)作用对刺参免疫指标的影响。刺参体腔液内几种免疫酶类活性受升温胁迫影响显著；而降温胁迫的影响并不显著。高盐胁迫可使刺参体腔液细胞吞噬活性、超氧化物歧化酶和过氧化氢酶活性显著变化，而对髓过氧化物酶和溶菌酶活性都没有显著影响。从这几种免疫指标变化情况来看，温度胁迫对刺参的影响要强于盐度胁迫，且高温胁迫的影响要强于低温胁迫。 3. 研究了温度(8、16、24℃)、盐度(20、25、35)和露空等急性胁迫(24h)作用对刺参体腔液儿茶酚胺类激素含量的影响。急性温度胁迫能够显著影响刺参体腔液内肾上腺素、去甲肾上腺素和多巴胺的含量；而盐度和露空胁迫对刺参体腔液肾上腺素含量、去甲肾上腺素含量以及多巴胺含量都没有产生显著影响。 4. 完成了刺参体腔液几种酶类变化的周年测定(2006年7月到2007年6月)。一年内刺参体腔液内几种酶类变化的转折点为9月份，10月份，1月份，2月份，4月份和5月份。刺参体腔液内几种酶类活性变化与温度和盐度等环境因素并不存在简单的相关关系，因此推测这些显著变化并不是单一温度、盐度等因素作用的结果，还可能与刺参生长、繁殖及其它环境因素的作用有关。 5. 开展了夏眠期间刺参免疫指标的现场研究(2006年7月到11月)。在此期间刺参体腔液细胞总数显著下降；刺参体腔液内免疫相关酶类均在8月份、9月份达到最高值；肾上腺素和去甲肾上腺素的含量均在8月下旬和11月下旬显著升高，而多巴胺含量则没有显著变化。刺参完全进入夏眠的8、9月份，体腔液内各项免疫指标也都具有显著变化，说明夏眠对刺参机体的免疫状态产生影响。 6. 探讨了刺参夏眠前后肠道基因表达的差异(2007年7月至12月)。抑制消减杂交实验结果表明接头连接效率大于50%，说明连接效率较高。消减效率分析结果表明，消减后进行cDNA保守序列扩增比未消减落后约10个循环才能明显看到扩增产物，说明对共同序列的消减效率比较高。利用抑制消减杂交技术成功构建了符合技术要求的夏眠刺参与未夏眠刺参肠道抑制消减杂交基因文库。
其他摘要	Deteriorations of germ plasm, environment and product quality in Aprostichopus japonicus culture become more and more serious. In view of this, studies on defense and neuroendocrine mechanism of A. japonicus were carried out. The interaction among environmental stress, stress hormone and immune functions was investigated systematically. On the base of physiology and molecular, we explored the mechanism of aestivation of A. japonicus. The main results are as follows: 1. The recent progress largely derived from immune characters, ecological immunity and aestivation of A. japonicus were reviewed in this paper. Furthermore, new thoughts on ecological immunity and aestivation of A. japonicus were suggested. 2. Temperature stress (0, 8, 16, 24, 32ºC) and salinity stress (20, 25, 35) on immune enzymes in coelome fluid of A. japonicus were undertaken in laboratory. Activities of immune enzymes were significantly affected by high-temperature stress, but no obvious changes in low-temperature stress. Superoxide dismutase and catalase activity were significantly affected by salinity change, but activities of myeloperoxidase and lysozyme demonstrated that A. japonicus tolerates limited salinity stress. High-temperature stress had greater effect on the immune responses of A. japonicus than low-temperature and salinity stresses. 3. Changes of catecholamine content in coelome fluid of A. japonicus caused by temperature stress (8, 16, 24ºC) and salinity stress (20, 25, 35) were estimated in laboratory. Activities of adrenalin, noradrenalin and dopamine in coelome fluid of A. japonicus were significantly affected by acute temperature stress, but not by salinity and exposure stress. 4. From July 2006 to June 2007, enzymes in coelome fluid of A. japonicus have been measured. Data showed that turning points were in September, October, January, February, April and May. In view of the life characters of A. japonicus, the significant changes of enzymes activity were associated with growth, reproduction and environment. 5. From July to November in 2006, the immune characters of A. japonicus were detected in field. Total coelomecyte count in coelome fluid of A. japonicus decreased significantly during aestivation. Activities of immune enzymes in coelome fluid of A. japonicus recorded high values during August and September which corresponds with the highest water temperature record during these months. Activities of adrenalin and noradrenalin increased significantly in the later August and November, but there was no obvious change of dopamine during aestivation. Data showed that the immune characters in coelome fluid of A. japonicus were affected during aestivation. 6. From July to November in 2007, gene differences in intestinal tract of A. japonicus during aestivation were cloned with suppression subtractive hybridization. Two percent gels electrophoresis to identify efficiency of adaptor ligation in suppression subtractive hybridization, more than 50% was efficient. Subtractive and unsubtractive cDNA as templates carried out PCR amplification to evaluate subtractive efficiency. The gels electrophoresis results showed that subtractive efficiency was efficient. Data showed that the subtractive library about intestine during aestivation was efficient.
页数	114
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/229
专题	海洋环流与波动重点实验室
推荐引用方式 GB/T 7714	王方雨. 刺参Apostichopus japonicus生态免疫与夏眠的基础研究[D]. 海洋研究所. 中国科学院海洋研究所,2008.

条目包含的文件
文件名称/大小	文献类型	版本类型	开放类型	使用许可
10001_20051800681203（1024KB）			限制开放	--	浏览