Marine Biotechnology

, Volume 21, Issue 2, pp 240–249 | Cite as

Expressions of Shell Matrix Protein Genes in the Pearl Sac and Its Correlation with Pearl Weight in the First 6 Months of Pearl Formation in Hyriopsis cumingii

  • Can Jin
  • Jing-Ying Zhao
  • Xiao-Jun LiuEmail author
  • Jia-Le LiEmail author
Original Article


Matrix proteins regulate crystal nucleation, morphology, and polymorphism during pearl biomineralization and have significant correlations with pearl quality traits in nucleated pearls. However, there is little information about the connection between pearl quality traits and matrix proteins in non-nucleated pearls. In this study, we analyzed CaCO3 deposition during the first month of non-nucleated pearl formation and examined the expression patterns of ten shell matrix protein genes (Hcperlucin, hic31, silkmapin, hic22, hic74, hic52, HcTyr, HcCA3, hic24 and Hc-upsalin) in the pearl sac of Hyriopsis cumingii. During pearl formation, CaCO3 crystals were initially deposited in a disorderly manner during days 12 and 15 of pearl formation. On days 18 and 21, CaCO3 crystals gradually nucleated on an organic membrane, and the pattern of crystal deposition changed markedly. Between days 24 and 30, crystals similar to nacre tablets were deposited; they then grew and formed connections in a more orderly fashion, eventually forming the nacreous layer. We observed high expression levels of shell matrix proteins during the phases of disordered or ordered CaCO3 deposition, suggesting they were involved in non-nucleated pearl formation. Furthermore, the expressions of nine matrix proteins were significantly correlated with pearl weight during the first 6 months after grafting. The prismatic-layer matrix protein hic31 and nacreous-layer matrix protein hic22 showed negative correlations with pearl weight, but the other seven nacreous-layer matrix proteins had significantly positive correlations with pearl weight. These results show the involvement of matrix proteins in pearl formation and in determination of quality traits.


Pearl formation Nacre Matrix protein Pearl weight Hyriopsis cumingii 



We thank International Science Editing ( for editing this manuscript.

Funding information

This work was financially supported by the National Natural Science Foundation of China (31672654), the Modern Agro-industry Technology Research System (CARS-49), and the Project of Shanghai Engineering and Technology Center for Promoting Ability (16DZ2281200).


  1. Addadi L, Joester D, Nudelman F, Weiner S (2006) Mollusk shell formation: a source of new concepts for understanding biomineralization processes. Chem Eur J 12:981–987CrossRefGoogle Scholar
  2. Arnaud-Haond S, Goyard E, Vonau V, Herbaut C, Prou J, Saulnier D (2007) Pearl formation: persistence of the graft during the entire process of biomineralization. Mar Biotechnol 9:113–116CrossRefPubMedGoogle Scholar
  3. Bai ZY, Lin JY, Ma KY, Wang GL, Niu DH, Li JL (2014) Identification of housekeeping genes suitable for gene expression analysis in the pearl mussel, Hyriopsis cumingii, during biomineralization. Mol Gen Genomics 289:717–725CrossRefGoogle Scholar
  4. Blay C, Parrad S, Cabral P, Aiho V, Ky CL (2016) Correlations between cultured pearl size parameters and PIF-177 biomarker expression in Pinctada margaritifera families reared in two contrasting environments. Estuar Coast Shelf Sci 182:254–260CrossRefGoogle Scholar
  5. Blay C, Planes S, Ky CL (2018) Cultured pearl surface quality profiling by the shell matrix protein gene expression in the biomineralised pearl sac tissue of Pinctada margaritifera. Mar Biotechnol 20:1–12CrossRefGoogle Scholar
  6. Checa AG, Rodríguez-Navarro AB (2005) Self-organisation of nacre in the shells of Pterioida (Bivalvia: Mollusca). Biomaterials 26:1071–1079CrossRefPubMedGoogle Scholar
  7. Chen XJ, Liu XJ, Bai ZY, Zhao LT, Li JL (2016) HcTyr and HcTyp-1 of Hyriopsis cumingii, novel tyrosinase and tyrosinase-related protein genes involved in nacre color formation. Comp Biochem Physiol B 204:1–8CrossRefPubMedGoogle Scholar
  8. Fang Z, Feng QL, Chi YZ, Xie LP, Zhang RQ (2008) Investigation of cell proliferation and differentiation in the mantle of Pinctada fucata (bivalve, Mollusca). Mar Biol 153:745–754CrossRefGoogle Scholar
  9. Gan HM, Austin C, Linton S (2018) Transcriptome-guided identification of carbohydrate active enzymes (CAZy) from the Christmas Island red crab, Gecarcoidea natalis and a vote for the inclusion of transcriptome-derived crustacean CAZys in comparative studies. Mar Biotechnol 20:654–665CrossRefPubMedGoogle Scholar
  10. Gardner LD, Mills D, Wiegand A, Leavesley D, Elizur A (2011) Spatial analysis of biomineralization associated gene expression from the mantle organ of the pearl oyster Pinctada maxima. BMC Genomics 12:455CrossRefPubMedPubMedCentralGoogle Scholar
  11. Grégoire C (1957) Topography of the organic components in mother-of-pearl. J Biophys Biochem Cytol 3:797–808CrossRefPubMedPubMedCentralGoogle Scholar
  12. Gu ZF, Yin XL, Yu CC, Zhan X, Shi YH, Wang AM (2016) Expression profiles of nine biomineralization genes and their relationship with pearl nacre thickness in the pearl oyster, Pinctada fucata martensii dunker. Aquac Res 47:1874–1884CrossRefGoogle Scholar
  13. Im S, Lee HN, Jung HS, Yang S, Park EJ, Mi SH, Jeong WJ, Choi DW (2017) Transcriptome-based identification of the desiccation response genes in marine red algae Pyropia tenera (Rhodophyta) and enhancement of abiotic stress tolerance by PtDRG2 in Chlamydomonas. Mar Biotechnol 19:232–245CrossRefPubMedGoogle Scholar
  14. Inoue N, Ishibashi R, Ishikawa T, Atsumi T, Aoki H, Komaru A (2010) Gene expression patterns and pearl formation in the Japanese pearl oyster (Pinctada fucata): a comparison of gene expression patterns between the pearl sac and mantle tissues. Aquaculture 308:S68–S74CrossRefGoogle Scholar
  15. Inoue N, Ishibashi R, Ishikawa T, Atsumi T, Aoki H, Komaru A (2011a) Can the quality of pearls from the Japanese pearl oyster (Pinctada fucata) be explained by the gene expression patterns of the major shell matrix proteins in the pearl sac? Mar Biotechnol 13:48–55CrossRefPubMedGoogle Scholar
  16. Inoue N, Ishibashi R, Ishikawa T, Atsumi T, Aoki H, Komaru A (2011b) Comparison of expression patterns of shell matrix protein genes in the mantle tissues between high- and low-quality pearl-producing recipients of the pearl oyster, Pinctada fucata. Zool Sci 28:32–36CrossRefPubMedGoogle Scholar
  17. Ky CL, Blay C, Sham-Koua M, Vanaa V, Lo C, Cabral P (2013) Family effect on cultured pearl quality in black-lipped pearl oyster Pinctada margaritifera and insights for genetic improvement. Aquat Living Resour 26:133–145CrossRefGoogle Scholar
  18. Ky CL, Quillien V, Broustal F, Soyez C, Devaux D (2018) Phenome of pearl quality traits in the mollusc transplant model Pinctada margaritifera. Sci Rep 8:2122CrossRefPubMedPubMedCentralGoogle Scholar
  19. Lin JY, Ma KY, Bai ZY, Li JL (2013) Molecular cloning and characterization of perlucin from the freshwater pearl mussel, Hyriopsis cumingii. Gene 526:210–216CrossRefPubMedGoogle Scholar
  20. Liu XJ, Li JL, Xiang L, Sun J, Zheng GL, Zhang GY, Wang HZ, Xie LP, Zhang RQ (2012) The role of matrix proteins in the control of nacreous layer deposition during pearl formation. Proc Biol Sci 279:1000–1007CrossRefPubMedGoogle Scholar
  21. Liu XJ, Dong SJ, Jin C, Bai ZY, Wang GL, Li JL (2015a) Silkmapin of Hyriopsis cumingii, a novel silk-like shell matrix protein involved in nacre formation. Gene 555:217–222CrossRefPubMedGoogle Scholar
  22. Liu XJ, Zeng SM, Dong SJ, Jin C, Li JL (2015b) A novel matrix protein hic31 from the prismatic layer of Hyriopsis cumingii displays a collagen-like structure. PLoS One.
  23. Liu XJ, Jin C, Wu LM, Dong SJ, Zeng SM, Li JL (2016) Isolation and cha racterization of a novel acidic matrix protein hic22 from the nacreous layer of the freshwater mussel, Hyriopsis cumingii. Genet Mol Res15(3).
  24. Liu XJ, Jin C, Wu LM, Dong SJ, Zeng SM, Li JL (2017a) Hic74, a novel alanine and glycine rich matrix protein related to nacreous layer formation in the mollusc Hyriopsis cumingii. Aquac Fish 2:119–123CrossRefGoogle Scholar
  25. Liu XJ, Pu JW, Zeng SM, Jin C, Dong SJ, Li JL (2017b) Hyriopsis cumingii hic52—a novel nacreous layer matrix protein with a collagen-like structure. Int J Biol Macromol 102:667–673CrossRefPubMedGoogle Scholar
  26. Liu XJ, Jin C, Guo W, Li JL (2018a) Identification and molecular characterization of Hc-upsalin, a novel matrix protein involved in nacreous-layer biomineralization in Hyriopsis cumingii. Thalassas.
  27. Liu XJ, Liu ZM, Jin C, Li HR, Li JL (2018b) A novel nacre matrix protein hic24 in Hyriopsis cumingii is essential for calcium carbonate nucleation and involved in pearl formation. Biotechnol Appl Biochem.
  28. Lowenstam HA (1981) Minerals formed by organisms. Science 211:1126–1131CrossRefPubMedGoogle Scholar
  29. Ma HY, Lee IS (2006) Characterization of vaterite in low quality freshwater-cultured pearls. Mater Sci Eng C Mater 26:721–723CrossRefGoogle Scholar
  30. Ma H, Su A, Zhang B, Li RK, Zhou L, Wang B (2009) Vaterite or aragonite observed in the prismatic layer of freshwater-cultured pearls from South China. Prog Nat Sci Mater 19:817–820CrossRefGoogle Scholar
  31. Ma HY, Li RK, Yang LX, Zhang BL, Shen MD, Mu SC, Wei QG (2011) A modified integrated model of the internal structure of Chinese cultured pearls. J Wuhan Univ Technol Mat Sci Edit 26:510–513CrossRefGoogle Scholar
  32. Ma YF, Qiao L, Feng QL (2013) Research progress on biomineralization mechanism of freshwater pearl. J Inorg Mater 28:109–116CrossRefGoogle Scholar
  33. Marie B, Joubert C, Tayalé A, Zanellacléon I, Belliard C, Piquemal D, Cochenneclaureau N, Marin F, Gueguen Y, Montagnani C (2012) Different secretory repertoires control the biomineralization processes of prism and nacre deposition of the pearl oyster shell. Proc Natl Acad Sci U S A 109:20986–20991CrossRefPubMedPubMedCentralGoogle Scholar
  34. Miyamoto H, Miyashita T, Okushima M, Nakano S, Morita T, Matsushiro A (1996) A carbonic anhydrase from the nacreous layer in oyster pearls. Proc Natl Acad Sci U S A 93:9657–9660CrossRefPubMedPubMedCentralGoogle Scholar
  35. Murr LE, Ramirez DA (2012) The microstructure of the cultured freshwater pearl. JOM 64:469–474CrossRefGoogle Scholar
  36. Olson IC, Blonsky AZ, Nobumichi T, Martin K, Boaz P, Romao CP, Mary Anne W, Gilbert PUPA (2013) Crystal nucleation and near-epitaxial growth in nacre. J Struct Biol 184:454–463CrossRefPubMedGoogle Scholar
  37. Roy NL, Jackson DJ, Marie B, Ramossilva P, Marin F (2014) The evolution of metazoan α-carbonic anhydrases and their roles in calcium carbonate biomineralization. Front Zool 11:75CrossRefGoogle Scholar
  38. Smith BL, Schäffer TE, Viani M, Thompson JB, Frederick NA, Kindt J, Belcher A, Stucky GD, Morse DE, Hansma PK (1999) Molecular mechanistic origin of the toughness of natural adhesives, fibres and composites. Nature 399:761–763CrossRefGoogle Scholar
  39. Takeuchi T, Endo K (2006) Biphasic and dually coordinated expression of the genes encoding major shell matrix proteins in the pearl oyster Pinctada fucata. Mar Biotechnol 8:52–61CrossRefPubMedGoogle Scholar
  40. Tayale A, Gueguen Y, Treguier C, Grand JL, Cochennec-Laureau N, Montagnani C, Ky CL (2012) Evidence of donor effect on cultured pearl quality from a duplicated grafting experiment on Pinctada margaritifera using wild donors. Aquat Living Resour 25:269–280CrossRefGoogle Scholar
  41. Tsalafouta A, Sarropoulou E, Papandroulakis N, Pavlidis M (2018) Characterization and expression dynamics of key genes involved in the gilthead sea bream (Sparus aurata) cortisol stress response during early ontogeny. Mar Biotechnol 20:611–622CrossRefPubMedGoogle Scholar
  42. Wang GL, Yuan YM, Li JL (2007) SSR analysis of genetic diversity and phylogenetic relationships among different populations of Hyriopsis cumingii from the five lakes of China. J Fish China 12:12–18Google Scholar
  43. Wang GL, Wang Q, Xu ZC, Wang YY, Li JL (2017) Identification, structural characterization and expression analysis of a novel carbonic anhydrase from freshwater mussel Hyriopsis cumingii. Gene 636:78–86CrossRefPubMedGoogle Scholar
  44. Yan ZG, Wei M, Liu ZT, Yang SW, Liu XJ, Sun J, Xie LP, Zhang RQ (2011) In vivo and in vitro biomineralization in the presence of the inner-shell film of pearl oyster. Acta Oceanol Sin 30:87–93CrossRefGoogle Scholar
  45. Yano M, Nagai K, Morimoto K, Miyamoto H (2007) A novel nacre protein N19 in the pearl oyster Pinctada fucata. Biochem Biophys Res Commun 362:158–163CrossRefPubMedGoogle Scholar
  46. Zhang LJ, He MX (2011) Quantitative expression of shell matrix protein genes and their correlations with shell traits in the pearl oyster Pinctada fucata. Aquaculture 314:73–79CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of AgricultureShanghaiChina
  2. 2.Class 1, 2016 Biological Science, College of Fisheries and Life ScienceShanghai Ocean UniversityShanghaiChina
  3. 3.National Demonstration Center for Experimental Fisheries Science EducationShanghai Ocean UniversityShanghaiChina
  4. 4.Shanghai Engineering Research Center of AquacultureShanghaiChina

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