| 摘要: |
| 为了深入了解舟形藻(Navicula sp.)生物膜诱导虾夷扇贝(Mizuhopecten yessoensis)幼虫附着变态的机理，实验选用舟形藻和虾夷扇贝幼虫作为研究对象，研究了4个浓度梯度舟形藻液(1A、10A、100A和1000A )下虾夷扇贝幼虫附着变态率以及幼虫响应舟形藻生物膜诱导的蛋白质组学差异。结果表明，稀释10倍后的藻液(1366个/ mm2)所形成的生物膜对幼虫诱导率最高为45%，与空白组(13%)差异极显著(P<0.01)，过高或过低浓度都导致诱导率降低。采用非标记定量技术(4D label free)蛋白质组学分析比较舟形藻生物膜和对照组幼虫发现，经舟形藻生物膜诱导后共316个蛋白表达显著上调或下调。如谷胱甘肽S转移酶、谷胱甘肽过氧化物酶、组织蛋白酶K、组织蛋白酶L、NAD(P)H 氧化酶(形成 H2O2)、软骨基质蛋白、胶原蛋白和铁蛋白等。以上这些差异蛋白表达的上下调均与贝类幼虫在附着变态过程中形态学变化息息相关。结合以上研究结果，舟形藻生物膜诱导虾夷扇贝幼虫附着变态的有效成分可能是其产生的胞外多糖，幼虫的体内的凝集素检测到舟形藻生物膜表面的多糖后，特异性与多糖结合，诱导幼虫完成附着和变态，研究结果可为虾夷扇贝人工培育提供理论依据。 |
| 关键词: 虾夷扇贝 舟形藻生物膜 变态 蛋白质组学 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| Metamorphosis of Mizuhopecten yessoensis larvae induced by Navicula sp. and its proteomic response |
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XU xiaoyan1, DU Meirong2, JIANG Zengjie2, JIANG Weiwei2, XIA Sudong1, QIN Xiaofang2, YAN Jin2, LYU Jianfu3
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1.Tianjin Agricultural University;2.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences;3.Harbin Engineering University
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| Abstract: |
| In order to understand the mechanism of the effect of film of Navicula sp. on the settlement and metamorphosis of Mizuhopecten yessoensis larvae, the settlement and metamorphosis rate of the larvae and the proteomic differences were investigated under four concentration gradients of Navicula solution (1A, 10A, 100A, and 1000A).The results showed that the highest metamorphosis rate of biofilm larvae (45%) was formed by the Navicula sp. of 10A (1,366/ mm2), which differed highly significantly (P<0.01) from that of the control (13%), and that too high or too low concentration resulted in a lower induction rate. Proteomic analysis using non-labeled quantitative technique (4D label free) to compare the proteomic study analysis of larvae from Navicula sp. and control group revealed that the expression of 198 proteins was significantly up-regulated and 118 proteins were significantly down-regulated by Navicula sp., and that the expression of cathepsin K, cathepsin L, cartilage matrix proteins, collagen, ferritin, glutathione S-transferase, glutathione peroxidase, and lectin were elevated, while the expression of proteins such as heat shock proteins 70 and NAD (P) H oxidase (forming H2O2)? proteins were decreased. Proteins such as histone K, histone L, and heat shock proteins are associated with apoptosis, indicating that there is substantial apoptosis during larval settlement and metamorphosis. Proteins such as cartilage matrix proteins, collagen and ferritin are associated with calcium carbonate shell formation, indicating that larvae induced by Navicula sp. film were in the process of completing the construction of adult shells. Proteins such as glutathione S-transferase, glutathione peroxidase, and NAD (P) H oxidase (forming H2O2) are associated with immune function, indicating the maintenance of cellular homeostasis during larval settlement and metamorphosis. C-type lectins and galectin bind to polysaccharides to trigger larval attachment and metamorphosis. The up-and down-regulation of these differential proteins was closely related to morphological changes in larvae during settlement and metamorphosis. The results of the above studies indicate that the effective component of the biofilm inducing the settlement and metamorphosis of the scallop larvae may be the extracellular polysaccharides produced by Navicula sp., and the lectin in the larvae"s body detects the polysaccharides on the surface of the film, and then binds with the polysaccharides specifically to induce the larvae to complete their settlement and metamorphosis. The aim is to provide a theoretical basis for artificial cultivation of bivalve seedlings. |
| Key words: Mizuhopecten yessoensis Navicula sp. bioflim metamorphosis proteomics |