scholarly journals Increased Leaf Nicotine Content by Targeting Transcription Factor Gene Expression in Commercial Flue-Cured Tobacco (Nicotiana tabacum L.)

Genes ◽  
2019 ◽  
Vol 10 (11) ◽  
pp. 930 ◽  
Author(s):  
Hai Liu ◽  
Tatyana I. Kotova ◽  
Michael P. Timko
Keyword(s):  
Transcription Factor ◽  
Nicotiana Tabacum ◽  
Gene Promoter ◽  
Bhlh Transcription Factor ◽  
Helix Loop Helix ◽  
Nicotiana Tabacum L ◽  
Meja Treatment ◽  
Transgenic Lines ◽  
Nicotine Biosynthesis ◽  
High Level

Nicotine, the most abundant pyridine alkaloid in cultivated tobacco (Nicotiana tabacum L.), is a potent inhibitor of insect and animal herbivory and a neurostimulator of human brain function. Nicotine biosynthesis is controlled developmentally and can be induced by abiotic and biotic stressors via a jasmonic acid (JA)-mediated signal transduction mechanism involving members of the APETALA 2/ethylene-responsive factor (AP2/ERF) and basic helix-loop-helix (bHLH) transcription factor (TF) families. AP2/ERF and bHLH TFs work combinatorically to control nicotine biosynthesis and its subsequent accumulation in tobacco leaves. Here, we demonstrate that overexpression of the tobacco NtERF32, NtERF221/ORC1, and NtMYC2a TFs leads to significant increases in nicotine accumulation in T2 transgenic K326 tobacco plants before topping. Up to 9-fold higher nicotine production was achieved in transgenics overexpressing NtERF221/ORC1 under the control of a constitutive GmUBI3 gene promoter compared to wild-type plants. The constitutive 2XCaMV35S promoter and a novel JA-inducible 4XGAG promoter were less effective in driving high-level nicotine formation. Methyljasmonic acid (MeJA) treatment further elevated nicotine production in all transgenic lines. Our results show that targeted manipulation of NtERF221/ORC1 is an effective strategy for elevating leaf nicotine levels in commercial tobacco for use in the preparation of reduced risk tobacco products for smoking replacement therapeutics.

scholarly journals The basic helix-loop-helix transcription factor TabHLH1 increases chlorogenic acid and luteolin biosynthesis in Taraxacum antungense Kitag

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Qun Liu ◽  
Li Li ◽  
Haitao Cheng ◽  
Lixiang Yao ◽  
Jie Wu ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Chlorogenic Acid ◽  
Gene Promoter ◽  
Bhlh Transcription Factor ◽  
Active Components ◽  
Mobility Shift ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix ◽  
Coa Ligase ◽  
Transgenic Lines

AbstractPolyphenols are the main active components of the anti-inflammatory compounds in dandelion, and chlorogenic acid (CGA) is one of the primary polyphenols. However, the molecular mechanism underlying the transcriptional regulation of CGA biosynthesis remains unclear. Hydroxycinnamoyl-CoA:quinate hydroxycinnamoyl transferase (HQT2) is the last rate-limiting enzyme in chlorogenic acid biosynthesis in Taraxacum antungense. Therefore, using the TaHQT2 gene promoter as a probe, a yeast one-hybrid library was performed, and a basic helix-loop-helix (bHLH) transcription factor, TabHLH1, was identified that shared substantial homology with Gynura bicolor DC bHLH1. The TabHLH1 transcript was highly induced by salt stress, and the TabHLH1 protein was localized in the nucleus. CGA and luteolin concentrations in TabHLH1-overexpression transgenic lines were significantly higher than those in the wild type, while CGA and luteolin concentrations in TabHLH1-RNA interference (RNAi) transgenic lines were significantly lower. Quantitative real-time polymerase chain reaction demonstrated that overexpression and RNAi of TabHLH1 in T. antungense significantly affected CGA and luteolin concentrations by upregulating or downregulating CGA and luteolin biosynthesis pathway genes, especially TaHQT2, 4-coumarate-CoA ligase (Ta4CL), chalcone isomerase (TaCHI), and flavonoid-3′-hydroxylase (TaF3′H). Dual-luciferase, yeast one-hybrid, and electrophoretic mobility shift assays indicated that TabHLH1 directly bound to the bHLH-binding motifs of proTaHQT2 and proTa4CL. This study suggests that TabHLH1 participates in the regulatory network of CGA and luteolin biosynthesis in T. antungense and might be useful for metabolic engineering to promote plant polyphenol biosynthesis.

Hxt encodes a basic helix-loop-helix transcription factor that regulates trophoblast cell development

Development ◽  
1995 ◽  
Vol 121 (8) ◽  
pp. 2513-2523 ◽  
Author(s):  
J.C. Cross ◽  
M.L. Flannery ◽  
M.A. Blanar ◽  
E. Steingrimsson ◽  
N.A. Jenkins ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Gene Promoter ◽  
Giant Cells ◽  
Placental Lactogen ◽  
Specific Gene ◽  
Bhlh Transcription Factor ◽  
Trophoblast Cells ◽  
Positive Role ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix

Trophoblast cells are the first lineage to form in the mammalian conceptus and mediate the process of implantation. We report the cloning of a basic helix-loop-helix (bHLH) transcription factor gene, Hxt, that is expressed in early trophoblast and in differentiated giant cells. A separate gene, Hed, encodes a related protein that is expressed in maternal deciduum surrounding the implantation site. Overexpression of Hxt in mouse blastomeres directed their development into trophoblast cells in blastocysts. In addition, overexpression of Hxt induced the differentiation of rat trophoblast (Rcho-1) stem cells as assayed by changes in cell adhesion and by activation of the placental lactogen-I gene promoter, a trophoblast giant cell-specific gene. In contrast, the negative HLH regulator, Id-1, inhibited Rcho-1 differentiation and placental lactogen-I transcription. These data demonstrate a role for HLH factors in regulating trophoblast development and indicate a positive role for Hxt in promoting the formation of trophoblast giant cells.

scholarly journals Overexpression of ThMYC4E Enhances Anthocyanin Biosynthesis in Common Wheat

2019 ◽  
Vol 21 (1) ◽  
pp. 137 ◽  
Author(s):  
Shuo Zhao ◽  
Xingyuan Xi ◽  
Yuan Zong ◽  
Shiming Li ◽  
Yun Li ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Common Wheat ◽  
Anthocyanin Biosynthesis ◽  
Pcr Analysis ◽  
Bhlh Transcription Factor ◽  
Anthocyanin Content ◽  
Metabolome Analysis ◽  
Helix Loop Helix ◽  
Transgenic Lines ◽  
Ethanol Extracts

The basic helix-loop helix (bHLH) transcription factor has been inferred to play an important role in blue and purple grain traits in common wheat, but to date, its overexpression has not been reported. In this study, the bHLH transcription factor ThMYC4E, the candidate gene controlling the blue grain trait from Th. Ponticum, was transferred to the common wheat JW1. The positive transgenic lines displayed higher levels of purple anthocyanin pigments in their grains, leaves and glumes. Stripping the glumes (light treatment) caused white grains to become purple in transgenic lines. RNA-Seq and qRT-PCR analysis demonstrated that the transcript levels of structural genes associated with anthocyanin biosynthesis were higher in transgenic wheat than the wild-type (WT), which indicated that ThMYC4E activated anthocyanin biosynthesis in the transgenic lines. Correspondingly, the anthocyanin contents in grains, roots, stems, leaves and glumes of transgenic lines were higher than those in the WT. Metabolome analysis demonstrated that the anthocyanins were composed of cyanidin and delphinidin in the grains of the transgenic lines. Moreover, the transgenic lines showed higher antioxidant activity, in terms of scavenging DPPH radicals, in the ethanol extracts of their grains. The overexpression of ThMYC4E sheds light on the traits related to anthocyanin biosynthesis in common wheat and provide a new way to improve anthocyanin content.

A novel bHLH transcription factor, NtbHLH1, modulates iron homeostasis in tobacco (Nicotiana tabacum L.)

2020 ◽  
Vol 522 (1) ◽  
pp. 233-239
Author(s):  
Yang-Yang Li ◽  
Xiao-Yan Sui ◽  
Jia-Shuo Yang ◽  
Xiao-Hua Xiang ◽  
Zun-Qiang Li ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Nicotiana Tabacum ◽  
Iron Homeostasis ◽  
Bhlh Transcription Factor ◽  
Nicotiana Tabacum L

scholarly journals bHLH Transcription Factor NtMYC2a Regulates Carbohydrate Metabolism during the Pollen Development of Tobacco (Nicotiana tabacum L. cv. TN90)

Plants ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 17
Author(s):  
Shiquan Bian ◽  
Tian Tian ◽  
Yongqiang Ding ◽  
Ning Yan ◽  
Chunkai Wang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Nicotiana Tabacum ◽  
Transgenic Plants ◽  
Carbohydrate Metabolism ◽  
Pollen Development ◽  
Starch Accumulation ◽  
Bhlh Transcription Factor ◽  
Pollen Maturation ◽  
Nicotiana Tabacum L

Basic helix-loop-helix (bHLH) transcription factor MYC2 regulates plant growth and development in many aspects through the jasmonic acid (JA) signaling pathway, while the role of MYC2 in plant carbohydrate metabolism has not been reported. Here, we generated NtMYC2a-overexpressing (NtMYC2a-OE) and RNA-interference-mediated knockdown (NtMYC2a-RI) transgenic plants of tobacco (Nicotiana tabacum L. cv. TN90) to investigate the role of NtMYC2a in carbohydrate metabolism and pollen development. Results showed that NtMYC2a regulates the starch accumulation and the starch-sugar conversion of floral organs, especially in pollen. The RT-qPCR analysis showed that the expression of starch-metabolic-related genes, AGPs, SS2 and BAM1, were regulated by NtMYC2a in the pollen grain, anther wall and ovary of tobacco plants. The process of pollen maturation was accelerated in NtMYC2a-OE plants and was delayed in NtMYC2a-RI plants, but the manipulation of NtMYC2a expression did not abolish the pollen fertility of the transgenic plants. Intriguingly, overexpression of NtMYC2a also enhanced the soluble carbohydrate accumulation in tobacco ovaries. Overall, our results demonstrated that the bHLH transcription factor NtMYC2a plays an important role in regulating the carbohydrate metabolism during pollen maturation in tobacco.

scholarly journals The Nicotiana tabacum Mediator subunit MED25 regulates nicotine biosynthesis through interacting with the basic helix-loop-helix (bHLH) transcription factor NtMYC2s

2021 ◽  
Author(s):  
Ge Bai ◽  
Yong Li ◽  
Da-Hai Yang ◽  
Tao Pang ◽  
Zhi-Yong Fan ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Acid Treatment ◽  
Plant Secondary Metabolites ◽  
Bhlh Transcription Factor ◽  
Wild Type ◽  
Helix Loop Helix ◽  
Transgenic Lines ◽  
In The Wild ◽  
Nicotine Biosynthesis

Nicotine is one of the most important secondary metabolites in tobacco, and its biosynthesis can be induced by topping and jasmonic acid treatment. NtMYC2s play pivotal roles in the regulation of nicotine. The mediator server as a bridge betwen the transcription factors and RNA polymerase in order to facilitates transcription and functions in plants. However, the role of mediator in the regulation of nicotine biosynthesis remains unknown. In this study, we firstly identify the NtMED25 through homologous analysis. NtMED25 interacts with NtMYC2s through the MD region. Interestingly, the nicotine content is decreased in the the knock-down transgenic lines of NtMED25, and the expression levels of two nicotine biosynthesis genes, NtQPT2 and NtPMT2, are also reduced when compared with that in the wild-type plants. Furthermore, NtMED25 enhances the binding of NtMYC2a/ NtMYC2b to the promoter of NtPMT2 and NtQPT2, and then facilitates the nicotine biosynthesis. Therefore, our study revealed the function of mediator in the regulation of nicotine, and provide the insight role on the transcriptional regulation of plant secondary metabolites.

scholarly journals FIT, a regulatory hub for iron deficiency and stress signaling in roots, and FIT-dependent and -independent gene signatures

2020 ◽  
Vol 71 (5) ◽  
pp. 1694-1705 ◽  
Author(s):  
Birte Schwarz ◽  
Petra Bauer
Keyword(s):  
Transcription Factor ◽  
Protein Interactions ◽  
Fe Deficiency ◽  
Bhlh Transcription Factor ◽  
Root Cells ◽  
Gene Signatures ◽  
Helix Loop Helix ◽  
Independent Gene ◽  
Fe Uptake ◽  
Fe Acquisition

Abstract Iron (Fe) is vital for plant growth. Plants balance the beneficial and toxic effects of this micronutrient, and tightly control Fe uptake and allocation. Here, we review the role of the basic helix–loop–helix (bHLH) transcription factor FIT (FER-LIKE FE DEFICIENCY-INDUCED TRANSCRIPTION FACTOR) in Fe acquisition. FIT is not only essential, it is also a central regulatory hub in root cells to steer and adjust the rate of Fe uptake by the root in a changing environment. FIT regulates a subset of root Fe deficiency (–Fe) response genes. Based on a combination of co-expression network and FIT-dependent transcriptome analyses, we defined a set of FIT-dependent and FIT-independent gene expression signatures and co-expression clusters that encode specific functions in Fe regulation and Fe homeostasis. These gene signatures serve as markers to integrate novel regulatory factors and signals into the –Fe response cascade. FIT forms a complex with bHLH subgroup Ib transcription factors. Furthermore, it interacts with key regulators from different signaling pathways that either activate or inhibit FIT function to adjust Fe acquisition to growth and environmental constraints. Co-expression clusters and FIT protein interactions suggest a connection of –Fe with ABA responses and root cell elongation processes that can be explored in future studies.

scholarly journals Tomato bHLH132 Transcription Factor Controls Growth and Defense and Is Activated by Xanthomonas euvesicatoria Effector XopD During Pathogenesis

2019 ◽  
Vol 32 (12) ◽  
pp. 1614-1622 ◽  
Author(s):  
Jung-Gun Kim ◽  
Mary Beth Mudgett
Keyword(s):  
Transcription Factor ◽  
Protease Activity ◽  
Transcriptional Profiling ◽  
Bhlh Transcription Factor ◽  
Transcription Activator ◽  
Tal Effectors ◽  
Sumo Protease ◽  
Tal Effector ◽  
Helix Loop Helix ◽  
Xanthomonas Euvesicatoria

Effector-dependent manipulation of host transcription is a key virulence mechanism used by Xanthomonas species causing bacterial spot disease in tomato and pepper. Transcription activator-like (TAL) effectors employ novel DNA-binding domains to directly activate host transcription, whereas the non-TAL effector XopD uses a small ubiquitin-like modifier (SUMO) protease activity to represses host transcription. The targets of TAL and non-TAL effectors provide insight to the genes governing susceptibility and resistance during Xanthomonas infection. In this study, we investigated the extent to which the X. euvesicatoria non-TAL effector strain Xe85-10 activates tomato transcription to gain new insight to the transcriptional circuits and virulence mechanisms associated with Xanthomonas euvesicatoria pathogenesis. Using transcriptional profiling, we identified a putative basic helix-loop-helix (bHLH) transcription factor, bHLH132, as a pathogen-responsive gene that is moderately induced by microbe-associated molecular patterns and defense hormones and is highly induced by XopD during X. euvesicatoria infection. We also found that activation of bHLH132 transcription requires the XopD SUMO protease activity. Silencing bHLH132 mRNA expression results in stunted tomato plants with enhanced susceptibility to X. euvesicatoria infection. Our work suggests that bHLH132 is required for normal vegetative growth and development as well as resistance to X. euvesicatoria. It also suggests new transcription-based models describing XopD virulence and recognition in tomato.

scholarly journals The Basic-Region Helix-Loop-Helix Transcription Factor DevR Significantly Affects Polysaccharide Metabolism inAspergillus oryzae

2019 ◽  
Vol 85 (8) ◽  
Author(s):  
Miao Zhuang ◽  
Zhi-Min Zhang ◽  
Long Jin ◽  
Bao-Teng Wang ◽  
Yasuji Koyama ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Aspergillus Oryzae ◽  
Bhlh Transcription Factor ◽  
Regulation Mechanism ◽  
Starch Metabolism ◽  
Poor Growth ◽  
Content Type ◽  
Helix Loop Helix ◽  
Amylase Production ◽  
Basic Region

ABSTRACTBasic-region helix-loop-helix (bHLH) proteins are a superfamily of transcription factors that are often involved in the control of growth and differentiation. Recently, it was reported that the bHLH transcription factor DevR is involved in both asexual and sexual development inAspergillus nidulansand regulates the conidial melanin production inAspergillus fumigatus. In this study, we identified and characterized anAspergillus oryzaegene that showed high similarity withdevRofA. nidulansandA. fumigatus(AodevR). In the AodevR-disrupted strain, growth was delayed and the number of conidia was decreased on Czapek-Dox (CD) minimal agar plates, but the conidiation was partially recovered by adding 0.6 M KCl. Simultaneously, the overexpression of AodevRwas induced and resulted in extremely poor growth when the carbon source changed from glucose to polysaccharide (dextrin) in the CD agar plate. Scanning electron microscopy (SEM) indicated that the overexpression of AodevRresulted in extremely thin aberrant hyphal morphology. Conversely, the deletion of AodevRresulted in thicker hyphae and in more resistance to Congo red relative to the control strain. Quantitative reverse transcriptase PCR (RT-PCR) further indicated that AoDevR significantly affects chitin and starch metabolism, and importantly, the overexpression of AodevRinhibited the expression of genes related to starch degradation. A yeast one-hybrid assay suggested that the DevR protein possibly interacted with the promoter ofamyR, which encodes a transcription factor involved in amylase production. Importantly, AoDevR is involved in polysaccharide metabolism and affects the growth of theA. oryzaestrain.IMPORTANCEAspergillus oryzaeis an industrially important filamentous fungus; therefore, a clear understanding of its polysaccharide metabolism and utilization is very important for its industrial utilization. In this study, we revealed that the basic-region helix-loop-helix (bHLH) transcription factor AoDevR is importantly involved in chitin and starch metabolism inA. oryzae. The overexpression of AodevRstrongly suppressed the expression of amylase-related genes. The results of a yeast one-hybrid assay suggested that the DevR protein potentially interacts with the promoter ofamyR, which encodes a transcription factor involved in amylase production and starch utilization. This study provides new insight for further revealing the regulation mechanism of amylase production inA. oryzae.

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