Overexpression of CDSP32 (GhTRX134) Cotton Gene Enhances Drought, Salt, and Oxidative Stress Tolerance in Arabidopsis

Upland cotton (Gossypium hirsutum L.) is the main natural fiber crop worldwide and is an essential source of seed oil and biofuel products. Many abiotic stresses, such as drought and salinity, constrain cotton production. Thioredoxins (TRXs) are a group of small ubiquitous proteins that are widely distributed among organisms. TRXs play a crucial role in regulating diverse functions during plant growth and development. In the present study, a novel GhTRX134 gene was characterized and overexpressed in Arabidopsis and silenced in cotton under drought stress. Furthermore, the proline content and enzyme activity levels were measured in transgenic plants and wild-type (Wt) plants under drought and salt stress. The results revealed that the overexpression of GhTRX134 enhanced abiotic stress tolerance. When GhTRX134 was silenced, cotton plants become more sensitive to drought. Taken together, these findings confirmed that the overexpression of GhTRX134 improved drought and salt tolerance in Arabidopsis plants. Therefore, the GhTRX134 gene can be transformed into cotton plants to obtain transgenic lines for more functional details.

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Knockdown of 60S ribosomal protein L14-2 reveals their potential regulatory roles to enhance drought and salt tolerance in cotton

Journal of Cotton Research ◽

10.1186/s42397-021-00102-7 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Margaret Linyerera SHIRAKU ◽

Richard Odongo MAGWANGA ◽

Xiaoyan CAI ◽

Joy Nyangasi KIRUNGU ◽

Yanchao XU ◽

...

Keyword(s):

Salt Stress ◽

Ribosomal Protein ◽

Large Scale ◽

Natural Fiber ◽

Substantial Reduction ◽

Regulatory Elements ◽

Scale Production ◽

Drought And Salt Tolerance ◽

Drought And Salt Stress ◽

Effects Of Drought

Abstract Background Cotton is a valuable economic crop and the main significant source of natural fiber for textile industries globally. The effects of drought and salt stress pose a challenge to strong fiber and large-scale production due to the ever-changing climatic conditions. However, plants have evolved a number of survival strategies, among them is the induction of various stress-responsive genes such as the ribosomal protein large (RPL) gene. The RPL gene families encode critical proteins, which alleviate the effects of drought and salt stress in plants. In this study, comprehensive and functional analysis of the cotton RPL genes was carried out under drought and salt stresses. Results Based on the genome-wide evaluation, 26, 8, and 5 proteins containing the RPL14B domain were identified in Gossypium hirsutum, G. raimondii, and G. arboreum, respectively. Furthermore, through bioinformatics analysis, key cis-regulatory elements related to RPL14B genes were discovered. The Myb binding sites (MBS), abscisic acid-responsive element (ABRE), CAAT-box, TATA box, TGACG-motif, and CGTCA-motif responsive to methyl jasmonate, as well as the TCA-motif responsive to salicylic acid, were identified. Expression analysis revealed a key gene, Gh_D01G0234 (RPL14B), with significantly higher induction levels was further evaluated through a reverse genetic approach. The knockdown of Gh_D01G0234 (RPL14B) significantly affected the performance of cotton seedlings under drought/salt stress conditions, as evidenced by a substantial reduction in various morphological and physiological traits. Moreover, the level of the antioxidant enzyme was significantly reduced in VIGS-plants, while oxidant enzyme levels increased significantly, as demonstrated by the higher malondialdehyde concentration level. Conclusion The results revealed the potential role of the RPL14B gene in promoting the induction of antioxidant enzymes, which are key in oxidizing the various oxidants. The key pathways need to be investigated and even as we exploit these genes in the developing of more stress-resilient cotton germplasms.

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De-novo transcriptome analysis unveils differentially expressed genes regulating drought and salt stress response in Panicum sumatrense

Scientific Reports ◽

10.1038/s41598-020-78118-3 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Rasmita Rani Das ◽

Seema Pradhan ◽

Ajay Parida

Keyword(s):

Gene Expression ◽

Salt Stress ◽

Abiotic Stress ◽

Stress Tolerance ◽

Differentially Expressed Genes ◽

Salinity Stress ◽

Abiotic Stress Tolerance ◽

Differentially Expressed ◽

Little Millet ◽

Drought And Salt Stress

AbstractScreening the transcriptome of drought tolerant variety of little millet (Panicum sumatrense), a marginally cultivated, nutritionally rich, susbsistent crop, can identify genes responsible for its hardiness and enable identification of new sources of genetic variation which can be used for crop improvement. RNA-Seq generated ~ 230 million reads from control and treated tissues, which were assembled into 86,614 unigenes. In silico differential gene expression analysis created an overview of patterns of gene expression during exposure to drought and salt stress. Separate gene expression profiles for leaf and root tissue revealed the differences in regulatory mechanisms operating in these tissues during exposure to abiotic stress. Several transcription factors were identified and studied for differential expression. 61 differentially expressed genes were found to be common to both tissues under drought and salinity stress and were further validated using qRT-PCR. Transcriptome of P. sumatrense was also used to mine for genic SSR markers relevant to abiotic stress tolerance. This study is first report on a detailed analysis of molecular mechanisms of drought and salinity stress tolerance in a little millet variety. Resources generated in this study can be used as potential candidates for further characterization and to improve abiotic stress tolerance in food crops.

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Genome-wide analysis of Myo-inositol oxygenase gene family in tomato reveals their involvement in ascorbic acid accumulation

10.21203/rs.2.14506/v1 ◽

2019 ◽

Author(s):

Shoaib Munir ◽

Muhammad Ali Mumtaz ◽

John Kojo Ahiakpa ◽

Genzhong Liu ◽

Wei Zheng ◽

...

Keyword(s):

Ascorbic Acid ◽

Abiotic Stress ◽

Gene Family ◽

Stress Tolerance ◽

Abiotic Stress Tolerance ◽

Light Response ◽

Genome Wide ◽

Genes Encoding ◽

Transgenic Lines ◽

Distribution Mapping

Abstract Background Ascorbic acid (Vitamin C, AsA) is an antioxidant metabolite involved in plant development and environmental stimuli. AsA biosynthesis has been well studied in plants and MIOX is a critical enzyme in plants AsA biosynthesis pathway. However, myo-inositol oxygenase (MIOX) gene family members and their involvement in AsA biosynthesis and response to abiotic stress remain unclear. Results In this study, five tomato genes encoding MIOX proteins and possessing MIOX motifs were identified. Structural analysis and distribution mapping showed that 5 MIOX genes contain different intron/exon patterns and unevenly distributed among four chromosomes. In addition, expression analyses indicated the remarkable expression of SlMIOX genes in different plant tissues. Furthermore, transgenic lines were obtained by over-expression of MIOX4 gene in tomato. The overexpression lines showed a significant increase in total ascorbate in leaves and red fruits compared to control. Expression analysis revealed that increased accumulation of AsA in MIOX4 overexpression lines is possible as a consequence of the multiple genes involved in AsA biosynthesis. Myo inositol (MI) feeding in leaf and fruit implied that AsA biosynthesis was mainly improved by Myo-inositol pathway in leaves and fruits. MIOX4 overexpression lines exhibited a betterlight response, abiotic stress tolerance, and AsA biosynthesis capacity. Conclusions These results showed that MIOX4 transgenic lines contribute to AsA biosynthesis, evident as better light response and improved oxidative stress tolerance. This study provides the first comprehensive analysis of MIOX gene family and their involvement in ascorbate biosynthesis in tomato.

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A Novel Sweetpotato WRKY Transcription Factor, IbWRKY2, Positively Regulates Drought and Salt Tolerance in Transgenic Arabidopsis

Biomolecules ◽

10.3390/biom10040506 ◽

2020 ◽

Vol 10 (4) ◽

pp. 506 ◽

Cited By ~ 6

Author(s):

Hong Zhu ◽

Yuanyuan Zhou ◽

Hong Zhai ◽

Shaozhen He ◽

Ning Zhao ◽

...

Keyword(s):

Salt Tolerance ◽

Ipomoea Batatas ◽

Transgenic Arabidopsis ◽

Abiotic Stress Tolerance ◽

Wrky Transcription Factor ◽

Oxygen Species ◽

Aba Signaling ◽

Ros Scavenging ◽

Drought And Salt Tolerance ◽

Transgenic Lines

WRKYs play important roles in plant growth, defense regulation, and stress response. However, the mechanisms through which WRKYs are involved in drought and salt tolerance have been rarely characterized in sweetpotato [Ipomoea batatas (L.) Lam.]. In this study, we cloned a WRKY gene, IbWRKY2, from sweetpotato and its expression was induced with PEG6000, NaCl, and abscisic acid (ABA). The IbWRKY2 was localized in the nucleus. The full-length protein exhibited transactivation activity, and its active domain was located in the N-terminal region. IbWRKY2-overexpressing Arabidopsis showed enhanced drought and salt tolerance. After drought and salt treatments, the contents of ABA and proline as well as the activity of superoxide dismutase (SOD) were higher in transgenic plants, while the malondialdehyde (MDA) and H2O2 contents were lower. In addition, several genes related to the ABA signaling pathway, proline biosynthesis, and the reactive oxygen species (ROS)-scavenging system, were significantly up-regulated in transgenic lines. These results demonstrate that IbWRKY2 confers drought and salt tolerance in Arabidopsis. Furthermore, IbWRKY2 was able to interact with IbVQ4, and the expression of IbVQ4 was induced by drought and salt treatments. These results provide clues regarding the mechanism by which IbWRKY2 contributes to the regulation of abiotic stress tolerance.

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Target of Rapamycin: function in abiotic stress tolerance in Arabidopsis and its involvement in a possible cross-talk with ribosomal proteins

10.1101/2020.01.15.907899 ◽

2020 ◽

Cited By ~ 1

Author(s):

Achala Bakshi ◽

Mazahar Moin ◽

M. S. Madhav ◽

Meher B. Gayatri ◽

Aramati B. M. Reddy ◽

...

Keyword(s):

Ribosomal Protein ◽

Stress Tolerance ◽

Protein Function ◽

Ribosomal Proteins ◽

Abiotic Stress Tolerance ◽

Small Subunit ◽

Target Of Rapamycin ◽

Chlorophyll Contents ◽

Transgenic Lines

AbstractThe Target of Rapamycin (TOR) protein kinase reprograms cellular metabolism under various environmental stresses. The overexpression of TOR in Arabidopsis resulted in increased plant growth including yield and biomass when compared with the wild type under both controlled and limited water conditions. In the present investigation, we report that Arabidopsis plants overexpressing TOR exhibited enhanced tolerance to the osmotic and salt stress treatments. Further to determine the role of TOR in abiotic stresses other than water limiting conditions, which were observed earlier in rice, we have treated high and medium TOR expressing Arabidopsis plants, ATR-1.4.27 and ATR-3.7.32 respectively, with stress-inducing chemical agents such as Mannitol (100 mM), NaCl (150 mM), Sorbitol (200 mM) and PEG (7%). Both the lines, ATR-1.4.27 and ATR-3.7.32 exhibited enhanced tolerance to these stresses. These lines also had increased proline and total chlorophyll contents under stress conditions compared with their corresponding WT counterparts. The upregulation of several osmotic stress inducible genes in Arabidopsis transgenic lines indicated the role of TOR in modulating multi-stress tolerance. In the present investigation, we have also analyzed the transcriptional upregulation of ribosomal protein large and small subunit (RPL and RPS) genes in AtTOR overexpressing rice transgenic lines, TR-2.24 and TR-15.1 generated earlier (Bakshi et al., 2017a), which indicated that TOR also positively regulates the transcription of ribosomal proteins (RP) along with the synthesis of rRNAs. Also, the observations from phosphoproteomic analysis in SALK lines of various Arabidopsis T-DNA insertion mutants of ribosomal proteins showed differential regulation in phosphorylation of p70kDa ribosomal protein S6K1 and comparative analysis of phosphorylation sites for RSK (Ribosomal S6 Kinases) in RPL6, RPL18, RPL23, RPL24 and RPS28C proteins of Arabidopsis, Interestingly, rice showed similarity in their peptide sequences and Ser/Thr positions. These results suggest that the phosphorylation of S6K1 is controlled by loss/ inhibition of ribosomal protein function to switch ‘on’/ ‘off’ the translational regulation for balanced growth and the pathways of both RPs and TOR are interlinked in a cyclic manner via phosphorylation of S6K1 as a modulatory step.

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AnDHN, a Dehydrin Protein From Ammopiptanthus nanus, Mitigates the Negative Effects of Drought Stress in Plants

Frontiers in Plant Science ◽

10.3389/fpls.2021.788938 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yibo Sun ◽

Linghao Liu ◽

Shaokun Sun ◽

Wangzhen Han ◽

Muhammad Irfan ◽

...

Keyword(s):

Abiotic Stress ◽

Stress Tolerance ◽

Abiotic Stress Tolerance ◽

Germination Rate ◽

Root System Architecture ◽

Ros Scavenging ◽

Negative Effects ◽

Ammopiptanthus Nanus ◽

Transgenic Lines ◽

Effects Of Drought

Dehydrins (DHNs) play crucial roles in a broad spectrum of abiotic stresses in model plants. However, the evolutionary role of DHNs has not been explored, and the function of DHN proteins is largely unknown in Ammopiptanthus nanus (A. nanus), an ancient and endangered legume species from the deserts of northwestern China. In this study, we isolated a drought-response gene (c195333_g1_i1) from a drought-induced RNA-seq library of A. nanus. Evolutionary bioinformatics showed that c195333_g1_i1 is an ortholog of Arabidopsis DHN, and we renamed it AnDHN. Moreover, DHN proteins may define a class of proteins that are evolutionarily conserved in all angiosperms that have experienced a contraction during the evolution of legumes. Arabidopsis plants overexpressing AnDHN exhibited morpho-physiological changes, such as an increased germination rate, higher relative water content (RWC), higher proline (PRO) content, increased peroxidase (POD) and catalase (CAT) activities, lower contents of malondialdehyde (MDA), H2O2 and O2–, and longer root length. Our results showed that the transgenic lines had improved drought resistance with deep root system architecture, excellent water retention, increased osmotic adjustment, and enhanced reactive oxygen species (ROS) scavenging. Furthermore, the transgenic lines also had enhanced salt and cold tolerance. Our findings demonstrate that AnDHN may be a good candidate gene for improving abiotic stress tolerance in crops.Key Message: Using transcriptome analysis in Ammopiptanthus nanus, we isolated a drought-responsive gene, AnDHN, that plays a key role in enhancing abiotic stress tolerance in plants, with strong functional diversification in legumes.

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Barley Plants Overexpressing Ferrochelatases (HvFC1 and HvFC2) Show Improved Photosynthetic Rates and Have Reduced Photo-Oxidative Damage under Drought Stress than Non-Transgenic Controls

Agronomy ◽

10.3390/agronomy10091351 ◽

2020 ◽

Vol 10 (9) ◽

pp. 1351

Author(s):

Dilrukshi S. K. Nagahatenna ◽

Boris Parent ◽

Everard J. Edwards ◽

Peter Langridge ◽

Ryan Whitford

Keyword(s):

Oxidative Stress ◽

Drought Stress ◽

Oxidative Damage ◽

Stress Tolerance ◽

Nuclear Gene ◽

Defense Responses ◽

Photosynthetic Performance ◽

Free Heme ◽

Transgenic Lines ◽

And Oxidative Stress

We investigated the roles of two Ferrochelatases (FCs), which encode the terminal enzyme for heme biosynthesis, in drought and oxidative stress tolerance in model cereal plant barley (Hordeum vulgare). Three independent transgenic lines ectopically overexpressing either barley FC1 or FC2 were selected and evaluated under well-watered, drought, and oxidative stress conditions. Both HvFC1 and HvFC2 overexpressing transgenics showed delayed wilting and maintained higher photosynthetic performance relative to controls, after exposure to soil dehydration. In each case, HvFC overexpression significantly upregulated the nuclear genes associated with detoxification of reactive oxygen species (ROS) upon drought stress. Overexpression of HvFCs, also suppressed photo-oxidative damage induced by the deregulated tetrapyrrole biosynthesis mutant tigrinad12. Previous studies suggest that only FC1 is implicated in stress defense responses, however, our study demonstrated that both FC1 and FC2 affect drought stress tolerance. As FC-derived free heme was proposed as a chloroplast-to-nuclear signal, heme could act as an important signal, stimulating drought responsive nuclear gene expression. This study also highlighted tetrapyrrole biosynthetic enzymes as potential targets for engineering improved crop performance, both in well-watered and water-limited environments.

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The WRKY Transcription Factor GmWRKY12 Confers Drought and Salt Tolerance in Soybean

International Journal of Molecular Sciences ◽

10.3390/ijms19124087 ◽

2018 ◽

Vol 19 (12) ◽

pp. 4087 ◽

Cited By ~ 21

Author(s):

Wen-Yan Shi ◽

Yong-Tao Du ◽

Jian Ma ◽

Dong-Hong Min ◽

Long-Guo Jin ◽

...

Keyword(s):

Abiotic Stress ◽

Salt Tolerance ◽

Stress Responses ◽

Abiotic Stress Tolerance ◽

Wrky Transcription Factor ◽

Rna Seq ◽

Soybean Seedlings ◽

Drought And Salt Tolerance ◽

Drought And Salt Stress ◽

Mda Content

WRKYs are important regulators in plant development and stress responses. However, knowledge of this superfamily in soybean is limited. In this study, we characterized the drought- and salt-induced gene GmWRKY12 based on RNA-Seq and qRT-PCR. GmWRKY12, which is 714 bp in length, encoded 237 amino acids and grouped into WRKY II. The promoter region of GmWRKY12 included ABER4, MYB, MYC, GT-1, W-box and DPBF cis-elements, which possibly participate in abscisic acid (ABA), drought and salt stress responses. GmWRKY12 was minimally expressed in different tissues under normal conditions but highly expressed under drought and salt treatments. As a nucleus protein, GmWRKY12 was responsive to drought, salt, ABA and salicylic acid (SA) stresses. Using a transgenic hairy root assay, we further characterized the roles of GmWRKY12 in abiotic stress tolerance. Compared with control (Williams 82), overexpression of GmWRKY12 enhanced drought and salt tolerance, increased proline (Pro) content and decreased malondialdehyde (MDA) content under drought and salt treatment in transgenic soybean seedlings. These results may provide a basis to understand the functions of GmWRKY12 in abiotic stress responses in soybean.

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Increasing seed thiamin content impacts stored carbon partitioning and subsequent seedling stress tolerance

10.1101/2021.08.03.454933 ◽

2021 ◽

Author(s):

Mohammad Yazdani ◽

James Aaron Davis ◽

Jeffrey F. Harper ◽

David K Shintani

Keyword(s):

Stress Tolerance ◽

Abiotic Stress Tolerance ◽

Carbon Partitioning ◽

Biotic And Abiotic Stresses ◽

Transgenic Seeds ◽

Essential Components ◽

Enhanced Expression ◽

Transgenic Lines ◽

Living Organisms ◽

Thiamin Pyrophosphate

Thiamin and thiamin pyrophosphate (TPP) are essential components for the function of enzymes involved in the metabolism of carbohydrates and amino acids in living organisms. In addition to its role as a cofactor, thiamin plays a key role in resistance against biotic and abiotic stresses in plants. Most of the studies used exogenous thiamin to enhance stress tolerance in plants. In this study, we achieved this objective by genetically engineering Arabidopsis thaliana and Camelina sativa for the seed-specific co-overexpression of the Arabidopsis thiamin biosynthetic genes Thi4, ThiC, and ThiE. Elevated thiamin content in the seeds of transgenic plants was accompanied by the enhanced expression levels of transcripts encoding thiamin cofactor-dependent enzymes. Furthermore, seed germination and root growth in thiamin over-producing lines were more tolerant to oxidative stress caused by salt and paraquat treatments. The transgenic seeds also accumulated more oil (up to16.4% in Arabidopsis and17.9% in C. sativa) and carbohydrate but less protein than the control seeds. The same results were also observed in TPP over-producing Arabidopsis plants generated by the seed-specific overexpression of TPK1. Together, our findings suggest that thiamin and TPP over-production in transgenic lines confer a boosted abiotic stress tolerance and alter the seed carbon partitioning as well.

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Genome-wide analysis of Myo-inositol oxygenase gene family in tomato reveals their involvement in ascorbic acid accumulation

10.21203/rs.2.14506/v3 ◽

2020 ◽

Author(s):

Shoaib Munir ◽

Muhammad Ali Mumtaz ◽

John Kojo Ahiakpa ◽

Genzhong Liu ◽

Weifang Chen ◽

...

Keyword(s):

Ascorbic Acid ◽

Abiotic Stress ◽

Gene Family ◽

Stress Tolerance ◽

Abiotic Stress Tolerance ◽

Light Response ◽

Genome Wide ◽

Genes Encoding ◽

Transgenic Lines ◽

Distribution Mapping

Abstract Background Ascorbic acid (Vitamin C, AsA) is an antioxidant metabolite involved in plant development and environmental stimuli. AsA biosynthesis has been well studied in plants, and MIOX is a critical enzyme in plants AsA biosynthesis pathway. However, Myo-inositol oxygenase (MIOX) gene family members and their involvement in AsA biosynthesis and response to abiotic stress remain unclear. Results In this study, five tomato genes encoding MIOX proteins and possessing MIOX motifs were identified. Structural analysis and distribution mapping showed that 5 MIOX genes contain different intron/exon patterns and unevenly distributed among four chromosomes. Besides, expression analyses indicated the remarkable expression of SlMIOX genes in different plant tissues. Furthermore, transgenic lines were obtained by over-expression of the MIOX4 gene in tomato. The overexpression lines showed a significant increase in total ascorbate in leaves and red fruits compared to control. Expression analysis revealed that increased accumulation of AsA in MIOX4 overexpression lines is possible as a consequence of the multiple genes involved in AsA biosynthesis. Myo inositol (MI) feeding in leaf and fruit implied that the Myo-inositol pathway improved the AsA biosynthesis in leaves and fruits. MIOX4 overexpression lines exhibited abetter light response, abiotic stress tolerance, and AsA biosynthesis capacity. Conclusions These results showed that MIOX4 transgenic lines contribute to AsA biosynthesis, evident as better light response and improved oxidative stress tolerance. This study provides the first comprehensive analysis of the MIOX gene family and their involvement in ascorbate biosynthesis in tomato.

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