不同温度条件下皱纹盘鲍幼鲍对低盐胁迫的响应及渗透压调节机制的初步研究

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	不同温度条件下皱纹盘鲍幼鲍对低盐胁迫的响应及渗透压调节机制的初步研究
	李俊元
学位类型	硕士
导师	刘晓
	2015-05-22
学位授予单位	中国科学院大学
学位授予地点	北京
学位专业	海洋生物学
关键词	皱纹盘鲍低盐温度耐受性渗透压调节
摘要	皱纹盘鲍（Haliotis discus hannai Ino）自然分布于西北太平洋沿岸，在我国自然分布于大连、烟台、威海、青岛、日照近岸浅海区。我国从20世纪80年代中期开始皱纹盘鲍的人工繁育和养殖，传统养殖区在大连、烟台和威海；从20世纪90年代初开始，皱纹盘鲍被南移到长江以南海域养殖，东海现已经成为皱纹盘鲍越冬基地和重要养殖区。我国南方地区具有降雨集中、降水量大的特点，因此养殖鲍受低盐胁迫的概率比在传统养殖区高得多。目前，有关皱纹盘鲍对环境因子的响应只限于单因素的研究，而温度、盐度对皱纹盘鲍的联合作用未见报道。本研究旨在探寻不同温度下，皱纹盘鲍幼鲍对低盐胁迫的响应及调节机制，包括不同温度下，皱纹盘鲍幼鲍对低盐胁迫的耐受力分析；温度、盐度对皱纹盘鲍幼鲍生长的影响；皱纹盘鲍幼鲍经长期低盐胁迫后，血淋巴各种离子及可溶性总蛋白含量的变化。现将主要成果总结如下： 1不同温度下幼鲍的盐度-死亡率动力学曲线实验设置6、10、14、18、22、24、26、28、30℃ 9个温度梯度，每个温度条件下设置7个盐度，分别为盐度16、18、20、22、24、26、30，每个温度盐度处理设置3个重复，每个重复30只幼鲍。培育90d后对照组（盐度30）在培育水温10～26℃范围内存活率在较高，在96.7%以上，在该温度范围内，不同盐度条件下幼鲍死亡状况如下：盐度24～30在水温10～26℃范围内存活率达到94%以上；盐度22，温度14、22、26℃培育的幼鲍分别于75、60、60d死亡率显著上升，培育90d后死亡率分别为7.8%、8.9%、26.7%；盐度20，温度14、18、22、26℃条件下，幼鲍死亡率分别在45、60、60、60d显著提高，培育90d后死亡率分别达到18.9%、21.1%、24.4%、34.8%；盐度18，温度10、14、18、22、26℃条件下幼鲍死亡率分别在75、45、45、30、30d显著提高，经90d胁迫处理死亡率分别达到18.9%、41.1%、65.6%、65.6%、64.4%。温度10~24℃，盐度16胁迫60d后，幼鲍死亡率均达到92%以上。以上结果表明：盐度24~30为幼鲍存活的适宜盐度，盐度22在10~22℃是适宜的，而在26℃条件下不适宜幼鲍生存；盐度18~20不适宜幼鲍的生存；盐度16为幼鲍的低盐耐受极限，在此盐度条件下幼鲍几乎全部死亡。 2 驯化养殖对幼鲍低盐耐受能力的影响设置5个温度梯度，分别为10、14、18、22、24℃，每个温度下设置低盐驯化组（盐度为22、20）及非驯化组（自然海水作对照）。经90d驯化后，将各处理组幼鲍置于盐度16海水中进行胁迫处理，结果表明盐度16海水中，经盐度22、20驯化的幼鲍在10、14、18℃条件下的死亡过程比对照组明显减慢；22、24℃条件下驯化对幼鲍死亡过程的减缓效果并不明显。 3 幼鲍在不同温度盐度下培育的生长状况实验设置8个温度梯度，分别为6、10、14、18、22、24、26、28℃，每个温度下设置三个盐度，为盐度30、26、24，每个温盐处理设3个重复，每个重复30只幼鲍。初始壳长在22.1~24.9mm，63d后测量幼鲍的壳长、壳宽及全湿重。结果表明，10～26℃范围内盐度30、26、24处理组幼鲍存活率均在96%以上，18~24℃为幼鲍生长的适宜温度范围，其中生长率最高点为温度22℃盐度26；在适宜生长的温度范围内适当降低盐度有利于幼鲍的生长。 4 低盐胁迫对幼鲍血淋巴生化组分的影响以温度26℃自然海水及温度26℃盐度24海水中预培养的皱纹盘鲍幼鲍为实验材料，将其进行低盐胁迫后，对幼鲍肌肉组织水分含量、血淋巴上清液中可溶性总蛋白、血蓝蛋白、牛磺酸、游离氨基酸及无机离子进行测定，结果表明：幼鲍肌肉组织含水量、血淋巴上清液中血蓝蛋白、游离氨基酸及牛磺酸含量均随着盐度的降低而呈现出上升的趋势。幼鲍血淋巴中Na⁺、Cl^-、Ca²⁺浓度随盐度降低显著下降，并于24h内达到稳定；K⁺浓度无显著变化。初步研究结果表明：血蓝蛋白、牛磺酸、游离氨基酸参与了幼鲍在低盐胁迫后的渗透压调节；幼鲍对Na⁺、Cl^-、Ca²⁺无调节能力，但对K⁺具有调节能力。
其他摘要	Pacific abalone (Haliotis discus hannai Ino) is naturally distributed along the Pacific Northwest and inhabits off the shore of Dalian, Yantai, Weihai, Qingdao, Rizhao City in neritic region of China. From 1980’s China initiated artificial breeding and cultivation of H. discus hannai which was conventionally carried out in Dalian, Yantai, and Weihai City. At the beginning of 1990’s, H. discus hannai was transferred southward to the south sea area of Yangtze River to feed and the East China Sea became a major area for H. discus hannai to overwinter and culture. Rainfall is concentrated and large in South China, so H. discus hannai is more susceptible to low salinity sea water there than in traditional mariculture area. By far, the researches on the response of H. discus hannai are limited to one single environmental factor, and the combined effects of temperature and salinity on H. discus hannai have not been reported. This thesis was aimed to explore the response of H. discus hannai exposed to low salinities at different temperatures and osmoregulatory mechanism of juvenile abalones acclimated to low salinity, including analysis of juvenile abalones’ tolerance to low salinity at different temperatures; the influence of temperature and salinity on abalone’s growth; influence of low salinity on biochemical composotions of juvenile abalones. The main results were summarized below: 1 salinity-mortality curve of juvenile abalones at different temperatures Juvenile abalones were studied using a two factorial experiment design with nine levels of temperature (6，10，14，18，22，24，26，28，30℃) and seven levels of salinity(16, 18, 20, 22, 24, 26, 30‰). Three replicate containers were used per treatment, with 30 juvenile abalones in each container. The survivals of 30‰ control groups at 10-26℃ after 90 days’ cultivation were above 96.7%, so we disscused the the mortality of juvenile abalones at different salinities between 10-26℃: the survival rates of salinity 24-30, 10-26℃groups were above 94%; mortalities at 22‰ and 14, 22, 26℃ increased significantly at 75^th , 60^thand 60^th day and reached 7.8%, 8.9% and 26.7% respectively after 90 days’ treatment; mortalities at 20‰ and 14, 18, 22, 26℃ increased significantly at 45 ^th, 60^th, 60^th, 60^th day and reached 18.9%, 21.1%, 24.4%, 34.8% respectively after 90 days’ treatment; mortalities at 18‰ and 10, 14, 18, 22, 26℃ increased significantly at 75^th, 45^th, 45^th, 30^th and 30^th day respectively and and reached 18.9%, 41.1%, 65.6%, 65.6%, 64.4% after 90 days’ treatment. Mortalities of the groups exposed to 16‰ at 10-24℃were above 92% after 60 days’ treatment. According to the results above, we can concluded that salinities between 24 to 30‰ were suitable for the survival of juvenile abalones, 22‰ and 10 to 22℃ were also suitable for abalones to survive but not at 22‰ and 26℃; salinities between 18 to 20‰ were not suitable for the survival of juvenile abalones; 16‰ was the tolerance limit of juvenile abalones in low salinities, at which almost all the abalones could not survive. 2 influence of adaption on juvenile abalones’ tolerance to low salinity Adaptive regimes were composed of five temperature levels (10, 14, 18, 22, 24℃) and three salinity levels (22 and 20‰ as adaptive groups; sea water as unadaptive groups). After 90 days’ adaption, all the groups were exposed to 16‰ and the result showed that adaptive groups at 10, 14 and 18℃ significantly postponed the death of juvenile abalones exposed 16‰, but the groups at 22, 24℃ could not. 3 the growth of juvenile abalones in different temperature and salinity Experiment was designed with eight levels of temperature (6,10, 14, 18, 22, 24, 26,28℃) and three levels of salinities (30, 26, 24‰), 90 individuals (3×30) were used in each treatment. The length of shells were controlled in the range of 22.1 to 24.9 mm. Shell length, shell width and total wet weight were measured after 63 days’ treatment. Survivals of the groups in 10-26℃ were above 96%. 18 to 24℃ were the most suitable temperature range for juvenile abalones’ growth, within which appropriately lowering salinity promoted abalone’s growth. 4 Influence of low salinity on biochemical compositions of juvenile abalones The juvenile abalones pre-cultured at sea water, 26℃ and salinity 24, 26℃ were stressed with low salinity , and water content of muscle tissue , soluble total protein , hemocyanin, taurine, free amino acid, and inorganic ions in supernatant hemolymph were measured. The result showed that water content in muscle tissue, hemocyanin, free amino acid and taurine increased when salinity decreased. Na⁺、Cl^-、Ca²⁺concentration declined significantly with the decreasing of sea water and stabilized within 24h, while K⁺ concentration did not varied significantly. Preliminary result suggested that hemocyanin, taurine, free amino acid functioned in osmoregulation of juvenile abalones in low salinity; juvenile abalones were actively capable of K⁺ regulation but not of Na⁺, Ca²⁺or Cl^-.
学科领域	主要研究方向 ; 海洋生物学
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/23758
专题	海洋生物技术研发中心
作者单位	中国科学院海洋研究所
第一作者单位	中国科学院海洋研究所
推荐引用方式 GB/T 7714	李俊元. 不同温度条件下皱纹盘鲍幼鲍对低盐胁迫的响应及渗透压调节机制的初步研究[D]. 北京. 中国科学院大学,2015.

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