Cloning and function analysis of BAG family genes in wheat

2016 ◽  
Vol 43 (5) ◽  
pp. 393
Author(s):  
Shiming Ge ◽  
Zhen Kang ◽  
Ying Li ◽  
Fuzhen Zhang ◽  
Yinzhu Shen ◽  
...  
Keyword(s):  
Heat Stress ◽  
Salinity Stress ◽  
Heat Tolerance ◽  
Expressed Sequence Tag ◽  
Function Analysis ◽  
Pcr Analysis ◽  
Subcellular Localisation ◽  
Hybrid Technique ◽  
Rt Pcr ◽  
Conserved Domain

By analysing the cDNA microarray of the salt tolerant mutant of wheat RH8706–49 under salinity stress, our results showed an expressed sequence tag fragment and acquired an unknown gene (designated as TaBAG) with a BAG conserved domain through electronic cloning and RT–PCR technology. The gene was registered into GenBank (No. FJ599765). After homologous alignment analysis, electronic cloning, and amplifying with RT–PCR, the other gene with a BAG conserved domain, TaBAG2, was obtained and registered into GenBank (No. GU471210). Quantitative PCR analysis demonstrated that TaBAG2 expression was induced by saline and heat stress. TaBAG gene expression under salinity stress increased remarkably but showed an insignificant response to heat stress. The adversity stress detection results showed that Arabidopsis overexpressing TaBAG and TaBAG2 exhibited an obvious salt tolerance increase. Under heat stress, Arabidopsis overexpressing TaBAG2 showed increased heat tolerance; however, the heat tolerance of Arabidopsis overexpressing TaBAG did not vary significantly under heat stress. Subcellular localisation results demonstrated that TaBAGs were mainly located in the cytoplasm and the cell nucleus. We applied fluorescence complementation and yeast two-hybrid technique to prove that TaBAG2 can obviously bond with TaHsp70 and TaCaMs. After the respective mutation of aspartic acid (D) and arginine (R) at high conservation in BAG domain of TaBAG2, the bonding interaction between TaBAG2 and TaHsp70 disappeared, indicating that the two amino acids were the key loci for the interaction between TaBAG2 and TaHsp70. Heat tolerance detection results demonstrated that the heat tolerance of Arabidopsis overexpressing and cotransfected with TaBAG2 and TaHsp70 was much higher than that of Arabidopsis overexpressing TaBAG2 and Arabidopsis overexpressing TaHSP70. This finding implies that the synergistic use of TaBAG2 and TaHSP70 can improve heat tolerance of plants.

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 Novel Insights Into Heat Tolerance Using Microbiome and Metabolomics Analyses in Dairy Cows Rumen Fluid

2021 ◽  
Author(s):  
Zichen Wang ◽  
Leizhen Liu ◽  
Fanglin Pang ◽  
Zhuo Zheng ◽  
Zhanwei Teng ◽  
...  
Keyword(s):  
Heat Stress ◽  
Heat Resistance ◽  
Dairy Cows ◽  
Propionic Acid ◽  
Heat Tolerance ◽  
Function Analysis ◽  
Rumen Fluid ◽  
Diagnostic Tools ◽  
Differential Heat ◽  
Rumen Microbiome

Abstract Background: Heat stress is a key issue of growing concern for livestock industry worldwide due to its negative effects not only on milk production, fertility, health, welfare, and economic returns of dairy cows, but also on the microbial communities in the rumen. However, the underlying relationship between rumen microbiome and its associated metabolism with heat tolerance in cow have not been extensively described yet. Therefore, the main objective of this study was to investigate differential heat resistance in Holstein cows using rumen microbiome and metabolome analyses.Methods: We performed both principal component analysis and membership function analysis to select 7 heat-tolerant (HT) and 7 heat-sensitive (HS) cows. The ruminal fluid samples of two groups were collected at two hours post feeding on 7th day of heat stress period, for analyses including rumen fermentation parameters, rumen microbiome and nontargeted metabolomics.Results: Under heat stress conditions, the HT cows had a significantly higher propionic acid content than the HS cows; whereas measures of the respiratory rate (RR), rectal temperature (RT), acetic,butyric acid and acetic acid to propionic acid ratio (A:P) in the HT cows were lower compared with the HS cows. Omics sequencing revealed that the relative abundance of Rikenellaceae_RC9_gut_group, Succiniclasticum, Ruminococcaceae_NK4A214_group and Christensenellaceae_R-7_group were significantly higher in the HT than HS cows; whereas Prevotella_1, Ruminococcaceae_UCG-014, and Shuttleworthia were significantly higher in the HS cows compared to HT cows. Substances mainly involved in carbohydrate metabolism, including glycerol, mannitol, and maltose, showed significantly higher content in the HT cows compared to that in the HS cows. Simultaneously, RR was significantly correlated with both differential microorganisms and distinct metabolites, suggesting three metabolites could be potential biomarkers for determining heat resistance that require further research.Conclusion: Overall, distinct changes in the rumen microbiome and metabolomics in the HT cows may be associated with better adaptability to heat stress. These findings suggest their use as diagnostic tools of heat tolerance in dairy cattle breeding schemes.

scholarly journals Phenotypical and Functional Characteristics of Human Regulatory T Cells during Ex Vivo Maturation from CD4+ T Lymphocytes

2021 ◽  
Vol 11 (13) ◽  
pp. 5776
Author(s):  
Varvara G. Blinova ◽  
Natalia S. Novachly ◽  
Sofya N. Gippius ◽  
Abdullah Hilal ◽  
Yulia A. Gladilina ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Ex Vivo ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Inflammatory Reactions ◽  
Effector Cells ◽  
Cycle Regulation ◽  
Further Development

Regulatory T cells (Tregs) participate in the negative regulation of inflammatory reactions by suppressing effector cells. In a number of autoimmune disorders, the suppressive function and/or the number of Tregs is compromised. The lack of active functioning Tregs can be restored with adoptive transfer of expanded ex vivo autologous Tregs. In our study, we traced the differentiation and maturation of Tregs CD4+CD25+FoxP3+CD127low over 7 days of cultivation from initial CD4+ T cells under ex vivo conditions. The resulting ex vivo expanded cell population (eTregs) demonstrated the immune profile of Tregs with an increased capacity to suppress the proliferation of target effector cells. The expression of the FoxP3 gene was upregulated within the time of expansion and was associated with gradual demethylation in the promotor region of the T cell-specific demethylation region. Real-time RT-PCR analysis revealed changes in the expression profile of genes involved in cell cycle regulation. In addition to FOXP3, the cells displayed elevated mRNA levels of Ikaros zinc finger transcription factors and the main telomerase catalytic subunit hTERT. Alternative splicing of FoxP3, hTERT and IKZF family members was demonstrated to be involved in eTreg maturation. Our data indicate that expanded ex vivo eTregs develop a Treg-specific phenotype and functional suppressive activity. We suggest that eTregs are not just expanded but transformed cells with enhanced capacities of immune suppression. Our findings may influence further development of cell immunosuppressive therapy based on regulatory T cells.

scholarly journals Genotype-by-environment interaction in Holstein heifer fertility traits using single-step genomic reaction norm models

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Rui Shi ◽  
Luiz Fernando Brito ◽  
Aoxing Liu ◽  
Hanpeng Luo ◽  
Ziwei Chen ◽  
...  
Keyword(s):  
Heat Stress ◽  
Dairy Cattle ◽  
Candidate Genes ◽  
Heat Tolerance ◽  
Reaction Norm ◽  
Single Step ◽  
Conception Rate ◽  
Critical Periods ◽  
Genotype By Environment ◽  
Fertility Traits

Abstract Background The effect of heat stress on livestock production is a worldwide issue. Animal performance is influenced by exposure to harsh environmental conditions potentially causing genotype-by-environment interactions (G × E), especially in highproducing animals. In this context, the main objectives of this study were to (1) detect the time periods in which heifer fertility traits are more sensitive to the exposure to high environmental temperature and/or humidity, (2) investigate G × E due to heat stress in heifer fertility traits, and, (3) identify genomic regions associated with heifer fertility and heat tolerance in Holstein cattle. Results Phenotypic records for three heifer fertility traits (i.e., age at first calving, interval from first to last service, and conception rate at the first service) were collected, from 2005 to 2018, for 56,998 Holstein heifers raised in 15 herds in the Beijing area (China). By integrating environmental data, including hourly air temperature and relative humidity, the critical periods in which the heifers are more sensitive to heat stress were located in more than 30 days before the first service for age at first calving and interval from first to last service, or 10 days before and less than 60 days after the first service for conception rate. Using reaction norm models, significant G × E was detected for all three traits regarding both environmental gradients, proportion of days exceeding heat threshold, and minimum temperature-humidity index. Through single-step genome-wide association studies, PLAG1, AMHR2, SP1, KRT8, KRT18, MLH1, and EOMES were suggested as candidate genes for heifer fertility. The genes HCRTR1, AGRP, PC, and GUCY1B1 are strong candidates for association with heat tolerance. Conclusions The critical periods in which the reproductive performance of heifers is more sensitive to heat stress are trait-dependent. Thus, detailed analysis should be conducted to determine this particular period for other fertility traits. The considerable magnitude of G × E and sire re-ranking indicates the necessity to consider G × E in dairy cattle breeding schemes. This will enable selection of more heat-tolerant animals with high reproductive efficiency under harsh climatic conditions. Lastly, the candidate genes identified to be linked with response to heat stress provide a better understanding of the underlying biological mechanisms of heat tolerance in dairy cattle.

Sign in / Sign up

Export Citation Format

Share Document