海洋酸化对福建牡蛎幼体基因表达的影响

发布时间：2018-04-11 01:07

本文选题：海洋酸化 + 福建牡蛎　；参考：《厦门大学》2014年硕士论文

【摘要】：随着人为排放到大气中的CO2持续增加,由CO2驱动的海洋酸化将导致海水pH值在本世纪显著下降,并很可能对海洋生物,特别是对钙质无脊椎动物及生态系统造成严重负面影响。越来越多的研究表明,海洋酸化会影响海洋钙质无脊椎动物的存活、发育和生理机能。然而,目前关于海洋酸化对福建牡蛎(Crassostrea angulata)幼体影响的研究报道还比较少。本文通过实验室内模拟未来海洋环境,研究了短期高C02暴露对我国重要经济水产养殖种类—福建牡蛎幼体发育的影响(对照组CO2浓度400ppm,实验组分别为1500ppm和3000ppm)。利用形态学和分子生物学方法对酸化暴露后的幼体进行相关研究,主要结果如下： 1.海洋酸化对福建牡蛎幼体贝壳生长的影响通过对福建牡蛎幼体贝壳形态参数的测量分析,发现1500ppm实验组较对照组贝壳形态无显著性差异,3000ppm实验组中幼体个体较对照组偏小,贝壳增长速度慢,畸形率升高。进一步采用扫描电镜联用EDS能谱法分析技术分析酸化暴露后D型幼体贝壳元素成分,发现海洋酸化对幼体钙元素沉积有一定的抑制作用,而且发现福建牡蛎幼体对1500ppm酸化程度有一定的耐受性。 2.海洋酸化条件F福建牡蛎幼体数字基因表达谱分析通过对正常条件下和海洋酸化条件下的福建牡蛎担轮幼体、D型初期幼体和D型2d幼体进行高通量测序,分别两两比较后获得差异基因5317个、74个和2641个。差异基因的GO功能显著性富集分析和Pathway显著性富集分析结果显示,海洋酸化主要抑制了胚胎发育、能量代谢、基础生物学过程和钙离子传导相关基因和通路的作用,从而导致幼体生长延迟。 3.海洋酸化对福建牡蛎幼体贝壳形成和生长相关基因的影响通过全胚原位杂交技术发现Tyr3、TGF-β信号通路和BMP信号通路均参与福建牡蛎幼体InCaS生物发生。在酸化条件下,TGF-β信号通路相关基因表达受到抑制,而Tyr3基因和BMP信号通路相关基因表达上调。形态学研究结果显示,在海洋酸化条件下福建牡蛎面盘幼体贝壳生长缓慢且个体偏小,畸形率高,因此,我们推测Tyr3和BMP信号通路相关基因上调表达后仍不能补偿海洋酸化对幼体造成的损伤,所以福建牡蛎在海洋酸化条件下贝壳虽仍能持续生长但大小形状都较对照组有变化。本研究通过扫描电镜联用EDS能谱法分析技术分析福建牡蛎幼体贝壳元素成分,发现海洋酸化对钙元素沉积有一定抑制作用；利用RNA-Seq技术探究海洋酸化对福建牡蛎幼体早期生长发育影响,结果显示,海洋酸化抑制早期胚胎发育、能量代谢和钙离子传导相关信号通路作用,从而导致幼体生长延迟；利用成熟的WISH方法探究Tyr3、TGF-β信号通路和BMP信号通路相关基因的功能,并用qRT-PCR技术研究了海洋酸化条件下它们的表达情况,结果显示,它们与InCaS生物发生和贝壳生长有关,在酸化条件下,TGF-β信号通路相关基因表达受到抑制,而Tyr3基因和BMP信号通路相关基因上调表达,结合形态学研究结果推测,海洋酸化条件下,某些基因和信号通路上调表达仍不能补偿酸化对幼体造成的损伤而导致幼体贝壳生长缓慢,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.

【学位授予单位】：厦门大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：X55;X174

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