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      Identification of Candidate Anthocyanin-Related Genes by Transcriptomic Analysis of ‘Furongli’ Plum ( Prunus salicina Lindl.) during Fruit Ripening Using RNA-Seq

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          Abstract

          Anthocyanins are important pigments and are responsible for red coloration in plums. However, little is known about the molecular mechanisms underlying anthocyanin accumulation in plum fruits. In this study, the RNA-seq technique was used to analyze the transcriptomic changes during fruit ripening in the red-fleshed plum ( Prunus salicina Lindl.) cultivar ‘Furongli’. Over 161 million high-quality reads were assembled into 52,093 unigenes and 49.4% of these were annotated using public databases. Of these, 25,681 unigenes had significant hits to the sequences in the NCBI Nr database, 17,203 unigenes showed significant similarity to known proteins in the Swiss-Prot database and 5816 and 8585 unigenes had significant similarity to existing sequences in the Kyoto Encyclopedia of Genes and Genomes and the Cluster of Orthologous Groups databases, respectively. A total of 3548 unigenes were differentially expressed during fruit ripening and 119 of these were annotated as involved in “biosynthesis of other secondary metabolites.” Biological pathway analysis and gene ontology term enrichment analysis revealed that 13 differentially expressed genes are involved in anthocyanin biosynthesis. Furthermore, transcription factors such as MYB and bHLH, which may control anthocyanin biosynthesis, were identified through coexpression analysis of transcription factors, and structural genes. Real-time qPCR analysis of candidate genes showed good correlation with the transcriptome data. These results contribute to our understanding of the molecular mechanisms underlying anthocyanin biosynthesis in plum flesh. The transcriptomic data generated in this study provide a basis for further studies of fruit ripening in plum.

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          Most cited references73

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          Retrotransposon-induced mutations in grape skin color.

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            An R2R3 MYB transcription factor associated with regulation of the anthocyanin biosynthetic pathway in Rosaceae

            Background The control of plant anthocyanin accumulation is via transcriptional regulation of the genes encoding the biosynthetic enzymes. A key activator appears to be an R2R3 MYB transcription factor. In apple fruit, skin anthocyanin levels are controlled by a gene called MYBA or MYB1, while the gene determining fruit flesh and foliage anthocyanin has been termed MYB10. In order to further understand tissue-specific anthocyanin regulation we have isolated orthologous MYB genes from all the commercially important rosaceous species. Results We use gene specific primers to show that the three MYB activators of apple anthocyanin (MYB10/MYB1/MYBA) are likely alleles of each other. MYB transcription factors, with high sequence identity to the apple gene were isolated from across the rosaceous family (e.g. apples, pears, plums, cherries, peaches, raspberries, rose, strawberry). Key identifying amino acid residues were found in both the DNA-binding and C-terminal domains of these MYBs. The expression of these MYB10 genes correlates with fruit and flower anthocyanin levels. Their function was tested in tobacco and strawberry. In tobacco, these MYBs were shown to induce the anthocyanin pathway when co-expressed with bHLHs, while over-expression of strawberry and apple genes in the crop of origin elevates anthocyanins. Conclusions This family-wide study of rosaceous R2R3 MYBs provides insight into the evolution of this plant trait. It has implications for the development of new coloured fruit and flowers, as well as aiding the understanding of temporal-spatial colour change.
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              White grapes arose through the mutation of two similar and adjacent regulatory genes.

              Most of the thousands of grapevine cultivars (Vitis vinifera L.) can be divided into two groups, red and white, based on the presence or absence of anthocyanin in the berry skin, which has been found from genetic experiments to be controlled by a single locus. A regulatory gene, VvMYBA1, which could activate anthocyanin biosynthesis in a transient assay, was recently shown not to be transcribed in white berries due to the presence of a retrotransposon in the promoter. We have found that the berry colour locus comprises two very similar genes, VvMYBA1 and VvMYBA2, located on a single bacterial artificial chromosome. Either gene can regulate colour in the grape berry. The white berry allele of VvMYBA2 is inactivated by two non-conservative mutations, one leads to an amino acid substitution and the other to a frame shift resulting in a smaller protein. Transient assays showed that either mutation removed the ability of the regulator to switch on anthocyanin biosynthesis. VvMYBA2 sequence analyses, together with marker information, confirmed that 55 white cultivars all contain the white berry allele, but not red berry alleles. These results suggest that all extant white cultivars of grape vines have a common origin. We conclude that rare mutational events occurring in two adjacent genes were essential for the genesis of the white grapes used to produce the white wines and white table grapes we enjoy today.
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                Author and article information

                Contributors
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                31 August 2016
                2016
                : 7
                : 1338
                Affiliations
                Fruit Research Institute, Fujian Academy of Agricultural Sciences Fuzhou, China
                Author notes

                Edited by: Claudio Bonghi, University of Padua, Italy

                Reviewed by: Chuang Ma, Northwest Agricultural and Forestry University, China; Antonio Ferrante, University of Milan, Italy

                This article was submitted to Crop Science and Horticulture, a section of the journal Frontiers in Plant Science

                Article
                10.3389/fpls.2016.01338
                5005409
                27630660
                cd5e6a68-04e2-4ace-98cd-cb59fb61e7d4
                Copyright © 2016 Fang, Zhou, Ye, Jiang and Pan.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 31 May 2016
                : 19 August 2016
                Page count
                Figures: 9, Tables: 4, Equations: 0, References: 105, Pages: 15, Words: 9817
                Categories
                Plant Science
                Original Research

                Plant science & Botany
                transcriptome,prunus salicina lindl.,anthocyanin biosynthesis,fruit ripening,biosynthetic enzyme,transcription factor

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