scholarly journals Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.)

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yuwei Cao ◽  
Mengmeng Bi ◽  
Panpan Yang ◽  
Meng Song ◽  
Guoren He ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Theoretical Basis ◽  
Anthocyanin Biosynthesis ◽  
Regulatory Proteins ◽  
Biosynthesis Pathway ◽  
Temporal Expression ◽  
Anthocyanin Pigmentation ◽  
Cdna Insert ◽  
R2r3 Myb ◽  
First Time

Abstract Background Anthocyanins, which belong to flavonoids, are widely colored among red-purple pigments in the Asiatic hybrid lilies (Lilium spp.). Transcription factor (TF) LhMYBSPLATTER (formerly known as LhMYB12-Lat), identified as the major kernel protein, regulating the anthocyanin biosynthesis pathway in ‘Tiny Padhye’ of Tango Series cultivars, which the pigmentation density is high in the lower half of tepals and this patterning is of exceptional ornamental value. However, the research on mechanism of regulating the spatial and temporal expression differences of LhMYBSPLATTER, which belongs to the R2R3-MYB subfamily, is still not well established. To explore the molecular mechanism of directly related regulatory proteins of LhMYBSPLATTER in the anthocyanin pigmentation, the yeast one-hybrid (Y1H) cDNA library was constructed and characterized. Results In this study, we describe a yeast one-hybrid library to screen transcription factors that regulate LhMYBSPLATTER gene expression in Lilium, with the library recombinant efficiency of over 98%. The lengths of inserted fragments ranged from 400 to 2000 bp, and the library capacity reached 1.6 × 106 CFU of cDNA insert, which is suitable to fulfill subsequent screening. Finally, seven prey proteins, including BTF3, MYB4, IAA6-like, ERF4, ARR1, ERF WIN1-like, and ERF061 were screened by the recombinant bait plasmid and verified by interaction with the LhMYBSPLATTER promoter. Among them, ERFs, AUX/IAA, and BTF3 may participate in the negative regulation of the anthocyanin biosynthesis pathway in Lilium. Conclusion A yeast one-hybrid library of lily was successfully constructed in the tepals for the first time. Seven candidate TFs of LhMYBSPLATTER were screened, which may provide a theoretical basis for the study of floral pigmentation.

scholarly journals Construction of Yeast One-hybrid Library and Screening of Transcription Factors Regulating LhMYB12-Lat Expression in Asiatic Hybrid Lilies (Lilium spp.)

2021 ◽  
Author(s):  
Yuwei Cao ◽  
Mengmeng Bi ◽  
Panpan Yang ◽  
Meng Song ◽  
Guoren He ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Theoretical Basis ◽  
Anthocyanin Biosynthesis ◽  
Regulatory Proteins ◽  
Biosynthesis Pathway ◽  
Temporal Expression ◽  
Anthocyanin Pigmentation ◽  
Cdna Insert ◽  
R2r3 Myb ◽  
First Time

Abstract Background: Anthocyanins, which belong to flavonoids, are widely colored among red-purple pigments in the Asiatic hybrid lilies (Lilium spp.). Transcription factor (TFs) LhMYB12-Lat, identified as the major kernel protein, regulating the anthocyanin biosynthesis pathway in ‘Tiny Padhye’ of Tango series cultivars, which the pigmentation density is high in the lower half of tepals and this patterning is of exceptional ornamental value. However, the research on mechanism of regulating the spatial and temporal expression differences of LhMYB12-Lat, which belongs to the R2R3-MYB subfamily, is still not well established. To explore the molecular mechanism of directly related regulatory proteins of LhMYB12-Lat in the anthocyanin pigmentation, the yeast one-hybrid (Y1H) cDNA library was constructed and characterized. Results: In this study, we describe a yeast one-hybrid library to screen transcription factors that regulate LhMYB12-Lat gene expression in Lilium, with the library recombinant efficiency of over 98%. The lengths of inserted fragments ranged from 400-2000 bp, and the library capacity reached 1.6 × 106 CFU of cDNA insert, which is suitable to fulfill subsequent screening. Finally, seven prey proteins, including BTF3, MYB4, IAA6-like, ERF4, ARR1, ERF WIN1-like, and ERF061 were screened by the recombinant bait plasmid and verified by interaction with the LhMYB12-Lat promoter. Among them, ERF, AUX/IAA, and BTF3 may participate in the negative regulation of the anthocyanin biosynthesis pathway in Lilium.Conclusion: A yeast one-hybrid library of lily was successfully constructed in the tepals for the first time. Seven candidate TFs of LhMYB12-Lat were screened, which may provide a theoretical basis for the study of floral pigmentation.

scholarly journals Metabolic Variations of Flavonoids in Leaves of T. media and T. mairei Obtained by UPLC-ESI-MS/MS

Molecules ◽  
2019 ◽  
Vol 24 (18) ◽  
pp. 3323 ◽  
Author(s):  
Tao Wang ◽  
Fengjiao Zhang ◽  
Weibing Zhuang ◽  
Xiaochun Shu ◽  
Zhong Wang
Keyword(s):  
Theoretical Basis ◽  
Anthocyanin Biosynthesis ◽  
Ultra Performance Liquid Chromatography ◽  
Phenylpropanoid Pathway ◽  
Total Flavonoids ◽  
Flavonoid Content ◽  
Esi Ms ◽  
Taxus Mairei ◽  
Novel Drugs ◽  
First Time

The needles of Taxus species contain a large number of bioactive compounds, such as flavonoids. In the present study, the total flavonoid content in leaves of Taxus media and Taxus mairei was 19.953 and 14.464 mg/g, respectively. A total of 197 flavonoid metabolites (70 flavones, 42 flavonols, 26 flavone C-glycosides, 20 flavanones, 15 anthocyanins, 13 isoflavones, 6 flavonolignans, and 5 proanthocyanidins) were identified for the first time by a widely targeted Ultra Performance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry (UPLC-ESI-MS/MS) method within the two Taxus species, containing 160 common metabolites, with 37 unique metabolites merely determined in T. mairei or T. media. Moreover, 42 differential flavonoid metabolites were screened in the two Taxus species, which showed specific metabolic patterns in isoflavonoid biosynthesis, anthocyanin biosynthesis, and flavone and flavonol biosynthesis pathways. Compared to T. mairei, a more activated phenylpropanoid pathway was found in T. media, which could be responsible for the higher content of total flavonoids in T. media. Our results provide new insights into the diversity of flavonoid metabolites between T. mairei and T. media, and provide a theoretical basis for the sufficient utilization of Taxus species and the development of novel drugs.

scholarly journals Chromatin structure shapes the search process of transcription factors

2016 ◽  
Author(s):  
Neslihan Avcu ◽  
Nacho Molina
Keyword(s):  
Transcription Factors ◽  
High Resolution ◽  
Diffusion Process ◽  
Protein Interactions ◽  
Chromatin Structure ◽  
3D Structure ◽  
Regulatory Proteins ◽  
Local Concentration ◽  
Multi Scale ◽  
First Time

The diffusion of regulatory proteins within the nucleus plays a crucial role in the dynamics of transcriptional regulation. The standard model assumes a 3D plus ID diffusion process: regulatory proteins either move freely in solution or slide on DNA. This model however does not considered the 3D structure of chromatin. Here we proposed a multi-scale stochastic model that integrates, for the first time, high-resolution information on chromatin structure as well as DNA-protein interactions. The dynamics of transcription factors was modeled as a slide plus jump diffusion process on a chromatin network based on pair-wise contact maps obtained from high-resolution Hi-C experiments. Our model allowed us to uncover the effects of chromatin structure on transcription factor occupancy profiles and target search times. Finally, we showed that binding sites clustered on few topological associated domains leading to a higher local concentration of transcription factors which could reflect an optimal strategy to efficiently use limited transcriptional resources.

scholarly journals Regulation of MYB Transcription Factors of Anthocyanin Synthesis in Lily Flowers

2021 ◽  
Vol 12 ◽  
Author(s):  
Xiaojuan Yin ◽  
Yibing Zhang ◽  
Li Zhang ◽  
Baohua Wang ◽  
Yidi Zhao ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Transient Expression ◽  
Anthocyanin Biosynthesis ◽  
Gene Promoter ◽  
Flower Color ◽  
Preliminary Evidence ◽  
Anthocyanin Synthesis ◽  
Myb Transcription Factors ◽  
Color Formation ◽  
R2r3 Myb

Flower color is the decisive factor that affects the commercial value of ornamental flowers. Therefore, it is important to study the regulation of flower color formation in lily to discover the positive and negative factors that regulate this important trait. In this study, MYB transcription factors (TFs) were characterized to understand the regulatory mechanism of anthocyanin biosynthesis in lily. Two R2R3-MYB TFs, LvMYB5, and LvMYB1, were found to regulate anthocyanin biosynthesis in lily flowers. LvMYB5, which has an activation motif, belongs to the SG6 MYB protein subgroup of Arabidopsis thaliana. Transient expression of LvMYB5 indicated that LvMYB5 can promote coloration in Nicotiana benthamiana leaves, and that expression of LvMYB5 increases the expression levels of NbCHS, NbDFR, and NbANS. VIGS experiments in lily petals showed that the accumulation of anthocyanins was reduced when LvMYB5 was silenced. Luciferase assays showed that LvMYB5 can promote anthocyanin synthesis by activating the ANS gene promoter. Therefore, LvMYB5 plays an important role in flower coloration in lily. In addition, the transient expression experiment provided preliminary evidence that LvMYB1 (an R2R3-MYB TF) inhibits anthocyanin synthesis in lily flowers. The discovery of activating and inhibitory factors related to anthocyanin biosynthesis in lily provides a theoretical basis for improving flower color through genetic engineering. The results of our study provide a new direction for the further study of the mechanisms of flower color formation in lilies.

scholarly journals Identification and Characterization of R2R3-MYB and bHLH Transcription Factors Regulating Anthocyanin Biosynthesis in Gentian Flowers

2008 ◽  
Vol 49 (12) ◽  
pp. 1818-1829 ◽  
Author(s):  
Takashi Nakatsuka ◽  
Katia Sanae Haruta ◽  
Chetsadaporn Pitaksutheepong ◽  
Yoshiko Abe ◽  
Yuko Kakizaki ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Anthocyanin Biosynthesis ◽  
R2r3 Myb ◽  
Bhlh Transcription Factors ◽  
Identification And Characterization

scholarly journals Identification of Two Novel R2R3-MYB Transcription factors, PsMYB114L and PsMYB12L, Related to Anthocyanin Biosynthesis in Paeonia suffruticosa

2019 ◽  
Vol 20 (5) ◽  
pp. 1055 ◽  
Author(s):  
Xinpeng Zhang ◽  
Zongda Xu ◽  
Xiaoyan Yu ◽  
Lanyong Zhao ◽  
Mingyuan Zhao ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Anthocyanin Biosynthesis ◽  
Critical Role ◽  
Flower Color ◽  
Anthocyanin Accumulation ◽  
Paeonia Suffruticosa ◽  
Myb Transcription Factors ◽  
Transgenic Lines ◽  
R2r3 Myb ◽  
R2r3 Myb Transcription Factors

Flower color is a charming phenotype with very important ornamental and commercial values. Anthocyanins play a critical role in determining flower color pattern formation, and their biosynthesis is typically regulated by R2R3-MYB transcription factors (TFs). Paeonia suffruticosa is a famous ornamental plant with colorful flowers. However, little is known about the R2R3-MYB TFs that regulate anthocyanin accumulation in P. suffruticosa. In the present study, two R2R3-MYB TFs, namely, PsMYB114L and PsMYB12L, were isolated from the petals of P. suffruticosa ‘Shima Nishiki’ and functionally characterized. Sequence analysis suggested that PsMYB114L contained a bHLH-interaction motif, whereas PsMYB12L contained two flavonol-specific motifs (SG7 and SG7-2). Subsequently, the in vivo function of PsMYB114L and PsMYB12L was investigated by their heterologous expression in Arabidopsis thaliana and apple calli. In transgenic Arabidopsis plants, overexpression of PsMYB114L and of PsMYB12L caused a significantly higher accumulation of anthocyanins, resulting in purple-red leaves. Transgenic apple calli overexpressing PsMYB114L and PsMYB12L also significantly enhanced the anthocyanins content and resulted in a change in the callus color to red. Meanwhile, gene expression analysis in A. thaliana and apple calli suggested that the expression levels of the flavonol synthase (MdFLS) and anthocyanidin reductase (MdANR) genes were significantly downregulated and the dihydroflavonol 4-reductase (AtDFR) and anthocyanin synthase (AtANS) genes were significantly upregulated in transgenic lines of PsMYB114L. Moreover, the expression level of the FLS gene (MdFLS) was significantly downregulated and the DFR (AtDFR/MdDFR) and ANS (AtANS/MdANS) genes were all significantly upregulated in transgenic lines plants of PsMYB12L. These results indicate that PsMYB114L and PsMYB12L both enhance anthocyanin accumulation by specifically regulating the expression of some anthocyanin biosynthesis-related genes in different plant species. Together, these results provide a valuable resource with which to further study the regulatory mechanism of anthocyanin biosynthesis in P. suffruticosa and for the breeding of tree peony cultivars with novel and charming flower colors.

scholarly journals Editing of the MYB genes in Brassica napus as a method to increase anthocyanin pigmentation and stress tolerance

2020 ◽  
Vol 224 ◽  
pp. 04022
Author(s):  
E V Mikhaylova ◽  
M. Y. Shein ◽  
A. Y. Artyukhin ◽  
A S Sukhareva ◽  
M. A. Panfilova ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Brassica Napus ◽  
Genome Editing ◽  
Anthocyanin Biosynthesis ◽  
Phytopathogenic Fungi ◽  
Anthocyanin Pigmentation ◽  
Dna Binding Domains ◽  
Binding Domains ◽  
Myb Genes ◽  
Genetic Constructs

Anthocyanin hyperaccumulation is an important agricultural trait, associated with resistance to abiotic stress, pests, phytopathogenic fungi and bacterial diseases. B. napus with increased anthocyanin pigmentation can be generated by genome editing. Many transcription factors of the MYB family are involved in stress response and anthocyanin biosynthesis. Genes AtMYB60, AtCPC and AtMYBL2 are negative regulators of anthocyanin biosynthesis in Arabidopsis, therefore the knockout of these genes can result in increased anthocyanin pigmentation. gRNA spacers were synthesized to target the orthologs of these genes, identified in Brassica napus. Resulting genetic constructs were introduced to the plant tissues by agroinfiltration. Transient expression of gRNAs targeting DNA-binding domains of MYB transcription factors along with Cas9 nuclease successfully promoted anthocyanin hyperaccumulation. These genetic constructs can be used for genome editing and production of new colored and stress tolerant varieties of oilseed rape.

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