A Soybean bZIP transcription factor GmbZIP71-4 Is Involved in Submergence Resistance in Transgenic Tobacco

doi:10.21203/rs.3.rs-3089812/v1

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A Soybean bZIP transcription factor GmbZIP71-4 Is Involved in Submergence Resistance in Transgenic Tobacco

https://doi.org/10.21203/rs.3.rs-3089812/v1

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Background

Although the role of the basic leucine zipper (bZIP) family of transcription factors in response to various abiotic stresses has been investigated, few studies have been conducted on their function in submergence stress.

Results

In this study, we localized a bZIP gene GmbZIP71-4 in the nucleus of soybean and constructed a GmbZIP71-4overexpressing tabocco line, which showed reduced submergence resistance due to the decreased abscisic acid (ABA) content. GO and KEGG pathway enrichment analysis based on chromatin immunoprecipitation assay sequencing (ChIP-seq) indicated that the most significant differences were the expression levels of the factors in plant hormone signal transduction, especially those in response to ABA. Electrophoretic Mobility Shift Assays (EMSA) demonstrated that GmbZIP71-4 bound to the promoter of GmABF2, which is consistent with the ChIP-qPCR result.

Conclusions

Our study showed that GmbZIP71-4 was a negative regulator of submergence stress tolerance. The findings in this work will set a solid foundation for the understanding of submergence resistance in plants.

GmbZIP71-4

transcription factor

abiotic stress

submergence resistant

ABA

Crop growth and yield are subject to a variety of abiotic stresses such as drought, salt, waterlogging, and low and high temperatures. In response to these stresses, plants have been developed many strategies such as positive or negative regulation of transcription factor expression.

Plant basic leucine zipper (bZIP) proteins are a class of transcription factors that play an important role in biological processes, such as plant growth [1], flower development [2], light signaling [3], seed germination [4], pathogen defense, and responses to various stresses [5–8]. bZIP proteins have been found in a variety of plants, such as Arabidopsis thaliana [9], rice [10], wheat [11], maize [12], soybean [13], apple [14] and camellia [15]. bZIP is composed of a basic region of 20 amino acids for specific DNA binding and a leucine zipper domain at the N-terminus[16–18]. The leucine zipper and the basic region involved in oligomerization are closely linked, and a leucine residue is present in every seven amino acids of the leucine zipper [19]. A bZIP dimer is formed by the interaction between the leucine zippers before it can bind to DNA sequences with the ACGT cis-element [20–21]. A number of DNA binding sites have been identified, such as A-box (TACGTA), C-box (GACGTC), G-box (CACGTG), and ABRE (ABA-responsive element)[22].

As an upstream regulator, bZIP proteins play an important role in the ABA signaling pathway by binding to the ABRE in the promoter region of ABA-induced genes under stresses. For instance, overexpression of wheat TabZIP60 and TaZIP14-B genes in Arabidopsis thaliana significantly improve tolerance to cold, salt and ABA stresses [23–24]. OsbZIP23 regulates the biosynthesis of ABA, and thus overexpression of OsbZIP23 can greatly improve salt and drought tolerance through the ABA signaling pathway in rice plants [25]. Similarly, studies have shown overexpression of a number of other bZIP genes, such as OsbZIP42, OsbZIP62 and OsbZIP66 significantly enhance drought tolerance by altering the ABA content and activating the ABA signaling pathway [26–28]. ZmbZIP72 is a member of the bZIP transcription factor family in maize; ZmbZIP72-overexpressing Arabidopsis plants exhibit elevated tolerance towards abiotic stresses such as drought, salt, ABA and osmotic stress [29]. Under drought, salt, high and low temperature stresses, ZmbZIP4 can be induced, which in turn positively regulates genes involved in ABA synthesis and increases the ABA content in ZmbZIP4-overexpressing plants [30]. Soybean bZIP transcription factors including GmbZIP44, GmbZIP46, GmbZIP62 and GmbZIP78 have been overexpressed in Arabidopsis, and the results show that the plants are tolerant to salt and low temperature stresses and exhibit low sensitivity to ABA, indicating that these genes negatively regulate the ABA signaling pathway during salt and low temperature stresses [31]. Overexpression of GmbZIP15 induces high sensitivity of soybean to drought and salt stresses, by reducing the transcriptional level of stress response genes [32]. Studies have shown that GmbZIP19 interacts with the promoter region of ABA-induced marker genes in soybean [33], and GmbZIP19-overexpressing A. thaliana is more sensitive to drought and salt stresses, indicating GmbZIP19 negatively regulates drought and salt stresses.

In this study, we identified a new bZIP gene, GmbZIP71-4, from soybean, which played an important role in submergence resistance in transgenic tobacco. Compared with the control, the transgenic plants overexpressing GmbZIP71-4 exhibited a decreased submergence resistance. GO and KEGG enrichment analysis of downstream target genes of GmbZIP71-4 showed that the significant difference between submergence treatment and the control was the expression levels of the factors in plant hormone signal transduction, especially those responding to abscisic acid. EMSA result showed that GmbZIP71-4-His fusion protein could bind to the GmABF2 cis-element, and this result was also confirmed by Chip-qPCR. Our study demonstrate that GmbZIP71-4 plays key roles in response to submergence stress.

Functional analysis of GmbZIP71-4

The overexpression vector of GmbZIP71-4 was constructed and transferred into tobacco. The T₂ generation transgenic lines were detected, and the positive lines were selected using the target primers (Table S2). The old leaves of the T₂ generation transgenic positive lines rotted and the new leaves were more severely stained than the control lines 48 h after the submergence treatment. The phenotypic identification showed that GmbZIP71-4 overexpression reduced the submergence resistance of the transgenic lines compared with that of the control lines in tobacco (Fig. 1A). The results of RT-qPCR showed that the expression of GmbZIP71 gene was very high in overexpressed tobacco transgenic plants, but not detected in control plants (Fig. 1B).

In addition, our result indicated that the transgenic lines remarkable decreased in activity of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and the content of malondialdehyde (MDA), soluble sugar and abscisic acid (ABA) also significantly decreased in the transgenic lines compared with that of the control (P < 0.01) (Fig. 2).

Subcellular localization of GmbZIP71-4

The subcellular location of GmbZIP71-4 was determined using the

p35S:GmbZIP71-4-GFP plasmid. The results showed that strong fluorescent signals of the fused GmbZIP71-4-GFP protein could be detected in the nuclear (Fig. 3). Because GmbZIP71-4 is proposed to be a transcription factor, the result is consistent with its function.

ChIP-Seq analysis

ChIP-Seq was performed to detect GmbZIP71-4-bound genes under submergence stresses. A total of 37.45–57.35 million raw reads were obtained from the root samples of soybean, from which, 36.33–55.33 million clean reads were selected after deleting the low quality reads. The total mapping ratio ranges from 85.61–95.66% as shown in Table S1. Pearson correlation analysis was performed, and the results showed that the biologically repeated samples exhibited very similar expression patterns of genes (Fig. S1).

Figure 4A showed the distribution of the differentially expressed peaks. It can be seen from the results that 46.83%, 32.01%, 9.51%, 6.97%, 3.49% and 1.19% of the peaks were found in the promoter region, intergenic regions, 5’-UTR, CDS, introns, and 3’-UTR, respectively. These results indicated that that the peaks were not enriched in just one particular region, but distributed throughout the whole genome of Glycine max Wm82.a2.v1 (Fig. 4B). A number of GmbZIP71-4-bound DNA motifs were identified by the HOMER software (http://homer.ucsd.edu/homer/ngs/peakMotifs.html), such as MA0551, MA0936 and MA0356 (Fig. 4C). To dissect the function the differentially expressed downstream of GmbZIP71-4-bound genes, gene ontology (GO) enrichment was performed, and the results show that genes in a variety of functional categories were enriched, such as sequence-specific DNA binding (GO:0043565), response to alcohol (GO:0097305), response to abscisic acid (GO:0009737), response to lipid (GO:0033993), and response to osmotic stress (GO:0006970) (Fig. 5A, Table S2). A number of significantly enriched pathways were identified by KEGG enrichment analysis, including MAPK signaling pathway-plant, inositol phosphate metabolism and plant hormone signal transduction (Fig. 5B, Table S3). Both GO and KEGG enrichment results enriched plant hormone, suggesting that GmbZIP71-4 gene maybe respond to submergence stress through hormone pathway.

Selection and identification of target genes bound by GmbZIP71-4

To further investigate the regulatory roles of GmbZIP71-4 in soybean under submergence stresses, we identified the downstream target genes of GmbZIP71-4 by GO and KEGG pathway enrichment analyses. The result showed that the most significant difference between the plants with the submergence treatment and the control plants was the hormone signal transduction (Fig. S2), such as ABA responsive element binding factors. Previous studies based on the transcriptome data (SRP181976) have shown that GmABF2 exhibits significant changes in expression under the submergence treatment compared with that of the control. Consequently, GmABF2 was selected for further studies.

Verification of the interaction between GmbZIP71-4 and GmABF2

The interaction between GmbZIP71-4 and GmABF2 was validated by EMSA. The His-tagged GmbZIP71-4 protein was expressed in the T7E strain and purified using the Ni-NTA resins. The recombinant GmbZIP71-4-His fusion protein binding to the motif MA0356 of the GmABF2 promoter was analyzed by EMSA. Even when the unlabeled competitive probe was 100-fold higher in concentration, no migrating band was detected. A mutant probe was designed, in which a small segment was mutated from TTTTTC to CCCCCA. No interaction between this mutant probe and GmbZIP71-4, indicating that GmABF2 cis-element was essential for GmbZIP71-4 binding (Fig. 6A).

ChIP-qPCR

The corresponding peak of GmABF2 was successfully validated by ChIP-qPCR analysis. The result displayed that the DNA fragment containing the MA0356 motif in the promoter of the GmABF2 gene was enriched in plants under the submergence treatment and the control plants (Fig. 6B). These results are consistent with that of EMSA, both confirmed the interaction between GmbZIP71-4 and GmABF2. Finally, we described the action mode of GmbZIP71-4 gene in regulating submergence-response. When submergence stress occurred, GmbZIP71-4 interacted with GmABF2 gene and influenced ABA content by participating in ABA signaling pathway to protect against submergence damage (Fig. 7).

GmbZIP71-4 is a negative regulator in submergence resistance

Many bZIP transcription factors are found to be positive regulators under abiotic stresses, such as TabZIP60, OsbZIP42, GmbZIP44, GmbZIP46, GmbZIP62 and GmbZIP78 [23, 27, 31]. However, recent studies have shown that bZIP proteins may also exhibit negative regulatory functions in response to stresses. For example, SlbZIP38 and CaATBZ1 negatively regulate the ABA signal pathway, which in turn decrease the tolerance to drought stress [34, 35]. CsbZIP18 can be induced by low temperature. However, overexpression of CsbZIP18 in Arabidopsis thaliana plants exhibits decreased sensitivity to ABA under freezing conditions, indicating CsbZIP18 negative regulates tolerance to low temperature through the ABA signaling pathway [36]. GmbZIP19 and GmbZIP15 are negative regulators of drought and salt stresses, and overexpression of these proteins result in reduced tolerance to drought and salt stresses [32, 33]. In this study, a bZIP transcription factor GmbZIP71-4 was identified, which was down-regulated in plants under the submergence treatment. When GmbZIP71-4 was overexpressed in tobacco, the submergence resistance of the transgenic lines decreased compared with the control lines, suggesting GmbZIP71-4 is a negative regulator of submergence resistance. GO and KEGG enrichment analysis of the downstream target genes of GmbZIP71-4 using ChIP-seq demonstrate that hormone signal transduction pathways, especially those in response to abscisic acid, were differentially regulated between the plants under the submergence treatment and the control plants.

GmbZIP71-4 regulates submergence stress through the ABA pathway

In general, the ABA content in plants can be rapidly increased in response to abiotic stresses such as drought, salt, low and high temperatures, and waterlogging, which in turn promotes the expression of numbers of enzymes and improves stress tolerance. For example, overexpression of ZmbZIP4 of maize can elevate the ABA content and increase plant resistance to abiotic stresses [30]. Similarly, the ABA content in IbbZIP1-overexpressing transgenic Arabidopsis plants is greatly increased as the genes involved in ABA biosynthesis are up-regulated under drought and salt stresses [37]. AhAREB1 is a member of the bZIP family of peanut; overexpression of AhAREB1 in Arabidopsis also results in up-regulated ABA synthesis-related genes and elevated ABA level, improving drought tolerance [38]. However, our study demonstrated that overexpression of GmbZIP71-4 in tobacco decreased the ABA content and thus reduced the resistance of plants to submergence stresses. Taken together, the GmbZIP71-4 gene isolated in this study was a negative regulatory gene of stress tolerance, which is consistent with many other studies.

GmbZIP71-4 responds to submergence stress by interacting with GmABF2

A number of bZIP proteins have been identified to regulate ABA-induced genes by binding to the promoter region of the downstream target genes under abiotic stresses. Studies have shown that GmbZIP19-overexpressing plants exhibit higher sensitivity towards drought and salt stresses; an ABA response gene AtABI5 was found to have elevated expression in GmbZIP19-overexpressing plants compared with the WT plants, indicating that GmbZIP19 regulates the ABA signaling pathway through the activation of AtABI5 [33]. Another soybean bZIP transcription factor, GmbZIP15, negatively regulates the expression of many stress response genes such as GmABF1, thus reducing the tolerance to abiotic stresses[32]. In our study, KEGG pathway enrichment analysis based on ChIP-seq between the plants under the submergence treatment and the control plants demonstrates that plant hormone signal transduction was greatly impacted. Based on previous transcriptome data (SRP181976), GmABF2 was identified, which could interact with GmbZIP71-4 as demonstrated by EMSA. ChIP-qPCR showed that the DNA fragment containing the MA0356 motif in the promoter of GmABF2 gene was enriched in both submergence treatment and control plants, which is consistent with the EMSA result.

Based on previously published transcriptome sequencing data, we identified a bZIP transcription factor GmbZIP71-4. GmbZIP71-4-overexpressing tobacco plants were constructed to investigate the function of GmbZIP71-4 in response to abiotic stresses, which showed that GmbZIP71-4 negatively regulated the ABA content and thus increased the sensitivity to abiotic stresses. In addition, the downstream target gene GmABF2 of GmbZIP71-4 was isolated through ChIP-Seq, and EMSA and ChIP-qPCR confirmed that GmbZIP71-4 interacted with GmABF2 by binding to the promoter region. Taken together, the results suggested that GmbZIP71-4 participated in the response to submergence stress through the ABA signaling pathway. In addition, the findings in our study provide a molecular basis for understanding the mechanisms in response to submergence stresses in plants.

Plants

Qihuang34, a soybean variety with strong submergence resistance, was employed for cloning the GmbZIP71-4 gene. The expressed sequence tag (EST) was obtained from the transcriptome sequence conducted by Lin et al. [39]. The function of GmbZIP71-4 was analyzed using Tobacco (Nicotiana benthamiana).

Evaluation of physiological parameters

The commercial kits (Suzhou Michy Biomedical Technology Co., Ltd) were used to evaluate the levels of catalase (CAT), peroxidase (SOD) and malondialdehyde (MDA).

Measurement of the ABA content

The roots of the transgenic tobacco plants were collected for the measurement of the ABA content. The analysis was performed on a QTRAP 6500 LC–MS/MS system, according to the method described previously [40]. Three replicates of each assay were performed.

RNA isolation and vector construction and plant transformation

The total RNA was obtained from the root of Qihuang34 using a RNA Easy Fast Plant Tissue Kit (TIANGEN, China), and the cDNA was synthesized using a FastKing RT Kit (TIANGEN, China). PRIMER5 was used to design the gene-specific primers. The overexpression vector of GmbZIP71-4 was constructed by inserting the whole CDS fragment of GmbZIP71-4 into the pCAMBIA1300 vector and then transformed into the tobacco plant.

Subcellular localization of GmbZIP71-4

Firstly, PCR was performed to obtain the full-length GmbZIP71-4 and a p35S: GmbZIP71-4-GFP construct was obtained by inserting the GmbZIP71-4 fragment into a 35S-promoter-driven GFP plasmid, which was then transformed to the tobacco plants. After 16–24 h, the protein expression pattern of GmbZIP71-4 was observed on a confocal laser scanning microscope (Nikon C1 Si, Japan).

Chromatin immunoprecipitation sequencing (ChIP-seq)

The plants were subject to the submergence treatment for 24 h after the first two true leaves were fully unfolded. Illumina sequencing libraries were constructed for both CK (untreated plants) and submergence treated plants and sent to Wuhan IGENEBOOK Biotechnology Co., Ltd (Wuhan, China) for ChIP assays [41]. The clean reads were obtained after removing the low-quality reads by Trimmomatic (version 0.38) [42], and then mapped onto the Williams82 a2v1 genome. The enriched regions were identified from the mapped reads by the MACS2 software with default parameters. Depending on the distance of the peaks to the genes, the peaks can be assigned to different regions of the genes [43], and the binding frequency of the peaks in different regions can be calculated. The gene structure of GmbZIP71-4 can be divided into five segments, a 2-kb upstream region of the transcription start site (TSS), introns, exons, and 3’ and 5’ untranslated regions (UTRs). Transcription factor prediction was performed by comparing the peak-associated protein sequences with those in the plant transcription factor database (http://bioinfo.life.hust.edu.cn/animaltfdb/). Motif occurrence for motifs with a maximum length of 12 was predicted using the HOMER v3 software with the default settings [44]. The GO terms were obtained with a p-value cutoff of 0.01 by a gene ontology enrichment analysis tool, EasyGO, developed by Zhou [45]. The overrepresented GO terms were detected by adjusting the p-values obtained from Fisher's exact test using false discovery rate for multiple comparisons. The KEGG enrichment analysis (http://www.genome.jp/kegg/) [46] was performed to annotate gene functions using the clusterProfiler tool in the R package [47].

ChIP-qPCR

Cross-linking between DNA and its associated protein was fixed by adding 1% formaldehyde for 10 min after the samples were washed twice with PBS buffer. After the reaction was quenched by 125 mM glycine, the samples were lysed. Soluble sheared chromatins were obtained by sonication on ice, and an anti-GmbZIP71-4 antibody was used to detected GmbZIP71-4 in the immunoprecipitation assays. The average DNA length of the soluble chromatins was about 200–500 bp. An IgG antibody (ab17870, Abcam) was used as a negative control. The immunoprecipitation system was incubated at 4°C overnight, and then incubated with 30 µL protein beads for another 3 h. Following described washing procedure, the immunoprecipitated DNA on the beads was eluted with 300 µL elution buffer (100 mM NaHCO₃, 1% SDS). RNase A (8 µg/mL) was added to the eluant at 65°C for 6 h to remove the RNA contaminant, and then proteinase K (345 µg/mL) was added to remove the protein contaminant overnight at 45°C. PCR was performed using the ChamQ SYBR Color qPCR Master Mix kit to amplify the immunoprecipitated DNA. The primers are listed in Table S2. The expression level was calculated using the 2^−ΔΔCt method and normalized by comparing with the input sample. The data are presented as the mean standard deviation of three replicates.

Electrophoresis mobility shift assay (EMSA)

The GmbZIP71-4 expressing vector was constructed and transformed into the T7E strain, which was incubated in LB medium at 37°C for 4 h. After IPTG (1 mM) was added to induce the expression of the GmbZIP71-4 protein, the cells were incubated again at 16°C for 16 h. The cells were harvested by centrifuging, and lysed by ultrasound. Ni-NTA affinity resins were used to purify the His-tagged GmbZIP71-4 protein from the cell lasate. After being washed with the washing buffer (PBS, pH 7.5) three times, gradient elution was performed using imidazole solutions with different concentrations. Based on the promoter sequence of GmABF2, a labelled probe and a mutant probe were synthesized and labeled with biotin at the 5’-end, which were used for the EMSA.

ABA Abscisic acid

ChIP-seq Chromatin immunoprecipitation assay sequencing

EMSA Electrophoretic Mobility Shift Assays

bZIP Basic leucine zipper

SOD Superoxide dismutase

CAT Catalase

POD Peroxidase

MDA Malondialdehyde

TSS Transcription start site

UTRs Untranslated regions

Ethics approval and consent to participate

Experimental research and studies on soybean plants comply with relevant institutional, national, and international guidelines, standards, and legislation. The soybean plants used were cultivated under controlled conditions and therefore did not posed a threat to other plant species.

Consent for publication

Not applicable.

Availability of data and materials

Chip-seq data has not been uploaded yet, and we can upload it if necessary.

Competing interests

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Funding

This work was financially supported by finance science and technology project of Hainan province, technical innovation special project of provincial research institutes (jscx202004).

Author’s Contributions

Y.L. and C.X. designed and supervised the research. Y.L. and J.X. participated in sowing and sampling. X.H. performed RNA isolation and molecular detection. Y.L, H.Z. and L.T. performed data mining, analysis and interpretation. Y.L., C.X. and X.H. wrote this manuscript together. D.C. and X.W. participated in the revision of the manuscript. All the authors discussed the results and contributed to the manuscript. All authors read and approved the final manuscript.

Acknowledgments

We are grateful to Hainan Key Laboratory of Crop Genetics and Breeding for providing the scientific research platform.

Authors information

Yanhui Lin

Institute of Food Crops, Hainan Academy of Agricultural Sciences/Hainan Key Laboratory of Crop Genetics and Breeding/Hainan Scientific Research Station of Crop Gene Resource and Germplasm Enhancement, Ministry of Agriculture, Haikou, 571100, China.

E-mail: [email protected]

Changjian Xia

Haikou Cigar Research Institute, Haikou, 571100, China.

E-mail: [email protected]

Xing Huo

Rice Research Institute, Guangdong Academy of Agricultural Sciences; Guangdong Key Laboratory of New Technology in Rice Breeding; Guangdong Rice Engineering Laboratory, Guangzhou 510640, China.

E-mail: [email protected]

Jing Xu

E-mail: [email protected]

Yapeng Li

Institute of Food Crops, Hainan Academy of Agricultural Sciences/Hainan Key Laboratory of Crop Genetics and Breeding /Hainan Scientific Research Station of Crop Gene Resource and Germplasm Enhancement, Ministry of Agriculture, Haikou, 571100, China. Sanya Research Institute of Hainan Academy of Agricultural Sciences, Sanya, 572000, China.

E-mail: [email protected]

Honglin Zhu

Institute of Food Crops, Hainan Academy of Agricultural Sciences/Hainan Key Laboratory of Crop Genetics and Breeding /Hainan Scientific Research Station of Crop Gene Resource and Germplasm Enhancement, Ministry of Agriculture, Haikou, 571100, China.

E-mail: [email protected]

Liqiong Tang

E-mail: [email protected]

Dexin Chen^*

Haikou Cigar Research Institute, Haikou, 571100, China.

E-mail: [email protected]

Xiaoning Wang^*

E-mail: [email protected]

Heinekamp T, Kuhlmann M, Lenk A, Strathmann A, Dröge-Laser W. The tobacco bZIP transcription factor BZI-1 binds to G-box elements in the promoters of phenylpropanoid pathway genes in vitro, but it is not involved in their regulation in vivo. Mol Genet Genomics. 2002;267(1):16–26.
Strathmann A, Kuhlmann M, Heinekamp T, Dröge-Laser W. BZI-1 specifically heterodimerises with the tobacco bZIP transcription factors BZI-2, BZI-3/TBZF and BZI-4, and is functionally involved in flower development. Plant J 2001, 28(4):397–408.
Mallappa C, Yadav V, Negi P, Chattopadhyay S. A basic leucine zipper transcription factor, G-box-binding factor 1, regulates blue light-mediated photomorphogenic growth in Arabidopsis. J Biol Chem. 2006;281(31):22190–9.
Toh S, McCourt P, Tsuchiya Y. HY5 is involved in strigolactone-dependent seed germination in Arabidopsis. Plant Signal Behav. 2012;7(5):556–8.
Rodriguez-Uribe L, O'Connell MA. A root-specific bZIP transcription factor is responsive to water deficit stress in tepary bean (Phaseolus acutifolius) and common bean (P. vulgaris). J Exp Bot. 2006;57(6):1391–8.
Sreenivasulu N, Sopory SK, Kavi Kishor PB. Deciphering the regulatory mechanisms of abiotic stress tolerance in plants by genomic approaches. Gene. 2007;388(1–2):1–13.
Zou M, Guan Y, Ren H, Zhang F, Chen F. A bZIP transcription factor, OsABI5, is involved in rice fertility and stress tolerance. Plant Mol Biol. 2008;66(6):675–83.
Amir Hossain M, Lee Y, Cho JI, Ahn CH, Lee SK, Jeon JS, Kang H, Lee CH, An G, Park PB. The bZIP transcription factor OsABF1 is an ABA responsive element binding factor that enhances abiotic stress signaling in rice. Plant Mol Biol. 2010;72(4–5):557–66.
Dröge-Laser W, Snoek BL, Snel B, Weiste C. The Arabidopsis bZIP transcription factor family-an update. Curr Opin Plant Biol. 2018;45(Pt A):36–49.
Nijhawan A, Jain M, Tyagi AK, Khurana JP. Genomic survey and gene expression analysis of the basic leucine zipper transcription factor family in rice. Plant Physiol. 2008;146(2):333–50.
Agarwal P, Baranwal VK, Khurana P. Genome-wide Analysis of bZIP Transcription Factors in wheat and Functional Characterization of a TabZIP under Abiotic Stress. Sci Rep. 2019;9(1):4608.
Wei K, Chen J, Wang Y, Chen Y, Chen S, Lin Y, Pan S, Zhong X, Xie D. Genome-wide analysis of bZIP-encoding genes in maize. DNA Res. 2012;19(6):463–76.
Zhang M, Liu Y, Shi H, Guo M, Chai M, He Q, Yan M, Cao D, Zhao L, Cai H, et al. Evolutionary and expression analyses of soybean basic Leucine zipper transcription factor family. BMC Genomics. 2018;19(1):159.
Li Y-Y, Meng D, Li M, Cheng L. Genome-wide identification and expression analysis of the bZIP gene family in apple (Malus domestica). Tree Genet Genomes. 2016;12(4):82.
Hou Y, Wu A, He Y, Li F, Wei C. Genome-wide characterization of the basic leucine zipper transcription factors in Camellia sinensis. Tree Genet Genomes. 2018;14(2):27.
Lee SC, Choi HW, Hwang IS, Choi DS, Hwang BK. Functional roles of the pepper pathogen-induced bZIP transcription factor, CAbZIP1, in enhanced resistance to pathogen infection and environmental stresses. Planta 2006, 224(5):1209–1225.
Ellenberger T. Getting a grip on DNA recognition: structures of the basic region leucine zipper, and the basic region helix-loop-helix DNA-binding domains. Curr Opin Struc Biol. 1994;4(1):12–21.
Hurst HC. Transcription factors 1: bZIP proteins. Protein Profile. 1994;1(2):123–68.
Landschulz WH, Johnson PF, McKnight SL. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins. Science. 1988;240(4860):1759–64.
Baxevanis AD, Vinson CR. Interactions of coiled coils in transcription factors: where is the specificity? Curr Opin Genet Dev. 1993;3(2):278–85.
Singh KB, Foley RC, Oñate-Sánchez L. Transcription factors in plant defense and stress responses. Curr Opin Plant Biol. 2002;5(5):430–6.
Izawa T, Foster R, Chua NH. Plant bZIP protein DNA binding specificity. J Mol Biol. 1993;230(4):1131–44.
Zhang L, Zhang L, Xia C, Zhao G, Liu J, Jia J, Kong X. A novel wheat bZIP transcription factor, TabZIP60, confers multiple abiotic stress tolerances in transgenic Arabidopsis. Physiol Plant. 2015;153(4):538–54.
Zhang L, Zhang L, Xia C, Gao L, Hao C, Zhao G, Jia J, Kong X. A Novel Wheat C-bZIP Gene, TabZIP14-B, Participates in Salt and Freezing Tolerance in Transgenic Plants. Front Plant Sci. 2017;8:710.
Xiang Y, Tang N, Du H, Ye H, Xiong L. Characterization of OsbZIP23 as a key player of the basic leucine zipper transcription factor family for conferring abscisic acid sensitivity and salinity and drought tolerance in rice. Plant Physiol. 2008;148(4):1938–52.
Yoon S, Lee D-K, Yu IJ, Kim YS, Choi YD, Kim J-K. Overexpression of the OsbZIP66 transcription factor enhances drought tolerance of rice plants. Plant Biotechnol Rep. 2017;11(1):53–62.
Joo J, Lee YH, Song SI. OsbZIP42 is a positive regulator of ABA signaling and confers drought tolerance to rice. Planta 2019, 249(5):1521–33.
Yang S, Xu K, Chen S, Li T, Xia H, Chen L, Liu H, Luo L. A stress-responsive bZIP transcription factor OsbZIP62 improves drought and oxidative tolerance in rice. BMC Plant Biol. 2019;19(1):260.
Ying S, Zhang DF, Fu J, Shi YS, Song YC, Wang TY, Li Y. Cloning and characterization of a maize bZIP transcription factor, ZmbZIP72, confers drought and salt tolerance in transgenic Arabidopsis. Planta. 2012;235(2):253–66.
Ma H, Liu C, Li Z, Ran Q, Xie G, Wang B, Fang S, Chu J, Zhang J. ZmbZIP4 Contributes to Stress Resistance in Maize by Regulating ABA Synthesis and Root Development. Plant Physiol. 2018;178(2):753–70.
Liao Y, Zou HF, Wei W, Hao YJ, Tian AG, Huang J, Liu YF, Zhang JS, Chen SY, Soybean. GmbZIP44, GmbZIP62 and GmbZIP78 genes function as negative regulator of ABA signaling and confer salt and freezing tolerance in transgenic Arabidopsis. Planta 2008, 228(2):225–240.
Zhang M, Liu Y, Cai H, Guo M, Chai M, She Z, Ye L, Cheng Y, Wang B, Qin Y. The bZIP Transcription Factor GmbZIP15 Negatively Regulates Salt- and Drought-Stress Responses in Soybean. Int J Mol Sci. 2020;21(20):7778.
He Q, Cai H, Bai M, Zhang M, Chen F, Huang Y, Priyadarshani S, Chai M, Liu L, Liu Y, et al. A soybean bZIP transcription factor GmbZIP19 confers multiple biotic and abiotic stress responses in plant. Int J Mol Sci. 2020;21(13):4071.
Joo H, Lim CW, Lee SC. Roles of pepper bZIP transcription factor CaATBZ1 and its interacting partner RING-type E3 ligase CaASRF1 in modulation of ABA signalling and drought tolerance. Plant J. 2019;100(2):399–410.
Pan Y, Hu X, Li C, Xu X, Su C, Li J, Song H, Zhang X, Pan Y. SlbZIP38, a Tomato bZIP Family Gene Downregulated by Abscisic Acid, Is a Negative Regulator of Drought and Salt Stress Tolerance. Genes (Basel) 2017, 8(12).
Yao L, Hao X, Cao H, Ding C, Yang Y, Wang L, Wang X. ABA-dependent bZIP transcription factor, CsbZIP18, from Camellia sinensis negatively regulates freezing tolerance in Arabidopsis. Plant Cell Rep. 2020;39(4):553–65.
Kang C, Zhai H, He S, Zhao N, Liu Q. A novel sweetpotato bZIP transcription factor gene, IbbZIP1, is involved in salt and drought tolerance in transgenic Arabidopsis. Plant Cell Rep. 2019;38(11):1373–82.
Li XY, Liu X, Yao Y, Li YH, Liu S, He CY, Li JM, Lin YY, Li L. Overexpression of Arachis hypogaea AREB1 gene enhances drought tolerance by modulating ROS scavenging and maintaining endogenous ABA content. Int J Mol Sci. 2013;14(6):12827–42.
Lin Y, Li W, Zhang Y, Xia C, Liu Y, Wang C, Xu R, Zhang L. Identification of Genes/Proteins Related to Submergence Tolerance by Transcriptome and Proteome Analyses in Soybean. Sci Rep. 2019;9(1):14688.
Chen W, Gong L, Guo Z, Wang W, Zhang H, Liu X, Yu S, Xiong L, Luo J. A novel integrated method for large-scale detection, identification, and quantification of widely targeted metabolites: application in the study of rice metabolomics. Mol Plant. 2013;6(6):1769–80.
Landt SG, Marinov GK, Kundaje A, Kheradpour P, Pauli F, Batzoglou S, Bernstein BE, Bickel P, Brown JB, Cayting P, et al. ChIP-seq guidelines and practices of the ENCODE and modENCODE consortia. Genome Res. 2012;22(9):1813–31.
Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014;30(15):2114–20.
Salmon-Divon M, Dvinge H, Tammoja K, Bertone P. PeakAnalyzer: genome-wide annotation of chromatin binding and modification loci. BMC Bioinformatics. 2010;11:415.
Hull RP, Srivastava PK, D'Souza Z, Atanur SS, Mechta-Grigoriou F, Game L, Petretto E, Cook HT, Aitman TJ, Behmoaras J. Combined ChIP-Seq and transcriptome analysis identifies AP-1/JunD as a primary regulator of oxidative stress and IL-1β synthesis in macrophages. BMC Genomics. 2013;14:92.
Zhou X, Su Z. EasyGO: Gene Ontology-based annotation and functional enrichment analysis tool for agronomical species. BMC Genomics. 2007;8:246.
Altermann E, Klaenhammer TR. PathwayVoyager: pathway mapping using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. BMC Genomics. 2005;6:60.
Yu G, Wang LG, Han Y, He QY. clusterProfiler: an R package for comparing biological themes among gene clusters. Omics 2012, 16(5):284–7.

No competing interests reported.

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A Soybean bZIP transcription factor GmbZIP71-4 Is Involved in Submergence Resistance in Transgenic Tobacco

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Abstract

Figures

Background

Results

ChIP-Seq analysis

ChIP-qPCR

Disscussion

Conclusions

Material and methods

Plants

Evaluation of physiological parameters

Measurement of the ABA content

RNA isolation and vector construction and plant transformation

Chromatin immunoprecipitation sequencing (ChIP-seq)

ChIP-qPCR

Electrophoresis mobility shift assay (EMSA)

Abbreviations

Declarations

References

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Supplementary Files

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