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Anthocyanins in Molecular Biology

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Biochemistry".

Deadline for manuscript submissions: closed (30 June 2022) | Viewed by 36209

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Dr. Sun-Hyung Lim

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School of Biotechnology, Division of Horticultural Biotechnology, Hankyong National University, Anseong 17579, Korea
Interests: metabolic engineering; genome editing; secondary metabolites; flavonoid biosynthetic mechanism; transcriptional factor

Special Issue Information

Dear Colleagues,

Due to their influence on the various colors found in flowers, fruits, and vegetables, as well as their strong antioxidant effects, plant biologists have been interested in the biosynthetic mechanisms of anthocyanins. Anthocyanin accumulation is determined by the balance between biosynthesis and degradation. Therefore, the understanding of the anthocyanin biosynthesis–degradation pathway and the regulation of anthocyanins will allow the development of novel horticultural crops enriched in these compounds, resulting in enhanced aesthetic properties and nutritional value. This Special Issue covers various aspects of biosynthetic and degradative mechanisms in anthocyanins, such as the identification of biosynthetic and degradation-related genes in anthocyanins via molecular biology, the investigation of anthocyanin degradation mechanisms, as well as metabolic engineering of anthocyanins in plants.

Dr. Sun-Hyung Lim
Guest Editor

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Keywords

Anthocyanins biosynthesis
Metabolic engineering
Anthocyanins degradation
Transcriptional factors for anthocyanin biosynthesis or degradation
Environmental regulation

Published Papers (5 papers)

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Research

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18 pages, 3922 KiB

Open AccessArticle

RsTTG1, a WD40 Protein, Interacts with the bHLH Transcription Factor RsTT8 to Regulate Anthocyanin and Proanthocyanidin Biosynthesis in Raphanus sativus

by Sun-Hyung Lim, Da-Hye Kim and Jong-Yeol Lee

Int. J. Mol. Sci. 2022, 23(19), 11973; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms231911973 - 09 Oct 2022

Cited by 6 | Viewed by 1747

Abstract

MBW complexes, consisting of MYB, basic helix–loop–helix (bHLH), and WD40 proteins, regulate multiple traits in plants, including anthocyanin and proanthocyanidin (PA) biosynthesis and the determination of epidermal cell fate. Here, a WD40 gene from Raphanus sativus, designated TRANSPARENT TESTA GLABRA 1 (RsTTG1), was cloned and functionally characterized. Heterologous expression of RsTTG1 in the Arabidopsis thaliana mutant ttg1-22 background restored accumulation of anthocyanin and PA in the mutant and rescued trichome development. In radish, RsTTG1 was abundantly expressed in all root and leaf tissues, independently of anthocyanin accumulation, while its MBW partners RsMYB1 and TRANSPARENT TESTA 8 (RsTT8) were expressed at higher levels in pigment-accumulating tissues. In yeast two-hybrid analysis, the full-length RsTTG1 protein interacted with RsTT8. Moreover, transient protoplast co-expression assays demonstrated that RsTTG1, which localized to both the cytoplasm and nucleus, moves from the cytoplasm to the nucleus in the presence of RsTT8. When co-expressed with RsMYB1 and RsTT8, RsTTG1 stably activated the promoters of the anthocyanin biosynthesis genes CHALCONE SYNTHASE (RsCHS) and DIHYDROFLAVONOL 4-REDUCTASE (RsDFR). Transient expression of RsTTG1 in tobacco leaves exhibited an increase in anthocyanin accumulation due to activation of the expression of anthocyanin biosynthesis genes when simultaneously expressed with RsMYB1 and RsTT8. These results indicate that RsTTG1 is a vital regulator of pigmentation and trichome development as a functional homolog of AtTTG1. Full article

(This article belongs to the Special Issue Anthocyanins in Molecular Biology)

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18 pages, 3300 KiB

Open AccessArticle

The R3-Type MYB Transcription Factor BrMYBL2.1 Negatively Regulates Anthocyanin Biosynthesis in Chinese Cabbage (Brassica rapa L.) by Repressing MYB–bHLH–WD40 Complex Activity

by JiYeon Kim, Da-Hye Kim, Jong-Yeol Lee and Sun-Hyung Lim

Int. J. Mol. Sci. 2022, 23(6), 3382; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23063382 - 21 Mar 2022

Cited by 17 | Viewed by 2677

Abstract

Chinese cabbage (Brassica rapa L.) leaves are purple in color due to anthocyanin accumulation and have nutritional and aesthetic value, as well as antioxidant properties. Here, we identified the R3 MYB transcription factor BrMYBL2.1 as a key negative regulator of anthocyanin biosynthesis. A Chinese cabbage cultivar with green leaves harbored a functional BrMYBL2.1 protein, designated BrMYBL2.1-G, with transcriptional repressor activity of anthocyanin biosynthetic genes. By contrast, BrMYBL2.1 from a Chinese cabbage cultivar with purple leaves carried a poly(A) insertion in the third exon of the gene, resulting in the insertion of multiple lysine residues in the predicted protein, designated BrMYBL2.1-P. Although both BrMYBL2.1 variants localized to the nucleus, only BrMYBL2.1-G interacted with its cognate partner BrTT8. Transient infiltration assays in tobacco leaves revealed that BrMYBL2.1-G, but not BrMYBL2.1-P, actively represses pigment accumulation by inhibiting the transcription of anthocyanin biosynthetic genes. Transient promoter activation assay in Arabidopsis protoplasts verified that BrMYBL2.1-G, but not BrMYBL2.1-P, can repress transcriptional activation of BrCHS and BrDFR, which was activated by co-expression with BrPAP1 and BrTT8. We determined that BrMYBL2.1-P may be more prone to degradation than BrMYBL2.1-G via ubiquitination. Taken together, these results demonstrate that BrMYBL2.1-G blocks the activity of the MBW complex and thus represses anthocyanin biosynthesis, whereas the variant BrMYBL2.1-P from purple Chinese cabbage cannot, thus leading to higher anthocyanin accumulation. Full article

(This article belongs to the Special Issue Anthocyanins in Molecular Biology)

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22 pages, 8956 KiB

Open AccessArticle

Novel Insights into Anthocyanin Metabolism and Molecular Characterization of Associated Genes in Sugarcane Rinds Using the Metabolome and Transcriptome

by Muhammad Junaid Rao, Mingzheng Duan, Mingchong Yang, Hongzeng Fan, Songhao Shen, Lihua Hu and Lingqiang Wang

Int. J. Mol. Sci. 2022, 23(1), 338; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms23010338 - 29 Dec 2021

Cited by 13 | Viewed by 2260

Abstract

Saccharum officinarum (sugarcane) is the fifth major cultivated crop around the world. Sugarcane rind is a promising source for anthocyanin pigments; however, limited information is available on the anthocyanin and its biosynthesis in sugarcane rinds. In this study, we have quantified 49 compounds including 6 flavonoids and 43 anthocyanins in the rind of 6 sugarcane cultivars by using LCMS/MS approach. Thirty of them were quantified for the first time in sugarcane. The 43 anthocyanins included 10 cyanidin (Cya), 11 pelargonidin (Pel), 9 peonidin (Peo), 5 malvidin (Mal), 4 delphinidin (Del), and 4 petunidin (Pet) metabolites. High contents of Cya derivatives were observed in the rind of YT71/210 (dark purple rind), such as cya-3-O-(6-O-malonyl)-glu 1283.3 µg/g and cya-3-O-glu 482.67 µg/g followed by ROC22 (red rind) 821.3 µg/g and 409 µg/g, respectively, whereas the YT93/159 (green rind) showed a minimum level of these compounds. Among six cultivars, ROC22 rind has high levels of Peo derivatives such as peo-3-O-glu (197 µg/g), peo-3-O-(6-O-malonyl)-glu (69 µg/g) and peo-3-O-(6-O-p-coumaryl)-glu (55.17 µg/g). The gene expression analysis revealed that some genes, including a MYB(t) gene, were highly associated with the color phenotype. Thus, we cloned and overexpressed the gene in Arabidopsis and found the pinkish brown color in the hypocotyl of all transgenic lines compared with the wild type. Hence, we have quantified a wide range of anthocyanins in major sugarcane cultivars, reported many new anthocyanins for the first time, and concluded that Cya and Peo derivatives are the major contributing factor of dissimilar colors in sugarcane. The finding and the verification of a novel MYB gene involved in anthocyanin biosynthesis have demonstrated that our study was very valuable for gene discovery and genetic improvement of sugarcane cultivars to harvest high anthocyanin contents. Full article

(This article belongs to the Special Issue Anthocyanins in Molecular Biology)

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19 pages, 5035 KiB

Open AccessArticle

Loss of the R2R3 MYB Transcription Factor RsMYB1 Shapes Anthocyanin Biosynthesis and Accumulation in Raphanus sativus

by Da-Hye Kim, Jundae Lee, JuHee Rhee, Jong-Yeol Lee and Sun-Hyung Lim

Int. J. Mol. Sci. 2021, 22(20), 10927; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms222010927 - 10 Oct 2021

Cited by 11 | Viewed by 2782

Abstract

The red or purple color of radish (Raphanus sativus L.) taproots is due to anthocyanins, which have nutritional and aesthetic value, as well as antioxidant properties. Moreover, the varied patterns and levels of anthocyanin accumulation in radish roots make them an interesting system for studying the transcriptional regulation of anthocyanin biosynthesis. The R2R3 MYB transcription factor RsMYB1 is a key positive regulator of anthocyanin biosynthesis in radish. Here, we isolated an allele of RsMYB1, named RsMYB1^Short, in radish cultivars with white taproots. The RsMYB1^Short allele carried a 4 bp insertion in the first exon causing a frame-shift mutation of RsMYB1, generating a truncated protein with only a partial R2 domain at the N-terminus. Unlike RsMYB1^Full, RsMYB1^Short was localized to the nucleus and the cytoplasm and failed to interact with their cognate partner RsTT8. Transient expression of genomic or cDNA sequences for RsMYB1^Short in radish cotyledons failed to induce anthocyanin accumulation, but that for RsMYB1^Full activated it. Additionally, RsMYB1^Short showed the lost ability to induce pigment accumulation and to enhance the transcript level of anthocyanin biosynthetic genes, while RsMYB1^Full promoted both processes when co-expressed with RsTT8 in tobacco leaves. As the result of the transient assay, co-expressing RsTT8 and RsMYB1^Full, but not RsMYB1^Short, also enhanced the promoter activity of RsCHS and RsDFR. We designed a molecular marker for RsMYB1 genotyping, and revealed that the RsMYB1^Short allele is common in white radish cultivars, underscoring the importance of variation at the RsMYB1 locus in anthocyanin biosynthesis in the radish taproot. Together, these results indicate that the nonsense mutation of RsMYB1 generated the truncated protein, RsMYB1^Short, that had the loss of ability to regulate anthocyanin biosynthesis. Our findings highlight that the frame shift mutation of RsMYB1 plays a key role in anthocyanin biosynthesis in the radish taproot. Full article

(This article belongs to the Special Issue Anthocyanins in Molecular Biology)

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Review

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16 pages, 1407 KiB

Open AccessReview

Light Induced Regulation Pathway of Anthocyanin Biosynthesis in Plants

by Yanyun Ma, Xu Ma, Xiang Gao, Weilin Wu and Bo Zhou

Int. J. Mol. Sci. 2021, 22(20), 11116; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms222011116 - 15 Oct 2021

Cited by 58 | Viewed by 25848

Abstract

Anthocyanins are natural pigments with antioxidant effects that exist in various fruits and vegetables. The accumulation of anthocyanins is induced by environmental signals and regulated by transcription factors in plants. Numerous evidence has indicated that among the environmental factors, light is one of the most signal regulatory factors involved in the anthocyanin biosynthesis pathway. However, the signal transduction of light and molecular regulation of anthocyanin synthesis remains to be explored. Here, we focus on the research progress of signal transduction factors for positive and negative regulation in light-dependent and light-independent anthocyanin biosynthesis. In particular, we will discuss light-induced regulatory pathways and related specific regulators of anthocyanin biosynthesis in plants. In addition, an integrated regulatory network of anthocyanin biosynthesis controlled by transcription factors is discussed based on the significant progress. Full article

(This article belongs to the Special Issue Anthocyanins in Molecular Biology)

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