發(fā)布時(shí)間：2018-04-19 19:23

  本文選題：底播蝦夷扇貝 + 適合度性狀��； 參考：《中國科學(xué)院研究生院(海洋研究所)》2016年博士論文

【摘要】：蝦夷扇貝(Patinopecten yessoensis)是一種重要的冷水性經(jīng)濟(jì)貝類之一。其自然分布區(qū)域位于西北太平洋,在韓國、日本以及中國均有大規(guī)模的人工養(yǎng)殖。目前蝦夷扇貝的養(yǎng)殖方式主要包括浮筏養(yǎng)殖和底播養(yǎng)殖。底播養(yǎng)殖由于不需要額外的養(yǎng)殖設(shè)施,可以減少養(yǎng)殖成本獲得更高的養(yǎng)殖效益。在中國,獐子島海區(qū)是蝦夷扇貝底播養(yǎng)殖的主要區(qū)域。2008年之前,長�？h蝦夷扇貝底播面積已經(jīng)達(dá)到122萬畝左右,如今長�？h獐子島蝦夷扇貝的確權(quán)海域達(dá)到322萬畝。但是隨著底播面積的增加,許多新的問題不斷出現(xiàn),比如養(yǎng)殖個(gè)體小型化,回捕率降低等問題嚴(yán)重影響著獐子島蝦夷扇貝底播產(chǎn)業(yè)的發(fā)展。2011年之后,獐子島蝦夷扇貝的畝產(chǎn)一直穩(wěn)定在80公斤左右,產(chǎn)量難以有大的提高。本研究以底播蝦夷扇貝的標(biāo)志物為抓手,研究底播蝦夷扇貝季節(jié)性生理、營養(yǎng)指標(biāo)及基因表達(dá)水平與生產(chǎn)性能的相關(guān)性,嘗試構(gòu)建現(xiàn)場及實(shí)驗(yàn)室水平的扇貝底播效率精確評估方法,探討影響扇貝底播效率的關(guān)鍵環(huán)境因子;開展免疫學(xué)及轉(zhuǎn)錄組學(xué)研究,嘗試探討蝦夷扇貝對高溫及溫度劇變等關(guān)鍵環(huán)境因子應(yīng)答及分子機(jī)制;通過構(gòu)建大規(guī)模家系,依據(jù)數(shù)量遺傳模型,探討關(guān)鍵脅迫環(huán)境因子(溫度)下蝦夷扇貝的適應(yīng)潛力,獲得適合底播養(yǎng)殖的蝦夷扇貝品種。以期對蝦夷扇貝的底播養(yǎng)殖提供有益的借鑒。1.底播蝦夷扇貝生產(chǎn)性能同環(huán)境因子、抗氧化代謝酶活性以及相對表達(dá)量關(guān)系的模型2012年12月-2014年12月期間同大連獐子島集團(tuán)股份有限公司合作,在獐子島海洋牧場選擇4個(gè)站位進(jìn)行影響底播蝦夷扇貝適合度性狀環(huán)境因子的調(diào)查工作。通過調(diào)查,我們發(fā)現(xiàn)底播蝦夷扇貝在每年的12月至次年6月的生長速度(殼長和重量)明顯高于6月至12月的生長速度(每年下半年幾乎停止生長)。通過線性回歸分析發(fā)現(xiàn)海水溫度、溶解氧含量、葉綠素含量、懸浮性顆粒物(SPM)以及顆粒性有機(jī)物(POM)等環(huán)境因子嚴(yán)重影響了底播蝦夷扇貝的生長、存活、性腺指數(shù)、肥滿度等生產(chǎn)性能。營養(yǎng)組分(糖原含量)以及幾種重要的免疫代謝相關(guān)酶活性(谷丙轉(zhuǎn)氨酶GPT、乳酸脫氫酶LDH、超氧化物歧化酶SOD以及總抗氧化能力TAOC)及其基因的相對表達(dá)量均會(huì)對蝦夷扇貝的生產(chǎn)性能產(chǎn)生影響。通過以上調(diào)查分析,我們建議蝦夷扇貝底播場地應(yīng)該選擇建在溫度偏低的地區(qū)以降低死亡率提高性腺指數(shù),選擇懸浮性顆粒物(spm)含量較低的地區(qū),以提高殼長生長率、重量生長率以及性腺指數(shù),選擇顆粒性有機(jī)物(pom)含量較高的地區(qū),以提高底播蝦夷扇貝的肥滿度。使用線性回歸分析,獲得了同蝦夷扇貝殼長生長率、重量生長率、性腺指數(shù)、肥滿度、死亡率具有顯著相關(guān)性的幾種重要的代謝以及抗氧化酶及基因表達(dá)量�；�于這些酶活性以及基因相對表達(dá)量數(shù)據(jù)首次利用主成分分析的方法構(gòu)建了預(yù)測底播蝦夷扇貝生產(chǎn)性能的預(yù)測模型,經(jīng)過pearson關(guān)聯(lián)分析可知,這些預(yù)測模型同相應(yīng)的生產(chǎn)性能之間具有顯著的相關(guān)性(p0.05)。這些模型是對蝦夷扇貝生長率、死亡率傳統(tǒng)調(diào)查方法的進(jìn)一步補(bǔ)充,對底播蝦夷扇貝養(yǎng)殖業(yè)的發(fā)展具有一定的促進(jìn)作用。2.蝦夷扇貝對高溫刺激的分子應(yīng)答機(jī)制在實(shí)驗(yàn)室條件下使用相應(yīng)的酶活試劑盒以及rnaseq技術(shù)分別從生理、轉(zhuǎn)錄組學(xué)層面對高溫導(dǎo)致蝦夷扇貝死亡的因素進(jìn)行探究。受到28℃高溫刺激以及恢復(fù)過程中,二齡貝的死亡率顯著高于一齡貝。cox模型分析結(jié)果顯示,高溫刺激后蝦夷扇貝的存活率同貝齡、gpt活性、p53含量、taoc活性以及hsp70含量變化均顯著相關(guān)(p0.05),其中g(shù)pt活性同扇貝的存活負(fù)相關(guān)(b0),其余因素同扇貝存活呈顯著正相關(guān)(b0)。此外,高溫導(dǎo)致蝦夷扇貝心跳紊亂以及心輸出量的減少,進(jìn)而減少血淋巴對機(jī)體的供氧能力,導(dǎo)致扇貝死亡。通過轉(zhuǎn)錄組學(xué)分析可知,在恢復(fù)24h時(shí)一齡貝中atp相關(guān)基因表達(dá)量下調(diào)導(dǎo)致蝦夷扇貝死亡;二齡貝中atp合成過程以及atp合成過程中電子傳遞過程導(dǎo)致了atp合成量降低更嚴(yán)重,導(dǎo)致了扇貝死亡率的升高。而且二齡貝在恢復(fù)24h時(shí)錯(cuò)配修復(fù)相關(guān)基因也顯著下調(diào),導(dǎo)致dna錯(cuò)配概率升高,在受到高溫刺激之后導(dǎo)致其死亡。再者,一齡貝在整個(gè)恢復(fù)過程中核糖體合成相關(guān)基因表達(dá)量的上升,在一定程度上減少了蝦夷扇貝的死亡,同時(shí)二齡貝在恢復(fù)48h條件下才能出現(xiàn)核糖體合成以及翻譯相關(guān)基因的顯著上升表達(dá),說明受到高溫刺激之后的恢復(fù)能力方面二齡貝明顯不如一齡貝。受到高溫刺激之后,相對于二齡貝hsp70等相關(guān)基因在一齡貝體內(nèi)表達(dá)量顯著升高,也在一定程度上解釋了二齡貝死亡率偏高的問題。3.蝦夷扇貝底播養(yǎng)殖性狀的遺傳評估利用獐子島集團(tuán)2011年和1985年自日本引進(jìn)的蝦夷扇貝為親本,通過家系育種方法于2013年2月培育73個(gè)家系,在2013年12選擇35個(gè)家系進(jìn)行蝦夷扇貝標(biāo)記底播實(shí)驗(yàn),在2014年12進(jìn)行統(tǒng)計(jì)計(jì)算其死亡率并同浮筏養(yǎng)殖的同一家系進(jìn)行比較,得到10個(gè)底播存活率顯著高于浮筏養(yǎng)殖的家系。同時(shí)使用AI-REML(或Quasi-REML)方法對2013年家系進(jìn)行遺傳參數(shù)評估。依據(jù)遺傳參數(shù)計(jì)算每個(gè)個(gè)體及家系的估計(jì)育種值(EBV),依據(jù)30%:70%經(jīng)濟(jì)加權(quán)后計(jì)算各個(gè)家系及個(gè)體的綜合育種值。共篩選21個(gè)家系作為選育F2家系的親本。在2015年利用篩選出的家系進(jìn)行雜交,利用底播以及浮筏養(yǎng)殖個(gè)體作為對照構(gòu)建2015年家系。選育組及對照組按常規(guī)沉箱及浮筏養(yǎng)殖流程分6個(gè)測定點(diǎn)進(jìn)行培育至2016年12月,目前該部分?jǐn)?shù)據(jù)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.

【學(xué)位授予單位】：中國科學(xué)院研究生院(海洋研究所)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S917.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 郝振林;劉京哲;唐雪嬌;湛W歐，

本文編號：1774391