Dataset Information

Over-Expression of the Heat-Responsive Wheat Gene TaHSP23.9 in Transgenic Arabidopsis Conferred Tolerance to Heat and Salt Stress.

ABSTRACT: The small heat shock proteins (sHSP) are stress-induced proteins with molecular weights ranging from 12 to 42 kDa that act as molecular chaperones to prevent the irreversible aggregation of denaturing proteins. In this study, we cloned the heat responsive gene TaHSP23.9 from wheat (Triticum aestivum) based on TMT-labeled quantitative proteomic analysis in our previous work and examined its function in the response of transgenic Arabidopsis to heat and salt stress. Amino acid alignment and phylogenetic tree analysis showed that TaHSP23.9 contained a typically conserved structure of the alpha-crystallin domain and is closely related to OsHSP23.2 in rice. Transient expression assays demonstrated that TaHSP23.9 is located on the endoplasmic reticulum. Quantitative real-time PCR demonstrated that TaHSP23.9 was expressed much more in filling grains under normal conditions and was significantly upregulated by heat and salt stress. Transgenic Arabidopsis plants that constitutively over-expressed TaHSP23.9 had no visible differences or adverse phenotypes compared with the wild type under normal conditions but exhibited enhanced tolerance to heat and salt stress under stress conditions. In addition, we found that the expression level of TaHSP23.9 was significantly higher in the heat-tolerant wheat varieties than in the heat-sensitive varieties. Our results suggest that TaHSP23.9 may function as a protein chaperone to positively regulate plant responses to heat and salt stress and could be developed as a molecular marker for screening heat-tolerant wheat varieties.

SUBMITTER: Wang J

PROVIDER: S-EPMC7069362 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Over-Expression of the Heat-Responsive Wheat Gene TaHSP23.9 in Transgenic Arabidopsis Conferred Tolerance to Heat and Salt Stress.

Wang Jun J Gao Xin X Dong Jun J Tian Xinyu X Wang Junzhe J Palta Jairo A JA Xu Shengbao S Fang Yan Y Wang Zhonghua Z

Frontiers in plant science 20200306

The small heat shock proteins (sHSP) are stress-induced proteins with molecular weights ranging from 12 to 42 kDa that act as molecular chaperones to prevent the irreversible aggregation of denaturing proteins. In this study, we cloned the heat responsive gene TaHSP23.9 from wheat (Triticum aestivum) based on TMT-labeled quantitative proteomic analysis in our previous work and examined its function in the response of transgenic Arabidopsis to heat and salt stress. Amino acid ...[more]

PMID: 32211001

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Project description:NAC (NAM, ATAF, and CUC) transcription factors play important roles in plant biological processes, including phytohormone homeostasis, plant development, and in responses to various environmental stresses.TaNAC29 was introduced into Arabidopsis using the Agrobacterium tumefaciens-mediated floral dipping method. TaNAC29-overexpression plants were subjected to salt and drought stresses for examining gene functions. To investigate tolerant mechanisms involved in the salt and drought responses, expression of related marker genes analyses were conducted, and related physiological indices were also measured. Expressions of genes were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR).A novel NAC transcription factor gene, designated TaNAC29, was isolated from bread wheat (Triticum aestivum). Sequence alignment suggested that TaNAC29 might be located on chromosome 2BS. TaNAC29 was localized to the nucleus in wheat protoplasts, and proved to have transcriptional activation activities in yeast. TaNAC29 was expressed at a higher level in the leaves, and expression levels were much higher in senescent leaves, indicating that TaNAC29 might be involved in the senescence process. TaNAC29 transcripts were increased following treatments with salt, PEG6000, H2O2, and abscisic acid (ABA). To examine TaNAC29 function, transgenic Arabidopsis plants overexpressing TaNAC29 were generated. Germination and root length assays of transgenic plants demonstrated that TaNAC29 overexpression plants had enhanced tolerances to high salinity and dehydration, and exhibited an ABA-hypersensitive response. When grown in the greenhouse, TaNAC29-overexpression plants showed the same tolerance response to salt and drought stresses at both the vegetative and reproductive period, and had delayed bolting and flowering in the reproductive period. Moreover, TaNAC29 overexpression plants accumulated lesser malondialdehyde (MDA), H2O2, while had higher superoxide dismutase (SOD) and catalase (CAT) activities under high salinity and/or dehydration stress.Our results demonstrate that TaNAC29 plays important roles in the senescence process and response to salt and drought stresses. ABA signal pathway and antioxidant enzyme systems are involved in TaNAC29-mediated stress tolerance mechanisms.

Project description:BackgroundSterol glycosyltrnasferases (SGT) are enzymes that glycosylate sterols which play important role in plant adaptation to stress and are medicinally important in plants like Withania somnifera. The present study aims to find the role of WsSGTL1 which is a sterol glycosyltransferase from W. somnifera, in plant's adaptation to abiotic stress.MethodologyThe WsSGTL1 gene was transformed in Arabidopsis thaliana through Agrobacterium mediated transformation, using the binary vector pBI121, by floral dip method. The phenotypic and physiological parameters like germination, root length, shoot weight, relative electrolyte conductivity, MDA content, SOD levels, relative electrolyte leakage and chlorophyll measurements were compared between transgenic and wild type Arabidopsis plants under different abiotic stresses--salt, heat and cold. Biochemical analysis was done by HPLC-TLC and radiolabelled enzyme assay. The promoter of the WsSGTL1 gene was cloned by using Genome Walker kit (Clontech, USA) and the 3D structures were predicted by using Discovery Studio Ver. 2.5.ResultsThe WsSGTL1 transgenic plants were confirmed to be single copy by Southern and homozygous by segregation analysis. As compared to WT, the transgenic plants showed better germination, salt tolerance, heat and cold tolerance. The level of the transgene WsSGTL1 was elevated in heat, cold and salt stress along with other marker genes such as HSP70, HSP90, RD29, SOS3 and LEA4-5. Biochemical analysis showed the formation of sterol glycosides and increase in enzyme activity. When the promoter of WsSGTL1 gene was cloned from W. somnifera and sequenced, it contained stress responsive elements. Bioinformatics analysis of the 3D structure of the WsSGTL1 protein showed functional similarity with sterol glycosyltransferase AtSGT of A. thaliana.ConclusionsTransformation of WsSGTL1 gene in A. thaliana conferred abiotic stress tolerance. The promoter of the gene in W.somnifera was found to have stress responsive elements. The 3D structure showed functional similarity with sterol glycosyltransferases.

Dataset Information

Over-Expression of the Heat-Responsive Wheat Gene TaHSP23.9 in Transgenic Arabidopsis Conferred Tolerance to Heat and Salt Stress.

Publications

Over-Expression of the Heat-Responsive Wheat Gene <i>TaHSP23.9</i> in Transgenic <i>Arabidopsis</i> Conferred Tolerance to Heat and Salt Stress.

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