PfmPif97-like Regulated by Pfm-miR-9b-5p Participates in Shell Formation in Pinctada Fucata Martensii

PLoS One. 2019 Dec 12;14(12):e0226367. doi: 10.1371/journal.pone.0226367. eCollection 2019.


Mollusk shell matrix proteins are important for the formation of organic frameworks, crystal nucleation, and crystal growth in Pinctada fucata martensii (P. f. martensii). MicroRNAs (miRNAs) are endogenous small non-coding RNAs that play important roles in many biological processes, including shell formation. In this study, we obtained the full-length sequence of Pif97-like gene in P. f. martensii (PfmPif97-like). PfmPif97-like was mainly distributed in mantle pallial and mantle edge. Correlation analysis indicated that the average shell thickness and weight showed a positive correlation with PfmPif97-like expression (P < 0.05). The inner surface of the nacreous layer and prismatic layer showed atypical growth when we knocked down the expression of PfmPif97-like by RNA interference (RNAi). We used a luciferase reporter assay to identify that miR-9b-5p of P. f. martensii (Pfm-miR-9b-5p) downregulated the expression of PfmPif97-like by interacting with the 3'-untranslated region (UTR) while we obtained the same result by injecting the Pfm-miR-9b-5p mimics in vivo. After injecting the mimics, we also observed abnormal growth in nacre layer and prismatic layer which is consistent with the result of RNAi. We proposed that PfmPif97-like regulated by Pfm-miR-9b-5p participates in shell formation of P. f. martensii. These findings provide important clues about the molecular mechanisms that regulate biomineralization in P. f. martensii.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animal Shells / physiology*
  • Animals
  • Base Sequence
  • Biomineralization
  • Extracellular Matrix Proteins / genetics
  • Extracellular Matrix Proteins / metabolism*
  • MicroRNAs / genetics*
  • Pinctada / genetics*


  • Extracellular Matrix Proteins
  • MicroRNAs

Grant support

This work was supported by Natural Science Foundation of Guangdong Province [Grant no. 2018A030310666]; National Natural Science Foundation of China [Grant no. 31672626]; Innovation Team Project from the Department of Education of Guangdong Province [Grant no. 2017KCXTD016]; Modern Agricultural Industrial System [CARS-049]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.