Functional characterization of a Glycine soja Ca2+ATPase in salt–alkaline stress responses

Abstract Background: WRKY transcription factors are a superfamily of regulators involved in diverse biological processes and stress responses in plants. However, knowledge is limited for WRKY family in camelina (Camelina sativa), an important Brassicaceae oil crop with strong tolerance against various stresses. Here, genome-wide characterization of WRKY proteins is performed to examine their gene-structures, phylogenetics, expressions, conserved motif organizations, and functional annotation to identify candidate WRKYs mediating regulation of stress resistance in camelina.Results: Total of 242 CsWRKY proteins encoded by 224 gene loci distributed uneven on chromosomes were identified, and classified into three groups via phylogenetic analysis according to their WRKY domains and zinc finger motifs. 15 CsWRKY gene loci generated 33 spliced variants. Orthologous WRKY gene pairs were identified, with 173 pairs in C. sativa and Arabidopsis genomes as well as 282 pairs for C. sativa and B. napus, respectively. 137 segmental duplication events were observed but no tandem duplication in camelina genome. Ten major conserved motifs were examined, with WRKYGQK as the most conserved and several variants existed in many CsWRKYs. Expression analysis revealed that half more CsWRKY genes were expressed constitutively, and a set of them had a tissue-specific expression. Notably, 11 CsWRKY genes exhibited significantly expression changes in plant seedlings under cold, salt, and drought stress, respectively, having preferentially inducible expression pattern in response to the stress.Conclusions: The present described a detail analysis of CsWRKY gen family and their expression profiled in twelve tissues and under several stress conditions. Segmental duplication is the major force for large expansion of this gene family, and a strong purifying pressure happened for CsWRKY proteins evolutionally. CsWRKY proteins play important roles for plant development, with differential functions in different tissues. Exceptionally, eleven CsWRKYs, particularly five alternative spliced isoforms were found to be the key players possibly in mediating plant response to various stresses. Overall, our results provide a foundation for understanding roles of CsWRKYs and the precise mechanism through which CsWRKYs regulate high stress resistance to stress as well as development of stress tolerance cultivars for Cruciferae crops.

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sRNA Target Prediction Organizing Tool (SPOT) Integrates Computational and Experimental Data To Facilitate Functional Characterization of Bacterial Small RNAs

mSphere ◽

10.1128/msphere.00561-18 ◽

2019 ◽

Vol 4 (1) ◽

Cited By ~ 4

Author(s):

Alisa M. King ◽

Carin K. Vanderpool ◽

Patrick H. Degnan

Keyword(s):

Experimental Data ◽

Stress Responses ◽

Small Rnas ◽

Metabolic Regulation ◽

Functional Characterization ◽

Target Prediction ◽

Structure Stability ◽

Accurate Identification ◽

Mrna Targets

Small RNAs (sRNAs) regulate gene expression in diverse bacteria by interacting with mRNAs to change their structure, stability, or translation. Hundreds of sRNAs have been identified in bacteria, but characterization of their regulatory functions is limited by difficulty with sensitive and accurate identification of mRNA targets. Thus, new robust methods of bacterial sRNA target identification are in demand. Here, we describe our small RNA target prediction organizing tool (SPOT), which streamlines the process of sRNA target prediction by providing a single pipeline that combines available computational prediction tools with customizable results filtering based on experimental data. SPOT allows the user to rapidly produce a prioritized list of predicted sRNA-target mRNA interactions that serves as a basis for further experimental characterization. This tool will facilitate elucidation of sRNA regulons in bacteria, allowing new discoveries regarding the roles of sRNAs in bacterial stress responses and metabolic regulation.

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Comprehensive genomic characterization of NAC transcription factor family and their response to salt and drought stress in peanut

BMC Plant Biology ◽

10.1186/s12870-020-02678-9 ◽

2020 ◽

Vol 20 (1) ◽

Author(s):

Cuiling Yuan ◽

Chunjuan Li ◽

Xiaodong Lu ◽

Xiaobo Zhao ◽

Caixia Yan ◽

...

Keyword(s):

Transcription Factor ◽

Drought Stress ◽

Stress Responses ◽

Expression Patterns ◽

Functional Characterization ◽

Nac Transcription Factor ◽

Rna Seq ◽

A Genome ◽

Salt And Drought Stress

Abstract Background Peanut is one of the most important oil crop species worldwide. NAC transcription factor (TF) genes play important roles in the salt and drought stress responses of plants by activating or repressing target gene expression. However, little is known about NAC genes in peanut. Results We performed a genome-wide characterization of NAC genes from the diploid wild peanut species Arachis duranensis and Arachis ipaensis, which included analyses of chromosomal locations, gene structures, conserved motifs, expression patterns, and cis-acting elements within their promoter regions. In total, 81 and 79 NAC genes were identified from A. duranensis and A. ipaensis genomes. Phylogenetic analysis of peanut NACs along with their Arabidopsis and rice counterparts categorized these proteins into 18 distinct subgroups. Fifty-one orthologous gene pairs were identified, and 46 orthologues were found to be highly syntenic on the chromosomes of both A. duranensis and A. ipaensis. Comparative RNA sequencing (RNA-seq)-based analysis revealed that the expression of 43 NAC genes was up- or downregulated under salt stress and under drought stress. Among these genes, the expression of 17 genes in cultivated peanut (Arachis hypogaea) was up- or downregulated under both stresses. Moreover, quantitative reverse transcription PCR (RT-qPCR)-based analysis revealed that the expression of most of the randomly selected NAC genes tended to be consistent with the comparative RNA-seq results. Conclusion Our results facilitated the functional characterization of peanut NAC genes, and the genes involved in salt and drought stress responses identified in this study could be potential genes for peanut improvement.

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Genome-wide identification and functional characterization of Camelina sativa WRKY gene family in response to abiotic stress

10.21203/rs.3.rs-36113/v2 ◽

2020 ◽

Author(s):

Yanan Song ◽

Hongli Cui ◽

Ying Shi ◽

Jinai Xue ◽

Chunli Ji ◽

...

Keyword(s):

Gene Family ◽

Stress Responses ◽

Stress Resistance ◽

Segmental Duplication ◽

Functional Characterization ◽

High Stress ◽

Camelina Sativa ◽

Genome Wide ◽

Large Expansion

Abstract Background: WRKY transcription factors are a superfamily of regulators involved in diverse biological processes and stress responses in plants. However, knowledge is limited for WRKY family in camelina (Camelina sativa), an important Brassicaceae oil crop with strong tolerance against various stresses. Here, genome-wide characterization of WRKY proteins is performed to examine their gene-structures, phylogenetics, expressions, conserved motif organizations, and functional annotation to identify candidate WRKYs mediating regulation of stress resistance in camelina. Results: Total of 242 CsWRKY proteins encoded by 224 gene loci distributed uneven on chromosomes were identified, and classified into three groups via phylogenetic analysis according to their WRKY domains and zinc finger motifs. 15 CsWRKY gene loci generated 33 spliced variants. Orthologous WRKY gene pairs were identified, with 173 pairs in C. sativa and Arabidopsis genomes as well as 282 pairs for C. sativa and B. napus, respectively. 137 segmental duplication events were observed but no tandem duplication in camelina genome. Ten major conserved motifs were examined, with WRKYGQK as the most conserved and several variants existed in many CsWRKYs. Expression analysis revealed that half more CsWRKY genes were expressed constitutively, and a set of them had a tissue-specific expression. Notably, 11 CsWRKY genes exhibited significantly expression changes in plant seedlings under cold, salt, and drought stress, respectively, having preferentially inducible expression pattern in response to the stress. Conclusions: The present described a detail analysis of CsWRKY gen family and their expression profiled in twelve tissues and under several stress conditions. Segmental duplication is the major force for large expansion of this gene family, and a strong purifying pressure happened for CsWRKY proteins evolutionally. CsWRKY proteins play important roles for plant development, with differential functions in different tissues. Exceptionally, eleven CsWRKYs, particularly five alternative spliced isoforms were found to be the key players possibly in mediating plant response to various stresses. Overall, our results provide a foundation for understanding roles of CsWRKYs and the precise mechanism through which CsWRKYs regulate high stress resistance to stress as well as development of stress tolerance cultivars for Cruciferae crops.

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Expression of the Malus sieversii NF-YB21 Encoded Gene Confers Tolerance to Osmotic Stresses in Arabidopsis thaliana

International Journal of Molecular Sciences ◽

10.3390/ijms22189777 ◽

2021 ◽

Vol 22 (18) ◽

pp. 9777

Author(s):

Chen Feng ◽

Yanyan Wang ◽

Yueting Sun ◽

Xiang Peng ◽

Xiang Zhang ◽

...

Keyword(s):

Stress Responses ◽

Functional Characterization ◽

Compatible Solutes ◽

Antioxidant Systems ◽

Drought Treatment ◽

Malus Sieversii ◽

Yield And Quality ◽

Genes Encoding ◽

Physiological Analysis

Drought is the main environmental factor that limits the yield and quality of apples (Malus × domestica) grown in arid and semi-arid regions. Nuclear factor Ys (NF-Ys) are important transcription factors involved in the regulation of plant growth, development, and various stress responses. However, the function of NF-Y genes is poorly understood in apples. Here, we identified 43 NF-Y genes in the genome of apples and conducted an initial functional characterization of the apple NF-Y. Expression analysis of NF-Y members in M. sieversii revealed that a large number of NF-Ys were highly expressed in the roots compared with the leaves, and a large proportion of NF-Y genes responded to drought treatment. Furthermore, heterologous expression of MsNF-YB21, which was significantly upregulated by drought, led to a longer root length and, thus, conferred improved osmotic and salt tolerance in Arabidopsis. Moreover, the physiological analysis of MsNF-YB21 overexpression revealed enhanced antioxidant systems, including antioxidant enzymes and compatible solutes. In addition, genes encoding catalase (AtCAT2, AtCAT3), superoxide dismutase (AtFSD1, AtFSD3, AtCSD1), and peroxidase (AtPER12, AtPER42, AtPER47, AtPER51) showed upregulated expression in the MsNF-YB21 overexpression lines. These results for the MsNF-Y gene family provide useful information for future studies on NF-Ys in apples, and the functional analysis of MsNF-YB21 supports it as a potential target in the improvement of apple drought tolerance via biotechnological strategies.

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Genome- and Transcriptome-Wide Characterization of bZIP Gene Family Identifies Potential Members Involved in Abiotic Stress Response and Anthocyanin Biosynthesis in Radish (Raphanus sativus L.)

International Journal of Molecular Sciences ◽

10.3390/ijms20246334 ◽

2019 ◽

Vol 20 (24) ◽

pp. 6334 ◽

Cited By ~ 3

Author(s):

Lianxue Fan ◽

Liang Xu ◽

Yan Wang ◽

Mingjia Tang ◽

Liwang Liu

Keyword(s):

Abiotic Stress ◽

Stress Responses ◽

Leucine Zipper ◽

Anthocyanin Biosynthesis ◽

Functional Characterization ◽

Abiotic Stress Response ◽

Basic Leucine Zipper ◽

Promoter Sequences ◽

Encoding Genes

Basic leucine zipper (bZIP) transcription factors play crucial roles in various abiotic stress responses as well as anthocyanin accumulation. Anthocyanins are most abundant in colorful skin radish, which exhibit strong antioxidant activity that offers benefits for human health. Here, a total of 135 bZIP-encoding genes were identified from radish genome. Synteny analysis showed that 104 radish and 63 Arabidopsis bZIP genes were orthologous. Transcriptome analysis revealed that 10 RsbZIP genes exhibited high-expression levels in radish taproot (RPKM>10). Specifically, RsbZIP010 exhibited down-regulated expression under Cd, Cr and Pb stresses, whereas RsbZIP031 and RsbZIP059 showed significant down-regulation under heat and salt stresses, respectively. RT-qPCR analysis indicated that RsbZIP011 and RsbZIP102 were significantly up-regulated in the tissues of radish with high anthocyanin contents. Furthermore, the promoter sequences of 39 anthocyanin-related genes were found to contain G-box or ACE-box elements that could be recognized by bZIP family members. Taken together, several RsbZIPs might be served as critical regulators in radish taproot under Cd, Cr, Pb, heat and salt stresses. RsbZIP011 and RsbZIP102 were the potential participants in anthocyanin biosynthesis pathway of radish. These results facilitate further investigation on functional characterization of bZIP genes in response to abiotic stress and anthocyanin biosynthesis in radish.

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Systematic analysis of JmjC gene family of Brassica napus and KDM5 subfamily involved in abiotic stress response

10.21203/rs.2.11104/v1 ◽

2019 ◽

Author(s):

Xinghui He ◽

Jiao Pan ◽

Boyu Liu ◽

Chengfang Tan ◽

Ying Ruan ◽

...

Keyword(s):

Stress Response ◽

Brassica Napus ◽

Stress Responses ◽

Retention Rate ◽

Sequence Similarity ◽

Expression Profiles ◽

Functional Characterization ◽

Homologous Gene ◽

Oilseed Crop

Abstract Background: Jumonji C (JmjC) proteins play an important role in plant development and stress response through the removal of lysine methylation from histones. Brassica napus, which originated from spontaneous hybridization between Brassica rapa and Brassica oleracea, is the most important oilseed crop after soybean, but evolutionary relationships and functions of JmjC proteins remain unclear. Results: 65 JmjC genes were identified from B. napus genome, 29 from B. rapa, and 23 from B. oleracea. These genes were grouped into seven clades according to conserved sequences, and their catalytic activities of demethylation were predicted. Group-KDM4/JHDM3 for H3K4/9/27/36, Group-KDM5A/B for H3K4, Group-JmjC domain-only A/B for H3K27/36, Group-KDM3/JHDM2 for H3K9, and Group-JMJD6 may be for arginine demethylases. B. napus inherited most of its JmjC genes from its parents. The average retention rate of B. napus JmjC gene from B. rapa (93.1%) and B. oleracea (82.6%) exceeded that of all homologous gene pairs (83.7%) across the whole B. napus genome. Thirteen new or duplicated JmjC genes have emerged in B. napus. Sequence similarity and domain organization analyses suggest that the functions of these genes might be diversified. Furthermore, KDM5 genes were examined under stress conditions due to H3K4 demethylation. Expression profiles indicated that the genes from B. napus are possibly involved in various stress responses. Conclusion: This study provides the first genome-wide characterization of JmjC genes in Brassica species. Its JmjC genes potentially have diverse functions, and its KDM5 genes might be involved in stress response. The results of this study facilitate the future functional characterization of the demethylation of JmjC family in Brassica crops.

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Investigation and Computational Analysis of the Sulfotransferase (SOT) Gene Family in Potato (Solanum tuberosum): Insights into Sulfur Adjustment for Proper Development and Stimuli Responses

Plants ◽

10.3390/plants10122597 ◽

2021 ◽

Vol 10 (12) ◽

pp. 2597

Author(s):

Sahar Faraji ◽

Parviz Heidari ◽

Hoorieh Amouei ◽

Ertugrul Filiz ◽

Abdullah ◽

...

Keyword(s):

Solanum Tuberosum ◽

Gene Family ◽

Stress Responses ◽

Active Sites ◽

Sulfur Metabolism ◽

Functional Characterization ◽

Phosphate Binding ◽

Protein Motifs ◽

A Genome

Various kinds of primary metabolisms in plants are modulated through sulfate metabolism, and sulfotransferases (SOTs), which are engaged in sulfur metabolism, catalyze sulfonation reactions. In this study, a genome-wide approach was utilized for the recognition and characterization of SOT family genes in the significant nutritional crop potato (Solanum tuberosum L.). Twenty-nine putative StSOT genes were identified in the potato genome and were mapped onto the nine S. tuberosum chromosomes. The protein motifs structure revealed two highly conserved 5′-phosphosulfate-binding (5′ PSB) regions and a 3′-phosphate-binding (3′ PB) motif that are essential for sulfotransferase activities. The protein–protein interaction networks also revealed an interesting interaction between SOTs and other proteins, such as PRTase, APS-kinase, protein phosphatase, and APRs, involved in sulfur compound biosynthesis and the regulation of flavonoid and brassinosteroid metabolic processes. This suggests the importance of sulfotransferases for proper potato growth and development and stress responses. Notably, homology modeling of StSOT proteins and docking analysis of their ligand-binding sites revealed the presence of proline, glycine, serine, and lysine in their active sites. An expression essay of StSOT genes via potato RNA-Seq data suggested engagement of these gene family members in plants’ growth and extension and responses to various hormones and biotic or abiotic stimuli. Our predictions may be informative for the functional characterization of the SOT genes in potato and other nutritional crops.

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A Tobacco Syringe Agroinfiltration-Based Method for a Phytohormone Transporter Activity Assay Using Endogenous Substrates

Frontiers in Plant Science ◽

10.3389/fpls.2021.660966 ◽

2021 ◽

Vol 12 ◽

Author(s):

Jiangzhe Zhao ◽

Min Ju ◽

Jiayun Qian ◽

Mengyuan Zhang ◽

Ting Liu ◽

...

Keyword(s):

Stress Responses ◽

Large Scale ◽

Functional Characterization ◽

Endogenous Hormones ◽

Activity Assay ◽

Long Distance ◽

Multiple Substrates ◽

Long Distance Transport ◽

Endogenous Substrates

Phytohormones are a group of small chemical molecules that play vital roles in plant development, metabolism, and stress responses. Phytohormones often have distinct biosynthesis and signaling perception sites, requiring long- or short-distance transportation. Unlike biosynthesis and signal transduction, phytohormone transport across cells and organs is poorly understood. The transporter activity assay is a bottleneck for the functional characterization of novel phytohormone transporters. In the present study, we report a tobacco syringe agroinfiltration and liquid chromatography tandem mass spectrometry (TSAL)-based method for performing a phytohormone transporter activity assay using endogenous hormones present in tobacco (Nicotiana benthamiana) leaves. A transporter activity assay using this method does not require isotope-labeled substrates and can be conveniently performed for screening multiple substrates by using endogenous hormones in tobacco leaves. The transporter activities of three known hormone transporters, namely AtABCG25 for abscisic acid, AtABCG16 for jasmonic acid, and AtPUP14 for cytokinin, were all successfully validated using this method. Using this method, cytokinins were found to be the preferred substrates of an unknown maize (Zea mays) transporter ZmABCG43. ZmABCG43 transporter activities toward cytokinins were confirmed in a cytokinin long-distance transport mutant atabcg14 through gene complementation. Thus, the TSAL method has the potential to be used for basic substrate characterization of novel phytohormone transporters or for the screening of novel transporters for a specific phytohormone on a large scale.

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