AP2 Domain Research Articles

UV-induced reactive oxygen species and transcriptional control of 3-deoxyanthocyanidin biosynthesis in black sorghum pericarp.

Black pericarp sorghum has notable value due to the biosynthesis of 3-deoxyanthocyanidins (3-DOAs), a rare class of bioactive polyphenols valued as antioxidant food additives and as bioactive compounds with cytotoxicity to human cancer cells. A metabolic and transcriptomic study was conducted to ascertain the cellular events leading to the activation of 3-DOA biosynthesis in black sorghum pericarp. Prolonged exposure of pericarp during grain maturation to high-fluence ultraviolet (UV) light resulted in elevated levels of reactive oxygen species (ROS) and the activation of 3-DOA biosynthesis in pericarp tissues. In conjunction with 3-DOA biosynthesis was the transcriptional activation of specific family members of early and late flavonoid biosynthesis pathway genes as well as the downstream activation of defense-related pathways. Promoter analysis of genes highly correlated with 3-DOA biosynthesis in black pericarp were enriched in MYB and HHO5/ARR-B motifs. Light microscopy studies of black pericarp tissues suggest that 3-DOAs are predominantly localized in the epicarp and are associated with the cell wall. A working model of UV-induced 3-DOA biosynthesis in black pericarp is proposed that shares features of plant immunity associated with pathogen attack or mechanical wounding. The present model depicts ROS accumulation, the transcriptional activation of receptor kinases and transcription factors (TFs) including NAC, WRKY, bHLH, AP2, and C2H2 Zinc finger domain. This study identified key biosynthetic and regulatory genes of 3-DOA accumulation in black pericarp and provided a deeper understanding of the gene networks and cellular events controlling this tissue-and genotype-specific trait.

Genome-wide identification and characterization of the AP2/ERF gene family in loblolly pine (Pinus taeda L.).

The loblolly pine (Pinus taeda L.) is one of the most profitable forest species worldwide owing to its quick growth, high wood yields, and strong adaptability. The AP2/ERF gene family plays a widespread role in the physiological processes of plant defense responses and the biosynthesis of metabolites. Nevertheless, there are no reports on this gene family in loblolly pine (P. taeda). In this study, a total of 303 members of the AP2/ERF gene family were identified. Through multiple sequence alignment and phylogenetic analysis, they were classified into four subfamilies, including AP2 (34), RAV (17), ERF (251), and Soloist (1). An analysis of the conservation domains, conserved motifs, and gene structure revealed that every PtAP2/ERF transcription factor (TF) had at least one AP2 domain. While evolutionary conservation was displayed within the same subfamilies, the distribution of conserved domains, conserved motifs, and gene architectures varied between subfamilies. Cis-element analysis revealed abundant light-responsive elements, phytohormone-responsive elements, and stress-responsive elements in the promoter of the PtAP2/ERF genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of potential target genes showed that the AP2/ERF gene family might play a critical role in plant growth and development, the response to environmental stresses, and metabolite biosynthesis. Utilizing quantitative real-time PCR (qRT-PCR), we examined the expression patterns of 10 randomly selected genes from Group IX after 6 h of treatments with mechanical injury, ethephon (Eth), and methyl jasmonate (MeJA). The AP2/ERF gene family in the loblolly pine was systematically analyzed for the first time in this study, offering a theoretical basis for exploring the functions and applications of AP2/ERF genes.

BrERF109 positively regulates the tolerances of drought and salt stress in Chinese cabbage

Ethylene response factors (ERFs) are plant-specific transcription factors and play essential roles in plants’ defense responses to abiotic stresses. In this study, we identified an ethylene response factor BrERF109, and explored its roles in response to drought and salt stress. The BrERF109 protein contained an AP2 domain and was localized in the nucleus. Expression analysis suggested that BrERF109 was mainly expressed in root and leaf, and highly induced by drought, salt, heat, and cold stresses. Ectopic expression of BrERF109 in Arabidopsis improved the tolerance to drought and salt stress in transgenic plants, accompanied by reduced ABA sensitivity, enhanced antioxidant enzyme activities and chlorophyll content, as well as induced the expression of canonical stress responsive genes. Silencing of BrERF109 in Chinese cabbage by virus-induced gene silencing (VIGS) led to plants’ susceptibility to drought and salt stress, and suppressed superoxide dismutase and peroxidase activities. Furthermore, BrLTP3, BrCIPK6, BrAZF2, and BrZAT10 genes containing GCC-box in their promoter regions were identified as downstream candidate targets of BrERF109. Overall, our study demonstrated that BrERF109 is a key positive regulator in response to drought and salt stress in Chinese cabbage, and serves as a valuable target gene for breeding new cultivars with enhanced tolerances to abiotic stresses in Brassica vegetables.

Genome-Wide Identification and Expression of the AP2/ERF Gene Family in Morus notabilis

The AP2/ERF gene family, referring to an exclusive class of transcription factors unique to plants, is involved in various biological processes, including plant growth and responses to environmental stresses like high salt and drought. In this study, the AP2/ERF gene in M. notabilis was comprehensively identified and bioinformatically analyzed based on the genomic data of M. notabilis. 106 members in the MnAP2/ERF gene family were identified in the M. notabilis genome and were categorized into five subfamilies: ERF, AP2, DREB, RAV, and Soloist, with the ERF subfamily representing 80.19% of the total. The MnAP2/ERF gene family was observed to be distributed on six chromosomes of M. notabilis. Members in the MnAP2/ERF gene family exhibited obvious differences in amino acid number, molecular weight, isoelectric point, and other properties. Approximately 68.87% of the MnAP2/ERF proteins were acidic, all exhibiting hydrophilic characteristics. Differences in conserved sequences and arrangement of AP2 domains were observed among distinct subfamilies, with genes in the same subfamily sharing similar conserved domain compositions. There were 47 genes without untranslated regions and 44 genes with two untranslated regions. The upstream functions of promoters were concentrated on light reactions and plant hormones. Evolutionarily, significant structural differences were identified, and 28 MnAP2/ERF gene family proteins could interact with each other. Moreover, 35 family genes were involved in 22 fragment repeat events, and 55 MnAP2/ERF and 84 AtAP2/ERF genes showed collinearity. The expression of the MnAP2/ERF gene family was significantly different in different parts, indicating that these gene family members were involved in different physiological activities. These results established a theoretical foundation for investigating the functional and evolutionary aspects of AP2/ERF gene family genes in M. notabilis, as well as exploring the root morphogenesis of M. notabilis. Additionally, this study contributes to a basis for the improvement of cultivar stress resistance of M. notabilis.

ERF111/ABR1: An AP2 Domain Transcription Factor Serving as a Hub for Multiple Signaling Pathways

ERF111/ABR1: An AP2 Domain Transcription Factor Serving as a Hub for Multiple Signaling Pathways

Transcription Factor MdPLT1 Involved Adventitious Root Initiation in Apple Rootstocks

The induction of adventitious roots is a key factor restricting the vegetative propagation of apple dwarf rootstocks. PLETHORA (PLT) transcription factors are involved in the regulation of plant stem cell niche and adventitious root development. In this study, we identified the PLT1 gene in apples by bioinformatics and analyzed its evolutionary relationship. The MdPLT1 gene was cloned from M9-T337 to verify its subcellular localization and analyze its function in transgenic tobacco. The MdPLT1 protein contained two conserved AP2 domains which may be similar to those of poplar the PtrPLT2a and PtrPLT2b with 85% support. The CDS sequence of the MdPLT1 gene was 1638 bp, encoding 545 amino acids. The transcription factor MdPLT1 was localized in the nucleus. The number of adventitious roots of tobacco plants overexpressing MdPLT1 significantly increased. In the adventitious roots of MdPLT1-overexpressed plants, the expression levels of genes related to the NtPINs family and the NtYUCCAs family were significantly increased. The results showed that MdPLT1 positively regulated adventitious root formation. This study provided a theoretical basis for the establishment of the fast vegetative propagation of apple dwarf rootstocks.

Molecular identification and functional characterization of a transcription factor GeRAV1 from Gelsemium elegans

BackgroundGelsemium elegans is a traditional Chinese medicinal plant and temperature is one of the key factors affecting its growth. RAV (related to ABI3/VP1) transcription factor plays multiple roles in higher plants, including the regulation of plant growth, development, and stress response. However, RAV transcription factor in G. elegans has not been reported.ResultsIn this study, three novel GeRAV genes (GeRAV1-GeRAV3) were identified from the transcriptome of G. elegans under low temperature stress. Phylogenetic analysis showed that GeRAV1-GeRAV3 proteins were clustered into groups II, IV, and V, respectively. RNA-sequencing (RNA-seq) and real-time quantitative PCR (qRT-PCR) analyses indicated that the expression of GeRAV1 and GeRAV2 was increased in response to cold stress. Furthermore, the GeRAV1 gene was successfully cloned from G. elegans leaf. It encoded a hydrophilic, unstable, and non-secretory protein that contained both AP2 and B3 domains. The amino acid sequence of GeRAV1 protein shared a high similarity of 81.97% with Camptotheca acuminata CaRAV. Subcellular localization and transcriptional self-activation experiments demonstrated that GeRAV1 was a nucleoprotein without self-activating activity. The GeRAV1 gene was constitutively expressed in the leaves, stems, and roots of the G. elegans, with the highest expression levels in roots. In addition, the expression of the GeRAV1 gene was rapidly up-regulated under abscisic acid (ABA), salicylic acid (SA), and methyl jasmonate (MeJA) stresses, suggesting that it may be involved in hormonal signaling pathways. Moreover, GeRAV1 conferred improved cold and sodium chloride tolerance in Escherichia coli Rosetta cells.ConclusionsThese findings provided a foundation for further understanding on the function and regulatory mechanism of the GeRAV1 gene in response to low-temperature stress in G. elegans.

Localization in vivo and in vitro confirms EnApiAP2 protein encoded by ENH_00027130 as a nuclear protein in Eimeria necatrix.

Apicomplexan AP2 family of proteins (ApiAP2) are transcription factors (TFs) that regulate parasite growth and development, but little is known about the ApiAP2 TFs in Eimeria spp. ENH_00027130 sequence is predicted to encode a Eimeria necatrix ApiAP2 protein (EnApiAP2). The cDNAs encoding full-length and truncated EnApiAP2 protein were cloned and sequenced, respectively. Then, the two cDNAs were cloned into the pET28a(+) expression vector and expressed expressed in Escherichia coli BL21. The mouse polyclonal antibody (pAb) and monoclonal antibody (mAb) against recombinant EnApiAP2 (rEnApiAP2) and EnApiAP2tr (rEnApiAP2tr) were prepared and used to localize the native EnApiAP2 protein in E. necatrix, respectively. Finally, the recombinant pEGFP-C1-ΔNLS-EnApiAP2s (knockout of a nuclear localization sequence, NLS) and pEGFP-C1-EnApiAP2 plasmid were constructed and transfected into DF-1 cells, respectively, to further observe subcellular localization of EnApiAP2 protein. The EnApiAP2 gene had a size of 5019 bp and encoded 1672 amino acids, containing a conserved AP2 domain with a secondary structure consisting of an α-helix and three antiparallel β-strands. The rEnApiAP2 and rEnApiAP2tr were predominantly expressed in the form of inclusion bodies, and could be recognized by the 6×His tag mAb and the serum of convalescent chickens after infection with E. necatrix, respectively. The native EnApiAP2 protein was detected in sporozoites (SZ) and second generation merozoites (MZ-2) extracts, with a size of approximately 210 kDa. A quantitative real-time PCR (qPCR) analysis showed that the transcription level of EnApiAP2 was significantly higher in SZ than in MZ-2, third generation merozoites (MZ-3) and gametocytes (P<0.01). EnApiAP2 protein was localized in the nuclei of SZ, MZ-2 and MZ-3 of E. necatrix. The protein of EnApiAP2 was localized in the nucleus of the DF-1 cells, whereas the ΔNLS-EnApiAP2 was expressed in the cytoplasm, which further confirmed that EnApiAP2 is nucleoprotein. EnApiAP2 protein encoded by ENH_00027130 sequence was localized in the nucleus of E. necatrix parasites, and relied on the NLS for migration to DF-1 cell nucleus. The function of EnApiAP2 need further study.

Transcription Factor GmERF105 Negatively Regulates Salt Stress Tolerance in Arabidopsis thaliana.

The Ethylene Response Factor (ERF) transcription factors form a subfamily of the AP2/ERF family that is instrumental in mediating plant responses to diverse abiotic stressors. Herein, we present the isolation and characterization of the GmERF105 gene from Williams 82 (W82), which is rapidly induced by salt, drought, and abscisic acid (ABA) treatments in soybean. The GmERF105 protein contains an AP2 domain and localizes to the nucleus. GmERF105 was selectively bound to GCC-box by gel migration experiments. Under salt stress, overexpression of GmERF105 in Arabidopsis significantly reduced seed germination rate, fresh weight, and antioxidant enzyme activity; meanwhile, sodium ion content, malonic dialdehyde (MDA) content, and reactive oxygen species (ROS) levels were markedly elevated compared to the wild type. It was further found that the transcription levels of CSD1 and CDS2 of two SOD genes were reduced in OE lines. Furthermore, the GmERF105 transgenic plants displayed suppressed expression of stress response marker genes, including KIN1, LEA14, NCED3, RD29A, and COR15A/B, under salt treatment. Our findings suggest that GmERF105 can act as a negative regulator in plant salt tolerance pathways by affecting ROS scavenging systems and the transcription of stress response marker genes.

Transcriptional variation in Babesia gibsoni (Wuhan isolate) between in vivo and in vitro cultures in blood stage

BackgroundBabesia gibsoni, the causative agent of canine babesiosis, belongs to the phylum Apicomplexa. The development of in vitro culture technology has driven research progress in various kinds of omics studies, including transcriptomic analysis of Plasmodium spp. between in vitro and in vivo environments, which has prompted the observation of diagnostic antigens and vaccine development. Nevertheless, no information on Babesia spp. could be obtained in this respect, which greatly hinders the further understanding of parasite growth and development in the blood stage.MethodsIn this study, considerable changes in the morphology and infectivity of continuous in vitro cultured B. gibsoni (Wuhan isolate) were observed compared to in vivo parasites. Based on these changes, B. gibsoni (Wuhan isolate) was collected from both in vivo and in vitro cultures, followed by total RNA extraction and Illumina transcriptome sequencing. The acquired differentially expressed genes (DEGs) were validated using qRT-PCR, and then functionally annotated through several databases. The gene with the greatest upregulation after in vitro culture was cloned from the genome of B. gibsoni (Wuhan isolate) and characterized by western blotting and indirect immunofluorescence assay for detecting the native form and cellular localization.ResultsThrough laboratory cultivation, multiple forms of parasites were observed, and the infectivity of in vitro cultured parasites in dogs was found to be lower. Based on these changes, Illumina transcriptome sequencing was conducted, showing that 377 unigenes were upregulated and 334 unigenes were downregulated. Notably, an AP2 transcription factor family, essential for all developmental stages of parasites, was screened, and the transcriptional changes in these family members were tested. Thus, the novel AP2 transcription factor gene (BgAP2-M) with the highest upregulated expression after in vitro adaptation was selected. This gene comprises an open reading frame (ORF) of 1989 base pairs encoding a full-length protein of 662 amino acids. BgAP2-M contains one AP2 domain and one ACDC conserved domain, which may be involved in the nuclear biology of parasites. The prepared polyclonal antibodies against the BgAP2-M peptides further detected a native size of ~ 73 kDa and were localized to the nuclei of B. gibsoni.ConclusionThis study presents a thorough transcriptome analysis of B. gibsoni in vivo and in vitro for the first time, contributing to a detailed understanding of the effects of environmental changes on the growth and development of parasites in the blood stage. Moreover, it also provides a deeper investigation for the different members of the ApiAP2 transcription factor family as various life stage regulators in Babesia spp.Graphical

Cloning and analysis of DlERF23 gene in flower induction

Irregular flowering is a serious problem in longan production. Identifying the flower induction-related genes and analyzing their regulation mechanism is the key to solve this problem. The APETALA2/ethylene responsive factor (AP2/ERF) superfamily members are transcription factors (TFs) that regulate diverse developmental processes, including flowering time, and stress responses in plants. However, there is still no research about AP2/ERF involved in the regulation of longan flower induction. In the present study, a AP2/ERF TF member DlERF23 was cloned from longan (Dimocarpus longan). It has a typical AP2 domain with the coding sequence (CDS) of DlERF23 is 552 bp in length and encodes 184 amino acids. The molecular weight of DlERF23 protein was 20.41 kda and the theoretical isoelectric point (PI) was 7.69. The amino acid sequence of DlERF23 protein had the highest similarity with CsERF23 (XP_006478313.1) of Citrus sinensis and CcERF23 (XP_006441807.2) of Citrus clementina. The results of qRT-PCR showed that the relative expression level of DlERF23 gene in pericarp was higher, followed by stem, leave, flower and flower bud. Meanwhile, DlERF23 gene significant down-regulated in the early stage of flower induction in ‘Sijimi’ (SJ) longan and up-regulated in the late stage of flower induction in ‘Shixia’ (SX). The results of transient expression of Arabidopsis protoplasts showed that the fluorescence signal was mainly concentrated in the nucleus. Moreover, overexpression of DlERF23 in Arabidopsis promoted early flowering. These results provide useful information for revealing the biological roles of DlERF23 in longan and increase our understanding of the AP2/ERF superfamily members in fruit trees.

Improved bioenergy value of residual rice straw by increased lipid levels from upregulation of fatty acid biosynthesis

BackgroundRice (Oryza sativa) straw is a common waste product that represents a considerable amount of bound energy. This energy can be used for biogas production, but the rate and level of methane produced from rice straw is still low. To investigate the potential for an increased biogas production from rice straw, we have here utilized WRINKLED1 (WRI1), a plant AP2/ERF transcription factor, to increase triacylglycerol (TAG) biosynthesis in rice plants. Two forms of Arabidopsis thaliana WRI1 were evaluated by transient expression and stable transformation of rice plants, and transgenic plants were analyzed both for TAG levels and biogas production from straw.ResultsBoth full-length AtWRI1, and a truncated form lacking the initial 141 amino acids (including the N-terminal AP2 domain), increased fatty acid and TAG levels in vegetative and reproductive tissues of Indica rice. The stimulatory effect of the truncated AtWRI1 was significantly lower than that of the full-length protein, suggesting a role for the deleted AP2 domain in WRI1 activity. Full-length AtWRI1 increased TAG levels also in Japonica rice, indicating a conserved effect of WRI1 in rice lipid biosynthesis. The bio-methane production from rice straw was 20% higher in transformants than in the wild type. Moreover, a higher producing rate and final yield of methane was obtained for rice straw compared with rice husks, suggesting positive links between methane production and a high amount of fatty acids.ConclusionsOur results suggest that heterologous WRI1 expression in transgenic plants can be used to improve the metabolic potential for bioenergy purposes, in particular methane production.

WRINKLED1 Positively Regulates Oil Biosynthesis in Carya cathayensis.

Hickory (Carya cathayensis Sarg.) is a kind of important woody oil tree species, and its nut has high nutritional value. Previous gene coexpression analysis showed that WRINKLED1 (WRI1) may be a core regulator during embryo oil accumulation in hickory. However, its specific regulatory mechanism on hickory oil biosynthesis has not been investigated. Herein, two hickory orthologs of WRI1 (CcWRI1A and CcWRI1B) containing two AP2 domains with AW-box binding sites and three intrinsically disordered regions (IDRs) but lacking the PEST motif in the C-terminus were characterized. They are nucleus-located and have self-activated ability. The expression of these two genes was tissue-specific and relatively high in the developing embryo. Notably, CcWRI1A and CcWRI1B can restore the low oil content, shrinkage phenotype, composition of fatty acid, and expression of oil biosynthesis pathway genes of Arabidopsis wri1-1 mutant seeds. Additionally, CcWRI1A/B were shown to modulate the expression of some fatty acid biosynthesis genes in the transient expression system of nonseed tissues. Transcriptional activation analysis further indicated that CcWRI1s directly activated the expression of SUCROSE SYNTHASE2 (SUS2), PYRUVATE KINASE β SUBUNIT 1 (PKP-β1), and BIOTIN CARBOXYL CARRIER PROTEIN2 (BCCP2) involved in oil biosynthesis. These results suggest that CcWRI1s can promote oil synthesis by upregulating some late glycolysis- and fatty acid biosynthesis-related genes. This work reveals the positive function of CcWRI1s in oil accumulation and provides a potential target for improving plant oil by bioengineering technology.

Genome-wide identification and expression analysis of DREB family genes in cotton

BackgroundDehydration responsive element-binding (DREB) transcription factors are widely present in plants, and involve in signalling transduction, plant growth and development, and stress response. DREB genes have been characterized in multiple species. However, only a few DREB genes have been studied in cotton, one of the most important fibre crops. Herein, the genome‑wide identification, phylogeny, and expression analysis of DREB family genes are performed in diploid and tetraploid cotton species.ResultsIn total, 193, 183, 80, and 79 putative genes containing the AP2 domain were identified using bioinformatics approaches in G. barbadense, G. hirsutum, G. arboretum, and G. raimondii, respectively. Phylogenetic analysis showed that based on the categorization of Arabidopsis DREB genes, 535 DREB genes were divided into six subgroups (A1–A6) by using MEGA 7.0. The identified DREB genes were distributed unevenly across 13/26 chromosomes of A and/or D genomes. Synteny and collinearity analysis confirmed that during the evolution, the whole genome duplications, segmental duplications, and/or tandem duplications occurred in cotton DREB genes, and then DREB gene family was further expanded. Further, the evolutionary trees with conserved motifs, cis-acting elements, and gene structure of cotton DREB gene family were predicted, and these results suggested that DREB genes might be involved in the hormone and abiotic stresses responses. The subcellular localization showed that in four cotton species, DREB proteins were predominantly located in the nucleus. Further, the analysis of DREB gene expression was carried out by real-time quantitative PCR, confirming that the identified DREB genes of cotton were involved in response to early salinity and osmotic stress.ConclusionsCollectively, our results presented a comprehensive and systematic understanding in the evolution of cotton DREB genes, and demonstrated the potential roles of DREB family genes in stress and hormone response.

AP2 domain structure and protein motif features in Azerbaijan local durum and bread wheat genotypes

AP2 domain structure and protein motif features in Azerbaijan local durum and bread wheat genotypes

Physiological Response of Selected Rice Accessions to Salinity and In Silico Analysis of DREB1A Gene Among Diploid Oryza Species

The responses of selected rice accessions to variant salt concentrations and in-silico analysis of DREB1A gene among diploid Oryza species were evaluated. Ten (10) rice accessions were selected based on their popularity in farmer's fields. Seedlings of each variety (one per pot) were watered with variant salt concentrations of 0mM, 100mM and 200mM for 21 days. The morpho-physiological characters (plant height, number of tillers, root length and dry weight) were evaluated using a standard evaluation system for rice. The reference sequences of OsDREB1A and AtDREB1A were used as queries to search against the 10 diploid Oryza species in the BLASTN of the PlantEnsembl database to reveal DREB1A orthologs. The retrieved DREB1A orthologs were used to compute the physicochemical properties of their proteins, gene motifs, intron-exon architecture and phylogenetic relationship. The studied accessions showed significant differences (p&lt;0.05) in morpho-physiological responses to salinity. The accessions Zaqama, Yar-Garnaki, Yar-Yuti, Samira and Chana-Beru performed better under salt stress and there was no significant difference (p&gt;0.05) between the control and salt-treated groups. Additionally, the in-silico analysis of DREB1A gene identified 10 orthologs with conserved single transcript, AP2 domain and unstable protein (characteristics of TFs) across the 10 diploid Oryza species. Phylogenetic analysis revealed 3 clusters of African rice and its progenitor, Asian rice and their relatives and O. brachyantha/O. punctata complex, similar to the evolution of rice species. Conclusively, salt stress affects rice in a concentration-dependent manner and DREB1A gene is a conserved plant transcription factor (TF) across diploid Oryza species.

DsDBF1, a Type A-5 DREB Gene, Identified and Characterized in the Moss Dicranum scoparium.

Plant dehydration-responsive element binding (DREB) transcription factors (TFs) play important roles during stress tolerance by regulating the expression of numerous genes involved in stresses. DREB TFs have been extensively studied in a variety of angiosperms and bryophytes. To date, no information on the identification and characterization of DREB TFs in Dicranum scoparium has been reported. In this study, a new DBF1 gene from D. scoparium was identified by cloning and sequencing. Analysis of the conserved domain and physicochemical properties revealed that DsDBF1 protein has a classic AP2 domain encoding a 238 amino acid polypeptide with a molecular mass of 26 kDa and a pI of 5.98. Subcellular prediction suggested that DsDBF1 is a nuclear and cytoplasmic protein. Phylogenetic analysis showed that DsDBF1 belongs to group A-5 DREBs. Expression analysis by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) revealed that DsDBF1 was significantly upregulated in response to abiotic stresses such as desiccation/rehydration, exposure to paraquat, CdCl2, high and freezing temperatures. Taken together, our data suggest that DsDBF1 could be a promising gene candidate to improve stress tolerance in crop plants, and the characterization of TFs of a stress tolerant moss such as D. scoparium provides a better understanding of plant adaptation mechanisms.

Genome-wide identification, comprehensive characterization of transcription factors, cis-regulatory elements, protein homology, and protein interaction network of DREB gene family in Solanum lycopersicum.

Tomato is a drought-sensitive crop which has high susceptibility to adverse climatic changes. Dehydration-responsive element-binding (DREB) are significant plant transcription factors that have a vital role in regulating plant abiotic stress tolerance by networking with DRE/CRT cis-regulatory elements in response to stresses. In this study, bioinformatics analysis was performed to conduct the genome-wide identification and characterization of DREB genes and promoter elements in Solanum lycopersicum. In genome-wide coverage, 58 SlDREB genes were discovered on 12 chromosomes that justified the criteria of the presence of AP2 domain as conserved motifs. Intron-exon organization and motif analysis showed consistency with phylogenetic analysis and confirmed the absence of the A3 class, thus dividing the SlDREB genes into five categories. Gene expansion was observed through tandem duplication and segmental duplication gene events in SlDREB genes. Ka/Ks values were calculated in ortholog pairs that indicated divergence time and occurrence of purification selection during the evolutionary period. Synteny analysis demonstrated that 32 out of 58 and 47 out of 58 SlDREB genes were orthologs to Arabidopsis and Solanum tuberosum, respectively. Subcellular localization predicted that SlDREB genes were present in the nucleus and performed primary functions in DNA binding to regulate the transcriptional processes according to gene ontology. Cis-acting regulatory element analysis revealed the presence of 103 motifs in 2.5-kbp upstream promoter sequences of 58 SlDREB genes. Five representative SlDREB proteins were selected from the resultant DREB subgroups for 3D protein modeling through the Phyre2 server. All models confirmed about 90% residues in the favorable region through Ramachandran plot analysis. Moreover, active catalytic sites and occurrence in disorder regions indicated the structural and functional flexibility of SlDREB proteins. Protein association networks through STRING software suggested the potential interactors that belong to different gene families and are involved in regulating similar functional and biological processes. Transcriptome data analysis has revealed that the SlDREB gene family is engaged in defense response against drought and heat stress conditions in tomato. Overall, this comprehensive research reveals the identification and characterization of SlDREB genes that provide potential knowledge for improving abiotic stress tolerance in tomato.

Genome-Wide Identification of AP2/ERF Transcription Factor Family and Functional Analysis of DcAP2/ERF#96 Associated with Abiotic Stress in Dendrobium catenatum.

APETALA2/Ethylene Responsive Factor (AP2/ERF) family plays important roles in reproductive development, stress responses and hormone responses in plants. However, AP2/ERF family has not been systematically studied in Dendrobium catenatum. In this study, 120 AP2/ERF family members were identified for the first time in D. catenatum, which were divided into four groups (AP2, RAV, ERF and DREB subfamily) according to phylogenetic analysis. Gene structures and conserved motif analysis showed that each DcAP2/ERF family gene contained at least one AP2 domain, and the distribution of motifs varied among subfamilies. Cis-element analysis indicated that DcAP2/ERF genes contained abundant cis-elements related to hormone signaling and stress response. To further identify potential genes involved in drought stress, 12 genes were selected to detect their expression under drought treatment through qRT-PCR analysis and DcAP2/ERF#96, a nuclear localized ethylene-responsive transcription factor, showed a strong response to PEG treatment. Overexpression of DcAP2/ERF#96 in Arabidopsis showed sensitivity to ABA. Molecular, biochemical and genetic assays indicated that DcAP2ERF#96 interacts with DREB2A and directly inhibits the expression of P5CS1 in response to the ABA signal. Taken together, our study provided a molecular basis for the intensive study of DcAP2/ERF genes and revealed the biological function of DcAP2ERF#96 involved in the ABA signal.

Genome-Wide Identification of ERF Transcription Factor Family and Functional Analysis of the Drought Stress-Responsive Genes in Melilotus albus.

As an important forage legume with high values in feed and medicine, Melilotus albus has been widely cultivated. The AP2/ERF transcription factor has been shown to play an important regulatory role in plant drought resistance, but it has not been reported in the legume forage crop M. albus. To digger the genes of M. albus in response to drought stress, we identified and analyzed the ERF gene family of M. albus at the genome-wide level. A total of 100 MaERF genes containing a single AP2 domain sequence were identified in this study, named MaERF001 to MaERF100, and bioinformatics analysis was performed. Collinearity analysis indicated that segmental duplication may play a key role in the expansion of the M. albus ERF gene family. Cis-acting element predictions suggest that MaERF genes are involved in various hormonal responses and abiotic stresses. The expression patterns indicated that MaERFs responded to drought stress to varying degrees. Furthermore, four up-regulated ERFs (MaERF008, MaERF037, MaERF054 and MaERF058) under drought stress were overexpressed in yeast and indicated their biological functions to confer the tolerance to drought. This work will advance the understanding of the molecular mechanisms underlying the drought response in M. albus. Further study of the promising potential candidate genes identified in this study will provide a valuable resource as the next step in functional genomics studies and improve the possibility of improving drought tolerance in M. albus by transgenic approaches.