| Coral Reefs is a kind of ecological system which is very special and important in the ocean. It keeps a high biological diversity and primary productivity, providing a breeding place for many marine life. It's known as the tropical rainforest in the ocean with the protection of the coastline, providing building materials and the discovery of new marine drugs and the important functions of the record for history of marine climate change. But in recent years, with the impact of global climate change and human activities causing environmental pollution and destruction, it results in coral reef biodiversity reduction,ecological function degradation. When the phenomenon has become increasingly prominent,the coral reef ecosystem is facing a serious threat. Therefore, to strengthen the coral reef biodiversity research, coral reef ecosystem function especially the effects on ecological environment, coral reef protection management and ecological restoration research have important significance to maintain the ecological balance of coral reefs. In this experiment,the coral Acropora Hyacinthus and Acropora humilis from coastal zone of Hainan province are as material at first to explore three different proteins extraction protocols. We evaluate the protein method by protein yield, protein spots number, 1-DE profile, 2-DE profile, and mass spectrometry compatibility rate to select which method is suitable for extracting the total protein of Acropora corals. Then we choose benzo pyrene (BaP) belong the polycyclic aromatic hydrocarbons (PAHs) as the stress factors to explore protein changes of Acropora humilis, which is aimed to reveal the toxicological mechanism of response to BaP stress at protein level. In this paper, the main four results are summarized as follows:(1) The contents of 16 PAHs in Dazhou Island and Mulan Bay seawater are 532.68 ng· L-1 and 82.13 ng · L-1, respectively. The PAHs detected in the seawater of Dazhou Island include low-ring (2-ring) to high-ring (6-ring) molecules, while PAHs detected in seawater of Mulan Bay are in the range of 4 rings and below. The contents of 16 PAHs in the Acropora hyacinthus body in Dazhou Island and the Acropora humilis body in Mulan Bay are 2096.60 ng·g-1 and 1213.46 ng·g-1, respectively, and both of their low and high rings are detected.(2 ) Among the three kinds of methods for protein extraction, the PBE phenol extraction method gets the largest protein yield(3.1-3.9 mg/g), the TRIzol reagent gets less and Kit methold gets the least(2.2 mg/g). From the 1-DE profile, the bands obtained from PBE phenol method, TRIzol reagent method and Kit method seem to be clear, but PBE phenol method gets the most number of bands. From 2-DE profile, three methods get a similar repeatability, but the PBE phenol extraction method gets the highest resolution. In the analysis of the total protein number and its specific spots, PBE phenol extraction method gets the most. In the identification of mass spectrometry, we find that the successful search rate of protein extracted by PBE phenol extraction method is the highest, which shows that it has a better mass spectrum compatibility. On the whole, it is concluded that the PBE method is more suitable for the study of coral protein extraction based on two-dimensional electrophoresis.(3 ) When Acropora humilis are exposured via different concentration of BaP (1 ?g/L,10 ?g/L, 100 ?g/L) five days, the control and the treatment don't cause changes of body morphology characterization of symbiotic coral. 2-DE is applied to display and compare all extractable proteins between the control and treatment among the dose-effect and time-effect.Quantitative image analysis using Image Master 2D Platinum 5.0 revealed that 48 proteins are exhibiting more than 2.0 fold differential expression. The differential expression proteins were excised from the gels, digested with trypsin, and the peptides are analyzed using MALDI-TOF /TOF MS coupled with database searches. In total, 39 proteins were successfully identified by MS/MS analysis, with 17 differential proteins up regulated and 22 differential proteins down regulated. From cell component, the cells, cell organelles, cellular structure, membrane structure from these differential proteins are changed. From molecular function, the function of protein binding, lipid binding, antigen binding, various enzymes,transcription factor activity, molecular structure are changed; Also these differential proteins participate in the cell metabolism, primary metabolism, organic matter, biosynthesis,metabolism and regulation of cell biological response and self regulation, cell communication,detoxification and autophagy from biological processes.(4 ) The actin cytoskeleton (protein No. 24), 14-3-3 protein (protein No. 49 and 50),and heat shock protein 71 kDa (protein No. 49) participate in vital Hippo signaling pathway and EB virus infection pathway in the KEGG metabolism pathways. These four proteins play an important role in anti stress and cell protection and repair, inhibition of apoptosis and other ways. The five proteins associated with photosynthesis (protein number 7, 10, 11, 12,13) are significantly down regulated in the analysis of differential proteins in symbiodinium,which reduces the photosynthesis of the algae. We regard these 9 important proteins as an important molecular marker for the effects of organic compounds on the stress to coral symbionts. |
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