Expression Of CaM And CaLP Genes Of Mantle And Visceral Mass In Hyriopsis Cumingii Cultured By Different Calcium Concentration

China’s pearl production ranked first in the world, but the pearl quality is not high, but low yield large pearls. In order to solve the problem of low quality of the Pearl, Hyriopsis cumingii of the visceral mass of pearl are explored, the study found that the visceral mass of large space, more conducive to large nucleated pearl cultivation.The study found that the gut space is large, more conducive to the cultivation of large nuclear pearl. Hyriopsis cumingii as our unique freshwater mussel, output of Pearl rounded shape, bright luster, excellent quality. The use of Hyriopsis cumingii as pearly mussel, from the mantle of the Hyriopsis cumingii location, visceral mass, gonad, lymphoid tissue acquisition organization and RNA was extracted, the transcriptome data measured. From Hyriopsis cumingii transcriptome library selected genes associated with the metabolism of calcium- calmodulin like protein（CALP） gene. The first validated transcriptome gene screening, experiments proved that the sequence of real and effective, can be directly for follow-up analysis. Analysis, it is found that four Ca LP genes that Ca LP₁ gene can encoding 152 amino acids, with four EF hand Ca2+ binding domain; Ca LP₂ gene encoding 163 amino acid, also has four EF hand Ca2+ binding domain; Ca LP₃ gene encoding 150 amino acids, with three EF hand Ca2+ binding domain; Ca LP₄ gene encoding 163 amino acid, with four EF hand Ca2+ binding domain. The Ca LP₁, Ca LP₃, Ca LP₄ homology with other species are more than 74%, with a highly conserved EF-hand region. Phylogenetic tree indicated that, Hyriopsis cumingii four genes and belong to the Pinctada fucata and Crassostrea gigas in the evolutionary tree gather for a close relationship.In order to explore the role of calmodulin（Ca M） gene and its gene in fresh water pearl oyster formation, study of 0, 0.5, 1.25, 2, 3 m M five different Ca2+ concentration of water environment of culturing Hyriopsis mussels, and after nucleus inserting 0 days, 20 days, 50 days, 90 days, 120 days a foreign film and dirty inside organizations sampling, using fluorescent quantitative method for detection of Ca M gene and four Ca LP gene expression. Results show that:（1） the Ca M gene: Ca M gene coat film and visceral mass at 0 m M Ca2+ concentration always in low expression level; 0.5 m M concentration decreased first and then increased and remained stable; 1.25 m M concentrations in 20 days in the visceral mass decreased, then it rises to the level of 0 days and remained stable; expression of under 2 m M concentrations in 20 days increased significantly at each concentration after the highest maintained high expression of state; 3 m M concentrations in 20 days after nucleus inserting expression increased, period after the expression of began to decrease（P < 0.05）. There was no significant difference in the 0 days after the nucleus, and the expression of the number of days after the day was significantly different. Under the same conditions, the expression of Ca M gene in visceral mass was higher than that in mantle. It was found that the concentration of 0.5 m M and 2 m M Ca2+ could promote the expression of Ca M gene, but the concentration of 0 m M and 3 m M could inhibit the expression of Ca M gene.（2） Ca LP₁ gene: in the absence of Ca LP₁, the Ca2+ gene will be reduced due to the lack of calcium, in the case of high concentrations of 2 m M and 3 m M will be promoted, showing a clear upward trend. And there was no significant inhibitory effect on 3 m M.（3） Ca LP₂ gene: Ca LP₂ in low concentration and high concentration are realized to reduce the trend. In the prophase of the nucleus, the expression of Ca LP₂ gene should be suppressed due to immunity. The expression of Ca LP₂ gene was much lower than that in the visceral mass, and the coat film was not expressed in many times.（4） Ca LP₃ gene: there was no significant difference in the expression of visceral mass and mantle in the nucleus of the 0 day. The expression of visceral mass in the lower concentration（0 m M, 0.5 m M） was higher than that of the mantle, and the expression of visceral mass was higher in the high concentration（2 m M, 3 m M） than in the mantle. The expression of 3 m M was stable, and the visceral mass was obviously increased.（5） Ca LP₄ gene: the coat film has not always showed a significant change in expression. In the visceral mass, at 20 days, the expression was high in low concentration, but the expression was inhibited at high concentration. At high concentrations（2 m M, 3 m M）, there was no significant change in the 20 days before and after 50 days, the expression level began to rise, there was a difference.In order to study different calcium concentrations in the culture environment of nacre deposition randomly selected 300 shell length of 8cm mussels visceral mass and mantle nucleated pearl nucleus inserting operation, after operation in nuclear in the environment of different Ca2+ concentration breeding 180 days, sampling analysis: Pearl diameter measurement and nacre deposition weight measurement shows that the 2 m M concentration of Ca is more suitable for pearl deposition, high concentration（3 m M） and low（0 m M, 0.5 m M） will cause the decline of nacre deposition. Observed in the nacre deposition appearance, mantle and visceral mass deposition have different characteristics. The coat film deposition is less than the visceral mass, and visceral mass of Pearl more rounded, mantle occurred in a bead deformity. The mantle of pearl color red pearl tender, smooth surface; and the deposition visceral mass of rough surface and the uneven pearl. The experimental results show that the peak value of the visceral mass is smaller than that of the outer mantle in the 1131 cm-1 and 1527 cm-1 of the carotenoid components. In the form of crystalline 155 cm-1, 706.5cm-1 and 1085.5cm-1 at the peak of visceral mass is less than the mantle.