scholarly journals Gateway cloning and in-planta transformation of drought stress responsive Ecmyb1 gene isolated from Eleusine coracana var.PRM 6107

2021 ◽  
Vol 22 (1) ◽  
pp. 205-211
Author(s):  
Megha Bhatt ◽  
Prafull Salvi ◽  
Pushpa Lohani
Keyword(s):  
Gene Expression ◽  
Arabidopsis Thaliana ◽  
Drought Stress ◽  
Stress Responses ◽  
Anthocyanin Biosynthesis ◽  
Regulatory Protein ◽  
Regulatory Elements ◽  
Growth And Yield ◽  
Yield Potential ◽  
Gateway Cloning

Drought is one of the key abiotic stress that critically influences the crops by restraining their growth and yield potential. Being sessile, plants tackle the detrimental effects of drought stress via modulating the cellular state by changing the gene expression. Such alteration of gene expression is essentially driven by the transcriptional syndicate. Transcription factors (TF) are the key regulatory protein that controls the expression of their target gene by binding to the cis-regulatory elements present in the promoter region. Myb-TFs ubiquitously present in all eukaryotes belong to one of the largest TF family, and play wide array of biological functions in plants including anthocyanin biosynthesis, vasculature system, cell signaling, seed maturation and abiotc stress responses. In the present study the full length Myb TF from Eleusine corocana was subcloned using Gateway cloning system and further transformed into Arabidopsis thaliana through floral dip method. Transgenic Arabidopsis thaliana plants harbouring Ecmyb1 gene were screened and grown in transgenic glasshouse under controlled conditions.

scholarly journals Molecular Cloning, Expression and Insilco Analysis of Drought Stress Inducible MYB Transcription Factor Encoding Gene From C4 Plant Eleusine Coracana

2021 ◽  
Author(s):  
MEGHA BHATT
Keyword(s):  
Gene Expression ◽  
Abiotic Stress ◽  
Drought Stress ◽  
Molecular Cloning ◽  
Stress Responses ◽  
Promoter Region ◽  
Regulatory Elements ◽  
Full Length ◽  
Growth And Yield ◽  
Yield Potential

Abstract Drought is one of the key abiotic stresses that critically influences the crops by restraining their growth and yield potential. Being sessile, plant tackle the detrimental effects of drought stress via modulating the cellular state by changing the gene expression. Such alteration of gene expression is essentially driven by the transcriptional syndicate. Transcription factors (TF) are the key regulatory protein that controls the expression of their target gene by binding to the cis-regulatory elements present in the promoter region. Myb-TF subiquitously present in all eukaryotes belong to one of the largest TF family, and play wide array of biological functions in plants including anthocyanin biosynthesis, vasculature system, cell signaling, seed maturation and abiotc stress responses. In the present study, isolation, and molecular cloning of full length Myb TF from Eleusine corocana has been performed. The isolated full-length coding sequence has 1053 bp and 350 aa was submitted to NCBI (Accession number MT312253). The transcript level of EcMYB increases under different abiotic stress treatment including dehydration, salinity, and high temperature stress. The promoter region of EcMyb1 was found to be enriched in stress-responsive cis-regulatory elements such as DRE, HSE, ABRE etc. In phylogenetic analysis, EcMyb1 appeared to have high homology with its monocot orthologs particularly Sateria italica, Hordeum vulgare, Saccharum barberi and Oryza sativa. The three-dimension protein structure was generated based on homology modeling and structural aspects were discussed. Further, Insilco analysis was conducted to explore the physiological properties, subcellular localization, potential post-translational modification sites (phosphorylation and glycosylation sites), and molecular and biological function of full-length protein. Overall, the expression profiling and Insilco analysis of EcMyb1 strongly indicated its potential role in abiotic stress response in Eleusine corocana.

scholarly journals Systematic Analysis of HD-ZIP Transcription Factors in Sesame Genome and Gene Expression Profiling of SiHD-ZIP Class I Entailing Drought Stress Responses at Early Seedling Stage

2021 ◽  
Author(s):  
Maryam Mehmood ◽  
Muhammad Jadoon Khan ◽  
Muhammad Jawad Khan ◽  
Nadeem Akhtar ◽  
Fizza Mughal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factors ◽  
Drought Stress ◽  
Stress Responses ◽  
Seedling Stage ◽  
Class I ◽  
Growth And Yield ◽  
Severe Drought ◽  
Oilseed Crop ◽  
Early Seedling

Abstract Sesame is a very ancient oilseed crop. Sesame sensitivity to drought stress at early seedling stage is one of the limiting factors affecting its growth and yield in the world. HD-ZIP transcription factors family is one of the most important families involved in drought stress responses in plants. In this study, total sixty one sesame HD-ZIP (SiHZ) proteins were identified in sesame, based on protein sequence homology with Arabidopsis and protein domain(s) architectures were predicted by Hidden Markov model (HMM). HD-ZIP proteins were then classified into four classes (HD-ZIP Class I-IV) according to the phylogenetic, conserved domain(s) motifs and gene structure analyses in sesame. Based on comparative phylogenetic analysis of sesame with Arabidopsis and maize HD-ZIP protein sequences, HD-ZIP Class I was subdivided into four subgroups α (SiHZ25, SiHZ43, SiHZ9 and SiHZ16), β1 (SiHZ10, SiHZ30, SiHZ32 and SiHZ26), β2 (SiHZ42 and SiHZ45) and (SiHZ17, SiHZ7 and SiHZ35). Twenty-one days old Sesame seedling were exposed to severe drought stress by withholding water for 7 days. Gene expression of 13 members of HD-ZIP Class I was performed in well- watered (control) and water stressed (treatment) seedlings. The results of gene expression analysis showed that, SiHZ7 (6.8 fold) and SiHZ35 (2.6 fold) from subgroup showed significantly high gene expression levels under drought stress in sesame seedlings. Thus, this study provides useful molecular information pinpointing the role SiHD-ZIP Class I in drought stress responses at early seedling stage and to develop sesame novel varieties with improved drought tolerance in sesame.

scholarly journals Alternative splicing landscapes in Arabidopsis thaliana across tissues and stress conditions highlight major functional differences with animals

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Guiomar Martín ◽  
Yamile Márquez ◽  
Federica Mantica ◽  
Paula Duque ◽  
Manuel Irimia
Keyword(s):  
Gene Expression ◽  
Arabidopsis Thaliana ◽  
Alternative Splicing ◽  
Stress Responses ◽  
Target Genes ◽  
Intron Retention ◽  
Single Gene ◽  
Exon Skipping ◽  
Environmental Response ◽  
Biotic Stresses

Abstract Background Alternative splicing (AS) is a widespread regulatory mechanism in multicellular organisms. Numerous transcriptomic and single-gene studies in plants have investigated AS in response to specific conditions, especially environmental stress, unveiling substantial amounts of intron retention that modulate gene expression. However, a comprehensive study contrasting stress-response and tissue-specific AS patterns and directly comparing them with those of animal models is still missing. Results We generate a massive resource for Arabidopsis thaliana, PastDB, comprising AS and gene expression quantifications across tissues, development and environmental conditions, including abiotic and biotic stresses. Harmonized analysis of these datasets reveals that A. thaliana shows high levels of AS, similar to fruitflies, and that, compared to animals, disproportionately uses AS for stress responses. We identify core sets of genes regulated specifically by either AS or transcription upon stresses or among tissues, a regulatory specialization that is tightly mirrored by the genomic features of these genes. Unexpectedly, non-intron retention events, including exon skipping, are overrepresented across regulated AS sets in A. thaliana, being also largely involved in modulating gene expression through NMD and uORF inclusion. Conclusions Non-intron retention events have likely been functionally underrated in plants. AS constitutes a distinct regulatory layer controlling gene expression upon internal and external stimuli whose target genes and master regulators are hardwired at the genomic level to specifically undergo post-transcriptional regulation. Given the higher relevance of AS in the response to different stresses when compared to animals, this molecular hardwiring is likely required for a proper environmental response in A. thaliana.

scholarly journals RETRACTED ARTICLE: The specific MYB binding sites bound by TaMYB in the GAPCp2/3 promoters are involved in the drought stress response in wheat

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Lin Zhang ◽  
Zhiqiang Song ◽  
Fangfang Li ◽  
Xixi Li ◽  
Haikun Ji ◽  
...  
Keyword(s):  
Gene Expression ◽  
Abiotic Stress ◽  
Drought Stress ◽  
Reporter Gene ◽  
Binding Sites ◽  
Promoter Sequence ◽  
Regulatory Elements ◽  
Cis Elements ◽  
Gus Reporter ◽  
Almost All

Abstract Background Drought stress is one of the major abiotic stresses that affects plant growth and productivity. The GAPCp genes play important roles in drought stress tolerance in multiple species. The aim of this experiment was to identify the core cis-regulatory elements that may respond to drought stress in the GAPCp2 and GAPCp3 promoter sequences. Results In this study, the promoters of GAPCp2 and GAPCp3 were cloned. The promoter activities were significantly improved under abiotic stress via regulation of Rluc reporter gene expression, while promoter sequence analysis indicated that these fragments were not almost identical. In transgenic Arabidopsis with the expression of the GUS reporter gene under the control of one of these promoters, the activities of GUS were strong in almost all tissues except the seeds, and the activities were induced after abiotic stress. The yeast one-hybrid system and EMSA demonstrated that TaMYB bound TaGAPCp2P/3P. By analyzing different 5′ deletion mutants of these promoters, it was determined that TaGAPCp2P (− 1312~ − 528) and TaGAPCp3P (− 2049~ − 610), including the MYB binding site, contained enhancer elements that increased gene expression levels under drought stress. We used an effector and a reporter to co-transform tobacco and found that TaMYB interacted with the specific MYB binding sites of TaGAPCp2P (− 1197~ − 635) and TaGAPCp3P (− 1456~ − 1144 and − 718~ − 610) in plant cells. Then, the Y1H system and EMSA assay demonstrated that these MYB binding sites in TaGAPCp2P (− 1135 and − 985) and TaGAPCp3P (− 1414 and − 665) were the target cis-elements of TaMYB. The deletion of the specific MYB binding sites in the promoter fragments significantly restrained the drought response, and these results confirmed that these MYB binding sites (AACTAAA/C) play vital roles in improving the transcription levels under drought stress. The results of qRT-PCR in wheat protoplasts transiently overexpressing TaMYB indicated that the expression of TaGAPCp2/3 induced by abiotic stress was upregulated by TaMYB. Conclusion The MYB binding sites (AACTAAA/C) in TaGAPCp2P/3P were identified as the key cis-elements for responding to drought stress and were bound by the transcription factor TaMYB.

scholarly journals Drought Stress Enhances Up-Regulation of Anthocyanin Biosynthesis in Grapevine leafroll-associated virus 3-Infected in vitro Grapevine (Vitis vinifera) Leaves

Plant Disease ◽  
2017 ◽  
Vol 101 (9) ◽  
pp. 1606-1615 ◽  
Author(s):  
Zhen-Hua Cui ◽  
Wen-Lu Bi ◽  
Xin-Yi Hao ◽  
Peng-Min Li ◽  
Ying Duan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Drought Stress ◽  
Vitis Vinifera ◽  
Anthocyanin Biosynthesis ◽  
Anthocyanin Accumulation ◽  
Expression Levels ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Regulated Gene Expression

Reddish-purple coloration on the leaf blades and downward rolling of leaf margins are typical symptoms of grapevine leafroll disease (GLD) in red-fruited grapevine cultivars. These typical symptoms are attributed to the expression of genes encoding enzymes for anthocyanins synthesis, and the accumulation of flavonoids in diseased leaves. Drought has been proven to accelerate development of GLD symptoms in virus-infected leaves of grapevine. However, it is not known how drought affects GLD expression nor how anthocyanin biosynthesis in virus-infected leaves is altered. The present study used HPLC to determine the types and levels of anthocyanins, and applied reverse transcription quantitative polymerase chain reaction (RT-qPCR) to analyze the expression of genes encoding enzymes for anthocyanin synthesis. Plantlets of Grapevine leafroll-associated virus 3 (GLRaV-3)-infected Vitis vinifera ‘Cabernet Sauvignon’ were grown in vitro under PEG-induced drought stress. HPLC found no anthocyanin-related peaks in the healthy plantlets with or without PEG-induced stress, while 11 peaks were detected in the infected plantlets with or without PEG-induced drought stress, but the peaks were significantly higher in infected drought-stressed plantlets. Increased accumulation of total anthocyanin compounds was related to the development of GLD symptoms in the infected plantlets under PEG stress. The highest level of up-regulated gene expression was found in GLRaV-3-infected leaves with PEG-induced drought stress. Analyses of variance and correlation of anthocyanin accumulation with related gene expression levels found that GLRaV-3-infection was the key factor in increased anthocyanin accumulation. This accumulation involved the up-regulation of two key genes, MYBA1 and UFGT, and their expression levels were further enhanced by drought stress.

scholarly journals Physiological and Proteomic Analysis Revealed the Response Mechanisms of two Different Grought-resistant Maize Varieties

2021 ◽  
Author(s):  
Hongjie Li ◽  
Mei Yang ◽  
Chengfeng Zhao ◽  
Yifan Wang ◽  
Renhe Zhang
Keyword(s):  
Drought Stress ◽  
Drought Tolerance ◽  
Stress Responses ◽  
Molecular Chaperones ◽  
Electron Flow ◽  
Photochemical Efficiency ◽  
Growth And Yield ◽  
Protective Mechanism ◽  
Drought Tolerant ◽  
Maize Varieties

Abstract Background: Drought stress seriously limits the seedling growth and yield of maize. Despite previous studies on drought resistance mechanisms by which maize cope with water deficient, the link between physiological and molecular variations are largely unknown. To reveal the complex regulatory mechanisms, comparative physiology and proteomic analyses were conducted to investigate the stress responses of two maize cultivars with contrasting tolerance to drought stress. Results: Physiological results showed that SD609 (drought-tolerant) maintains higher photochemical efficiency by enhancing CEF (cyclic electron flow) protective mechanism and antioxidative enzymes activities. Proteomics analysis revealed a total of 198 and 102 proteins were differentially expressed in SD609 and SD902, respectively. Further enrichment analysis indicated that drought-tolerant ‘SD609’ increased the expression of proteins related to photosynthesis, antioxidants/detoxifying enzymes, molecular chaperones and metabolic enzymes. The up-regulation proteins related to PSII repair and photoprotection mechanisms resulted in more efficient photochemical capacity in tolerant variety under moderate drought. However, the drought-sensitive ‘SD902’ only induced molecular chaperones and sucrose synthesis pathways, and failed to protect the impaired photosystem. Further analysis indicated that proteins related to the electron transport chain, redox homeostasis and heat shock proteins (HSPs) could be important in protecting plants from drought stress. Conclusions: Our experiments explored the mechanism of drought tolerance, and obtained detailed information about the interconnection of physiological research and protein research. In summary, our findings could provide new clues into further understanding of drought tolerance mechanisms in maize.

scholarly journals Generation of guard cell RNA-seq transcriptomes during progressive drought and recovery using an adapted INTACT protocol for Arabidopsis thaliana shoot tissue

2021 ◽  
Author(s):  
Anna van Weringh ◽  
Asher Pasha ◽  
Eddi Esteban ◽  
Paul J. Gamueda ◽  
Nicholas J. Provart
Keyword(s):  
Gene Expression ◽  
Arabidopsis Thaliana ◽  
Drought Stress ◽  
Guard Cell ◽  
Crop Production ◽  
Leaf Tissue ◽  
Cell Types ◽  
Severe Drought ◽  
Data Sets ◽  
Rna Seq

Drought is an important environmental stress that limits crop production. Guard cells (GC) act to control the rate of water loss. To better understand how GCs change their gene expression during a progressive drought we generated guard cell-specific RNA-seq transcriptomes during mild, moderate, and severe drought stress. We additionally sampled re-watered plants that had experienced severe drought stress. These transcriptomes were generated using the INTACT system to capture the RNA from GC nuclei. We optimized the INTACT protocol for Arabidopsis thaliana leaf tissue, incorporating fixation to preserve RNA during nuclear isolation. To be able to identify gene expression changes unique to GCs, we additionally generated transcriptomes from all cell types, using a 35S viral promoter to capture the nuclei of all cell types in leaves. These data sets highlight shared and unique gene expression changes between GCs and the bulk leaf tissue. The timing of gene expression changes is different between GCs and other cell types: we found that only GCs had detectable gene expression changes at the earliest drought time point. The drought responsive GC and leaf RNA-seq transcriptomes are available in the Arabidopsis ePlant at the Bio-Analytic Resource for Plant Biology website.

scholarly journals Comparative Genome-wide Analysis and Expression Profiling of Histone Acetyltransferases and Histone Deacetylases Involved in the Response to Drought in Wheat

2020 ◽  
Author(s):  
Hua Li ◽  
Huajie Liu ◽  
Xinxin Pei ◽  
Hongyu Chen ◽  
Xiao Li ◽  
...  
Keyword(s):  
Drought Stress ◽  
Stress Responses ◽  
Histone Deacetylases ◽  
Gene Expression Analysis ◽  
Regulatory Elements ◽  
Wheat Genome ◽  
Histone Acetyltransferases ◽  
Promoter Regions ◽  
Conserved Domain ◽  
Genome Wide

Abstract Background: Histone acetyltransferases (HATs) and histone deacetylases (HDACs) contribute to plant growth, development, and stress responses. A number of HAT and HDAC genes have been identified in several plants. However, wheat HATs and HDACs have not been comprehensively characterized. In this study, we identified TaHATs and TaHDACs in the wheat genome using bioinformatics tools. Result: In total, 30 TaHAT genes and 53 TaHDAC genes were detected in the wheat genome. As described in other plants, TaHATs were classified into four subfamilies (i.e., GNAT, p300/CBP, MYST, and TAFII250) and TaHDACs were divided into three subfamilies (i.e., RPD3/HDA1, HD2, and SIR2). Phylogenetic and conserved domain analyses showed that TaHATs and TaHDACs are highly similar to those in Arabidopsis and rice; however, divergence and expansion from Arabidopsis and rice were also observed. We detected many stress-related cis-regulatory elements in the promoter regions of these genes (i.e., ABRE, STRE, MYB et al.). Further, based on a comparative expression analyses of three varieties with different degrees of drought resistance under drought stress, we found that TaHAG2, TaHAG3, TaHAC2, TaHDA18, TaHDT1, and TaHDT2 are likely regulate drought stress in wheat. Conclusions: In this study, TaHATs and TaHDACs from the wheat genome were identified. Three TaHATs and three TaHDACs were very likely to regulate drought stress based on a promoter analysis and gene expression analysis. These results provide a foundation for further research on the regulation of acetylation in wheat and its role in the response to drought stress.

scholarly journals The Maize WRKY Transcription Factor ZmWRKY40 Confers Drought Resistance in Transgenic Arabidopsis

2018 ◽  
Vol 19 (9) ◽  
pp. 2580 ◽  
Author(s):  
Chang-Tao Wang ◽  
Jing-Na Ru ◽  
Yong-Wei Liu ◽  
Jun-Feng Yang ◽  
Meng Li ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Transcription Factors ◽  
Drought Stress ◽  
Stress Response ◽  
Abiotic Stresses ◽  
Transgenic Arabidopsis ◽  
Regulatory Elements ◽  
Wrky Transcription Factor ◽  
Growth And Yield ◽  
Abiotic Stress Response

Abiotic stresses restrict the growth and yield of crops. Plants have developed a number of regulatory mechanisms to respond to these stresses. WRKY transcription factors (TFs) are plant-specific transcription factors that play essential roles in multiple plant processes, including abiotic stress response. At present, little information regarding drought-related WRKY genes in maize is available. In this study, we identified a WRKY transcription factor gene from maize, named ZmWRKY40. ZmWRKY40 is a member of WRKY group II, localized in the nucleus of mesophyll protoplasts. Several stress-related transcriptional regulatory elements existed in the promoter region of ZmWRKY40. ZmWRKY40 was induced by drought, high salinity, high temperature, and abscisic acid (ABA). ZmWRKY40 could rapidly respond to drought with peak levels (more than 10-fold) at 1 h after treatment. Overexpression of ZmWRKY40 improved drought tolerance in transgenic Arabidopsis by regulating stress-related genes, and the reactive oxygen species (ROS) content in transgenic lines was reduced by enhancing the activities of peroxide dismutase (POD) and catalase (CAT) under drought stress. According to the results, the present study may provide a candidate gene involved in the drought stress response and a theoretical basis to understand the mechanisms of ZmWRKY40 in response to abiotic stresses in maize.

scholarly journals Evolution of Anthocyanin Biosynthesis in Maize Kernels: The Role of Regulatory and Enzymatic Loci

Genetics ◽  
1996 ◽  
Vol 143 (3) ◽  
pp. 1395-1407 ◽  
Author(s):  
Michael A Hanson ◽  
Brandon S Gaut ◽  
Adrian O Stec ◽  
Susan I Fuerstenberg ◽  
Major M Goodman ◽  
...  
Keyword(s):  
Protein Function ◽  
Sequence Data ◽  
Anthocyanin Biosynthesis ◽  
Regulatory Protein ◽  
Regulatory Elements ◽  
Kernel Development ◽  
Genetic Tests ◽  
Anthocyanin Pathway ◽  
Regulatory Loci ◽  
Functional Alleles

Abstract Understanding which genes contribute to evolutionary change and the nature of the alterations in them are fundamental challenges in evolution. We analyzed regulatory and enzymatic genes in the maize anthocyanin pathway as related to the evolution of anthocyanin-pigmented kernels in maize from colorless kernels of its progenitor, teosinte. Genetic tests indicate that teosinte possesses functional alleles at all enzymatic loci. At two regulatory loci, most teosintes possess alleles that encode functional proteins, but ones that are not expressed during kernel development and not capable of activating anthocyanin biosynthesis there. We investigated nucleotide polymorphism at one of the regulatory loci, c1. Several observations suggest that c1 has not evolved in a strictly neutral manner, including an exceptionally low level of polymorphism and a biased representation of haplotypes in maize. Curiously, sequence data show that most of our teosinte samples possess a promoter element necessary for the activation of the anthocyanin pathway during kernel development, although genetic tests indicate that teosinte c1 alleles are not active during kernel development. Our analyses suggest that the evolution of the purple kernels resulted from changes in cis regulatory elements at regulatory loci and not changes in either regulatory protein function nor the enzymatic loci.

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