scholarly journals Heterologous Expression of Poplar WRKY18/35 Paralogs in Arabidopsis Reveals Their Antagonistic Regulation on Pathogen Resistance and Abiotic Stress Tolerance via Variable Hormonal Pathways

2020 ◽  
Vol 21 (15) ◽  
pp. 5440 ◽  
Author(s):  
Li Guo ◽  
Chaofeng Li ◽  
Yuanzhong Jiang ◽  
Keming Luo ◽  
Changzheng Xu
Keyword(s):  
Transcription Factors ◽  
Transgenic Plants ◽  
Abiotic Stress Tolerance ◽  
Germination Rate ◽  
Pathogen Resistance ◽  
Pcr Analysis ◽  
Wild Type ◽  
Biotic And Abiotic Stresses ◽  
Qrt Pcr ◽  
Root Analysis

WRKY transcription factors (WRKY TFs) are one of the largest protein families in plants, and most of them play vital roles in response to biotic and abiotic stresses by regulating related signaling pathways. In this study, we isolated two WRKY TF genes PtrWRKY18 and PtrWRKY35 from Populustrichocarpa and overexpressed them in Arabidopsis. Expression pattern analyses showed that PtrWRKY18 and PtrWRKY35 respond to salicylic acid (SA), methyl JA (MeJA), abscisic acid (ABA), B. cinereal, and P. syringae treatment. The transgenic plants conferred higher B. cinerea tolerance than wild-type (WT) plants, and real-time quantitative (qRT)-PCR assays showed that PR3 and PDF1.2 had higher expression levels in transgenic plants, which was consistent with their tolerance to B. cinereal. The transgenic plants showed lower P. syringae tolerance than WT plants, and qRT-PCR analysis (PR1, PR2, and NPR1) also corresponded to this phenotype. Germination rate and root analysis showed that the transgenic plants are less sensitive to ABA, which leads to the reduced tolerance to osmotic stress and the increase of the death ratio and stomatal aperture. Compared with WT plants, a series of ABA-related genes (RD29A, ABO3, ABI4, ABI5, and DREB1A) were significantly down-regulated in PtrWRKY18 and PtrWRKY35 overexpression plants. All of these results demonstrated that the two WRKY TFs are multifunctional transcription factors in plant resistance.

scholarly journals Characterization and Functional Analysis of FaHsfC1b from Festuca arundinacea Conferring Heat Tolerance in Arabidopsis

2018 ◽  
Vol 19 (9) ◽  
pp. 2702 ◽  
Author(s):  
Lili Zhuang ◽  
Wei Cao ◽  
Jian Wang ◽  
Jingjin Yu ◽  
Zhimin Yang ◽  
...  
Keyword(s):  
Heat Stress ◽  
Transgenic Plants ◽  
Heat Tolerance ◽  
Festuca Arundinacea ◽  
Photochemical Efficiency ◽  
Grass Species ◽  
Pcr Analysis ◽  
Plant Tolerance ◽  
Qrt Pcr ◽  
Class C

Heat transcription factors (Hsfs) belong to a large gene family classified into A, B, and C groups, with classes A and B Hsfs being well-characterized and known for their roles in plant tolerance to abiotic stresses. The functions and roles of Class C Hsfs are not well-documented. The objectives of this study were to characterize a class C Hsf gene (FaHsfC1b) cloned from tall fescue (Festuca arundinacea), a perennial grass species, and to determine the physiological functions of FaHsfC1b in regulating heat tolerance by overexpressing FaHsfC1b in Arabidopsis thaliana. Full length cDNA of FaHsfC1b was cloned and the sequence alignment showed that it had high similarity to OsHsfC1b with typical DNA binding domain, hydrophobic oligomerization domain, and a nucleus localization signal. Transient expression with FaHsfC1b-eGFP in protoplasts of Arabidopsis leaves indicated its nucleus localization. qRT-PCR analysis showed that FaHsfC1b responded to heat, osmotic, salt, and cold stress in leaves and roots during 48-h treatment. Physiological analysis showed that FaHsfC1b overexpression enhanced plant survival rate, chlorophyll content, and photochemical efficiency, while it resulted in decreases in electrolyte leakage, H2O2 and O2− content under heat stress. qRT-PCR showed that endogenous HsfC1 was induced in transgenic plants and the expression levels of heat protection protein genes, including several HSPs, AtGalSyn1, AtRof1, and AtHSA32, as well as ABA-synthesizing gene (NCED3) were significantly upregulated in transgenic plants overexpressing FaHsfC1b under heat stress. Our results first demonstrate that HsfC1b plays positive roles in plant tolerance to heat stress in association with the induction and upregulation of heat-protective genes. HsfC1b may be used as a candidate gene for genetic modification of cool-season plant species for improving heat tolerance.

scholarly journals Effects of Certain cis-Regulatory Elements on Stage-Specific vitellogenin Expression in the Bombyx mori (Lepidoptera: Bombycidae)

2020 ◽  
Vol 20 (3) ◽  
Author(s):  
Guanwang Shen ◽  
Hongling Liu ◽  
Ying Lin ◽  
Dongxu Xing ◽  
Yujing Zhang ◽  
...  
Keyword(s):  
Amino Acids ◽  
Transcription Factors ◽  
Bombyx Mori ◽  
Regulatory Network ◽  
Theoretical Basis ◽  
Regulatory Elements ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Stage Specificity ◽  
E Box

Abstract Bombyx mori vitellogenin (BmVg) is highly upregulated during pupation, and the 20-hydroxyecdysone and amino acids may regulate stage-specific BmVg expression. However, previous studies showed that other factors may also affect stage-specific BmVg expression. Here, we characterized effective BmVg transcription factors by identifying the corresponding cis-regulatory elements (CREs). We prepared transgenic B. mori, in which DsRed was driven by various lengths of BmVg promoter. qRT-PCR analysis showed that DsRed expression driven by a 1.0-kb BmVg promoter (VgP1.0K) was consistent with endogenous BmVg. VgP1.0K specificity was closer to the endogenous BmVg promoter than that of VgP0.8K. These results suggest that CREs affecting stage-specific BmVg expression were localized to the 1.0-kb BmVg promoter. We investigated the effects of certain CREs that could influence the stage specificity of BmVg promoter on BmVg expression in transgenic B. mori. The relative DsRed expression was significantly reduced in transgenic female B. mori and the peak in DsRed expression was delayed after E-box CRE mutation. These results demonstrate that the E-box element enhanced BmVg expression and also affected stage-specific BmVg expression. Moreover, the relative DsRed expression was significantly increased in transgenic female of B. mori after 3×BD CRE mutation in BmVg promoter. However, the stage specificity of the mutated promoter was consistent with that of the endogenous BmVg promoter. The 3×BD element downregulated BmVg but had no effect on stage-specific BmVg expression. The present study promoted the process of elucidating the regulatory network for stage-specific BmVg expression and furnished a theoretical basis for the application of BmVg promoter.

scholarly journals Functional analysis of CgWRKY57 from Cymbidium goeringii in ABA response

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e10982
Author(s):  
Huanhuan Liu ◽  
Lianping Wang ◽  
Xijun Jing ◽  
Yue Chen ◽  
Fengrong Hu
Keyword(s):  
Abiotic Stress ◽  
Transgenic Plants ◽  
Real Time ◽  
Growth And Development ◽  
Transmembrane Domain ◽  
Germination Rate ◽  
Wild Type ◽  
Abiotic Stress Response ◽  
Group Iii ◽  
Cymbidium Goeringii

Background The orchid is one of the top ten Chinese flowers and has high ornamental value and elegant color. However, orchids are vulnerable to abiotic stresses during their growth and development, and the molecular mechanism of the abiotic stress response in orchids is unclear. WRKY proteins belong to a transcription factor family that plays important roles in biotic stress, abiotic stress, growth and development in plants, but little is known about the WRKY family in Cymbidium goeringii. Methods The specific fragment of the CgWRKY57 gene of C. goeringii was analyzed by bioinformatics. The expression of the CgWRKY57 gene of C. goeringii under 4 °C, 42 °C water and ABA stress as well as different tissues was detected by real-time fluorescence quantitative PCR. CgWRKY57 gene was overexpressed in wild type Arabidopsis thaliana by inflorescence infection method, and the function of transgenic lines under ABA stress was analyzed. Results CgWRKY57 was cloned from C. goeringii and found to encode 303 amino acids. The CgWRKY57 protein is an acidic, nonsecreted hydrophilic protein without a signal peptide or transmembrane domain. The CgWRKY57 protein is located to the nucleus and may function intracellularly according to its predicted subcellular localization. A domain analysis and homology comparison showed that the CgWRKY57 protein has a “WRKYGQK” domain and belongs to Group III of the WRKY family, and a phylogenetic analysis demonstrated that CgWRKY57 is closely related to OsWRKY47. CgWRKY57 was expressed in the roots, stems, leaves and floral organs of C. goeringii, and its expression level was highest in the roots according to real-time qPCR analysis. There were significant differences in CgWRKY57 expression under 4 °C, 42 °C ABA and water stress treatments, and its expression changed greatly under ABA stress. The expression of CgWRKY57 in transgenic plants was significantly higher than that in wild type plants under ABA stress, and the root length and germination rate were reduced in transgenic plants compared to wild type plants. Conclusions These results indicate that CgWRKY57 overexpression is responsive to ABA stress, and they provide a foundation for future analyses of the biological functions of the WRKY family in C. goeringii.

scholarly journals NtMYB3, an R2R3-MYB from Narcissus, Regulates Flavonoid Biosynthesis

2019 ◽  
Vol 20 (21) ◽  
pp. 5456 ◽  
Author(s):  
Muhammad Anwar ◽  
Weijun Yu ◽  
Hong Yao ◽  
Ping Zhou ◽  
Andrew C. Allan ◽  
...  
Keyword(s):  
Transgenic Tobacco ◽  
Flavonoid Biosynthesis ◽  
Pcr Analysis ◽  
Wild Type ◽  
Narcissus Tazetta ◽  
Over Expression ◽  
Qrt Pcr ◽  
Red Color ◽  
R2r3 Myb ◽  
R2r3 Myb Transcription Factors

R2R3-MYB transcription factors play important roles in the regulation of plant flavonoid metabolites. In the current study, NtMYB3, a novel R2R3-MYB transcriptional factor isolated from Chinese narcissus (Narcissus tazetta L. var. chinensis), was functionally characterized. Phylogenetic analysis indicated that NtMYB3 belongs to the AtMYB4-like clade, which includes repressor MYBs involved in the regulation of flavonoid biosynthesis. Transient assays showed that NtMYB3 significantly reduced red pigmentation induced by the potato anthocyanin activator StMYB-AN1 in agro-infiltrated leaves of tobacco. Over-expression of NtMYB3 decreased the red color of transgenic tobacco flowers, with qRT-PCR analysis showing that NtMYB3 repressed the expression levels of genes involved in anthocyanin and flavonol biosynthesis. However, the proanthocyanin content in flowers of transgenic tobacco increased as compared to wild type. NtMYB3 showed expression in all examined narcissus tissues; the expression level in basal plates of the bulb was highest. A 968 bp promoter fragment of narcissus FLS (NtFLS) was cloned, and transient expression and dual luciferase assays showed NtMYB3 repressed the promoter activity. These results reveal that NtMYB3 is involved in the regulation of flavonoid biosynthesis in narcissus by repressing the biosynthesis of flavonols, and this leads to proanthocyanin accumulation in the basal plate of narcissus.

scholarly journals Molecular Cloning and Functional Analysis of GmLACS2-3 Reveals Its Involvement in Cutin and Suberin Biosynthesis along with Abiotic Stress Tolerance

2021 ◽  
Vol 22 (17) ◽  
pp. 9175
Author(s):  
Asma Ayaz ◽  
Haodong Huang ◽  
Minglü Zheng ◽  
Wajid Zaman ◽  
Donghai Li ◽  
...  
Keyword(s):  
Glycine Max ◽  
Abiotic Stresses ◽  
Abiotic Stress Tolerance ◽  
Expression Patterns ◽  
Wild Type ◽  
Biotic And Abiotic Stresses ◽  
Aerial Parts ◽  
Chain Acyl ◽  
Important Crop Plant

Cutin and wax are the main precursors of the cuticle that covers the aerial parts of plants and provide protection against biotic and abiotic stresses. Long-chain acyl-CoA synthetases (LACSs) play diversified roles in the synthesis of cutin, wax, and triacylglycerol (TAG). Most of the information concerned with LACS functions is obtained from model plants, whereas the roles of LACS genes in Glycine max are less known. Here, we have identified 19 LACS genes in Glycine max, an important crop plant, and further focused our attention on 4 LACS2 genes (named as GmLACS2-1, 2, 3, 4, respectively). These GmLACS2 genes display different expression patterns in various organs and also show different responses to abiotic stresses, implying that these genes might play diversified functions during plant growth and against stresses. To further identify the role of GmLACS2-3, greatly induced by abiotic stresses, we transformed a construct containing its full length of coding sequence into Arabidopsis. The expression of GmLACS2-3 in an Arabidopsis atlacs2 mutant greatly suppressed its phenotype, suggesting it plays conserved roles with that of AtLACS2. The overexpression of GmLACS2-3 in wild-type plants significantly increased the amounts of cutin and suberin but had little effect on wax amounts, indicating the specific role of GmLACS2-3 in the synthesis of cutin and suberin. In addition, these GmLACS2-3 overexpressing plants showed enhanced drought tolerance. Taken together, our study deepens our understanding of the functions of LACS genes in different plants and also provides a clue for cultivating crops with strong drought resistance.

scholarly journals Sucrose Enhances Anthocyanin Accumulation in Torenia by Promoting Expression of Anthocyanin Biosynthesis Genes

Horticulturae ◽  
2021 ◽  
Vol 7 (8) ◽  
pp. 219
Author(s):  
Aung Htay Naing ◽  
Junping Xu ◽  
Kyeung Il Park ◽  
Mi Young Chung ◽  
Chang Kil Kim
Keyword(s):  
Transcription Factors ◽  
Transgenic Plants ◽  
Plant Growth ◽  
Anthocyanin Biosynthesis ◽  
Anthocyanin Accumulation ◽  
Wild Type ◽  
Anthocyanin Production

We examined the effects of different sucrose concentrations (3%, 5%, and 7%) on anthocyanin accumulation and plant growth in wild type (WT) and transgenic (T2) torenia cultivar “Kauai Rose” overexpressing the anthocyanin regulatory transcription factors B-Peru + mPAP1 or RsMYB1. Sucrose increased anthocyanin production in both WT and transgenic plants, with higher anthocyanin production in transgenic plants compared to WT plants. Higher sucrose concentrations increased production of anthocyanin in transgenic and WT plants, with increased anthocyanin production associated with increased expression of anthocyanin biosynthesis genes. Higher sucrose concentrations reduced growth of WT and transgenic plants. Our results indicate that sucrose enhances anthocyanin production in torenia by regulating anthocyanin biosynthesis genes.

Citrus sinensis CBF1 Functions in Cold Tolerance by Modulating Putrescine Biosynthesis Through Regulation of ARGININE DECARBOXYLASE

2021 ◽  
Author(s):  
Jie Song ◽  
Hao Wu ◽  
Feng He ◽  
Jing Qu ◽  
Yue Wang ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Stress Response ◽  
Transgenic Plants ◽  
Cold Stress ◽  
Cold Tolerance ◽  
Citrus Sinensis ◽  
Sweet Orange ◽  
Arginine Decarboxylase ◽  
Wild Type ◽  
Shed Light

Abstract C-repeat (CRT) binding factors (CBFs) are well known to act as crucial transcription factors that function in cold stress response. Arginine decarboxylase (ADC)-mediated putrescine biosynthesis has been reported to be activated in plants exposed to cold conditions, but it remains elusive whether CBFs can regulate ADC expression and putrescine accumulation. In this study, we show that cold up-regulated ADC gene (CsADC) and elevation of endogenous putrescine content in sweet orange (Citrus sinensis). Promoter of CsADC contains two CRT sequences that are canonical elements recognized by CBFs. Sweet orange genome contains four CBFs (CsCBF1-4), in which CsCBF1 was significantly induced by cold. CsCBF1, located in the nucleus, was demonstrated to bind directly and specifically to the promoter of CsADC and acted as a transcriptional activator. Overexpression of CsCBF1 led to notable elevation of CsADC and putrescine level in sweet orange transgenic plants, along with remarkably enhanced cold tolerance, relative to the wild type (WT). However, pretreatment with D-arginine, an ADC inhibitor, caused prominent reduction of endogenous putrescine level in the overexpressing lines, accompanied by greatly compromised cold tolerance. Taken together, these results demonstrate that CBF1 of sweet orange directly regulates ADC expression and modulates putrescine synthesis for orchestrating the cold tolerance. Our findings shed light into the transcriptional regulation of putrescine accumulation through targeting the ADC gene in the presence of cold stress. Meanwhile, this study illustrates a new mechanism underlying the CBF-mediated cold stress response.

scholarly journals Overexpression of an evolutionarily conserved drought-responsive sugarcane gene enhances salinity and drought resilience

2019 ◽  
Vol 124 (4) ◽  
pp. 691-700 ◽  
Author(s):  
Kevin Begcy ◽  
Eduardo D Mariano ◽  
Carolina G Lembke ◽  
Sonia Marli Zingaretti ◽  
Glaucia M Souza ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Oxidative Damage ◽  
Germination Rate ◽  
Co2 Concentration ◽  
Dry Mass ◽  
Wild Type ◽  
Gas Exchange Parameters ◽  
Physiological Level ◽  
Tobacco Seeds ◽  
Genes Encoding

Abstract Background and Aims Improving drought adaptation is more pressing for crops such as sugarcane, rice, wheat and maize, given the high dependence of these crops on irrigation. One option for enhancing adaptation to water limitation in plants is by transgenic approaches. An increasing number of genes that are associated with mechanisms used by plants to cope with water scarcity have been discovered. Genes encoding proteins with unknown functions comprise a relevant fraction of the genes that are modulated by drought. We characterized a gene in response to environmental stresses to gain insight into the unknown fraction of the sugarcane genome. Scdr2 (Sugarcane drought-responsive 2) encodes a small protein and shares highly conserved sequences within monocots, dicots, algae and fungi. Methods Plants overexpressing the Scdr2 sugarcane gene were examined in response to salinity and drought. Measurements of the gas exchange parameters, germination rate, water content, dry mass and oxidative damage were performed. Seeds as well as juvenile plants were used to explore the resilience level of the transgenic plants when compared with wild-type plants. Key Results Overexpression of Scdr2 enhanced germination rates in tobacco seeds under drought and salinity conditions. Juvenile transgenic plants overexpressing Scdr2 and subjected to drought and salinity stresses showed higher photosynthesis levels, internal CO2 concentration and stomatal conductance, reduced accumulation of hydrogen peroxide in the leaves, no penalty for photosystem II and faster recovery after submission to both stress conditions. Respiration was not strongly affected by both stresses in the Scdr2 transgenic plants, whereas wild-type plants exhibited increased respiration rates. Conclusions Scdr2 is involved in the response mechanism to abiotic stresses. Higher levels of Scdr2 enhanced resilience to salinity and drought, and this protection correlated with reduced oxidative damage. Scdr2 confers, at the physiological level, advantages to climate limitations. Therefore, Scdr2 is a potential target for improving sugarcane resilience to abiotic stress.

scholarly journals Transcriptome-Based Analysis of Dof Family Transcription Factors and Their Responses to Abiotic Stress in Tea Plant (Camellia sinensis)

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
Hui Li ◽  
Wei Huang ◽  
Zhi-Wei Liu ◽  
Yong-Xin Wang ◽  
Jing Zhuang
Keyword(s):  
Transcription Factors ◽  
Abiotic Stress ◽  
Camellia Sinensis ◽  
Stress Resistance ◽  
Abiotic Stress Tolerance ◽  
Plant Stress ◽  
Tea Plant ◽  
Interaction Networks ◽  
Qrt Pcr ◽  
Dof Transcription Factors

Tea plant (Camellia sinensis (L.) O. Kuntze) is affected by abiotic stress during its growth and development. DNA-binding with one finger (Dof) transcription factors (TFs) play important roles in abiotic stress tolerance of plants. In this study, a total of 29 putative Dof TFs were identified based on transcriptome of tea plant, and the conserved domains and common motifs of these CsDof TFs were predicted and analyzed. The 29 CsDof proteins were divided into 7 groups (A, B1, B2, C1, C2.1, C2.2, and D2), and the interaction networks of Dof proteins in C. sinensis were established according to the data in Arabidopsis. Gene expression was analyzed in “Yingshuang” and “Huangjinya” under four experimental stresses by qRT-PCR. CsDof genes were expressed differentially and related to different abiotic stress conditions. In total, our results might suggest that there is a potential relationship between CsDof factors and tea plant stress resistance.

scholarly journals Transcriptomic analysis reveals key transcription factors associated to drought tolerance in a wild papaya (Carica papaya) genotype

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0245855
Author(s):  
Humberto Estrella-Maldonado ◽  
Amaranta Girón Ramírez ◽  
Gabriela Fuentes Ortiz ◽  
Santy Peraza-Echeverría ◽  
Octavio Martínez-de la Vega ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Water Deficit ◽  
Carica Papaya ◽  
Differential Expression Analysis ◽  
Transcriptomic Analysis ◽  
Cdna Libraries ◽  
Pcr Analysis ◽  
Water Deficit Stress ◽  
Wild Genotype ◽  
Qrt Pcr

Most of the commercial papaya genotypes show susceptibility to water deficit stress and require high volumes of irrigation water to yield properly. To tackle this problem, we have collected wild native genotypes of Carica papaya that have proved to show better physiological performance under water deficit stress than the commercial cultivar grown in Mexico. In the present study, plants from a wild Carica papaya genotype and a commercial genotype were subjected to water deficit stress (WDS), and their response was characterized in physiological and molecular terms. The physiological parameters measured (water potential, photosynthesis, Fv/Fm and electrolyte leakage) confirmed that the papaya wild genotype showed better physiological responses than the commercial one when exposed to WDS. Subsequently, RNA-Seq was performed for 4 cDNA libraries in both genotypes (susceptible and tolerant) under well-watered conditions, and when they were subjected to WDS for 14 days. Consistently, differential expression analysis revealed that after 14 days of WDS, the wild tolerant genotype had a higher number of up-regulated genes, and a higher number of transcription factors (TF) that were differentially expressed in response to WDS, than the commercial genotype. Thus, six TF genes (CpHSF, CpMYB, CpNAC, CpNFY-A, CpERF and CpWRKY) were selected for further qRT-PCR analysis as they were highly expressed in response to WDS in the wild papaya genotype. qRT-PCR results confirmed that the wild genotype had higher expression levels (REL) in all 6 TF genes than the commercial genotype. Our transcriptomic analysis should help to unravel candidate genes that may be useful in the development of new drought-tolerant cultivars of this important tropical crop.

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