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Transcriptional factors, metabolic engineering, anthocyanin,

Sandra Fresquet-Corrales, Edelín Roque, Alejandro Sarrión-Perdigones, Maricruz Rochina, María P López-Gresa, Huertas M Díaz-Mula, José M Bellés, Francisco Tomás-Barberán, José P Beltrán, Luis A Cañas
Proanthocyanidins (PAs), or condensed tannins, are powerful antioxidants that remove harmful free oxygen radicals from cells. To engineer the anthocyanin and proanthocyanidin biosynthetic pathways to de novo produce PAs in two Nicotiana species, we incorporated four transgenes to the plant chassis. We opted to perform a simultaneous transformation of the genes linked in a multigenic construct rather than classical breeding or retransformation approaches. We generated a GoldenBraid 2.0 multigenic construct containing two Antirrhinum majus transcription factors (AmRosea1 and AmDelila) to upregulate the anthocyanin pathway in combination with two Medicago truncatula genes (MtLAR and MtANR) to produce the enzymes that will derivate the biosynthetic pathway to PAs production...
2017: PloS One
J Andrew Jones, Victoria R Vernacchio, Shannon M Collins, Abhijit N Shirke, Yu Xiu, Jacob A Englaender, Brady F Cress, Catherine C McCutcheon, Robert J Linhardt, Richard A Gross, Mattheos A G Koffas
Fermentation-based chemical production strategies provide a feasible route for the rapid, safe, and sustainable production of a wide variety of important chemical products, ranging from fuels to pharmaceuticals. These strategies have yet to find wide industrial utilization due to their inability to economically compete with traditional extraction and chemical production methods. Here, we engineer for the first time the complex microbial biosynthesis of an anthocyanin plant natural product, starting from sugar...
June 6, 2017: MBio
Teresa Cordero, Mohamed A Mohamed, Juan-José López-Moya, José-Antonio Daròs
Potato virus Y (PVY) is a major threat to the cultivation of potato and other solanaceous plants. By inserting a cDNA coding for the Antirrhinum majus Rosea1 transcription factor into a PVY infectious clone, we created a biotechnological tool (PVY-Ros1) that allows infection by this relevant plant virus to be tracked by the naked eye with no need for complex instrumentation. Rosea1 is an MYB-type transcription factor whose expression activates the biosynthesis of anthocyanin pigments in a dose-specific and cell-autonomous manner...
2017: Frontiers in Microbiology
Kasturi Dasgupta, Roger Thilmony, Ed Stover, Maria Luiza Oliveira, James Thomson
The level of anthocyanins in plants vary widely among cultivars, developmental stages and environmental stimuli. Previous studies have reported that the expression of various MYBs regulate anthocyanin pigmentation during growth and development. Here we examine the activity of 3 novel R2R3-MYB transcription factor (TF) genes, PamMybA.1, PamMybA.3 and PamMybA.5 from Prunus americana. The anthocyanin accumulation patterns mediated by CaMV double35S promoter (db35Sp) controlled expression of the TFs in transgenic tobacco were compared with citrus-MoroMybA, Arabidopsis-AtMybA1 and grapevine-VvMybA1 transgenics during their entire growth cycles...
April 3, 2017: GM Crops & Food
Xianzhi He, Yong Li, Darlene Lawson, De-Yu Xie
In this study, we investigate the metabolic engineering of anthocyanins in two dark tobacco crops (Narrow Leaf Madole and KY171) and evaluate the effects on physiological features of plant photosynthesis. Arabidopsis PAP1 (production of anthocyanin pigment 1) gene (AtPAP1) encodes a R2R3-type MYB transcript factor that is a master component of regulatory complexes controlling anthocyanin biosynthesis. AtPAP1 was introduced to Narrow Leaf Madole and KY171 plants. Multiple transgenic plants developed red/purple pigmentation in different tissues...
January 2017: Physiologia Plantarum
Wenjun Huang, A B M Khaldun, Haiyan Lv, Liuwen Du, Chanjuan Zhang, Ying Wang
A R2R3-MYB transcription factor EsAN2 was isolated from Epimedium sagittatum and functionally characterized to regulate the anthocyanin biosynthetic pathway. Epimedium plants are used widely both as traditional Chinese medicinal herbs and ornamental perennials. Anthocyanins, acting as major contributors to plant color diversity, their biosynthesis are regulated by a series of transcription factors, including MYB, bHLH and WD40 protein. Previously, a MYB transcription factor involved in regulation of the anthocyanin pathway from Epimedium sagittatum, EsMYBA1 has been isolated, but was found to be expressed mostly in leaves...
April 2016: Plant Cell Reports
Wenjun Huang, Shaohua Zeng, Gong Xiao, Guoyan Wei, Sihong Liao, Jianjun Chen, Wei Sun, Haiyan Lv, Ying Wang
Herba epimedii (Epimedium), a traditional Chinese medicine, has been widely used as a kidney tonic and antirheumatic medicine for thousands of years. In Epimedium, flavonoids have been demonstrated to be the main bioactive components (BCs). However, the molecular biosynthetic and regulatory mechanisms of flavonoid-derived BCs remain obscure. In this study, we isolated 12 structural genes and two putative transcription factors (TFs) in the flavonoid pathway. Phytochemical analysis showed that the total content of four representative BCs (epimedin A, B, C, and icariin) decreased slightly or dramatically in two lines of Epimedium sagittatum during leaf development...
2015: Frontiers in Plant Science
Shujuan Zhao, Jinjia Zhang, Ronghui Tan, Li Yang, Xiaoyu Zheng
Tanshinones are valuable natural diterpenoids from danshen (Salvia miltiorrhiza Bunge). Here, it was demonstrated that maize transcription factor C1 improved the accumulation of tanshinones by comprehensively upregulating the pathway genes, especially SmMDC and SmPMK in danshen hairy roots, yielding total tanshinones up to 3.59mg g(-1) of dry weight in line C1-6, a 3.4-fold increase compared with the control. Investigation of 2024bp of the SmMDC promoter fragment revealed that C1-mediated upregulation of terpenoid genes was possibly due to the direct interaction of C1 with its recognition sequences...
December 2015: Journal of Experimental Botany
Meiliang Zhou, Li Wei, Zhanmin Sun, Lihua Gao, Yu Meng, Yixiong Tang, Yanmin Wu
Proanthocyanidins (PA), also known as condensed tannins, contribute to important forage legumes traits including disease resistance and forage quality. PA in forage plants has both positive and negative effects on feed digestibility and animal performance. The analytical methods and their applicability in measuring the contents of PA in forage plants are essential to studies on their nutritional effects. In spite of important breakthroughs in our understanding of the PA biosynthesis, important questions still remain to be answered such as the PA polymerization and transport...
May 2015: Applied Microbiology and Biotechnology
Kui Lin-Wang, Tony K McGhie, Mindy Wang, Yuhui Liu, Benjamin Warren, Roy Storey, Richard V Espley, Andrew C Allan
The woodland strawberry, Fragaria vesca is a model fruit for a number of rosaceous crops. We have engineered altered concentrations of anthocyanin in F. vesca, to determine the impact on plant growth and fruit quality. Anthocyanin concentrations were significantly increased by over-expression or decreased by knock-down of the R2R3 MYB activator, MYB10. In contrast, a potential bHLH partner for MYB10 (bHLH33) did not affect the anthocyanin pathway when knocked down using RNAi constructs. Metabolic analysis of fruits revealed that, of all the polyphenolics surveyed, only cyanidin, and pelargonidin glucoside, and coumaryl hexose were significantly affected by over-expression and knock down of MYB10...
2014: Frontiers in Plant Science
Tomoko Mitsunami, Masahiro Nishihara, Ivan Galis, Kabir Md Alamgir, Yuko Hojo, Kohei Fujita, Nobuhiro Sasaki, Keichiro Nemoto, Tatsuya Sawasaki, Gen-ichiro Arimura
Anthocyanin pigments and associated flavonoids have demonstrated antioxidant properties and benefits for human health. Consequently, current plant bioengineers have focused on how to modify flavonoid metabolism in plants. Most of that research, however, does not consider the role of natural biotic stresses (e.g., herbivore attack). To understand the influence of herbivore attack on the metabolic engineering of flavonoids, we examined tobacco plants overexpressing the Arabidopsis PAP1 gene (encoding an MYB transcription factor), which accumulated anthocyanin pigments and other flavonoids/phenylpropanoids...
2014: PloS One
Ming-Zhu Shi, De-Yu Xie
Arabidopsis thaliana is the first model plant, the genome of which has been sequenced. In general, intensive studies on this model plant over the past nearly 30 years have led to many new revolutionary understandings in every single aspect of plant biology. Here, we review the current understanding of anthocyanin biosynthesis in this model plant. Although the investigation of anthocyanin structures in this model plant was not performed until 2002, numerous studies over the past three decades have been conducted to understand the biosynthesis of anthocyanins...
2014: Recent Patents on Biotechnology
Yuan Zhang, Ya-Ping Yan, Yu-Cui Wu, Wen-Ping Hua, Chen Chen, Qian Ge, Zhe-Zhi Wang
To produce beneficial phenolic acids for medical and commercial purposes, researchers are interested in improving the normally low levels of salvianolic acid B (Sal B) produced by Salvia miltiorrhiza. Here, we present a strategy of combinational genetic manipulation to enrich the precursors available for Sal B biosynthesis. This approach, involving the lignin pathway, requires simultaneous, ectopic expression of an Arabidopsis Production of Anthocyanin Pigment 1 transcription factor (AtPAP1) plus co-suppression of two endogenous, key enzyme genes: cinnamoyl-CoA reductase (SmCCR) and caffeic acid O-methyltransferase (SmCOMT)...
January 2014: Metabolic Engineering
Jian Qiu, Fenghua Gao, Guoan Shen, Chonghui Li, Xiaoyan Han, Qiao Zhao, Dexiu Zhao, Xuejun Hua, Yongzhen Pang
The rare wild species of snow lotus Saussurea involucrata is a commonly used medicinal herb with great pharmacological value for human health, resulting from its uniquely high level of phenylpropanoid compound production. To gain information on the phenylpropanid biosynthetic pathway genes in this critically important medicinal plant, global transcriptome sequencing was performed. It revealed that the phenylpropanoid pathway genes were well represented in S. involucrata. In addition, we introduced two key phenylpropanoid pathway inducing transcription factors (PAP1 and Lc) into this medicinal plant...
2013: PloS One
Richard A Dixon, Chenggang Liu, Ji Hyung Jun
Monomeric anthocyanins and polymeric proanthocyanidins (condensed tannins) contribute to important plant traits such as flower and fruit pigmentation, fruit astringency, disease resistance and forage quality. Recent advances in our understanding of the transcriptional control mechanisms that regulate anthocyanin and condensed tannin formation in plants suggest new approaches for the engineering of quality traits associated with these molecules. In particular, MYB family transcription factors are emerging as central players in the coordinated activation of sets of genes specific for the anthocyanin and tannin pathways...
April 2013: Current Opinion in Biotechnology
Yue Feng, Cong-Mei Cao, Meenu Vikram, Sunghun Park, Hye Jin Kim, Jong Chan Hong, Luis Cisneros-Zevallos, Hisashi Koiwa
Inducible gene expression is a powerful tool to study and engineer genes whose overexpression could be detrimental for the host organisms. However, only limited systems have been adopted in plant biotechnology. We have developed an osmotically inducible system using three components of plant origin, RD29a (Responsive to Dehydration 29A) promoter, CBF3 (C-repeat Binding Factor 3) transcription factor and cpl1-2 (CTD phosphatase-like 1) mutation. The osmotic stress responsible RD29a promoter contains the CBF3 binding sites and thus RD29A-CBF3 feedforward cassette enhances induction of RD29a promoter under stress...
2011: PloS One
Tiziana Crifò, Ivana Puglisi, Goffredo Petrone, Giuseppe Reforgiato Recupero, Angela Roberta Lo Piero
The productivity and the geographical distribution of most commercially important Citrus varieties are markedly affected by environmental low temperatures. As gene engineering has been shown to be a favourable alternative to produce germplasm with improved cold tolerance, a broad group of cold regulated genes have been previously identified from several Citrus spp. By contrast, little information regarding the cold stress response of pigmented sweet orange varieties is available although they might provide a pivotal contribution to define the whole events occurring in cold exposed Citrus fruits...
May 1, 2011: Gene
Zhijian T Li, Sadanand A Dhekney, Dennis J Gray
We report the development of a convenient plant-based reporter system to analyze promoters and facilitate selection of genetically engineered plants. The VvMybA1 gene of grapevine (Vitis vinifera L.) regulates the last metabolic step of anthocyanin biosynthesis and its ectopic expression leads to anthocyanin production in otherwise non-pigmented cells. To develop an anthocyanin-based quantitative reporter system, the VvMybA1 gene was isolated from V. vinifera 'Merlot' and placed under control of three promoters to test its ability to distinguish different activity levels...
October 2011: Transgenic Research
Ming-Zhu Shi, De-Yu Xie
We report metabolic engineering of Arabidopsis red cells and genome-wide gene expression analysis associated with anthocyanin biosynthesis and other metabolic pathways between red cells and wild-type (WT) cells. Red cells of A. thaliana were engineered for the first time from the leaves of production of anthocyanin pigment 1-Dominant (pap1-D). These red cells produced seven anthocyanin molecules including a new one that was characterized by LC-MS analysis. Wild-type cells established as a control did not produce anthocyanins...
April 2011: Planta
Sitakanta Pattanaik, Joshua R Werkman, Que Kong, Ling Yuan
Regulation of gene expression is largely coordinated by a complex network of interactions between transcription factors (TFs), co-factors, and their cognate cis-regulatory elements in the genome. TFs are multidomain proteins that arise evolutionarily through protein domain shuffling. The modular nature of TFs has led to the idea that specific modules of TFs can be re-designed to regulate desired gene(s) through protein engineering. Utilization of designer TFs for the control of metabolic pathways has emerged as an effective approach for metabolic engineering...
2010: Methods in Molecular Biology
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