scholarly journals The Basic Helix-Loop-Helix Transcription Factor SmbHLH1 Represses Anthocyanin Biosynthesis in Eggplant

2021 ◽  
Vol 12 ◽  
Author(s):  
Zhaofei Duan ◽  
Shiyu Tian ◽  
Guobin Yang ◽  
Min Wei ◽  
Jing Li ◽  
...  
Keyword(s):  
Plant Species ◽  
Anthocyanin Biosynthesis ◽  
Solanum Melongena ◽  
Amino Acid Sequences ◽  
Negative Regulator ◽  
Bimolecular Fluorescence Complementation ◽  
Luciferase Assay ◽  
Anthocyanin Accumulation ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix

Many basic helix-loop-helix transcription factors (TFs) have been reported to promote anthocyanin biosynthesis in numerous plant species, but little is known about bHLH TFs that inhibit anthocyanin accumulation. In this study, SmbHLH1 from Solanum melongena was identified as a negative regulator of anthocyanin biosynthesis. However, SmbHLH1 showed high identity with SmTT8, which acts as a SmMYB113-dependent positive regulator of anthocyanin-biosynthesis in plants. Overexpression of SmbHLH1 in eggplant caused a dramatic decrease in anthocyanin accumulation. Only the amino acid sequences at the N and C termini of SmbHLH1 differed from the SmTT8 sequence. Expression analysis revealed that the expression pattern of SmbHLH1 was opposite to that of anthocyanin accumulation. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays showed that SmbHLH1 could not interact with SmMYB113. Dual-luciferase assay demonstrated that SmbHLH1 directly repressed the expression of SmDFR and SmANS. Our results demonstrate that the biological function of bHLHs in anthocyanin biosynthesis may have evolved and provide new insight into the molecular functions of orthologous genes from different plant species.

scholarly journals Identification and Characterization of MYB-bHLH-WD40 Regulatory Complex Members Controlling Anthocyanidin Biosynthesis in Blueberry Fruits Development

Genes ◽  
2019 ◽  
Vol 10 (7) ◽  
pp. 496 ◽  
Author(s):  
Zhao ◽  
Li ◽  
Zhu ◽  
Chang ◽  
Li ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Anthocyanin Biosynthesis ◽  
Expression Patterns ◽  
Differentially Expressed Gene ◽  
Amino Acid Sequences ◽  
Bimolecular Fluorescence Complementation ◽  
Myb Transcription Factor ◽  
Helix Loop Helix ◽  
Reverse Transcription Pcr ◽  
Regulatory Complex

Anthocyanins is the main representative of flavonoids in blueberry fruits. The anthocyanins biosynthetic pathway has been extensively studied in numerous model plants and fruit crops at biochemical, genetic, and molecular levels. However, the mechanisms by which the MYB transcription factor/basic helix-loop-helix (bHLH) domain protein/WD-repeat (MYB-bHLH-WD40) complexes regulate anthocyanin biosynthesis in blueberry is still limited. In the present study, we identified 11 MYB, 7 bHLH, and 6 WD40 genes in blueberry fruits, using amino acid sequences of homologous MYB-bHLH-WD40 complexes in Arabidopsis, apple, grape, and strawberry. To understand these mechanisms, the expression patterns of MYB-bHLH-WD40 genes were examined and validated using differentially expressed gene (DEG) analysis and quantitative real-time reverse transcription PCR (qRT-PCR), respectively. The expression patterns of MYB-bHLH-WD40 genes positively correlated with anthocyanin accumulation and color development in blueberry fruits. Consistent with the effects of other transcriptional regulators, the VcMYBL1::GFP, VcbHLH1::GFP, and VcWDL2::GFP fusion proteins were only observed in the nucleus. The protein-protein interactions (PPIs) and bimolecular fluorescence complementation (BiFC) assay suggested a possible link between VcbHLHL1 and VcMYBL1. Finally, a model was proposed and discussed for how the expression of the MYB-bHLH-WD40 complexes can promote anthocyanin biosynthesis in blueberry fruits. To our knowledge, this study was the first to evaluate MYB-bHLH-WD40 complexes in blueberry fruits, and it provides a foundation to dissect the function of the mechanism.

scholarly journals Genome-wide analysis of basic helix–loop–helix superfamily members related to anthocyanin biosynthesis in eggplant (Solanum melongena L.)

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7768 ◽  
Author(s):  
Shiyu Tian ◽  
Lujun Li ◽  
Min Wei ◽  
Fengjuan Yang
Keyword(s):  
Woody Plants ◽  
Tissue Specificity ◽  
Anthocyanin Biosynthesis ◽  
Solanum Melongena ◽  
Defense Responses ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix ◽  
Genome Wide ◽  
Solid Foundation ◽  
Functional Analyses

The basic helix–loop–helix (bHLH) superfamily is considered the second largest transcription factor (TF) family. It plays regulatory roles in the developmental processes of plants and in their defense responses. In recent years, many bHLH superfamily genes have been identified and characterized in herbaceous and woody plants. However, the comprehensive genomic and functional analyses of these genes in eggplant (Solanum melongena L.) have not been reported. In this study, 121 bHLH TFs were identified in the recently released eggplant genome. The phylogeny, gene structure and conserved motifs of the SmbHLH gene were comprehensively studied. Subsequently, the phylogenetic relationship between the bHLH of eggplant and the bHLH of other species was analyzed, and the proteins were classified into 17 subfamilies. Among these protein sequences, 16 subgroups were clustered into the functional clades of Arabidopsis. Two candidate genes (SmbHLH1, SmbHLH117) that may be involved in anthocyanin biosynthesis were screened. The tissue specificity or differential expression of the bHLH genes in different tissues and under various light and temperature conditions suggested the differential regulation of tissue development and metabolism. This study not only provides a solid foundation for the functional dissection of the eggplant bHLH gene family but may also be useful for the future synthesis of anthocyanins in eggplant.

scholarly journals SlBBX20 interacts with the COP9 signalosome subunit SlCSN5-2 to regulate anthocyanin biosynthesis by activating SlDFR expression in tomato

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Dan Luo ◽  
Cheng Xiong ◽  
Aihua Lin ◽  
Chunli Zhang ◽  
Wenhui Sun ◽  
...  
Keyword(s):  
Growth Regulation ◽  
Anthocyanin Biosynthesis ◽  
Plant Stress ◽  
Underlying Mechanism ◽  
Bimolecular Fluorescence Complementation ◽  
Cop9 Signalosome ◽  
Anthocyanin Accumulation ◽  
Related Transcription Factor ◽  
Hybrid Screening

AbstractAnthocyanins play vital roles in plant stress tolerance and growth regulation. Previously, we reported that the photomorphogenesis-related transcription factor SlBBX20 regulates anthocyanin accumulation in tomato. However, the underlying mechanism remains unclear. Here, we showed that SlBBX20 promotes anthocyanin biosynthesis by binding the promoter of the anthocyanin biosynthesis gene SlDFR, suggesting that SlBBX20 directly activates anthocyanin biosynthesis genes. Furthermore, we found by yeast two-hybrid screening that SlBBX20 interacts with the COP9 signalosome subunit SlCSN5-2, and the interaction was confirmed by bimolecular fluorescence complementation and coimmunoprecipitation assays. SlCSN5 gene silencing led to anthocyanin hyperaccumulation in the transgenic tomato calli and shoots, and SlCSN5-2 overexpression decreased anthocyanin accumulation, suggesting thSlCSN5-2 enhanced the ubiquitination of SlBBX20 and promoted the degradation of SlBBX20 in vivo. Consistently, silencing the SlCSN5-2 homolog in tobacco significantly increased the accumulation of the SlBBX20 protein. Since SlBBX20 is a vital regulator of photomorphogenesis, the SlBBX20-SlCSN5-2 module may represent a novel regulatory pathway in light-induced anthocyanin biosynthesis.

scholarly journals The Isolation and Identification of Anthocyanin-Related GSTs in Chrysanthemum

Horticulturae ◽  
2021 ◽  
Vol 7 (8) ◽  
pp. 231
Author(s):  
Yajing Li ◽  
Xiaofen Liu ◽  
Fang Li ◽  
Lili Xiang ◽  
Kunsong Chen
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Vital Role ◽  
Luciferase Assay ◽  
Amino Acid Residues ◽  
Anthocyanin Accumulation ◽  
Isolation And Identification ◽  
Specific Amino Acid ◽  
Protein Sequence Alignment ◽  
Glutathione S Transferases ◽  
Transient Overexpression

Anthocyanin is the crucial pigment for the coloration of red chrysanthemum flowers, which synthesizes in the cytosol and is transported to the vacuole for stable storage. In general, glutathione S-transferases (GSTs) play a vital role in this transport. To date, there is no functional GST reported in chrysanthemums. Here, a total of 94 CmGSTs were isolated from the chrysanthemum genome, with phylogenetic analysis suggesting that 16 members of them were clustered into the Phi subgroup which was related to anthocyanin transport. Among them, the expression of CmGST1 was positively correlated with anthocyanin accumulation. Protein sequence alignment revealed that CmGST1 included anthocyanin-related GST-specific amino acid residues. Further transient overexpression experiments in tobacco leaves showed that CmGST1 could promote anthocyanin accumulation. In addition, a dual-luciferase assay demonstrated that CmGST1 could be regulated by CmMYB6, CmbHLH2 and CmMYB#7, which was reported to be related to anthocyanin biosynthesis. Taken together, we suggested that CmGST1 played a key role in anthocyanin transport and accumulation in chrysanthemums.

HighAN1variability and interaction with basic helix-loop-helix co-factors related to anthocyanin biosynthesis in potato leaves

2014 ◽  
Vol 80 (3) ◽  
pp. 527-540 ◽  
Author(s):  
Vincenzo D'Amelia ◽  
Riccardo Aversano ◽  
Giorgia Batelli ◽  
Immacolata Caruso ◽  
Mar Castellano Moreno ◽  
...  
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix

scholarly journals FoSTUA, Encoding a Basic Helix-Loop-Helix Protein, Differentially Regulates Development of Three Kinds of Asexual Spores, Macroconidia, Microconidia, and Chlamydospores, in the Fungal Plant Pathogen Fusarium oxysporum

2004 ◽  
Vol 3 (6) ◽  
pp. 1412-1422 ◽  
Author(s):  
Toshiaki Ohara ◽  
Takashi Tsuge
Keyword(s):  
Fusarium Oxysporum ◽  
Fluorescent Protein ◽  
Negative Regulator ◽  
Vascular Wilt ◽  
Wild Type ◽  
Low Frequencies ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix ◽  
Chlamydospore Formation ◽  
Green Fluorescent

ABSTRACT The soil-borne fungus Fusarium oxysporum causes vascular wilt of a wide variety of plant species. F. oxysporum produces three kinds of asexual spores, macroconidia, microconidia, and chlamydospores. Falcate macroconidia are formed generally from terminal phialides on conidiophores and rarely from intercalary phialides on hyphae. Ellipsoidal microconidia are formed from intercalary phialides on hyphae. Globose chlamydospores with thick walls are developed by the modification of hyphal and conidial cells. Here we describe FoSTUA of F. oxysporum, which differentially regulates the development of macroconidia, microconidia, and chlamydospores. FoSTUA encodes a basic helix-loop-helix protein with similarity to Aspergillus nidulans StuA, which has been identified as a transcriptional regulator controlling conidiation. Nuclear localization of FoStuA was verified by using strains expressing FoStuA-green fluorescent protein fusions. The FoSTUA-targeted mutants exhibited normal microconidium formation in cultures. However, the mutants lacked conidiophores and produced macroconidia at low frequencies only from intercalary phialides. Thus, FoSTUA appears to be necessary to induce conidiophore differentiation. In contrast, chlamydospore formation was dramatically promoted in the mutants. These data demonstrate that FoStuA is a positive regulator and a negative regulator for the development of macroconidia and chlamydospores, respectively, and is dispensable for microconidium formation in cultures. The disease-causing ability of F. oxysporum was not affected by mutations in FoSTUA. However, the mutants produced markedly fewer macroconidia and microconidia in infected plants than the wild type. These results suggest that FoSTUA also has an important role for microconidium formation specifically in infected plants.

scholarly journals The RNA Directed DNA Methylation (RdDM) Pathway Regulates Anthocyanin Biosynthesis in Crabapple (Malus cv. spp.) Leaves by Methylating the McCOP1 Promoter

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2466
Author(s):  
Yifan Xing ◽  
Ziyi Xie ◽  
Weilei Sun ◽  
Yuying Sun ◽  
Zhenyun Han ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Methylation Level ◽  
Bisulfite Sequencing ◽  
Anthocyanin Biosynthesis ◽  
Apple Fruit ◽  
Bimolecular Fluorescence Complementation ◽  
Rna Expression ◽  
Anthocyanin Accumulation ◽  
Transcript Levels ◽  
Direct Target

The synthesis of anthocyanin pigments in plants is known to be regulated by multiple mechanisms, including epigenetic regulation; however, the contribution of the RNA-directed DNA methylation (RdDM) pathway is not well understood. Here, we used bisulfite sequencing and Real Time (RT)-quantitative (q) PCR to analyze the methylation level of the promoter of constitutively photomorphogenic 1 (McCOP1) from Malus cv. spp, a gene involved in regulating anthocyanin biosynthesis. The CHH methylation level of the McCOP1 promoter was negatively correlated with McCOP1 RNA expression, and inhibiting DNA methylation caused decreased methylation of the McCOP1 promoter and asymmetric cytosine CHH methylation. We observed that the McCOP1 promoter was a direct target of the RdDM pathway argonaute RISC component 4 (McAGO4) protein, which bound to a McCOP1 promoter GGTTCGG site. Bimolecular fluorescence complementation (BIFC) analysis showed that RNA-directed DNA methylation (McRDM1) interacted with McAGO4 and another RdDM protein, domains rearranged methyltransferase 2 (McDRM2), to regulate the CHH methylation of the McCOP1 promoter. Detection of CHH methylation and COP1 gene expression in the Arabidopsis thalianaatago4, atdrm2 and atrdm1 mutants showed that RDM1 is the effector of the RdDM pathway. This was confirmed by silencing McRDM1 in crabapple leaves or apple fruit, which resulted in a decrease in McCOP1 CHH methylation and an increase in McCOP1 transcript levels, as well as in anthocyanin accumulation. In conclusion, these results show that the RdDM pathway is involved in regulating anthocyanin accumulation through CHH methylation of the McCOP1 promoter.

scholarly journals Induction of Anthocyanin Accumulation in Crabapple (Malus cv.) Leaves by Low Temperatures

HortScience ◽  
2015 ◽  
Vol 50 (5) ◽  
pp. 640-649 ◽  
Author(s):  
Ji Tian ◽  
Zhen-yun Han ◽  
Li-ru Zhang ◽  
Ting-Ting Song ◽  
Jie Zhang ◽  
...  
Keyword(s):  
Low Temperatures ◽  
Anthocyanin Biosynthesis ◽  
Anthocyanin Accumulation ◽  
Human Diet ◽  
Helix Loop Helix ◽  
Leaf Coloration ◽  
Regulatory Complex ◽  
R2r3 Myb ◽  
Reverse Transcript ◽  
Anthocyanin Biosynthetic Genes

Anthocyanins are protective pigments that accumulate in plant organs such as fruits and leaves, and are nutritionally valuable components of the human diet. There is thus considerable interest in the factors that regulate synthesis. Malus crabapple leaves are rich sources of these compounds, and in this study we analyzed leaf coloration, anthocyanin levels, and the expression levels of anthocyanin biosynthetic and regulatory genes in three crabapple cultivars (Royalty, Prairifire, and Flame) following various temperature treatments. We found that low temperatures (LTs) promoted anthocyanin accumulation in ‘Royalty’ and ‘Prairifire’, leading to red leaves, but not in ‘Flame’, which accumulated abundant colorless flavonols and retained green colored leaves. Quantitative reverse transcript PCR (RT-PCR) analyses indicated that the expression of several anthocyanin biosynthetic genes was induced by LTs, as were members of the R2R3-MYB, basic helix–loop–helix (bHLH) and WD40 transcription factor families that are thought to act in a complex. We propose that anthocyanin biosynthesis is differentially regulated in the three cultivars by LTs via the expression of members of this anthocyanin regulatory complex.

scholarly journals Coordinated Regulation of Biosynthetic and Regulatory Genes Coincides with Anthocyanin Accumulation in Developing Eggplant Fruit

2015 ◽  
Vol 140 (2) ◽  
pp. 129-135 ◽  
Author(s):  
John R. Stommel ◽  
Judith M. Dumm
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Molecular Genetic ◽  
Solanum Melongena ◽  
Regulatory Genes ◽  
Model Systems ◽  
Anthocyanin Content ◽  
Anthocyanin Accumulation ◽  
Transcript Levels ◽  
Dark Violet ◽  
Marketable Fruit

Violet to black pigmentation of eggplant (Solanum melongena L.) fruit is caused by anthocyanin accumulation. Model systems demonstrate the role of regulatory genes in the control of anthocyanin biosynthesis. Anthocyanin structural gene transcription requires the expression of at least one member of each of three transcription factor families: MYB, MYC, and WD. To determine the molecular genetic basis for anthocyanin pigmentation in eggplant fruit, we used real-time polymerase chain reaction (PCR) to evaluate the expression of anthocyanin biosynthetic (Chs, Dfr, Ans) and regulatory (Myc, MybB, MybC, Wd) genes in S. melongena genotypes that produce fruit with dark violet (‘Classic’) or white (‘Ghostbuster’) coloration, respectively. Transcript levels and anthocyanin content were evaluated in fruit at various stages of development ranging from small post-anthesis fruit to full-sized marketable fruit. Anthocyanin content increased 9-fold in developing violet-colored ‘Classic’ fruit, whereas low but detectable concentrations were found in white ‘Ghostbuster’ fruit. Chs, Dfr, and Ans as well as MybC and Myc transcript levels were significantly higher in ‘Classic’ in comparison with ‘Ghostbuster’ fruit at comparable stages of fruit development with greatest differences observed for Ans transcript levels. MybC and Myc transcript levels increased in developing ‘Classic’ fruit coincident with increasing anthocyanin content. MybB and Wd transcript levels were not coordinated with changes in biosynthetic transcript levels or anthocyanin concentration.

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