Populus Trichocarpa Research Articles

Search for motifs, associated with response to abscisic acid in promoters of genes of Quercus robur and Populus trichocarpa plants

Search for motifs, associated with response to abscisic acid in promoters of genes of <i>Quercus robur</i> and <i>Populus trichocarpa</i> plants

Genome-wide identification and expression analysis of EPF/EPFL gene family in Populus trichocarpa.

The Epidermal Patterning Factor/EPF-like (EPF/EPFL) family encodes a specific type of secreted protein in plants and plays an important role in plant growth and development, especially in the process of morphogenesis. To investigate the characteristics of EPF/EPFL gene family members and their regulatory functions in stomatal development of Populus trichocarpa, a total of 15 EPF/EPFL family genes were identified. Then the gene structure, chromosome location, phylogenetic relationship, protein conserved domain and gene expression profile were analyzed. According to phylogenetic analysis, PtEPF/EPFL can be classified into four groups. The gene structure and protein conservation motifs within the EPF family indicate the high conservation of the PtEPF/EPFL sequence. The promoter region of PtEPF/EPFL was found to contain cis-elements in response to stress and plant hormones. In addition, RT-qPCR results indicated that the PtEPF/EPFL have a differentially expressed in different tissues. Under drought stress treatment, a substantial upregulation was observed in the majority of PtEPF/EPFL members, suggesting their potential involvement in drought response. These results provide a theoretical basis for future exploration of the characteristics and functions of more PtEPF/EPFL genes.

Genome-wide identification of poplar GSTU gene family and its PtrGSTU23 and PtrGSTU40 to improve salt tolerance in poplar

Glutathione S-transferases (GSTs) are abundant proteins encoded by a highly differentiated family of ancient genes. GSTs play an important role in plant cell detoxification and growth and development. However, their precise function in growth and response to abiotic stresses in poplar remains unclear. We have identified 81 GST genes from Populus trichocarpa, which have been categorized into 8 subfamilies. Among these subfamilies, the Tau subfamily stands out as the largest, consisting of 58 members. We isolated two Tau-like Glutathione S-transferases encoding genes, PtrGSTU23 and PtrGSTU40, and investigated the biological functions of these two genes in transgenic poplar to regulate salt tolerance in poplar. Under salt stress, reactive oxygen species (ROS) accumulate in plants, and overexpression of PtrGSTU23/40 in 84 K poplar (Populus alba × Populus glandulosa) could play a crucial role in regulating ROS scavenging by controlling the activity of plant GST enzyme. This regulation of GST enzyme activity enhanced the tolerance of poplar to salt stress. Ultimately, our findings contribute to the availability of genetic resources that can be utilized in the breeding of salt tolerance traits within fast-growing tree species.

Genome-Wide Identification and Expression Analysis of Auxin Response Factor (ARF) Gene Family in Betula pendula

As the key transcription factors regulating auxin responsive genes expression, auxin response factors (ARFs) play critical roles in diverse aspects of plant growth and development. Betula pendula is a valuable ornamental tree, and the information on ARF gene family of B. pendula is needed for better understanding. The publication of the genome sequence of B. pendula enable to analyze the bioinformatics information and expression pattern of BpeARF gene family on the genome-wide basis. In this study, physical and chemical properties, chromosome location, phylogenetic relationship, gene structure, conserved domain, motif composition, and cis-acting element of BpeARF gene family were analyzed, and expression patterns of BpeARF genes were investigated using completely random design in different tissues and under exogenous NAA and drought treatments. A total of 17 BpeARF genes was identified from B. pendula genome, which were unevenly distributed on 13 chromosomes and encoded adequate proteins ranging from 613 to 1135 amino acids in length. Three BpeARF gene pairs were formed by segmental duplication, and the Ka/Ks values of these BpeARF gene pairs were less than 1. According to the phylogenetic relationship among B. pendula, Betula platyphylla, Populus trichocarpa, and Arabidopsis thaliana, the BpeARF genes were divided into four classes, and the intron/exon structure, conserved domain, and motif composition showed high similarity among the BpeARF genes within the same class. The cis-acting elements in the promoter regions of BpeARF genes were related to tissue development, hormone response, and stress resistance. Quantitative real-time PCR exhibited diverse expression patterns of BpeARF genes in different tissues and in response to exogenous auxin treatment and drought stress. The expressions of one, ten, seven, and three BpeARF genes were the high levels in buds, young leaves, stems, and roots, respectively. Under exogenous NAA treatment, six BpeARF genes in stems and roots were upregulated expression at all timepoints. Under drought stress, BpeARF7 and BpeARF15 were upregulated in stems and roots, and BpeARF5 and BpeARF6 were downregulated in leaves, stems, and roots. Our results provided valuable information for the classification and putative functions of BpeARF gene family, which may be helpful for selecting candidate genes and verifying gene function in the genetic engineering of birch trees in further research.

A COMPASS histone H3K4 trimethyltransferase pentamer transactivates drought tolerance and growth/biomass production in Populus trichocarpa.

Histone H3 lysine-4 trimethylation (H3K4me3) activating drought-responsive genes in plants for drought adaptation has long been established, but the underlying regulatory mechanisms are unknown. Here, using yeast two-hybrid, bimolecular fluorescence complementation, biochemical analyses, transient and CRISPR-mediated transgenesis in Populus trichocarpa, we unveiled in this adaptation a regulatory interplay between chromatin regulation and gene transactivation mediated by an epigenetic determinant, a PtrSDG2-1-PtrCOMPASS (complex proteins associated with Set1)-like H3K4me3 complex, PtrSDG2-1-PtrWDR5a-1-PtrRbBP5-1-PtrAsh2-2 (PtrSWRA). Under drought conditions, a transcription factor PtrAREB1-2 interacts with PtrSWRA, forming a PtrSWRA-PtrAREB1-2 pentamer, to recruit PtrSWRA to specific promoter elements of drought-tolerant genes, such as PtrHox2, PtrHox46, and PtrHox52, for depositing H3K4me3 to promote and maintain activated state of such genes for tolerance. CRISPR-edited defects in the pentamer impaired drought tolerance and elevated expression of PtrHox2, PtrHox46, or PtrHox52 improved the tolerance as well as growth in P. trichocarpa. Our findings revealed the identity of the underlying H3K4 trimethyltransferase and its interactive arrangement with the COMPASS for catalysis specificity and efficiency. Furthermore, our study uncovered how the H3K4 trimethyltransferase-COMPASS complex is recruited to the effector genes for elevating H3K4me3 marks for improved drought tolerance and growth/biomass production in plants.

Structure and Expression Analysis of PtrSUS, PtrINV, PtrHXK, PtrPGM, and PtrUGP Gene Families in Populus trichocarpa Torr. and Gray.

Exogenous nitrogen and carbon can affect plant cell walls, which are composed of structural carbon. Sucrose synthase (SUS), invertase (INV), hexokinase (HXK), phosphoglucomutase (PGM), and UDP-glucose pyrophosphorylase (UGP) are the key enzymes of sucrose metabolism involved in cell wall synthesis. To understand whether these genes are regulated by carbon and nitrogen to participate in structural carbon biosynthesis, we performed genome-wide identification, analyzed their expression patterns under different carbon and nitrogen treatments, and conducted preliminary functional verification. Different concentrations of nitrogen and carbon were applied to poplar (Populus trichocarpa Torr. and Gray), which caused changes in cellulose, lignin, and hemicellulose contents. In poplar, 6 SUSs, 20 INVs, 6 HXKs, 4 PGMs, and 2 UGPs were identified. Moreover, the physicochemical properties, collinearity, and tissue specificity were analyzed. The correlation analysis showed that the expression levels of PtrSUS3/5, PtrNINV1/2/3/5/12, PtrCWINV3, PtrVINV2, PtrHXK5/6, PtrPGM1/2, and PtrUGP1 were positively correlated with the cellulose content. Meanwhile, the knockout of PtrNINV12 significantly reduced the cellulose content. This study could lay the foundation for revealing the functions of SUSs, INVs, HXKs, PGMs, and UGPs, which affected structural carbon synthesis regulated by nitrogen and carbon, proving that PtrNINV12 is involved in cell wall synthesis.

Engineered Methylococcus capsulatus Bath for efficient methane conversion to isoprene

Isoprene has numerous industrial applications, including rubber polymer and potential biofuel. Microbial methane-based isoprene production could be a cost-effective and environmentally benign process, owing to a reduced carbon footprint and economical utilization of methane. In this study, Methylococcus capsulatus Bath was engineered to produce isoprene from methane by introducing the exogenous mevalonate (MVA) pathway. Overexpression of MVA pathway enzymes and isoprene synthase from Populus trichocarpa under the control of a phenol-inducible promoter substantially improved isoprene production. M. capsulatus Bath was further engineered using a CRISPR-base editor to disrupt the expression of soluble methane monooxygenase (sMMO), which oxidizes isoprene to cause toxicity. Additionally, optimization of the metabolic flux in the MVA pathway and culture conditions increased isoprene production to 228.1 mg/L, the highest known titer for methanotroph-based isoprene production. The developed methanotroph could facilitate the efficient conversion of methane to isoprene, resulting in the sustainable production of value-added chemicals.

Dynamic changes to the plant cuticle include the production of volatile cuticular wax-derived compounds.

The cuticle is a hydrophobic structure that seals plant aerial surfaces from the surrounding environment. To better understand how cuticular wax composition changes over development, we conducted an untargeted screen of leaf surface lipids from black cottonwood (Populus trichocarpa). We observed major shifts to the lipid profile across development, from a phenolic and terpene-dominated profile in young leaves to an aliphatic wax-dominated profile in mature leaves. Contrary to the general pattern, levels of aliphatic cis-9-alkenes decreased in older leaves following their accumulation. A thorough examination revealed that the decrease in cis-9-alkenes was accompanied by a concomitant increase in aldehydes, one of them being the volatile compound nonanal. By applying exogenous alkenes to P. trichocarpa leaves, we show that unsaturated waxes in the cuticle undergo spontaneous oxidative cleavage to generate aldehydes and that this process occurs similarly in other alkene-accumulating systems such as balsam poplar (Populus balsamifera) leaves and corn (Zea mays) silk. Moreover, we show that the production of cuticular wax-derived compounds can be extended to other wax components. In bread wheat (Triticum aestivum), 9-hydroxy-14,16-hentriacontanedione likely decomposes to generate 2-heptadecanone and 7-octyloxepan-2-one (a caprolactone). These findings highlight an unusual route to the production of plant volatiles that are structurally encoded within cuticular wax precursors. These processes could play a role in modulating ecological interactions and open the possibility for engineering bioactive volatile compounds into plant waxes.

Correction to: Genome‑wide identification of NRAMP family genes in Populus trichocarpa and their roles in transport of heavy metals

Correction to: Genome‑wide identification of NRAMP family genes in Populus trichocarpa and their roles in transport of heavy metals

Genome-wide identification and characterization of the RZFP gene family and analysis of its expression pattern under stress in Populus trichocarpa

Forest trees face many abiotic stressors during their lifetime, including drought, heavy metals, high salinity, and chills, affecting their quality and yield. The RING-type ubiquitin ligase E3 is an invaluable component of the ubiquitin-proteasome system (UPS) and participates in plant growth and environmental interactions. Interestingly, only a few studies have explored the RING ZINC FINGER PROTEIN (RZFP) gene family. This study identified eight PtrRZFPs genes in the Populus genome, and their molecular features were analyzed. Gene structure analysis revealed that all PtrRZFPs genes contained >10 introns. Evolutionarily, the RZFPs were separated into four categories, and segmental replication events facilitated their amplification. Notably, many stress-related elements have been identified in the promoters of PtrRZFPs using Cis-acting element analysis. Moreover, some PtrRZFPs were significantly induced by drought and sorbitol, revealing their potential roles in regulating stress responses. Particularly, overexpression of the PtrRZFP1 gene in poplars conferred excellent drought tolerance; however, PtrRZFP1 knockdown plants were drought-sensitive. We identified the potential upstream transcription factors of PtrRZFPs and revealed the possible biological functions of RZFP1/4/7 in resisting osmotic and salt stress, laying the foundation for subsequent biological function studies and providing genetic resources for genetic engineering breeding for drought resistance in forest trees. This study offers crucial information for the further exploration of the functions of RZFPs in poplars.

Regulation of secondary growth by poplar BLADE-ON-PETIOLE genes in Arabidopsis.

BLADE-ON-PETIOLE (BOP) genes are essential regulators of vegetative and reproductive development in land plants. First characterized in Arabidopsis thaliana (Arabidopsis), members of this clade function as transcriptional co-activators by recruiting TGACG-motif binding (TGA) basic leucine zipper (bZIP) transcription factors. Highly expressed at organ boundaries, these genes are also expressed in vascular tissue and contribute to lignin biosynthesis during secondary growth. How these genes function in trees, which undergo extensive secondary growth to produce wood, remains unclear. Here, we investigate the functional conservation of BOP orthologs in Populus trichocarpa (poplar), a widely-used model for tree development. Within the poplar genome, we identified two BOP-like genes, PtrBPL1 and PtrBPL2, with abundant transcripts in stems. To assess their functions, we used heterologous assays in Arabidopsis plants. The promoters of PtrBPL1 and PtrBPL2, fused with a β-glucuronidase (GUS) reporter gene showed activity at organ boundaries and in secondary xylem and phloem. When introduced into Arabidopsis plants, PtrBPL1 and PtrBPL2 complemented leaf and flower patterning defects in bop1 bop2 mutants. Notably, Arabidopsis plants overexpressing PtrBPL1 and PtrBPL2 showed defects in stem elongation and the lignification of secondary tissues in the hypocotyl and stem. Finally, PtrBPL1 and PtrBPL2 formed complexes with TGA bZIP proteins in yeast. Collectively, our findings suggest that PtrBPL1 and PtrBPL2 are orthologs of Arabidopsis BOP1 and BOP2, potentially contributing to secondary growth regulation in poplar trees. This work provides a foundation for functional studies in trees.

ENFOQUES GENÓMICOS Y TRANSCRIPTÓMICOS PARA ESTUDIAR ÁRBOLES MADERABLES: PERSPECTIVAS PARA EL ESTUDIO DE CEDRO ROJO (Cedrela odorata L.)

&lt;p&gt;&lt;strong&gt;Introduction&lt;/strong&gt;: The high-throughput genomic and transcriptomic approach has been developed and implemented to address the main challenges that the timber forestry sector faces, such as population growth, climate change, deforestation and the loss of forest ecosystem services. &lt;strong&gt;Objective:&lt;/strong&gt; To carry out a bibliographic review focused on the genomes and transcriptomes of timber trees reported in the databases, with special attention to red cedar (&lt;em&gt;Cedrela odorata&lt;/em&gt; L.), due to its importance as precious wood in Mexico. &lt;strong&gt;Methodology:&lt;/strong&gt; A literature review was carried out, directed at studying timber trees with genomic and transcriptomic strategies in different databases such as Pubmed, Scopus, Google Scholar, ScienceDirect, Wiley Online Library, MDPI and Scielo to identify the timber species that have been reported genomes and transcriptomes. The structure of the review was the genomics of timber trees, the transcriptomics of wood, and the potential species for study due to their importance and finally, the databases for consultation. Subsequently, a bibliometric study was carried out with the bibliometrix library in R Studio. &lt;strong&gt;Main results&lt;/strong&gt;: The first genome to be assembled at the chromosome level was the black cottonwood. Among the timber trees, the genomes of black cottonwood, desert poplar, eucalyptus and oak with a length of 392, 496, 691 and 789 Mb have been reported. Through study of the transcriptome, it has been possible to identify genes related to formation of the wood in a hybrid poplar (&lt;em&gt;Populus alba&lt;/em&gt; L.&lt;em&gt; × P. glandulosa&lt;/em&gt;) and &lt;em&gt;P. tremula&lt;/em&gt; L. and with drought tolerance in &lt;em&gt;Pinus massoniana&lt;/em&gt; and &lt;em&gt;Pinus pinaster&lt;/em&gt; Aiton. In red cedar (&lt;em&gt;Cedrela odorata&lt;/em&gt; L.), the transcriptome was obtained by sequencing a single leaf, identifying 52,181 gene models. In the NCBI, EMBL-EBI, TreeGenes, PLAZA databases and the hardwood genomics website it is possible to find information related to the genomics and transcriptomics of timber species. &lt;strong&gt;Implications:&lt;/strong&gt; More research is required in omics in timber, particularly in red cedar, since the search on these topics yielded little information. &lt;strong&gt;Conclusion&lt;/strong&gt;: Through the bibliographic review in databases, the timber trees that have a described genome and transcriptome were identified. This information can be used for the assembly and annotation of new genomes, identification of genes and molecular markers, among other applications.&lt;/p&gt;

Genome-wide identification of NRAMP family genes in Populus trichocarpa and their roles in transport of heavy metals

Genome-wide identification of NRAMP family genes in Populus trichocarpa and their roles in transport of heavy metals

Heterologous expression of PtAAS1 reveals the metabolic potential of the common plant metabolite phenylacetaldehyde for auxin synthesis in planta.

Aromatic aldehydes and amines are common plant metabolites involved in several specialized metabolite biosynthesis pathways. Recently, we showed that the aromatic aldehyde synthase PtAAS1 and the aromatic amino acid decarboxylase PtAADC1 contribute to the herbivory-induced formation of volatile 2-phenylethanol and its glucoside 2-phenylethyl-β-D-glucopyranoside in Populus trichocarpa. To unravel alternative metabolic fates of phenylacetaldehyde and 2-phenylethylamine beyond alcohol and alcohol glucoside formation, we heterologously expressed PtAAS1 and PtAADC1 in Nicotiana benthamiana and analyzed plant extracts using untargeted LC-qTOF-MS and targeted LC-MS/MS analysis. While the metabolomes of PtAADC1-expressing plants did not significantly differ from those of control plants, expression of PtAAS1 resulted in the accumulation of phenylacetic acid (PAA) and PAA-amino acid conjugates, identified as PAA-aspartate and PAA-glutamate. Herbivory-damaged poplar leaves revealed significantly induced accumulation of PAA-Asp, while levels of PAA remained unaltered upon herbivory. Transcriptome analysis showed that members of auxin-amido synthetase GH3 genes involved in the conjugation of auxins with amino acids were significantly upregulated upon herbivory in P. trichocarpa leaves. Overall, our data indicates that phenylacetaldehyde generated by poplar PtAAS1 serves as a hub metabolite linking the biosynthesis of volatile, non-volatile herbivory-induced specialized metabolites, and phytohormones, suggesting that plant growth and defense can be balanced on a metabolic level.

Interactions with microbial consortia have variable effects in organic carbon and production of exometabolites among genotypes of Populus trichocarpa.

Poplar is a short-rotation woody crop frequently studied for its significance as a sustainable bioenergy source. The successful establishment of a poplar plantation partially depends on its rhizosphere-a dynamic zone governed by complex interactions between plant roots and a plethora of commensal, mutualistic, symbiotic, or pathogenic microbes that shape plant fitness. In an exploratory endeavor, we investigated the effects of a consortium consisting of ectomycorrhizal fungi and a beneficial Pseudomonas sp. strain GM41 on plant growth (including height, stem girth, leaf, and root growth) and as well as growth rate over time, across four Populus trichocarpa genotypes. Additionally, we compared the level of total organic carbon and plant exometabolite profiles across different poplar genotypes in the presence of the microbial consortium. These data revealed no significant difference in plant growth parameters between the treatments and the control across four different poplar genotypes at 7 weeks post-inoculation. However, total organic carbon and exometabolite profiles were significantly different between the genotypes and the treatments. These findings suggest that this microbial consortium has the potential to trigger early signaling responses in poplar, influencing its metabolism in ways crucial for later developmental processes and stress tolerance.

Genome-wide analysis of the PtHMGR gene family and functional validation of PtHMGR5 improving drought tolerance in Populus trichocarpa

Terpenoids are one of the main types of secondary metabolites in plants and are crucial for plant growth, development and defense. 3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) is a rate-limiting enzyme for terpene biosynthesis in the MVA pathways. In this study, six PtHMGR genes (PtHMGR1-PtHMGR6) were identified in Populus trichocarpa and distributed on six chromosomes. Collinear analysis of PtHMGR genes suggested that six replication gene pairs were acquired by segmental duplication events in the Populus genome. In addition, the promoters of PtHMGRs contained abundant light-response elements, hormone-response elements and abiotic stress-response elements. Expression profile analysis demonstrated that the PtHMGR genes were induced by various abiotic stresses and hormone treatments. Furthermore, transgenic expression of PtHMGR5 in Populus significantly reduced membrane damage and increased drought (mannitol) tolerance. In conclusion, our findings provide valuable information regarding PtHMGR genes, which is of great significance for the cultivation of improved plants in response to drought (mannitol) stress.

Genome-Wide Identification and Expression Analysis of the Copper Transporter (COPT/Ctr) Gene Family in Kandelia obovata, a Typical Mangrove Plant.

The copper transporter (COPT/Ctr) gene family plays a critical part in maintaining the balance of the metal, and many diverse species depend on COPT to move copper (Cu) across the cell membrane. In Arabidopsis thaliana, Oryza sativa, Medicago sativa, Zea mays, Populus trichocarpa, Vitis vinifera, and Solanum lycopersicum, a genome-wide study of the COPT protein family was performed. To understand the major roles of the COPT gene family in Kandelia obovata (Ko), a genome-wide study identified four COPT genes in the Kandelia obovata genome for the first time. The domain and 3D structural variation, phylogenetic tree, chromosomal distributions, gene structure, motif analysis, subcellular localization, cis-regulatory elements, synteny and duplication analysis, and expression profiles in leaves and Cu were all investigated in this research. Structural and sequence investigations show that most KoCOPTs have three transmembrane domains (TMDs). According to phylogenetic research, these KoCOPTs might be divided into two subgroups, just like Populus trichocarpa. KoCOPT gene segmental duplications and positive selection pressure were discovered by universal analysis. According to gene structure and motif analysis, most KoCOPT genes showed consistent exon-intron and motif organization within the same group. In addition, we found five hormones and four stress- and seven light-responsive cis-elements in the KoCOPTs promoters. The expression studies revealed that all four genes changed their expression levels in response to copper (CuCl2) treatments. In summary, our study offers a thorough overview of the Kandelia obovata COPT gene family's expression pattern and functional diversity, making it easier to characterize each KoCOPT gene's function in the future.

Preliminary estimates of genetic parameters and familial selection for non-native poplars show good potential for genetic gains on growth, cold hardiness, trunk quality and Sphaerulina musiva susceptibility

Genetic parameters for growth, trunk quality, and susceptibility to frost and Sphaerulina musiva attack were estimated from 34 half-sib families of hybrid poplar from the crossing of non-native parents, Populus maximowiczii A. Henry, and Populus trichocarpa Torr. & Gray, 3 and 6 years after planting. The use of spatial analysis proved to be the best method for quantitative growth data. The proportion of the among-family variance to the total (phenotypic) variance as well as the high heritabilities of growth and susceptibility to frost and Spaherulina musiva showed a high potential for selection for these traits while the quality traits were under low genetic control. Some families showed gains for several traits, suggesting the possibility of developing a selection index to obtain superior families that show gain for not only growth but quality and adaptive traits as well. Type B correlations were high, suggesting that families responded in the same way regardless of the site. High type A correlation between growth traits at 3 and 6 years showed early selection potential, although these relationships should be confirmed with future measurements to evaluate this effect at maturity. These results can be integrated into the strategy for improving hybrid poplar parental populations and, in the longer term, will make it possible to optimize the selection of individuals with traits of interest for the operational deployment of hybrid poplar clones.

Drought and Salinity Tolerance of the PtWRKY33 Gene in Populus

In this study, the PtWRKY33 gene sequence was cloned by PCR from cDNA, and systematic evolutionary analysis was performed. qRT–PCR was performed to measure the expression of the PtWRKY33 gene in the roots, stems and leaves of Populus tremuloides under drought and saline stresses, and overexpression vectors were constructed and genetically transformed into tobacco. The results showed that the WRKY transcription factors of Populus trichocarpa, Populus euphratica and Populus alba were in the same branch and were closely related to each other. Treating poplar seedlings with 20% PEG6000 solution to simulate drought stress showed that expression of the PtWRKY33 gene was induced and increased 1.89 times, 3.45 times and 11.6 times in leaves, stems and roots. The relative expression of the PtWRKY33 gene was slower to respond to 150 mM NaCl treatment but was nonetheless induced. At 48 h, the increase was 1–3 times in leaves, stems and roots, respectively. NaHCO3 (60 mM) was used to treat poplar seedlings with alkaline salt stress; the results indicated that the PtWRKY33 gene was sensitive to NaHCO3 stress treatment, and that its relative expression was significantly increased and increased 10.51 times (24 h), 6.56 times (6 h) and 5.16 times (24 h) in leaves, stems and roots, respectively. In this study, we found that NaCl and NaHCO3 stress treatments were able to induce an increase in the expression of the NtSOD1 and NtAPX2 genes using qRT–PCR, and the significant increase in expression under the treatments compared with WT may be caused by overexpression of the PtWRKY33 gene. Overexpression of the PtWRKY33 gene in tobacco enhanced the antioxidant stress capacity and improved the salinity tolerance of transgenic tobacco. These results indicate that the PtWRKY33 gene is a key gene for improved salinity-tolerant growth, which is important for future molecular breeding of tree resistance.

Populus trichocarpa PtHSFA4a Enhances Heat Tolerance by Regulating Expression of APX1 and HSPs

Heat stress can severely inhibit plant growth and reproduction, resulting in heavy financial and crop yield losses. Heat shock transcription factors (HSFs) play an important role in regulating plant responses to abiotic stress. However, compared with the in-depth study of HSF gene function in herbaceous species, reports on the regulatory mechanism of the response of HSFs to heat stress in trees are scarce. Here, we demonstrated that PtHSFA4a is induced by high temperatures in Populus trichocarpa leaves. Intense GUS activity was detected in the leaves of PtHSFA4a promoter-GUS reporter transgenic line under heat stress. Ectopic expression of PtHSFA4a in Arabidopsis thaliana enhanced heat stress tolerance, which reduced malondialdehyde and reactive oxygen species levels. RT-qPCR revealed that the expression of key heat stress-related genes (that is, AtMBF1c, AtZAT12, AtAPX1, AtHSA32, and AtHSPs) was upregulated in PtHSFA4a transgenic plants. Additionally, PtHSFA4a directly bind to the promoters of AtAPX1 and AtHSPs under heat stress to enhance heat tolerance by upregulating the antioxidant defense system and maintaining protein folding homeostasis in A. thaliana leaves. These findings provide novel insights into the molecular mechanisms underlying PtHSFA4a-mediated regulation of plant responses to heat stress.