Heterologous expression of rice RNA-binding glycine-rich (RBG) gene OsRBGD3 in transgenic Arabidopsis thaliana confers cold stress tolerance

2019 ◽  
Vol 46 (5) ◽  
pp. 482 ◽  
Author(s):  
Sangram K. Lenka ◽  
Amit K. Singh ◽  
Senthilkumar K. Muthusamy ◽  
Shuchi Smita ◽  
Viswanathan Chinnusamy ◽  
...  
Keyword(s):  
Low Temperature ◽  
Cold Stress ◽  
Stress Tolerance ◽  
Rna Binding ◽  
Transgenic Arabidopsis ◽  
Rice Cultivar ◽  
Early Flowering ◽  
Rna Recognition Motif ◽  
New Genes ◽  
Drought Tolerant

Imparting cold stress tolerance to crops is a major challenge in subtropical agriculture. New genes conferring cold tolerance needs to be identified and characterised for sustainable crop production in low-temperature stress affected areas. Here we report functional characterisation of OsRBGD3, classified previously as a class D glycine-rich RNA recognition motif (RRM) containing proteins from a drought-tolerant Indica rice cultivar N22. The gene was isolated by screening yeast one-hybrid library using the minimal promoter region of the OsMYB38 that is necessary for cold stress-responsive expression. OsRBGD3 exhibited cold, drought and salt stress inductive expression in a drought tolerant N22 rice cultivar as compared with susceptible variety IR64. OsRBGD3 was found to be localised to both nuclear and cytoplasmic subcellular destinations. Constitutive overexpression of the OsRBGD3 in transgenic Arabidopsis conferred tolerance to cold stress. ABA sensitivity was also observed in transgenic lines suggesting the regulatory role of this gene in the ABA signalling pathway. OsRBGD3 overexpression also attributed to significant root development and early flowering in transgenics. Hence, OsRBGD3 could be an important target for developing cold tolerant early flowering rice and other crops’ genotypes for increasing production in low temperature affected areas.

scholarly journals Overexpression of Cotton a DTX/MATE Gene Enhances Drought, Salt, and Cold Stress Tolerance in Transgenic Arabidopsis

2019 ◽  
Vol 10 ◽  
Author(s):  
Pu Lu ◽  
Richard Odongo Magwanga ◽  
Joy Nyangasi Kirungu ◽  
Yangguang Hu ◽  
Qi Dong ◽  
...  
Keyword(s):  
Cold Stress ◽  
Stress Tolerance ◽  
Transgenic Arabidopsis

Gene-Coexpression Network Analysis Identifies Specific Modules and Hub Genes Related to Cold Stress in Rice

2021 ◽  
Author(s):  
Zhichi Zeng ◽  
Sichen Zhang ◽  
Wenyan Li ◽  
Baoshan Chen ◽  
Wenlan Li
Keyword(s):  
Network Analysis ◽  
Low Temperature ◽  
Cold Stress ◽  
Cold Tolerance ◽  
Temperature Stress ◽  
Low Temperature Stress ◽  
Rice Cultivars ◽  
Hub Genes ◽  
New Genes ◽  
Short Period

Abstract Background: When plants are subjected to cold stress, they undergo a series of molecular and physiological changes to protect themselves from injury. Indica cultivars can usually withstand only mild cold stress in a relatively short period. Hormone-mediated defence response plays an important role in cold stress. Weighted gene co-expression network analysis (WGCNA) is a very useful tool for studying the correlation between genes, identifying modules with high phenotype correlation, and identifying Hub genes in different modules. Many studies have elucidated the molecular mechanisms of cold tolerance in different plants, but little information about the recovery process after cold stress is available.Results: To understand the molecular mechanism of cold tolerance in rice, we performed comprehensive transcriptome analyses during cold treatment and recovery stage in two cultivars of near-isogenic lines (9311 and DC907). Twelve transcriptomes in two rice cultivars were determined. A total of 2509 new genes were predicted by fragment splicing and assembly, and 7506 differentially expressed genes were identified by pairwise comparison. A total of 26 modules were obtained by expression-network analysis, 12 of which were highly correlated with cold stress or recovery treatment. We further identified candidate Hub genes associated with specific modules and analysed their regulatory relationships based on coexpression data. Results showed that various plant-hormone regulatory genes acted together to protect plants from physiological damage under short-term low-temperature stress. We speculated that this may be common in rice. Under long-term cold stress, rice improved the tolerance to low-temperature stress by promoting autophagy, sugar synthesis, and metabolism.Conclusion: Through WGCNA analysis at the transcriptome level, we provided a potential regulatory mechanism for the cold stress and recovery of rice cultivars and identified candidate central genes. Our findings provided an important reference for the future cultivation of rice strains with good tolerance.

scholarly journals A Genetic Screen for Novel Components of the Ras/Mitogen-Activated Protein Kinase Signaling Pathway That Interact With the yan Gene of Drosophila Identifies split ends, a New RNA Recognition Motif-Containing Protein

Genetics ◽  
2000 ◽  
Vol 154 (2) ◽  
pp. 695-712 ◽  
Author(s):  
Ilaria Rebay ◽  
Fangli Chen ◽  
Francis Hsiao ◽  
Peter A Kolodziej ◽  
Bing H Kuang ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Rna Binding ◽  
Mapk Pathway ◽  
Ectopic Expression ◽  
Mapk Signaling ◽  
Genetic Screen ◽  
Rna Recognition Motif ◽  
Rtk Signaling ◽  
New Genes ◽  
Split Ends

Abstract The receptor tyrosine kinase (RTK) signaling pathway is used reiteratively during the development of all multicellular organisms. While the core RTK/Ras/MAPK signaling cassette has been studied extensively, little is known about the nature of the downstream targets of the pathway or how these effectors regulate the specificity of cellular responses. Drosophila yan is one of a few downstream components identified to date, functioning as an antagonist of the RTK/Ras/MAPK pathway. Previously, we have shown that ectopic expression of a constitutively active protein (yanACT) inhibits the differentiation of multiple cell types. In an effort to identify new genes functioning downstream in the Ras/MAPK/yan pathway, we have performed a genetic screen to isolate dominant modifiers of the rough eye phenotype associated with eye-specific expression of yanACT. Approximately 190,000 mutagenized flies were screened, and 260 enhancers and 90 suppressors were obtained. Among the previously known genes we recovered are four RTK pathway components, rolled (MAPK), son-of-sevenless, Star, and pointed, and two genes, eyes absent and string, that have not been implicated previously in RTK signaling events. We also isolated mutations in five previously uncharacterized genes, one of which, split ends, we have characterized molecularly and have shown to encode a member of the RRM family of RNA-binding proteins.

A Study of Arabidopsis Cold Stress Tolerance Improvement via AthHOS1-Targeting HOS1-AmiRNA Approach

2021 ◽  
Author(s):  
Marzieh Karimi ◽  
Behrouz Shiran ◽  
Mohammad Rabei ◽  
Hossein Fallahi ◽  
Bojana Banović Đeri
Keyword(s):  
Gene Expression ◽  
Arabidopsis Thaliana ◽  
Circadian Rhythm ◽  
Comparative Analysis ◽  
Low Temperature ◽  
Transgenic Plants ◽  
Cold Stress ◽  
Stress Tolerance ◽  
Low Temperatures ◽  
Wild Type

Abstract In this study the artificial microRNAs (amiRNAs) technology targeting HOS1 gene was tested for its applicability for the improvement of cold stress tolerance in Landsberg-0 (Ler-0) ecotype of Arabidopsis thaliana. The chosen approach was designed to suppress AtHOS1 gene expression through the overexpression of amiRNA-HOS1. The effect of AtHOS1-amiRNA overexpression to transgenic plants’ response to cold stress was determined by Real Time PCR. The expression levels of amiRNA and its target, AtHOS1 gene, were observed in 3-week old seedlings of T3 generation and in wild-type plants after 6h, 12h, 24h, 48h and 96h of their exposure to cold stress (4ºC). Comparative analysis revealed that AtHOS1-amiRNA negatively regulated AtHOS1 in transgenic plants upon plants lengthen exposure (for 48h and 96h) to low temperature (Pearson’s correlation coefficient of -0.407; P < 0.05). Even though prolonged cold stress caused extended up regulation of AtHOS1 in wild type plants, in transgenic plants AtHOS1-amiRNA suppression disturbed expected AtHOS1 circadian rhythm by preventing further AtHOS1 up regulation. Moreover, transgenic plants showed AtHOS1 down regulation 96h after the cold stress onset, due to sufficient overexpression of AtHOS1-amiRNA, which allowed cold signaling amplification in transgenic plants. As a result of that, cold-acclimated transformed plants displayed 17% higher freezing tolerance (-1°C to -8°C) in comparison to wild type plants, demonstrating the success of chosen approach in improving Arabidopsis tolerance to low temperatures, at least in Ler-0 ecotype.

scholarly journals OsCRP1, a Ribonucleoprotein Gene, Regulates Chloroplast mRNA Stability That Confers Drought and Cold Tolerance

2021 ◽  
Vol 22 (4) ◽  
pp. 1673
Author(s):  
Seung Woon Bang ◽  
Ho Suk Lee ◽  
Su-Hyun Park ◽  
Dong-Keun Lee ◽  
Jun Sung Seo ◽  
...  
Keyword(s):  
Cold Stress ◽  
Stress Tolerance ◽  
Cold Tolerance ◽  
Molecular Mechanisms ◽  
Rna Binding ◽  
Mrna Levels ◽  
Wild Type ◽  
Rna Immunoprecipitation ◽  
Chloroplast Rna ◽  
Chloroplast Mrna

Chloroplast ribonucleoproteins (cpRNPs) are nuclear-encoded and highly abundant proteins that are proposed to function in chloroplast RNA metabolism. However, the molecular mechanisms underlying the regulation of chloroplast RNAs involved in stress tolerance are poorly understood. Here, we demonstrate that CHLOROPLAST RNA-BINDING PROTEIN 1 (OsCRP1), a rice (Oryza sativa) cpRNP gene, is essential for stabilization of RNAs from the NAD(P)H dehydrogenase (NDH) complex, which in turn enhances drought and cold stress tolerance. An RNA-immunoprecipitation assay revealed that OsCRP1 is associated with a set of chloroplast RNAs. Transcript profiling indicated that the mRNA levels of genes from the NDH complex significantly increased in the OsCRP1 overexpressing compared to non-transgenic plants, whereas the pattern in OsCRP1 RNAi plants were opposite. Importantly, the OsCRP1 overexpressing plants showed a higher cyclic electron transport (CET) activity, which is essential for elevated levels of ATP for photosynthesis. Additionally, overexpression of OsCRP1 resulted in significantly enhanced drought and cold stress tolerance with higher ATP levels compared to wild type. Thus, our findings suggest that overexpression of OsCRP1 stabilizes a set of mRNAs from genes of the NDH complex involved in increasing CET activity and production of ATP, which consequently confers enhanced drought and cold tolerance.

scholarly journals OsmiR528 Enhances Cold Stress Tolerance by Repressing Expression of Stress Response-related Transcription Factor Genes in Plant Cells

2019 ◽  
Vol 20 (2) ◽  
pp. 100-114 ◽  
Author(s):  
Wei Tang ◽  
Wells A. Thompson
Keyword(s):  
Transcription Factor ◽  
Stress Response ◽  
Low Temperature ◽  
Cold Stress ◽  
Stress Tolerance ◽  
Molecular Mechanisms ◽  
Plant Cells ◽  
Temperature Tolerance ◽  
Low Temperature Tolerance ◽  
Related Transcription Factor

Background: MicroRNAs participate in many molecular mechanisms and signaling transduction pathways that are associated with plant stress tolerance by repressing expression of their target genes. However, how microRNAs enhance tolerance to low temperature stress in plant cells remains elusive. Objective: In this investigation, we demonstrated that overexpression of the rice microRNA528 (OsmiR528) increases cell viability, growth rate, antioxidants content, ascorbate peroxidase (APOX) activity, and superoxide dismutase (SOD) activity and decreases ion leakage rate and thiobarbituric acid reactive substances (TBARS) under low temperature stress in Arabidopsis (Arabidopsis thaliana), pine (Pinus elliottii), and rice (Oryza sativa). Methods: To investigate the potential mechanism of OsmiR528 in increasing cold stress tolerance, we examined expression of stress-associated MYB transcription factors OsGAMYB-like1, OsMYBS3, OsMYB4, OsMYB3R-2, OsMYB5, OsMYB59, OsMYB30, OsMYB1R, and OsMYB20 in rice cells by qRT-PCR. Results: Our experiments demonstrated that OsmiR528 decreases expression of transcription factor OsMYB30 by targeting a F-box domain containing protein gene (Os06g06050), which is a positive regulator of OsMYB30. In OsmiR528 transgenic rice, reduced OsMYB30 expression results in increased expression of BMY genes OsBMY2, OsBMY6, and OsBMY10. The transcript levels of the OsBMY2, OsBMY6, and OsBMY10 were elevated by OsMYB30 knockdown, but decreased by Os- MYB30 overexpression in OsmiR528 transgenic cell lines, suggesting that OsmiR528 increases low temperature tolerance by modulating expression of stress response-related transcription factor. Conclusion: Our experiments provide novel information in increasing our understanding in molecular mechanisms of microRNAs-associated low temperature tolerance and are valuable in plant molecular breeding from monocotyledonous, dicotyledonous, and gymnosperm plants.

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