本文選題：海洋酸化 + 福建牡蠣�。� 參考：《廈門大學(xué)》2014年碩士論文

【摘要】：隨著人為排放到大氣中的CO2持續(xù)增加,由CO2驅(qū)動(dòng)的海洋酸化將導(dǎo)致海水pH值在本世紀(jì)顯著下降,并很可能對(duì)海洋生物,特別是對(duì)鈣質(zhì)無(wú)脊椎動(dòng)物及生態(tài)系統(tǒng)造成嚴(yán)重負(fù)面影響。越來(lái)越多的研究表明,海洋酸化會(huì)影響海洋鈣質(zhì)無(wú)脊椎動(dòng)物的存活、發(fā)育和生理機(jī)能。然而,目前關(guān)于海洋酸化對(duì)福建牡蠣(Crassostrea angulata)幼體影響的研究報(bào)道還比較少。本文通過(guò)實(shí)驗(yàn)室內(nèi)模擬未來(lái)海洋環(huán)境,研究了短期高C02暴露對(duì)我國(guó)重要經(jīng)濟(jì)水產(chǎn)養(yǎng)殖種類—福建牡蠣幼體發(fā)育的影響(對(duì)照組CO2濃度400ppm,實(shí)驗(yàn)組分別為1500ppm和3000ppm)。利用形態(tài)學(xué)和分子生物學(xué)方法對(duì)酸化暴露后的幼體進(jìn)行相關(guān)研究,主要結(jié)果如下： 1.海洋酸化對(duì)福建牡蠣幼體貝殼生長(zhǎng)的影響 通過(guò)對(duì)福建牡蠣幼體貝殼形態(tài)參數(shù)的測(cè)量分析,發(fā)現(xiàn)1500ppm實(shí)驗(yàn)組較對(duì)照組貝殼形態(tài)無(wú)顯著性差異,3000ppm實(shí)驗(yàn)組中幼體個(gè)體較對(duì)照組偏小,貝殼增長(zhǎng)速度慢,畸形率升高。進(jìn)一步采用掃描電鏡聯(lián)用EDS能譜法分析技術(shù)分析酸化暴露后D型幼體貝殼元素成分,發(fā)現(xiàn)海洋酸化對(duì)幼體鈣元素沉積有一定的抑制作用,而且發(fā)現(xiàn)福建牡蠣幼體對(duì)1500ppm酸化程度有一定的耐受性。 2.海洋酸化條件F福建牡蠣幼體數(shù)字基因表達(dá)譜分析 通過(guò)對(duì)正常條件下和海洋酸化條件下的福建牡蠣擔(dān)輪幼體、D型初期幼體和D型2d幼體進(jìn)行高通量測(cè)序,分別兩兩比較后獲得差異基因5317個(gè)、74個(gè)和2641個(gè)。差異基因的GO功能顯著性富集分析和Pathway顯著性富集分析結(jié)果顯示,海洋酸化主要抑制了胚胎發(fā)育、能量代謝、基礎(chǔ)生物學(xué)過(guò)程和鈣離子傳導(dǎo)相關(guān)基因和通路的作用,從而導(dǎo)致幼體生長(zhǎng)延遲。 3.海洋酸化對(duì)福建牡蠣幼體貝殼形成和生長(zhǎng)相關(guān)基因的影響 通過(guò)全胚原位雜交技術(shù)發(fā)現(xiàn)Tyr3、TGF-β信號(hào)通路和BMP信號(hào)通路均參與福建牡蠣幼體InCaS生物發(fā)生。在酸化條件下,TGF-β信號(hào)通路相關(guān)基因表達(dá)受到抑制,而Tyr3基因和BMP信號(hào)通路相關(guān)基因表達(dá)上調(diào)。形態(tài)學(xué)研究結(jié)果顯示,在海洋酸化條件下福建牡蠣面盤幼體貝殼生長(zhǎng)緩慢且個(gè)體偏小,畸形率高,因此,我們推測(cè)Tyr3和BMP信號(hào)通路相關(guān)基因上調(diào)表達(dá)后仍不能補(bǔ)償海洋酸化對(duì)幼體造成的損傷,所以福建牡蠣在海洋酸化條件下貝殼雖仍能持續(xù)生長(zhǎng)但大小形狀都較對(duì)照組有變化。 本研究通過(guò)掃描電鏡聯(lián)用EDS能譜法分析技術(shù)分析福建牡蠣幼體貝殼元素成分,發(fā)現(xiàn)海洋酸化對(duì)鈣元素沉積有一定抑制作用；利用RNA-Seq技術(shù)探究海洋酸化對(duì)福建牡蠣幼體早期生長(zhǎng)發(fā)育影響,結(jié)果顯示,海洋酸化抑制早期胚胎發(fā)育、能量代謝和鈣離子傳導(dǎo)相關(guān)信號(hào)通路作用,從而導(dǎo)致幼體生長(zhǎng)延遲；利用成熟的WISH方法探究Tyr3、TGF-β信號(hào)通路和BMP信號(hào)通路相關(guān)基因的功能,并用qRT-PCR技術(shù)研究了海洋酸化條件下它們的表達(dá)情況,結(jié)果顯示,它們與InCaS生物發(fā)生和貝殼生長(zhǎng)有關(guān),在酸化條件下,TGF-β信號(hào)通路相關(guān)基因表達(dá)受到抑制,而Tyr3基因和BMP信號(hào)通路相關(guān)基因上調(diào)表達(dá),結(jié)合形態(tài)學(xué)研究結(jié)果推測(cè),海洋酸化條件下,某些基因和信號(hào)通路上調(diào)表達(dá)仍不能補(bǔ)償酸化對(duì)幼體造成的損傷而導(dǎo)致幼體貝殼生長(zhǎng)緩慢,畸形率升高。
[Abstract]:With anthropogenic emissions to the atmosphere of CO2 continued to increase, driven by CO2 ocean acidification will cause seawater pH value decreased significantly in this century, and is likely to marine organisms, especially caused a serious negative impact on the calcareous invertebrates and ecosystem. More and more studies show that ocean acidification will affect the survival of marine calcareous invertebrates. Developmental and physiological function. However, at present about ocean acidification on oyster (Crassostrea angulata) in Fujian reported the influence of larvae is still relatively small. Through laboratory simulation of the future of the marine environment, to research the high short-term effects of C02 exposure on the development of China's economically important aquaculture species in Fujian oyster larvae (control group CO2 concentration 400ppm the experimental group, respectively 1500ppm and 3000ppm). The research of larval acidification after exposure by morphological and molecular biological methods, the main results As follows:
Effect of 1. ocean acidification on the growth of shellfish of oyster larvae in Fujian
Through the measurement of Fujian oyster larvae shell morphology parameter analysis, found the 1500ppm experimental group than in the control group has no difference of morphology of the shell, 3000ppm in the experimental group than the control group of individual larvae, shell growth slow, malformation rate was higher. Further by means of scanning electron microscopy combined with EDS analysis technology to analyze the acidification after exposure to type D larvae shell element spectrum method, found that ocean acidification has inhibitory effects on larval calcium deposition, and found that the Fujian oyster larvae had certain tolerance to 1500ppm acidification.
2. marine acidification conditions F analysis of digital gene expression profiles in the juvenile oysters of Fujian
Based on the normal conditions and ocean acidification conditions of Fujian oyster trochophora, D type and D type 2D early larvae larvae by high-throughput sequencing, respectively, compared to 22 for differential gene 5317, 74 and 2641. The difference of GO gene function significant enrichment analysis and Pathway significant enrichment analysis results show that ocean acidification mainly inhibits embryonic development, energy metabolism, basic biological processes and calcium related genes and pathways, leading to larval growth delay.
Effects of 3. ocean acidification on the formation and growth related genes of the shell formation and growth of oyster larvae in Fujian
Through the whole embryo in situ hybridization showed that Tyr3, TGF- beta and BMP signal pathways are involved in InCaS biogenesis. Fujian oyster larvae in acid conditions, TGF- beta signaling pathway related gene expression was inhibited, and Tyr3 gene and BMP signaling pathway related gene expression. Results showed that morphology, in ocean acidification conditions Fujian oyster veliger shell growth is slow and the individual small, deformity rate is high, therefore, we speculate that the expression of Tyr3 and BMP signaling pathway related genes up-regulated after still cannot compensate for the damage caused by ocean acidification of larvae, the Fujian ocean acidification conditions in oyster shells can still continue to grow in size and shape but compared with the control group change.
This study by scanning electron microscopy combined with EDS analysis of Fujian oyster larvae shell element technology spectrum method, found that ocean acidification has inhibitory effect on calcium deposition; using RNA-Seq technology to explore the ocean acidification on Fujian oyster larvae early growth effect, results show that the inhibition of early embryonic development of ocean acidification, energy metabolism and calcium ion conduction the related signal pathway, leading to larval growth delay; exploring the Tyr3 using the WISH method of mature, TGF- beta signaling pathway and BMP signaling pathway related genes, and to study the expression of their ocean acidification conditions by qRT-PCR technology showed that they and InCaS biogenesis and shell growth, in the condition of acidification TGF-, beta signaling pathway related gene expression was inhibited, while the expression of Tyr3 gene and BMP signaling pathway related genes up-regulated, combined with morphological research It is speculated that under the condition of marine acidification, the up regulation of some genes and signaling pathways still can not compensate for the damage caused by acidification to juveniles, which leads to slower growth of larvae and higher malformation rate.

【學(xué)位授予單位】：廈門大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：X55;X174

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