温度对底播虾夷扇贝适合度性状影响的研究

发布时间：2018-04-19 19:23

本文选题：底播虾夷扇贝 + 适合度性状　；参考：《中国科学院研究生院(海洋研究所)》2016年博士论文

【摘要】：虾夷扇贝(Patinopecten yessoensis)是一种重要的冷水性经济贝类之一。其自然分布区域位于西北太平洋,在韩国、日本以及中国均有大规模的人工养殖。目前虾夷扇贝的养殖方式主要包括浮筏养殖和底播养殖。底播养殖由于不需要额外的养殖设施,可以减少养殖成本获得更高的养殖效益。在中国,獐子岛海区是虾夷扇贝底播养殖的主要区域。2008年之前,长海县虾夷扇贝底播面积已经达到122万亩左右,如今长海县獐子岛虾夷扇贝的确权海域达到322万亩。但是随着底播面积的增加,许多新的问题不断出现,比如养殖个体小型化,回捕率降低等问题严重影响着獐子岛虾夷扇贝底播产业的发展。2011年之后,獐子岛虾夷扇贝的亩产一直稳定在80公斤左右,产量难以有大的提高。本研究以底播虾夷扇贝的标志物为抓手,研究底播虾夷扇贝季节性生理、营养指标及基因表达水平与生产性能的相关性,尝试构建现场及实验室水平的扇贝底播效率精确评估方法,探讨影响扇贝底播效率的关键环境因子;开展免疫学及转录组学研究,尝试探讨虾夷扇贝对高温及温度剧变等关键环境因子应答及分子机制;通过构建大规模家系,依据数量遗传模型,探讨关键胁迫环境因子(温度)下虾夷扇贝的适应潜力,获得适合底播养殖的虾夷扇贝品种。以期对虾夷扇贝的底播养殖提供有益的借鉴。1.底播虾夷扇贝生产性能同环境因子、抗氧化代谢酶活性以及相对表达量关系的模型2012年12月-2014年12月期间同大连獐子岛集团股份有限公司合作,在獐子岛海洋牧场选择4个站位进行影响底播虾夷扇贝适合度性状环境因子的调查工作。通过调查,我们发现底播虾夷扇贝在每年的12月至次年6月的生长速度(壳长和重量)明显高于6月至12月的生长速度(每年下半年几乎停止生长)。通过线性回归分析发现海水温度、溶解氧含量、叶绿素含量、悬浮性颗粒物(SPM)以及颗粒性有机物(POM)等环境因子严重影响了底播虾夷扇贝的生长、存活、性腺指数、肥满度等生产性能。营养组分(糖原含量)以及几种重要的免疫代谢相关酶活性(谷丙转氨酶GPT、乳酸脱氢酶LDH、超氧化物歧化酶SOD以及总抗氧化能力TAOC)及其基因的相对表达量均会对虾夷扇贝的生产性能产生影响。通过以上调查分析,我们建议虾夷扇贝底播场地应该选择建在温度偏低的地区以降低死亡率提高性腺指数,选择悬浮性颗粒物(spm)含量较低的地区,以提高壳长生长率、重量生长率以及性腺指数,选择颗粒性有机物(pom)含量较高的地区,以提高底播虾夷扇贝的肥满度。使用线性回归分析,获得了同虾夷扇贝壳长生长率、重量生长率、性腺指数、肥满度、死亡率具有显著相关性的几种重要的代谢以及抗氧化酶及基因表达量。基于这些酶活性以及基因相对表达量数据首次利用主成分分析的方法构建了预测底播虾夷扇贝生产性能的预测模型,经过pearson关联分析可知,这些预测模型同相应的生产性能之间具有显著的相关性(p0.05)。这些模型是对虾夷扇贝生长率、死亡率传统调查方法的进一步补充,对底播虾夷扇贝养殖业的发展具有一定的促进作用。2.虾夷扇贝对高温刺激的分子应答机制在实验室条件下使用相应的酶活试剂盒以及rnaseq技术分别从生理、转录组学层面对高温导致虾夷扇贝死亡的因素进行探究。受到28℃高温刺激以及恢复过程中,二龄贝的死亡率显著高于一龄贝。cox模型分析结果显示,高温刺激后虾夷扇贝的存活率同贝龄、gpt活性、p53含量、taoc活性以及hsp70含量变化均显著相关(p0.05),其中gpt活性同扇贝的存活负相关(b0),其余因素同扇贝存活呈显著正相关(b0)。此外,高温导致虾夷扇贝心跳紊乱以及心输出量的减少,进而减少血淋巴对机体的供氧能力,导致扇贝死亡。通过转录组学分析可知,在恢复24h时一龄贝中atp相关基因表达量下调导致虾夷扇贝死亡;二龄贝中atp合成过程以及atp合成过程中电子传递过程导致了atp合成量降低更严重,导致了扇贝死亡率的升高。而且二龄贝在恢复24h时错配修复相关基因也显著下调,导致dna错配概率升高,在受到高温刺激之后导致其死亡。再者,一龄贝在整个恢复过程中核糖体合成相关基因表达量的上升,在一定程度上减少了虾夷扇贝的死亡,同时二龄贝在恢复48h条件下才能出现核糖体合成以及翻译相关基因的显著上升表达,说明受到高温刺激之后的恢复能力方面二龄贝明显不如一龄贝。受到高温刺激之后,相对于二龄贝hsp70等相关基因在一龄贝体内表达量显著升高,也在一定程度上解释了二龄贝死亡率偏高的问题。3.虾夷扇贝底播养殖性状的遗传评估利用獐子岛集团2011年和1985年自日本引进的虾夷扇贝为亲本,通过家系育种方法于2013年2月培育73个家系,在2013年12选择35个家系进行虾夷扇贝标记底播实验,在2014年12进行统计计算其死亡率并同浮筏养殖的同一家系进行比较,得到10个底播存活率显著高于浮筏养殖的家系。同时使用AI-REML(或Quasi-REML)方法对2013年家系进行遗传参数评估。依据遗传参数计算每个个体及家系的估计育种值(EBV),依据30%:70%经济加权后计算各个家系及个体的综合育种值。共筛选21个家系作为选育F2家系的亲本。在2015年利用筛选出的家系进行杂交,利用底播以及浮筏养殖个体作为对照构建2015年家系。选育组及对照组按常规沉箱及浮筏养殖流程分6个测定点进行培育至2016年12月,目前该部分数据正在采集中。
[Abstract]:Patinopecten yessoensis is one of the most important cold water shellfish. Its natural distribution area is located in the Northwest Pacific Ocean, in Korea, Japan and China. The cultivation methods of scallop mainly include floating raft culture and bottom sowing culture. Bottom sowing culture is not required for extra breeding. In China, Changhai County is the main area of sowing and breeding of scallop and scallop in Changhai County.2008 years ago. The sowing area of scallop scallop in Changhai County has reached about 1 million 220 thousand mu. Now, the sea area of Changhai County is 3 million 220 thousand mu. In addition, many new problems have emerged, such as the miniaturization of the individual culture and the reduction of the recapture rate, which seriously affect the development of the bottom sowing industry of the scallop scallop. After.2011 years, the yield of the scallop of the scallop has been stable at about 80 kilograms, and the yield is difficult to be greatly improved. Hand, study the seasonal physiology, nutrition index, gene expression level and production performance of scallop scallop, try to construct the accurate evaluation method of scallop sowing efficiency in the field and laboratory level, explore the key environmental factors that affect the sowing efficiency of scallop, and carry out the study of immunology and transcriptional studies to try to explore the scallop scallop. The response and molecular mechanism of key environmental factors, such as high temperature and temperature upheaval, were constructed in large scale families, based on the quantitative genetic model, to explore the adaptation potential of the scallop scallop under the critical environmental factors (temperature), and to obtain the varieties suitable for the bottom sowing of the scallop. In order to provide a useful reference for the bottom sowing of the scallop, the.1. bottom sowing was provided. The relationship between the environmental factors, the activity of antioxidant enzymes and the relative expression amount of the production performance of scallop's scallop scallop, in December 2012 -2014 year and December, in cooperation with the Limited by Share Ltd of Dalian water deer island group, selected 4 stations in the sea pasture of the deer son island to investigate the environmental factors affecting the suitability of the bottom sowing of the scallop scallop. We found that the growth rate (shell length and weight) of the scallop from December to June of the next year was significantly higher than that of June to December (almost ceased to grow in the second half of the year). The sea temperature, dissolved oxygen content, chlorophyllin content, suspended particulate matter (SPM) and particulate organic matter (PO) were found by linear regression analysis. M) and other environmental factors seriously affected the growth, survival, sex gland index, fullness and other production properties of the bottom sowing shrimp scallop. Nutritional components (glycogen content) and several important activities of immuno metabolic related enzymes (alanine transaminase GPT, lactate dehydrogenase LDH, superoxide dismutase SOD, and total antioxidant capacity TAOC) and relative expression of their genes According to the above investigation, we suggest that the bottom sowing site of the scallop's scallop should be built in a low temperature area to reduce the death rate and increase the gonadal index, and to select the areas with low SPM content to increase the growth rate of the shell, the weight growth rate and the gonadal index. Select the area with high content of granular organic matter (POM) in order to improve the fullness of the bottom sowing scallop. By linear regression analysis, several important metabolism, antioxidant enzymes and gene expressions of the long rate, weight growth rate, gonadal index, manure index, fatness and mortality were obtained. Enzyme activity and gene relative expression data were used to predict the production performance of scallop scallop by principal component analysis for the first time. Through Pearson correlation analysis, the prediction models have significant correlation with corresponding production performance (P0.05). These models are the growth rate and death of scallop scallop. Further supplement to the traditional investigation method, it has a certain promoting effect on the development of shrimp scallop cultivation in the bottom sowing, the molecular response mechanism of.2. scallop to the high temperature stimulation in the laboratory conditions, using the corresponding enzyme activity kit and the rnaseq technology from the physiological and transcriptional layer to the cause of the death of the scallop. The mortality of two age shellfish was significantly higher than that of the one age.Cox model in the process of high temperature stimulation and recovery at 28 C. The results showed that the survival rate of the scallop was significantly correlated with the age of shellfish, GPT activity, p53 content, TAOC activity and HSP70 content (P0.05), and GPT activity was negatively correlated with the survival of scallop. (B0), the remaining factors have a significant positive correlation with the survival of the scallop (B0). In addition, high temperature leads to the disturbance of heartbeat and the decrease of cardiac output, which reduces the oxygen supply capacity of the body and causes the death of scallops. Through the transcriptional analysis, the down-regulation of ATP related genes in the first instar shellfish in the first instar of 24h leads to the scallop of the scallop. Death; the ATP synthesis process in the two age shellfish and the electron transfer process in the process of ATP synthesis lead to the decrease of the ATP synthesis, which leads to the increase of the scallop mortality. Moreover, the related genes of the mismatch repair of the two age shellfish also decrease significantly in the recovery of 24h, leading to the increase of the DNA mismatch probability and the death after being stimulated by high temperature. In the whole recovery process, the expression of ribosome related genes was increased, and the death of the scallop was reduced to a certain extent. At the same time, the two age shellfish could produce ribosome synthesis and significantly increase the expression of related genes under the condition of recovery of 48h, indicating that the recovery ability of two instar bainite after the high temperature stimulation was in the age of bainite. Compared with two age shellfish HSP70 and other related genes in the first age shellfish, the expression of the two age shellfish increased significantly, and to some extent explained the problem of the high mortality rate of two age shellfish. The genetic evaluation of the bottom sowing of the scallop scallop,.3., was used for the introduction of the prawn fan introduced from Japan in 2011 and 1985. As a parent, 73 families were bred in February 2013 by family breeding. In 2013, 35 families were selected to carry out the labelled bottom sowing experiment of scallop's scallop. In 2014 12, the mortality rate was calculated and compared with the same family in the same family with floating raft culture. The survival rate of 10 bottom sowing was significantly higher than that of floating raft culture. At the same time, AI-R was used. The EML (or Quasi-REML) method was used to evaluate the genetic parameters of the family in 2013. Based on the genetic parameters, the estimated breeding values of each individual and family (EBV) were calculated, and the comprehensive breeding values of families and individuals were calculated according to 30%: 70% economic weighting. 21 families were selected as parents for the selection of F2 families. In 2015, the selected families were selected. The breeding group and the control group were divided into 6 Determination points according to the conventional caisson and floating raft culture process to December 2016, and the data were being collected in the 2015.

【学位授予单位】：中国科学院研究生院(海洋研究所)
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S917.4

【参考文献】