本文選題：弧菌 + 厚殼貽貝��； 參考：《上海海洋大學》2017年碩士論文

【摘要】：厚殼貽貝(Mytilus coruscus)是我國主要的經(jīng)濟養(yǎng)殖貝類,同時也是我國海洋污損生物的主要類群之一,在我國沿海有廣大的分布區(qū)域。厚殼貽貝主要通過附著在附著基(網(wǎng)繩、潮間帶巖石、堤岸設施表等)的表面生活,附著過程對于厚殼貽貝來說是極其重要生命歷程,當環(huán)境因素發(fā)生不利變化時,厚殼貽貝會通過自身行為,切斷足絲并重新選擇附著基再次附著,因此研究附著基對厚殼貽貝附著的影響對于增進厚殼貽貝的養(yǎng)殖產(chǎn)業(yè)和防御厚殼貽貝在污損生物方面的作用均具有重要的意義。以往的研究發(fā)現(xiàn)自然微生物被膜是厚殼貽貝附著的高效天然誘導物,且作為自然微生物被膜的重要成員之一的海洋細菌,是影響厚殼貽貝稚貝附著的重要因素。同時以往的研究結果也顯示,海洋細菌中具有極大比例的海洋弧菌(Vibrio),海洋弧菌是最早定植在自然微生物被膜表面的細菌類群之一。因此,本文以海洋弧菌和厚殼貽貝為研究對象,探究了海洋弧菌微生物被膜對厚殼貽貝稚貝的附著的影響,并分析了海洋弧菌微生物被膜的代謝產(chǎn)物中蛋白及糖的差異。主要包括下列內容:1.不同來源海洋弧菌形成的微生物被膜對厚殼貽貝稚貝附著的影響為探討廣泛存在于近海環(huán)境中的海洋弧菌和貝類附著的相互作用關系,本研究通過從自然微生物被膜和厚殼貽貝成體腸道內分離了海洋弧菌,測定其種屬及親緣性,調查了這些不同來源弧菌形成的微生物被膜與厚殼貽貝稚貝附著的調控作用。結果表明,測試弧菌形成的微生物被膜密度隨著初始細菌密度的增加呈現(xiàn)不同程度的增加。源于自然微生物被膜和貽貝腸道內的10株弧菌均能顯著促進厚殼貽貝稚貝的附著,不同菌株形成微生物被膜的誘導活性存在顯著差異,稚貝在Vibrio sp.17微生物被膜上的附著率最高,為67%±2%,附著率變化范圍為17%-67%。微生物被膜對稚貝附著誘導活性與被膜密度呈顯著相關性,其中弧菌Vibrio crassostreae ECSMB14106的相關性系數(shù)最高,為0.8992。此外,微生物被膜的誘導活性與弧菌的來源無關。本實驗初步探明了海洋弧菌對厚殼貽貝稚貝附著的影響,有助于進一步理解微生物被膜調控厚殼貽貝的附著分子機理。2.不同稚貝誘導活性的弧菌微生物被膜生物學特性分析實驗選取10~8 cells/ml的密度下形成的微生物被膜,結果發(fā)現(xiàn)誘導活性最高的菌株為Vibrio sp.17,其次為V.crassostreae ECSMB14106、第三為Vibrio sp.13、最低為Vibrio sp.19。本研究應用了共聚焦顯微技術、電子顯微鏡掃描技術、SDS-PAGE蛋白分析技術以及胞外多糖染色技術分析了四株弧菌形成微生物被膜的生物學特性。共聚焦結果發(fā)現(xiàn)四株細菌形成的被膜膜厚存在顯著差異,膜厚與稚貝附著相關。掃描電鏡結果發(fā)現(xiàn)菌株Vibrio sp.17和V.crassostreae ECSMB14106的細菌形態(tài)呈短圓桿狀,菌株Vibrio sp.19呈長桿狀,而菌株Vibrio sp.13生長聚集成花瓣狀。SDS-PAGE蛋白技術分析菌膜總蛋白發(fā)現(xiàn),四株細菌的菌膜總蛋白種類不同,其中Vibrio sp.17的蛋白條帶最多,有31個條帶。而胞外多糖染色結果發(fā)現(xiàn),Vibrio sp.17的多糖含量最高。本研究從多個角度分析了誘導活性不同的同種細菌形成的微生物被膜的生物學特性,發(fā)現(xiàn)了其膜厚、蛋白差異、胞外多糖含量之間的規(guī)律,為探究影響厚殼貽貝稚貝附著的主要誘導因子奠定了一定基礎。
[Abstract]:Mytilus coruscus is the main economic culturing shellfish in China, and it is also one of the main groups of marine fouling organisms in China. There are broad distribution areas in the coastal areas of China. The thick shell mussels are mainly surface living by attaching to the attachment base (net, intertidal rock, dike setting and so on), and the attachment process for thick shell mussels It is an extremely important life course. When the environmental factors are disadvantageous, the thick shell mussels will cut off the foot silk and re select the attachments to attach again through their own behavior. Therefore, the effect of the attachment group on the adhesion of the thick shell mussels has the effect on promoting the culture industry of the thick shell mussel and preventing the fouling of the thick shell mussels. It is of great significance. Previous studies have found that natural microorganism membrane is an efficient natural inducer attached to mussels, and marine bacteria, one of the important members of the natural microorganism membrane, is an important factor affecting the attachment of juvenile shells of mussels. Vibrio (Vibrio), Vibrio marine vibrio is one of the earliest bacteria colonized on the surface of natural microbes. Therefore, the effects of Vibrio marine Vibrio and thick shell mussel on the attachment of juvenile marine Vibrio mussels were investigated, and the proteins in the metabolites of the membrane of Vibrio marine Vibrio were analyzed. The main contents of the difference are as follows: 1. the influence of the microorganism formed by different sources of Vibrio marine Vibrio on the attachment of the juvenile shells of mussels is to explore the interaction relationship between marine Vibrio and shellfish, which is widely existed in the near sea environment. This study separates the ocean from the natural microorganism membrane and the thick shell mussels into the intestinal tract. Vibrio, determination of its species and affinity, investigated the regulation of the adhesion of microbes formed by different sources of Vibrio and juvenile shellfish of mussels. The results showed that the membrane density of microbes formed by Vibrio was increased in varying degrees with the increase of initial bacterial density. The 10 strains of Vibrio can significantly promote the attachment of the juvenile shellfish of the mussel. There are significant differences in the induction activity of the biofilm of different strains. The attachment rate of the juvenile shells on the Vibrio sp.17 membrane is the highest, which is 67% + 2%, and the adhesion rate of the adhesion rate is significantly related to the adhesion induced activity of the 17%-67%. microbe to the juvenile shellfish and the density of the membrane. Among them, the correlation coefficient of Vibrio Vibrio crassostreae ECSMB14106 is the highest, which is 0.8992.. The induction activity of the microorganism membrane is not related to the source of Vibrio. This experiment preliminarily explored the effect of Vibrio marine Vibrio on the attachment of the juvenile shells of mussels mussels, which is helpful to further understand the molecular mechanism of the adhesion of the thick shell mussels by the microorganism membrane to understand the molecular mechanism of.2.. The biological characteristics of Vibrio microbes with different juveniles were analyzed by the membrane biological characteristics of 10~8 cells/ml. The results showed that the highest inducible activity was Vibrio sp.17, followed by V.crassostreae ECSMB14106, third was Vibrio sp.13, and the lowest was Vibrio sp.19.. The confocal microscopy was used in this study. Techniques, electron microscope scanning, SDS-PAGE protein analysis and extracellular polysaccharide staining were used to analyze the biological characteristics of the biofilm of four Vibrio strains. The confocal results showed that there were significant differences in the membrane thickness of the four bacteria, and the thickness of the membrane was closely related to the attachment of the larvae. The scanning electron microscope showed that the strain Vibrio sp.17 and V were found. The bacterial morphology of.Crassostreae ECSMB14106 was short and rod-shaped, and the strain Vibrio sp.19 was long rod like, and the strain Vibrio sp.13 grew to integrate the integrated petal.SDS-PAGE protein to analyze the total protein of the mycelia. The total protein species of the four strains of bacteria were different, of which the egg white bands of Vibrio sp.17 were the most and 31 bands were stained, and the extracellular polysaccharide was stained. The results showed that the polysaccharide content of Vibrio sp.17 was the highest. In this study, the biological characteristics of the membrane were analyzed from several angles, and the film thickness, protein difference and the content of extracellular polysaccharide were found. Basics.
【學位授予單位】：上海海洋大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S917.4

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