| Oriental river prawn Macrobrachium nipponense is a freshwater economic aquaculture specie,which is widely distributed in fresh or brackish water areas such as rivers,lakes and reservoirs in China,Japan,Vietnam,and other Eastern and Southern-eastern Asian countries.The growth trait of oriental river prawn is an important economic trait.The selection breeding of growth trait is an important way to improve the culture yields and economic benefits.The muscle growth of M.nipponenese is a typical form of rapid increase in body length and weight.In this study,muscle growth-related genes including troponin I(TNI),myosin light chain(MLC)and myosin heavy chain(MHC),were identified from the transcriptome,genome and other genomic databases of M.nipponense by using bioinformatics technology.Based on the systematic study of their structure,tissue expression and phylogeny,their expression in response to salinity stress was analyzed.The aim is to provide molecular basis for the selection breeding of muscle growth and salt tolerance traits of oriental river prawn.The main findings are as follows:(1)Structure,expression analysis of Tn I gene and response to salinity stress in oriental river prawnThe total length of Tn I gene c DNA is 1489 bp,including 1332 bp ORF coding region,encoding 444 amino acids.The protein structure of the gene contained 4 αspirals,without β fold.Phylogenetic analysis showed that Tn I gene was closely related to Decapoda shrimps and crabs in the phylogenetic tree.Tn I gene was expressed in eight tissues of oriental river prawn,with highest expression in muscle tissue and the lowest expression in hepatopancreas.The results showed that the expression of Tn I gene increased gradually when the salinity increased from 0 to 14,decreased when the salinity increased from 14 to 21,and the expression was the lowest when salinity increased to 21,which was significantly lower than that at salinity 0.(2)Structure,expression analysis of MLC gene and response to salinity stress in oriental river prawnIn this study,MLC genes were found on chromosomes 16,30 and 41,named MLC16,MLC30 and MLC41 genes respectively.The total length of MLC16 gene c DNA is 486 bp,including 423 bp ORF coding region,encoding 141 amino acids.Through the prediction and analysis of protein tertiary structure,it was found that the protein structure of the gene contained 5 β folding and 8 α spiral.The MLC16 gene is closely related to mollusks in the phylogenetic tree.The expression of MLC16 gene in eight tissues of oriental river prawn was analyzed by q RT-PCR,in which the expression in muscle tissue was the highest and in gill tissue was the lowest.The results showed that the results showed that the expression of MLC16 gene increased gradually when the salinity increased from 0 to 7,decreased when the salinity increased from 7 to 21,and the expression was the lowest when salinity increased to21,which was lower than that at salinity 0.The total length of MLC30 gene c DNA is 1024 bp,including 828 bp ORF coding region,encoding 276 amino acids.Through the prediction and analysis of protein tertiary structure,it was found that the protein structure of the gene contained4 β folding and 8 α spiral.The MLC30 gene is closely related to decapoda and molluscs in the phylogenetic tree.The expression of MLC30 gene in eight tissues of oriental river prawn was analyzed by q RT-PCR,in which the expression was the highest in muscle tissue and the lowest in abdominal ganglion.The results showed that the expression of MLC30 gene increased gradually when the salinity increased from 0 to 7,decreased when the salinity increased from 7 to 21,and the expression was the lowest when salinity increased to 21,which was significantly lower than that at salinity 0.The total length of MLC41 gene c DNA is 413 bp,including 360 bp ORF coding region,encoding 120 amino acids.Through the prediction and analysis of protein tertiary structure,it was found that the protein structure of the gene contained 5 β fold and 7 α spiral.The MLC41 gene is closely related to decapoda and molluscs in the phylogenetic tree.The expression of MLC41 gene in seven tissues of oriental river prawn was analyzed by q RT-PCR.The expression in muscle tissue was the highest,but not in gill tissue.The results showed that the expression of MLC41 gene increased gradually when the salinity increased from 0 to 7,decreased when the salinity increased from 7 to 21,and the expression was the lowest when salinity increased to21,which was significantly lower than that at salinity 0.(3)Structure,expression analysis of MHC gene and response to salinity stress in oriental river prawnIn this study,MHC genes were found on chromosomes 7 and 17,named MHC7 and MHC17 genes respectively.The full length of MHC7 gene c DNA is 2379 bp,including 2109 bp ORF coding region,encoding 703 amino acids.Through the prediction and analysis of protein tertiary structure,it was found that the protein structure of the gene contained 8 α spiral,not included β fold.The MHC7 gene is closely related to decapoda in the phylogenetic tree.The expression characteristics of MHC7 gene in eight tissues of oriental river prawn were analyzed by q RT-PCR.The results showed that the expression of MHC7 gene was the highest in heart tissue and very low in stomach,gill,intestine,heart and hepatopancreas.The full length of MHC17 gene c DNA is 6021 bp,including 5811 bp ORF coding region,encoding 1937 amino acids.Through the prediction and analysis of protein tertiary structure,it was found that the protein structure of the gene contained16 α spiral,not included β fold.The MHC17 gene is closely related to decapoda in the phylogenetic tree.And the q RT-PCR showed that the gene was most expressed in the muscle tissue of oriental river prawn,but not in the eyestalk.The results showed that the expression of MHC17 gene increased gradually when the salinity increased from 0to 7,decreased when the salinity increased from 7 to 21,and the expression was the lowest when salinity increased to 21,which was significantly lower than that at salinity 0. |
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