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      An R3-MYB repressor, BnCPC forms a feedback regulation with MBW complex to modulate anthocyanin biosynthesis in Brassica napus

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          Abstract

          Background

          Anthocyanins are metabolites of phenylpropanoid pathway, and involves in diverse processes of plant development and adaptation, which are regulated by the MYB-bHLH-WD40 (MBW) protein complexes. Many R2R3-MYB activators have been well characterized, but the MYB repressors in anthocyanin biosynthesis were recognized recently, which are also important in modulating phenylpropanoid metabolism in plants. The regulatory mechanism of anthocyanin biosynthesis in oil crop Brassica napus remains to be revealed.

          Results

          In this study, we identified an anthocyanin repressor BnCPC in B. napus. BnCPC encoded a typical R3-MYB protein containing a conserved [D/E]Lx2[R/K]x3Lx6Lx3R motif for interaction with bHLH proteins. Overexpression of BnCPC in B. napus inhibited anthocyanin accumulation, especially under anthocyanin inducible conditions. Protein–protein interaction and dual-luciferase assays confirmed that BnCPC could compete with BnPAP1 to interact with bHLHs (BnTT8 and BnEGL3), and repress the expression of anthocyanin biosynthetic genes (e.g., BnDFR) that activated by MBW complexes. Moreover, we found BnCPC inhibited the MBW complex-induced BnCPC activity.

          Conclusions

          Overall, this research demonstrated that BnCPC repressed anthocyanin biosynthesis by affecting the formation of MBW complex, and formed a feedback loop to regulate anthocyanin accumulation in B. napus.

          Supplementary Information

          The online version contains supplementary material available at 10.1186/s13068-022-02227-6.

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          Most cited references81

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          Kenneth Livak, Thomas D. Schmittgen (2001)
          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
          Article 0 comments Cited 24392 times – based on 0 reviews     Review now
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            Transient expression vectors for functional genomics, quantification of promoter activity and RNA silencing in plants

            Roger Hellens, Andrew Allan, Ellen N Friel … (2006)
            Background We describe novel plasmid vectors for transient gene expression using Agrobacterium, infiltrated into Nicotiana benthamiana leaves. We have generated a series of pGreenII cloning vectors that are ideally suited to transient gene expression, by removing elements of conventional binary vectors necessary for stable transformation such as transformation selection genes. Results We give an example of expression of heme-thiolate P450 to demonstrate effectiveness of this system. We have also designed vectors that take advantage of a dual luciferase assay system to analyse promoter sequences or post-transcriptional regulation of gene expression. We have demonstrated their utility by co-expression of putative transcription factors and the promoter sequence of potential target genes and show how orthologous promoter sequences respond to these genes. Finally, we have constructed a vector that has allowed us to investigate design features of hairpin constructs related to their ability to initiate RNA silencing, and have used these tools to study cis-regulatory effect of intron-containing gene constructs. Conclusion In developing a series of vectors ideally suited to transient expression analysis we have provided a resource that further advances the application of this technology. These minimal vectors are ideally suited to conventional cloning methods and we have used them to demonstrate their flexibility to investigate enzyme activity, transcription regulation and post-transcriptional regulatory processes in transient assays.
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              Plant genetics. Early allopolyploid evolution in the post-Neolithic Brassica napus oilseed genome.

              Boulos Chalhoub, France Denoeud, Shengyi Liu … (2014)
              Oilseed rape (Brassica napus L.) was formed ~7500 years ago by hybridization between B. rapa and B. oleracea, followed by chromosome doubling, a process known as allopolyploidy. Together with more ancient polyploidizations, this conferred an aggregate 72× genome multiplication since the origin of angiosperms and high gene content. We examined the B. napus genome and the consequences of its recent duplication. The constituent An and Cn subgenomes are engaged in subtle structural, functional, and epigenetic cross-talk, with abundant homeologous exchanges. Incipient gene loss and expression divergence have begun. Selection in B. napus oilseed types has accelerated the loss of glucosinolate genes, while preserving expansion of oil biosynthesis genes. These processes provide insights into allopolyploid evolution and its relationship with crop domestication and improvement. Copyright © 2014, American Association for the Advancement of Science.
              Article 0 comments Cited 479 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Youping Wang: wangyp@yzu.edu.cn
                Jinjin Jiang: jjjiang@yzu.edu.cn
                Journal
                Journal ID (nlm-ta): Biotechnol Biofuels Bioprod
                Journal ID (iso-abbrev): Biotechnol Biofuels Bioprod
                Title: Biotechnology for Biofuels and Bioproducts
                Publisher: BioMed Central (London )
                ISSN (Electronic): 2731-3654
                Publication date (Electronic): 29 November 2022
                Publication date PMC-release: 29 November 2022
                Publication date Collection: 2022
                Volume: 15
                Electronic Location Identifier: 133
                Affiliations
                [1 ]GRID grid.268415.c, Jiangsu Provincial Key Laboratory of Crop Genetics and Physiology, , Yangzhou University, ; Yangzhou, 225009 China
                [2 ]Joint International Research Laboratory of Agriculture and Agri-Product Safety, The Ministry of Education of China, Yangzhou, 225009 China
                [3 ]GRID grid.263761.7, ISNI 0000 0001 0198 0694, School of Biological and Food Engineering, , Suzhou University, ; Suzhou, 234000 China
                Article
                Publisher ID: 2227
                DOI: 10.1186/s13068-022-02227-6
                PMC ID: 9706894
                PubMed ID: 36447291
                SO-VID: 773cb138-0ce4-4485-8a29-39bd4d3b80b9
                Copyright © © The Author(s) 2022
                License:

                Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

                History
                Date received : 13 September 2022
                Date accepted : 9 November 2022
                Funding
                Funded by: the National Natural Science Foundations of China
                Award ID: 31972963, 32201744
                Award ID: 31972963, 32201744
                Funded by: the National Key Research and Development Program of China
                Award ID: 2018YFE0108000
                Categories
                Subject: Research
                Custom metadata
                issue-copyright-statement © The Author(s) 2022

                Keywords: brassica napus,anthocyanin,bncpc,mbw complex,repressor,bndfr
                Data availability:
                Keywords: brassica napus, anthocyanin, bncpc, mbw complex, repressor, bndfr

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