Studies On The Immunity-related Enzymes Activities And The Genes Expression Characteristics In Hyriopsis Schlegerlii And Hyriopsis Cumingii

Acid phosphatase,alkaline phosphatase,phenoloxidase,lysozyme and catalase were analysed in Hyriopsis schlegerlii through biochemistry methods.The immunity-related gene SOD,CAT,GST,QM and alpha- Macroglobulins were reported in H.schlegerlii and Hyriopsis cumingiiii.The expression characteristics of these genes were investigated when the freshwater mussels were infected by Aeromonas hydrophila,and the expression in eleven tissues were studied.1.The results of the activities of immunity-related enzymesa).The activities of Aeromonas hydrophila-resistant and Bacillus subtilis-resistant in H.schlegerlii were0.83±0.01U and 0.34±0.01U,respectively,while theses in H.cumingiiii were 0.57±0.03U and 0.26±0.01U.The activities of Escherichia coli-resistant and Staphylococcus aureus-resistant were not detected in both H.schlegerlii and H.cumingiiii.b).The activities of ACP and ALP existed in both cell-free haemolymph and haemocytes in different age H.schlegerlii,both of the two enzymes in cell-free haemolymph was significantly larger than that of haemocyte(P＜0.05).In both cell-free haemolymph and haemocyte,The ACP and ALP demonstrated significant diffenence among various age animals(P＜0.05).The lysozyme activity increased with age increased,and in four-year-old was 2-fold of one-year individuals,to 0.93±0.05U(P＜0.05).However,LYZ in cell-free haemolymph was unchanged in all four groups.The mean of lysozyme activity was 0.44U.The catalase activity indicated that there existed apparent difference in cell-free haemolymph and haemocyte(P＜0.05).The PO activities of the haemocyte were stronger than that of cell-free haemolymph in all groups.2.Cloning of immunity-related genesa).The GST partial cDNA sequence were acquired in H.schlegerlii and H.cumingiiii. After Aeromonas hydrophila infection,the expression intensity in H.schlegerlii increased soon,Aeromonas hydrophila had the induced effect on GST mRNA expression.There was different trend in H.cumingiiii,whose GST mRNA decreased at first and then increased.There was high expression in liver.b).There was a protein kinase c phosphorylation site and the ribosome protein L10 labelling sequence in induced QM amino sequence in present work.In the semi-quantity RT-PCR experiment,QM mRNA expression intensity was less in one-year-old animals while that of the two-year-old ones was the highest.After Aeromonas hydrophila infection,the expression intensity in H.schlegerlii decreased soon and then increased gradually,while that of H.cumingii always hold with low levels.QM mRNA was detected in labellum,foot and liver in both of two bivalves.c).The catalase gene expression in different age was coincided with the measurement of the catalase enzyme activity,After bacterial injection,3 hours samples suggested that CAT mRNA expression intensity in H.schlegerlii was higher than H.cumingiiii.All of eleven tissues existed CAT mRNA.d).In H.schlegerlii,the SOD mRNA were not detected in both one-year and four-year animals.SOD mRNA was detected in healthy group,and was not detected after bacterial infection.In H.cumingiiii,SOD mRNA was detected in 3 and 6 hours samples.In both of the freshwater mussel,SOD mRNA was detected in labellum and kidney.e).In H.schlegerlii,ot2-M mRNA expression decreasd gradually with age increased.After bacterial infection,the expression intensity decreased at first and then increased,while the expression intensity kept with high level in H.cumingiiii.α2-M mRNA only detected in several tissues of H.schlegerlii while in all tissues of H.cumingiiii...