Mitf Involved in Innate Immunity by Activating Tyrosinase-Mediated Melanin Synthesis in Pteria penguin

The microphthalmia-associated transcription factor (MITF) is an important transcription factor that plays a key role in melanogenesis, cell proliferation, survival and immune defense in vertebrate. However, its function and function mechanism in bivalve are still rarely known. In this research, first, a Mitf gene was characterized from Pteria penguin (P. penguin). The PpMitf contained an open reading frame of 1,350 bp, encoding a peptide of 449 deduced amino acids with a highly conserved basic helix-loop-helix-leucine zipper (bHLH-LZ) domain. The PpMITF shared 55.7% identity with amino acid sequence of Crassostrea gigas (C. gigas). Tissue distribution analysis revealed that PpMitf was highly expressed in mantle and hemocytes, which were important tissues for color formation and innate immunity. Second, the functions of PpMitf in melanin synthesis and innate immunity were identified. The PpMitf silencing significantly decreased the tyrosinase activity and melanin content, indicating PpMitf involved in melanin synthesis of P. penguin. Meanwhile, the PpMitf silencing clearly down-regulated the expression of PpBcl2 (B cell lymphoma/leukemia-2 gene) and antibacterial activity of hemolymph supernatant, indicating that PpMitf involved in innate immunity of P. penguin. Third, the function mechanism of PpMitf in immunity was analyzed. The promoter sequence analysis of tyrosinase (Tyr) revealed two highly conserved E-box elements, which were specifically recognized by HLH-LZ of MITF. The luciferase activities analysis showed that Mitf could activate the E-box in Tyr promoter through highly conserved bHLH-LZ domain, and demonstrated that PpMitf involved in melanin synthesis and innate immunity by regulating tyrosinase expression. Finally, melanin from P. penguin, the final production of Mitf-Tyr-melanin pathway, was confirmed to have direct antibacterial activity. The results collectively demonstrated that PpMitf played a key role in innate immunity through activating tyrosinase-mediated melanin synthesis in P. penguin.

Spatio-Temporal Variation in Growth Performance and Condition of the Winged Pearl Oyster Pteria penguin

Environmental conditions can strongly influence the growth performance of pearl oysters and affect pearl farm production schedules. Growth and condition index (CI) of two age cohorts of Pteria penguin were measured for 13 months to investigate differences in growth performance between four culture sites within the northern (Vava’u) and southern (Tongatapu) island groups of the Kingdom of Tonga. Environmental conditions were also measured at culture sites and used to explore potential effects on oyster growth and condition. Between island groups, growth performance of P. penguin was superior at northern sites and was most strongly related to higher water temperatures at these sites. Within the southern island group, growth performance varied significantly between sites and may be driven by differences in wave energy. Monthly growth rates (GM) of P. penguin also showed significant temporal variation related to age and environmental conditions. This study demonstrated significant variation in the growth performance of P. penguin at latitudinal and local scales and suggests that in oligotrophic marine environments with minimal terrestrial inputs, such as Tonga, water temperature and wave exposure may be the primary environmental conditions influencing the growth performance of P. penguin. This study therefore recommends that optimal culture sites for P. penguin in Tonga are characterized primarily by warmer water temperatures (25–30°C) and low wave exposure (<15 joules m2 day–1). Culture of P. penguin at sites with more suitable environmental conditions enables pearl production to begin up to 34.2 % (6.5 months) earlier than at less-suitable sites and this may greatly influence mabé pearl farm profitability and feasibility.

Diversified Biomineralization Roles of Pteria penguin Pearl Shell Lectins as Matrix Proteins

Previously, we isolated jacalin-related lectins termed PPL2, PPL3 (PPL3A, 3B and 3C) and PPL4 from the mantle secretory fluid of Pteria penguin (Mabe) pearl shell. They showed the sequence homology with the plant lectin family, jacalin-related β-prism fold lectins (JRLs). While PPL3s and PPL4 shared only 35%–50% homology to PPL2A, respectively, they exhibited unique carbohydrate binding properties based on the multiple glycan-binding profiling data sets from frontal affinity chromatography analysis. In this paper, we investigated biomineralization properties of these lectins and compared their biomineral functions. It was found that these lectins showed different effects on CaCO3 crystalization, respectively, although PPL3 and PPL2A showed similar carbohydrate binding specificities. PPL3 suppressed the crystal growth of CaCO3 calcite, while PPL2A increased the number of contact polycrystalline calcite composed of more than one crystal with various orientations. Furthermore, PPL4 alone showed no effect on CaCO3 crystalization; however, PPL4 regulated the size of crystals collaborated with N-acetyl-D-glucosamine and chitin oligomer, which are specific in recognizing carbohydrates for PPL4. These observations highlight the unique functions and molecular evolution of this lectin family involved in the mollusk shell formation.

Ketebalan Lapisan Bagian Puncak dan Dasar Mutiara Kerang Mabe (Pteria penguin)

Tujuan penelitian ini adalah mengamati karakteristik lapisan bagian puncak (top) dan dasar (base) mutiara blister (Pteria penguin) berdasarkan ukuran cangkang. Penelitian ini dilaksanakan selama empat bulan pada lokasi budidaya mutiara mabe di Perairan Palabusa Selat Buton Sulawesi Tenggara. Hewan uji dua kelompok berdasarkan ukuran cangkang masing-masing 40 ekor. Kelompok pertama (K1) berukuran rata-rata tinggi 72,41 mm, panjang 87,89 mm, dan tebal 25,32 mm; Kelompok kedua (K2) berukuran rata-rata tinggi 89,03 mm, panjang 105,87 mm dan tebal 29,57 mm. Parameter yang diukur adalah ketebalan lapisan mutiara pada bagian atas, puncak (top) dan ketebalan lapisan pada bagian dasar (base), dan morfometrik cangkang karang. Hasil penelitian menunjukkan hubungan positif terhadap lapisan base dan top dengan ukuran cangkang antara panjang, tinggi, dan tebal. Ketebalan lapisan mutiara pada bagian top K2 yaitu 0.23±0.06 mm, lebih tinggi dari K1 yaitu 0.22±0.05 mm, demikian juga ketebalan lapisan mutiara bagian base pada kerang K2 yaitu 0.32±0.09 mm lebih tinggi dari pada K1 yaitu 0.30±0.06 mm. Makin tebal ukuran cangkang, cenderung menghasilkan mutiara yang lebih tebal.