scholarly journals Transcriptome-Wide Identification of WRKY Transcription Factor and Functional Characterization of RgWRKY37 Involved in Acteoside Biosynthesis in Rehmannia glutinosa

2021 ◽  
Vol 12 ◽  
Author(s):  
Fengqing Wang ◽  
Xinrong Li ◽  
Xin Zuo ◽  
Mingming Li ◽  
Chunyan Miao ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Zinc Finger ◽  
Hairy Roots ◽  
Sequence Data ◽  
Functional Characterization ◽  
Transcript Abundance ◽  
Wrky Transcription Factor ◽  
Regulation Mechanism ◽  
Rehmannia Glutinosa ◽  
Transcriptome Sequence Data

WRKYs play important roles in plant metabolism, but their regulation mechanism in Rehmannia glutinosa remains elusive. In this study, 37 putative WRKY transcription factors (TFs) with complete WRKY domain from R. glutinosa transcriptome sequence data were identified. Based on their conserved domains and zinc finger motif, the R. glutinosa WRKY TFs were divided into five groups. Structural feature analysis shows that the 37 RgWRKY proteins contain WRKYGQK/GKK domains and a C2H2/C2HC-type zinc finger structure. To identify the function of RgWRKY members involved in acteoside biosynthesis, transcriptional profiles of 37 RgWRKYs in hairy roots under salicylic acid (SA), methyl jasmonate (MeJA), and hydrogen peroxide (H2O2) treatments were systematically established using RNA-seq analysis. Based on the correlationship between the expression levels of RgWRKY genes and acteoside content, RgWRKY7, RgWRKY23, RgWRKY34, RgWRKY35, and RgWRKY37 were suggested to be involved in acteoside biosynthesis in R. glutinosa, and RgWRKY37 was selected for gene functional research. Overexpression of RgWRKY37 increased the content of acteoside and total phenylethanoid glycosides (PhGs) in hairy roots and enhanced the transcript abundance of seven enzyme genes involved in the acteoside biosynthesis pathway. These results strongly suggest the involvement of the WRKY transcription factor in the regulation of acteoside biosynthesis.

scholarly journals Expression Analyses of Soybean VOZ Transcription Factors and the Role of GmVOZ1G in Drought and Salt Stress Tolerance

2020 ◽  
Vol 21 (6) ◽  
pp. 2177 ◽  
Author(s):  
Bo Li ◽  
Jia-Cheng Zheng ◽  
Ting-Ting Wang ◽  
Dong-Hong Min ◽  
Wen-Liang Wei ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Transcription Factors ◽  
Salt Stress ◽  
Abiotic Stress ◽  
Stress Tolerance ◽  
Zinc Finger ◽  
Hairy Roots ◽  
Yeast Cells ◽  
Salt Stress Tolerance ◽  
Drought And Salt Stress

Vascular plant one-zinc-finger (VOZ) transcription factor, a plant specific one-zinc-finger-type transcriptional activator, is involved in regulating numerous biological processes such as floral induction and development, defense against pathogens, and response to multiple types of abiotic stress. Six VOZ transcription factor-encoding genes (GmVOZs) have been reported to exist in the soybean (Glycine max) genome. In spite of this, little information is currently available regarding GmVOZs. In this study, GmVOZs were cloned and characterized. GmVOZ genes encode proteins possessing transcriptional activation activity in yeast cells. GmVOZ1E, GmVOZ2B, and GmVOZ2D gene products were widely dispersed in the cytosol, while GmVOZ1G was primarily located in the nucleus. GmVOZs displayed a differential expression profile under dehydration, salt, and salicylic acid (SA) stress conditions. Among them, GmVOZ1G showed a significantly induced expression in response to all stress treatments. Overexpression of GmVOZ1G in soybean hairy roots resulted in a greater tolerance to drought and salt stress. In contrast, RNA interference (RNAi) soybean hairy roots suppressing GmVOZ1G were more sensitive to both of these stresses. Under drought treatment, soybean composite plants with an overexpression of hairy roots had higher relative water content (RWC). In response to drought and salt stress, lower malondialdehyde (MDA) accumulation and higher peroxidase (POD) and superoxide dismutase (SOD) activities were observed in soybean composite seedlings with an overexpression of hairy roots. The opposite results for each physiological parameter were obtained in RNAi lines. In conclusion, GmVOZ1G positively regulates drought and salt stress tolerance in soybean hairy roots. Our results will be valuable for the functional characterization of soybean VOZ transcription factors under abiotic stress.

scholarly journals Transcription factor SmWRKY1 positively promote the biosynthesis of tanshinones inSalvia miltiorrhiza

2017 ◽  
Author(s):  
Wenzhi Cao ◽  
Yao Wang ◽  
Min Shi ◽  
Xiaolong Hao ◽  
Weiwei Zhao ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Dry Weight ◽  
Wrky Transcription Factor ◽  
Mep Pathway ◽  
Regulation Mechanism ◽  
Phylogenetic Tree Analysis ◽  
Multiple Sequence ◽  
Over Expression ◽  
Localization Analysis ◽  
Transgenic Lines

AbstractTanshinones, one group of bioactive diterpenes, were widely used in the treatment of cardiovascular diseases. WRKYs play important roles in plant metabolism, but their regulation mechanism in S. miltiorrhiza remains elusive. In this study, one WRKY transcription factor SmWRKY1 was isolated and characterized from S. miltiorrhiza. Multiple sequence alignment and phylogenetic tree analysis showed SmWRKY1 shared high homology with other plant WRKYs such as CrWRKY1. SmWRKY1 were predominantly expressed in leaves and stems, and was responsive to salicylic acid (SA), methyl jasmonate (MeJA) and nitric oxide (NO) treatment. Subcellular localization analysis found that SmWRKY1 was localized in the nucleus. Over-expression of SmWRKY1 significantly elevated the transcripts of genes involved in MEP pathway especially 1-deoxy-D-xylulose 5-phosphate synthase (SmDXS) and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (SmDXR), resulted in over 6 folds increase in tanshinones production in transgenic lines (up to 13.731mg/g dry weight (DW)) compared with the control lines. Dual-luciferase (Dual-LUC) assay showed that SmWRKY1 can positively regulate SmDXR expression by binding to its promoter. Our work revealed that SmWRKY1 participated in the regulation of tanshinones biosynthesis and acted as a positive regulator through activating SmDXR in the MEP pathway, thus discloses a new insight to further excavate the regulation mechanism of tanshinones biosynthesis.

scholarly journals Isolation, Expression, and Promoter Analysis ofGbWRKY2: A Novel Transcription Factor Gene fromGinkgo biloba

2015 ◽  
Vol 2015 ◽  
pp. 1-17 ◽  
Author(s):  
Yong-Ling Liao ◽  
Yong-Bao Shen ◽  
Jie Chang ◽  
Wei-Wei Zhang ◽  
Shui-Yuan Cheng ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Zinc Finger ◽  
Stress Responses ◽  
Genomic Dna ◽  
Single Copy ◽  
Wrky Transcription Factor ◽  
Full Length ◽  
Flanking Sequence ◽  
Reading Frame ◽  
Full Length Cdna

WRKY transcription factor is involved in multiple life activities including plant growth and development as well as biotic and abiotic responses. We identified 28WRKYgenes from transcriptome data ofGinkgo bilobaaccording to conserved WRKY domains and zinc finger structure and selected three WRKY genes, which areGbWRKY2, GbWRKY16, andGbWRKY21, for expression pattern analysis.GbWRKY2was preferentially expressed in flowers and strongly induced by methyl jasmonate. Here, we cloned the full-length cDNA and genomic DNA ofGbWRKY2. The full-length cDNA ofGbWRKY2was 1,713 bp containing a 1,014 bp open reading frame encoding a polypeptide of 337 amino acids. TheGbWRKY2genomic DNA had one intron and two exons. The deducedGbWRKY2contained one WRKY domain and one zinc finger motif.GbWRKY2was classified into Group II WRKYs. Southern blot analysis revealed thatGbWRKY2was a single copy gene inG. biloba. Manycis-acting elements related to hormone and stress responses were identified in the 1,363 bp-length 5′-flanking sequence ofGbWRKY2, including W-box, ABRE-motif, MYBCOREs, and PYRIMIDINE-boxes, revealing the molecular mechanism of upregulated expression ofGbWRKY2by hormone and stress treatments. Further functional characterizations in transiently transformed tobacco leaves allowed us to identify the region that can be considered as the minimal promoter.

scholarly journals Transcription Factor OpWRKY3 Is Involved in the Development and Biosynthesis of Camptothecin and Its Precursors in Ophiorrhiza pumila Hairy Roots

2019 ◽  
Vol 20 (16) ◽  
pp. 3996 ◽  
Author(s):  
Can Wang ◽  
Chao Wu ◽  
Yao Wang ◽  
Chenhong Xie ◽  
Min Shi ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Hairy Roots ◽  
Indole Alkaloid ◽  
Wrky Transcription Factor ◽  
Significant Feature ◽  
Total Production ◽  
Phylogenetic Tree Analysis ◽  
Reading Frame ◽  
Group Iii ◽  
Localization Analysis

The plant Ophiorrhiza pumila produces camptothecin (CPT), a kind of terpene indole alkaloid (TIAs) that has been widely used in treatment of cancer. Tryptophan-arginine-lysine-tyrosine (WRKY) transcription factors have been reported to play important roles in plant metabolism and development. In this study, a novel WRKY transcription factor named OpWRKY3 was isolated from O. pumila, with full-length open reading frame (ORF) of 1128 bp, encoding 375 amino acids. Phylogenetic tree analysis revealed that OpWRKY3 shared the highest homology with VvWRKY30, and it is a significant feature belonging to group III. OpWRKY3 was responsive to various treatments, including gibberellin (GA3), methyl jasmonate (MJ), acetylsalicylic acid (ASA), salicylic acid (SA), and abscisic acid (ABA). Besides, OpWRKY3 is expressed predominantly in stems. Subcellular localization analysis showed that OpWRKY3 localized in the nucleus. The biomass of OpWRKY3-SRDX transgenic hairy roots (S line) was visibly suppressed, while there were slight changes between overexpression of the OpWRKY3 line (OE line) and the control. In addition, the concentration and total production of camptothecin precursors including loganin and secologanin were significantly changed in both OE and S lines while total production of CPT was significantly changed in most transgenic lines. Thus, the present work revealed that OpWRKY3 may act as a regulator in the growth and development of O. pumila, and in production of camptothecin and its precursors.

scholarly journals Isolation and Functional Characterization of the Promoters of Miltiradiene Synthase Genes, TwTPS27a and TwTPS27b, and Interaction Analysis with the Transcription Factor TwTGA1 from Tripterygium wilfordii

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 418
Author(s):  
Yanbo Huo ◽  
Bin Zhang ◽  
Ling Chen ◽  
Jing Zhang ◽  
Xing Zhang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Target Genes ◽  
Interaction Analysis ◽  
Functional Characterization ◽  
Gus Activity ◽  
Core Region ◽  
Regulation Mechanism ◽  
Tripterygium Wilfordii ◽  
Potential Core ◽  
Responsive Element

Miltiradiene synthase (MS) genes, TwTPS27a and TwTPS27b, are the key diterpene synthase genes in the biosynthesis of triptolide, which is an important medicinally active diterpenoid in Tripterygium wilfordii. However, the mechanism underlying the regulation of key genes TwTPS27a/b in triptolide biosynthesis remains unclear. In this study, the promoters of TwTPS27a (1496 bp) and TwTPS27b (1862 bp) were isolated and analyzed. Some hormone-/stress-responsive elements and transcription factor (TF) binding sites were predicted in both promoters, which might be responsible for the regulation mechanism of TwTPS27a/b. The β-glucuronidase (GUS) activity analysis in promoter deletion assays under normal and methyl jasmonate (MeJA) conditions showed that the sequence of −921 to −391 bp is the potential core region of the TwTPS27b promoter. And the TGACG-motif, a MeJA-responsive element found in this core region, might be responsible for MeJA-mediated stress induction of GUS activity. Moreover, the TGACG-motif is also known as the TGA TF-binding site. Yeast one-hybrid and GUS transactivation assays confirmed the interaction between the TwTPS27a/b promoters and the TwTGA1 TF (a MeJA-inducible TGA TF upregulating triptolide biosynthesis in T. wilfordii), indicating that TwTPS27a/b are two target genes regulated by TwTGA1. In conclusion, our results provide important information for elucidating the regulatory mechanism of MS genes, TwTPS27a and TwTPS27b, as two target genes of TwTGA1, in jasmonic acid (JA)-inducible triptolide biosynthesis.

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