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      Mantle Branch-Specific RNA Sequences of Moon Scallop Amusium pleuronectes to Identify Shell Color-Associated Genes

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          Abstract

          Amusium pleuronectes (Linnaeus) that secretes red- and white-colored valves in two branches of mantle tissues is an excellent model for shell color research. High-throughput transcriptome sequencing and profiling were applied in this project to reveal the detailed molecular mechanism of this phenotype differentiation. In this study, 50,796,780 and 54,361,178 clean reads were generated from the left branch (secreting red valve, RS) and right branch (secreting white valve, WS) using the Illumina Hiseq 2000 platform. De novo assembly generated 149,375 and 176,652 unigenes with an average length of 764 bp and 698 bp in RS and WS, respectively. Kyoto encyclopedia of genes and genomes (KEGG) metabolic pathway analysis indicated that the differentially expressed genes were involved in 228 signaling pathways, and 43 genes were significantly enriched (P<0.01). Nineteen of 20 differentially expressed vitellogenin genes showed significantly high expression in RS, which suggested that they probably played a crucial role in organic pigment assembly and transportation of the shell. Moreover, 687 crystal formation-related (or biomineralization-related) genes were detected in A. pleuronectes, among which 144 genes exhibited significant difference between the two branches. Those genes could be classified into shell matrix framework participants, crystal nucleation and growth-related elements, upstream regulation factors, Ca level regulators, and other classifications. We also identified putative SNP and SSR markers from these samples which provided the markers for genetic diversity analysis, genetic linkage, QTL analysis. These results provide insight into the complexity of shell color differentiation in A. pleuronectes so as valuable resources for further research.

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          Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.).

          A software tool was developed for the identification of simple sequence repeats (SSRs) in a barley ( Hordeum vulgare L.) EST (expressed sequence tag) database comprising 24,595 sequences. In total, 1,856 SSR-containing sequences were identified. Trimeric SSR repeat motifs appeared to be the most abundant type. A subset of 311 primer pairs flanking SSR loci have been used for screening polymorphisms among six barley cultivars, being parents of three mapping populations. As a result, 76 EST-derived SSR-markers were integrated into a barley genetic consensus map. A correlation between polymorphism and the number of repeats was observed for SSRs built of dimeric up to tetrameric units. 3'-ESTs yielded a higher portion of polymorphic SSRs (64%) than 5'-ESTs did. The estimated PIC (polymorphic information content) value was 0.45 +/- 0.03. Approximately 80% of the SSR-markers amplified DNA fragments in Hordeum bulbosum, followed by rye, wheat (both about 60%) and rice (40%). A subset of 38 EST-derived SSR-markers comprising 114 alleles were used to investigate genetic diversity among 54 barley cultivars. In accordance with a previous, RFLP-based, study, spring and winter cultivars, as well as two- and six-rowed barleys, formed separate clades upon PCoA analysis. The results show that: (1) with the software tool developed, EST databases can be efficiently exploited for the development of cDNA-SSRs, (2) EST-derived SSRs are significantly less polymorphic than those derived from genomic regions, (3) a considerable portion of the developed SSRs can be transferred to related species, and (4) compared to RFLP-markers, cDNA-SSRs yield similar patterns of genetic diversity.
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            An acidic matrix protein, Pif, is a key macromolecule for nacre formation.

            The mollusk shell is a hard tissue consisting of calcium carbonate crystals and an organic matrix. The nacre of the shell is characterized by a stacked compartment structure with a uniformly oriented c axis of aragonite crystals in each compartment. Using a calcium carbonate-binding assay, we identified an acidic matrix protein, Pif, in the pearl oyster Pinctada fucata that specifically binds to aragonite crystals. The Pif complementary DNA (cDNA) encoded a precursor protein, which was posttranslationally cleaved to produce Pif 97 and Pif 80. The results from immunolocalization, a knockdown experiment that used RNA interference, and in vitro calcium carbonate crystallization studies strongly indicate that Pif regulates nacre formation.
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              Insect cuticular sclerotization: a review.

              Different regions of an insect cuticle have different mechanical properties, partly due to different degrees of stabilization and hardening occurring during the process of sclerotization, whereby phenolic material is incorporated into the cuticular proteins. Our understanding of the chemistry of cuticular sclerotization has increased considerably since Mark Pryor in 1940 suggested that enzymatically generated ortho-quinones react with free amino groups, thereby crosslinking the cuticular proteins. The results obtained since then have confirmed the essential features of Pryor's suggestion, and the many observations and experiments, which have been obtained, have led to a detailed and rather complex picture of the sclerotization process, as described in this review. However, many important questions still remain unanswered, especially regarding the precise regional and temporal regulation of the various steps in the process. (c) 2009 Elsevier Ltd. All rights reserved.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                23 October 2015
                2015
                : 10
                : 10
                : e0141390
                Affiliations
                [1 ]Fishery College, Guangdong Ocean University, Zhanjiang, China
                [2 ]Laboratory of Marine Pearl Culture, Zhanjiang, China
                [3 ]Environment Protection Monitoring Station, Environmental Protection Agency of Zhanjiang, Zhanjiang, China
                Institute of Oceanology, Chinese Academy of Sciences, CHINA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: RLH QHW. Performed the experiments: RLH QHW ZZ XXZ. Analyzed the data: RLH ZZ XXZ. Contributed reagents/materials/analysis tools: QHW XXD. Wrote the paper: RLH XXZ YJ YWD. Revised the manuscript: YJ.

                Article
                PONE-D-15-28721
                10.1371/journal.pone.0141390
                4619886
                26496197
                acfe8a78-90ff-4c80-8e03-22d3d8801b21
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 6 July 2015
                : 6 October 2015
                Page count
                Figures: 4, Tables: 0, Pages: 15
                Funding
                This work was supported by the National Natural Science Foundation, Award Number: 31372526, Recipient:Yuewen Deng; the National Natural Science Foundation, Award Number: 31272635, Recipient: Xiaodong Du; the Project of Science and Technology of Zhanjiang, Award Number: 2014B101, Recipient: Ronglian Huang; and the Project of Enhancing School with Innovation of Guangdong Ocean University, Award Number: GDOU2014050207, Recipient: Qingheng Wang.
                Categories
                Research Article
                Custom metadata
                Data are archived at the NCBI Sequence Read Archive (SRA) under accession number SRA297257.

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