Full-Length Transcriptome Analysis Identifies Candidate Molecular Features Associated with Golden-Lip Coloration in Pinctada fucata martensii
Yong Liu, Shuai Chen, Hongmei Yang, Weilin Du, Wenhao Yu, Yuewen Deng, Zhe ZhengShell and pearl coloration is an economically important trait in Pinctada fucata martensii, with the golden-lip phenotype associated with commercially valuable golden nacre. In this study, full-length transcriptome sequencing was used to characterize transcript-abundance and transcript-structure patterns in pooled mantle samples from golden- and silver-lip oysters, with one pooled Iso-Seq library generated for each tissue/color condition from three individuals. Among the analyzed mantle samples, the largest transcript-profile differences between the color-line libraries were observed in mantle pallial tissue. The ferritin heavy-chain gene fth1-a showed moderately higher normalized transcript abundance in the Y_MP library than in W_MP but did not meet the predefined twofold screening threshold for library-level abundance differences. RT-qPCR analysis of additional independent individuals showed significantly higher fth1-a expression in golden-lip oysters. A custom anti-PmFTH1-A antibody was evaluated by Western blotting, and immunofluorescence analysis detected PmFTH1-A immunoreactivity within the decalcified shell organic matrix. Candidate alternative-splicing and alternative-polyadenylation events were identified in transcripts associated with carotenoid-related processes and biomineralization. COL1A1 showed higher transcript abundance in Y_MP, and RT-qPCR analysis of additional individuals showed significantly higher COL1A1 expression in the golden-lip group. Additional library-level differences involved transcripts associated with immune and stress-response processes. Overall, this exploratory study identifies candidate transcriptional and transcript-structural differences between the analyzed golden- and silver-lip samples and provides independent experimental support for higher fth1-a and COL1A1 expression in golden-lip oysters. These candidates require functional investigation and validation in independently replicated populations before any evaluation as molecular markers for selective breeding.