Transgenic and gene-editing technologies are essential for gene functional analysis and crop improvement. However, the pleiotropic effects and unknown mechanisms of morphogenic genes have hindered their broader application. In this study, we employed the one-step de novo shoot organogenesis (DNSO) method, and demonstrated that overexpression of the morphogenic gene Arabidopsis thanalia GROWTH-REGULATING FACTOR 5 (AtGRF5) significantly enhanced genetic transformation efficiency in cucurbit crops by promoting callus proliferation and increasing dense cells during regeneration. High-resolution time-series transcriptomics and single-cell RNA sequencing revealed that AtGRF5 overexpression induced auxin-related genes and expanded stem cell populations during cucumber DNSO. Using DNA-affinity purification sequencing (DAP-seq) in combination with spatiotemporal differential gene expression analysis, we identified CsIAA19 as a key downstream target of AtGRF5, with its modulation playing a pivotal role in regeneration. Rescuing CsIAA19 in AtGRF5-overexpressing explant reversed the enhanced callus proliferation and regeneration. To address growth defects caused by AtGRF5 overexpression, we developed an abscisic acid-inducible AtGRF5 expression system, significantly improving transformation and gene-editing efficiency across diverse genotypes while minimizing pleiotropic effects. In summary, this research provides mechanistic insights into AtGRF5-mediated transformation and offers a practical solution to overcome challenges in cucurbit crop genetic modification.

Synthetic microbial communities (SynComs) are a promising tool for making full use of the beneficial functions imparted by whole bacterial consortia. However, the complexity of reconstructed SynComs often limits their application in sustainable agriculture. Furthermore, inter-strain interactions are often neglected during SynCom construction. Here, we propose a strategy for constructing a simplified and functional SynCom (sfSynCom) by using elite helper strains that significantly improve the beneficial functions of the core symbiotic strain, here Bradyrhizobium elkanii BXYD3, to sustain the growth of soybean (Glycine max). We first identified helper strains that significantly promote nodulation and nitrogen fixation in soybean mediated by BXYD3. Two of these helper strains assigned to the Pantoea taxon produce acyl homoserine lactones, which significantly enhanced the colonization and infection of soybean by BXYD3. Finally, we constructed a sfSynCom from these core and helper strains. This sfSynCom based on the core-helper strategy was more effective at promoting nodulation than inoculation with BXYD3 alone and achieved effects comparable to those of a complex elite SynCom previously constructed on the basis of potential beneficial functions between microbes and plants alone. Our results suggest that considering interactions between strains as well as those between strains and the host plant might allow construction of sfSynComs.

Saline–alkaline soils are a major environmental problem that limit plant growth and crop productivity. Plasma membrane H⁺-ATPases and the salt overly sensitive (SOS) signaling pathway play important roles in plant responses to saline–alkali stress. However, little is known about the functional genes and mechanisms regulating the transcription of H⁺-ATPases and SOS pathway genes under saline–alkali stress. In the present study, we identified that the plant AT-rich sequence and zinc-binding (TaPLATZ2) transcription factor are involved in wheat response to saline–alkali stress by directly suppressing the expression of TaHA2/TaSOS3. The knockdown of TaPLATZ2 enhances salt and alkali stress tolerance, while overexpression of TaPLATZ2 leads to salt and alkali stress sensitivity in wheat. In addition, TaWRKY55 directly upregulated the expression of TaPLATZ2 during saline–alkali stress. Through knockdown and overexpression of TaWRKY55 in wheat, TaWRKY55 was shown to negatively modulate salt and alkali stress tolerance. Genetic analyses confirmed that TaPLATZ2 functions downstream of TaWRKY55 in response to salt and alkaline stresses. These findings provide a TaWRKY55–TaPLATZ2–TaHA2/TaSOS3 regulatory module that regulates wheat responses to saline–alkali stress.

The phytohormone jasmonates (JAs) regulate plant growth and defense responses. The reproductive organs of flowers are devastated by insect herbivores. However, the molecular mechanisms of floral defense remain largely unknown. Here, we found that the Arabidopsis JA receptor CORONATINE INSENSITIVE1 (COI1) and its substrates JA ZIM-domain (JAZ) repressors, and the mediator subunit MEDIATOR25-based MED25–MYC–MYB (MMM) complexes, including MYC2/3/4/5 and MYB28/29/76, mediated floral defense against the insects Helicoverpa armigera, Spodoptera exigua, and Spodoptera frugiperda. The flower-specific IIIa bHLH factors ABORTED MICROSPORES (AMS) and DYSFUNCTIONAL TAPETUM 1 (DYT1) were JAZ-interaction proteins. They interacted with members of the MMM complexes, inhibited the transcriptional activity of MYC2 and MYB28, and repressed floral defense against insects. AMS and DYT1 recruited the flower-specific MYB21/24, and these MYBs interacted with members of MMM complexes, inhibited the MYC2–MYB28 function, and suppressed floral defense against insects. Our study revealed that the JA–COI1–JAZ–MMM pathway mediated flower defense, and the AMS/DYT1–MYB21/24 module antagonized the MMM complexes to repress floral defense against insects.

Lodging reduces grain yield and quality in cereal crops. Lodging resistance is affected by the strength of the culm, which is influenced by the culm diameter, culm wall thickness, and cell wall composition. To explore the genetic architecture of culm diameter in rice (Oryza sativa), we conducted a genome-wide association study (GWAS). We identified STRONG CULM 2 (STRONG2), which encodes the mannan synthase CSLA5, and showed that plants that overexpressed this gene had increased culm diameter and improved lodging resistance. STRONG2 appears to increase the levels of cell wall components, such as mannose and cellulose, thereby enhancing sclerenchyma development in stems. SNP14931253 in the STRONG2 promoter contributes to variation in STRONG2 expression in natural germplasms and the transcription factor MYB61 directly activates STRONG2 expression. Furthermore, STRONG2 overexpressing plants produced significantly more grains per panicle and heavier grains than the wild-type plants. These results demonstrate that the MYB61–STRONG2 module positively regulates culm diameter and lodging resistance, information that could guide breeding efforts for improved yield in rice.

Besides playing a crucial role in plant immunity via the nonexpressor of pathogenesis-related (NPR) proteins, increasing evidence shows that salicylic acid (SA) can also regulate plant root growth. However, the transcriptional regulatory network controlling this SA response in plant roots is still unclear. Here, we found that NPR1 and WRKY45, the central regulators of SA response in rice leaves, control only a reduced sector of the root SA signaling network. We demonstrated that SA attenuates root growth via a novel NPR1/WRKY45-independent pathway. Furthermore, using regulatory network analysis and mutant characterization, we identified a set of new NPR1/WRKY45-independent regulators that conservedly modulate the root development and root-associated microbiota composition in both Oryza sativa (monocot) and Arabidopsis thaliana (dicot) in response to SA. Our results established the SA signaling as a central element regulating plant root functions under ecologically relevant conditions. These results provide new insights to understand how regulatory networks control plant responses to abiotic and biotic stresses.

Reactive oxygen species (ROS) plays critical roles in modulating plant growth and stress response and its homeostasis is fine tuned using multiple peroxidases. H₂O₂, a major kind of ROS, is removed rapidly and directly using three catalases, CAT1, CAT2, and CAT3, in Arabidopsis. Although the activity regulations of catalases have been well studied, their degradation pathway is less clear. Here, we report that CAT2 and CAT3 protein abundance was partially controlled using the 26S proteasome. To further identify candidate proteins that modulate the stability of CAT2, we performed yeast-two-hybrid screening and recovered several clones encoding a protein with RING and vWA domains, CIRP1 (CAT2 Interacting RING Protein 1). Drought and oxidative stress downregulated CIRP1 transcripts. CIRP1 harbored E3 ubiquitination activity and accelerated the degradation of CAT2 and CAT3 by direct interaction and ubiquitination. The cirp1 mutants exhibited stronger drought and oxidative stress tolerance, which was opposite to the cat2 and cat3 mutants. Genetic analysis revealed that CIRP1 acts upstream of CAT2 and CAT3 to negatively regulate drought and oxidative stress tolerance. The increased drought and oxidative stress tolerance of the cirp1 mutants was due to enhanced catalase (CAT) activities and alleviated ROS levels. Our data revealed that the CIRP1–CAT2/CAT3 module plays a vital role in alleviating ROS levels and balancing growth and stress responses in Arabidopsis.

Coniferous forests are under severe threat of the rapid anthropogenic climate warming. Abies (firs), the fourth-largest conifer genus, is a keystone component of the boreal and temperate dark-coniferous forests and harbors a remarkably large number of relict taxa. However, the uncertainty of the phylogenetic and biogeographic history of Abies significantly impedes our prediction of future dynamics and efficient conservation of firs. In this study, using 1,533 nuclear genes generated from transcriptome sequencing and a complete sampling of all widely recognized species, we have successfully reconstructed a robust phylogeny of global firs, in which four clades are strongly supported and all intersectional relationships are resolved, although phylogenetic discordance caused mainly by incomplete lineage sorting and hybridization was detected. Molecular dating and ancestral area reconstruction suggest a Northern Hemisphere high-latitude origin of Abies during the Late Cretaceous, but all extant firs diversified during the Miocene to the Pleistocene, and multiple continental and intercontinental dispersals took place in response to the late Neogene climate cooling and orogenic movements. Notably, four critically endangered firs endemic to subtropical mountains of China, including A. beshanzuensis, A. ziyuanensis, A. fanjingshanensis and A. yuanbaoshanensis from east to west, have different origins and evolutionary histories. Moreover, three hotspots of species richness, including western North America, central Japan, and the Hengduan Mountains, were identified in Abies. Elevation and precipitation, particularly precipitation of the coldest quarter, are the most significant environmental factors driving the global distribution pattern of fir species diversity. Some morphological traits are evolutionarily constrained, and those linked to elevational variation (e.g., purple cone) and cold resistance (e.g., pubescent branch and resinous bud) may have contributed to the diversification of global firs. Our study sheds new light on the spatiotemporal evolution of global firs, which will be of great help to forest management and species conservation in a warming world.

In C₃ plants, photorespiration is an energy expensive pathway that competes with photosynthetic CO₂ assimilation and releases CO₂ into the atmosphere, potentially reducing C₃ plant productivity by 20%-50%. Consequently, reducing the flux through photorespiration has been recognized as a major way to improve C₃ crop photosynthetic carbon fixation and productivity. While current research efforts in engineering photorespiration are mainly based on the modification of chloroplast glycolate metabolic steps, only limited studies have explored optimizations in other photorespiratory metabolic steps. Here, we engineered an imGS bypass within the rice mitochondria to bypass the photorespiratory glycine toward glycine betaine, thereby, improving the photosynthetic carbon fixation in rice. The imGS transgenic rice plants exhibited significant accumulation of glycine betaine, reduced photorespiration, and elevated photosynthesis and photosynthate levels. Additionally, the introduction of imGS bypass into rice leads to an increase in the number of branches and grains per panicle which may be related to cytokinin and gibberellin signaling pathways. Taken together, these results suggest diverting mitochondrial glycine from photorespiration toward glycine betaine synthesis can effectively enhance carbon fixation and panicle architecture in rice, offering a promising strategy for developing functional mitochondrial photorespiratory bypasses with the potential to enhance plant productivity.

The calcineurin B-like protein (CBL)-CBL-interacting protein kinase (CIPK) Ca²⁺ sensors play crucial roles in the plant's response to drought stress. However, there have been few reports on the synergistic regulation of drought stress by CBL-CIPK and abscisic acid (ABA) core signaling components. In this study, we discovered that ZmCIPK33 positively regulates drought resistance in maize. ZmCIPK33 physically interacts with and is enhanced by phosphorylation from ZmSnRK2.10. Drought stress can activate ZmCIPK33, which is partially dependent on ZmSnRK2.10. ZmCIPK33 in combination with ZmSnRK2.10 can activate the slow anion channel ZmSLAC1 in Xenopus laevis oocytes independently of CBLs, whereas ZmCIPK33 or ZmSnRK2.10 alone is unable to do so. Furthermore, ZmCIPK33 phosphorylates ZmPP2C11 at Ser60, which leads to a reduction in the interaction between ZmPP2C11 and ZmEAR1 (the ortholog of Arabidopsis Enhancer of ABA co-Receptor 1) and weakens the phosphatase activity of ZmPP2C11, consequently, enhancing the activity of ZmSnRK2.10 in an in vitro assay and in the in-gel assay of the zmcipk33 mutant. Our findings provide novel insights into the molecular mechanisms underlying the reciprocal enhancement of Ca²⁺ and ABA signaling under drought stress in maize.

Insects secret chemosensory proteins (CSPs) into plant cells as potential effector proteins during feeding. The molecular mechanisms underlying how CSPs activate plant immunity remain largely unknown. We show that CSPs from six distinct insect orders induce dwarfism when overexpressed in Nicotiana benthamiana. Agrobacterium-mediated transient expression of Nilaparvata lugens CSP11 (NlCSP11) triggered cell death and plant dwarfism, both of which were dependent on ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1), N requirement gene 1 (NRG1) and SENESCENCE-ASSOCIATED GENE 101 (SAG101), indicating the activation of effector-triggered immunity (ETI) in N. benthamiana. Overexpression of NlCSP11 led to stronger systemic resistance against Pseudomonas syringae DC3000 lacking effector HopQ1-1 and tobacco mosaic virus, and induced higher accumulation of salicylic acid (SA) in uninfiltrated leaves compared to another effector XopQ that is recognized by a Toll-interleukin-1 receptor (TIR) domain nucleotide-binding leucine-rich repeat receptor (TNL) called ROQ1 in N. benthamiana. Consistently, NlCSP11-induced dwarfism and systemic resistance, but not cell death, were abolished in N. benthamiana transgenic line expressing the SA-degrading enzyme NahG. Through large-scale virus-induced gene silencing screening, we identified a TNL protein that mediates the recognition of CSPs (RCSP), including aphid effector MP10 that triggers resistance against aphids in N. benthamiana. Co-immunoprecipitation, bimolecular fluorescence complementation and AlphaFold2 prediction unveiled an interaction between NlCSP11 and RCSP. Interestingly, RCSP does not contain the conserved catalytic glutamic acid in the TIR domain, which is required for TNL function. Our findings point to enhanced ETI and systemic resistance by a TNL protein via hyperactivation of the SA pathway. Moreover, RCSP is the first TNL identified to recognize an insect effector.

Mitogen-activated protein kinase (MAPK) cascades play a fundamental role in plant immunity by transducing external signals inside plant cells. Here, we defined a wheat MAPK cascade, composed of the mitogen-activated protein kinase kinase (MAPKK) TaMKK2 and its downstream MAPK TaMAPK6, which phosphorylates the core immune regulator TaSGT1 (suppressor of G2 allele of Skp1), resulting in enhanced nuclear entry of TaSGT1, thereby conferring resistance against the devastating wheat pathogen Puccinia striiformis f. sp. tritici (Pst). Hence, we identified a TaMKK2-TaMAPK6-TaSGT1 signaling cascade that contributes to wheat stripe rust resistance. During infection, Pst secrets a haustorium-associated secreted protein 215 (HASP215), that targets TaMKK2 and interferes with the interaction of TaMKK2 with TaMAPK6 to suppress TaMAPK6 phosphorylation and activation, thereby leading to reduced capacity of TaMAPK6 to phosphorylate TaSGT1. Consequently, inhibition of TaMAPK6-mediated TaSGT1 phosphorylation resulted in decreased nuclear translocation of TaSGT1 and suppressed plant immunity. Our work elucidates the positive function of TaMKK2-TaMAPK6 cascade in wheat immunity by regulating the immune component TaSGT1, and its regulation by the rust effector HASP215, providing new insights into the MAPK cascade on crop immunity and the pathogenicity mechanism of obligate biotrophic fungus.

Leaf senescence can be triggered by various abiotic stresses. Among these, heat stress emerges as a pivotal environmental factor, particularly in light of the predicted rise in global temperatures. However, the molecular mechanism underlying heat-induced leaf senescence remains largely unexplored. As a cool-season grass species, tall fescue (Festuca arundinacea) is an ideal and imperative material for investigating heat-induced leaf senescence because heat stress easily triggers leaf senescence to influence its forage yield and turf quality. Here, we investigated the role of FaNAC047 in heat-induced leaf senescence. Overexpression of FaNAC047 promoted heat-induced leaf senescence in transgenic tall fescue that was evidenced by a more seriously destructive photosystem and higher accumulation of reactive oxygen species (ROS), whereas knockdown of FaNAC047 delayed leaf senescence. Further protein-DNA interaction assays indicated that FaNAC047 directly activated the transcriptions of NON-YELLOW COLORING 1 (FaNYC1), NYC1-like (FaNOL), and STAY-GREEN (FaSGR) but directly inhibited Catalases 2 (FaCAT2) expression, thereby promoting chlorophyll degradation and ROS accumulation. Subsequently, protein-protein interaction assays revealed that FaNAC047 physically interacted with FaNAC058 to enhance its regulatory effect on FaNYC1, FaNOL, FaSGR, and FaCAT2. Additionally, FaNAC047 could transcriptionally activate FaNAC058 expression to form a regulatory cascade, driving senescence progression. Consistently, the knockdown of FaNAC058 significantly delayed heat-induced leaf senescence. Collectively, our results reveal that FaNAC047-FaNAC058 module coordinately mediates chlorophyll degradation and ROS production to positively regulate heat-induced leaf senescence. The findings illustrate the molecular network of heat-induced leaf senescence for breeding heat-resistant plants.

Glycosylation, a prevalent post-translational modification in eukaryotic secreted and membrane-associated proteins, plays a pivotal role in diverse physiological and pathological processes. Although UDP-N-acetylglucosamine (UDP-GlcNAc) is essential for this modification, the specific glycosylation mechanisms during plant leaf senescence and defense responses remain poorly understood. In our research, we identified a novel rice mutant named rbb1 (resistance to blast and bacterial blight1), exhibiting broad-spectrum disease resistance. This mutant phenotype results from a loss-of-function mutation in the gene encoding glucosamine-6-phosphate acetyltransferase, an important enzyme in D-glucosamine 6-phosphate acetylation. The rbb1 mutant demonstrates enhanced defense responses, evident in increased resistance to rice blast and bacterial blight, along with the upregulation of defense-response genes. Various biochemical markers indicate an activated defense mechanism in the rbb1 mutant, such as elevated levels of reactive oxygen species and malondialdehyde, reduced enzyme activity and UDP-GlcNAc content, and decreased expression of N-glycan and O-glycan modifying proteins. Moreover, proteome analysis of N-glycosylation modifications reveals alterations in the N-glycosylation of several disease-resistance-related proteins, with a significant reduction in Prx4 and Prx13 in rbb1-1. Additionally, the knockout of Prx4 or Prx13 also enhances resistance to Xanthomonas oryzae pv. oryzae (Xoo) and Magnaporthe oryzae (M. oryzae). This study uncovers a novel mechanism of defense response in rice, suggesting potential targets for the development of disease-resistant varieties.

Diatoms rely on fucoxanthin chlorophyll a/c-binding proteins (FCPs) for light harvesting and energy quenching under marine environments. Here we report two cryo-electron microscopic structures of photosystem I (PSI) with either 13 or five fucoxanthin chlorophyll a/c-binding protein Is (FCPIs) at 2.78 and 3.20 Å resolutions from Thalassiosira pseudonana grown under high light (HL) conditions. Among them, five FCPIs are stably associated with the PSI core, these include Lhcr3, RedCAP, Lhcq8, Lhcf10, and FCP3. The eight additional Lhcr-type FCPIs are loosely associated with the PSI core and detached under the present purification conditions. The pigments of this centric diatom showed a higher proportion of chlorophylls a, diadinoxanthins, and diatoxanthins; some of the chlorophyll as and diadinoxanthins occupy the locations of fucoxanthins found in the huge PSI-FCPI from another centric diatom Chaetoceros gracilis grown under low-light conditions. These additional chlorophyll as may form more energy transfer pathways and additional diadinoxanthins may form more energy dissipation sites relying on the diadinoxanthin-diatoxanthin cycle. These results reveal the assembly mechanism of FCPIs and corresponding light-adaptive strategies of T. pseudonana PSI-FCPI, as well as the convergent evolution of the diatom PSI-FCPI structures.

Plant regeneration is the process during which differentiated tissues or cells can reverse or alter their developmental trajectory to repair damaged tissues or form new organs. In the plant regeneration process, the cell wall not only functions as a foundational barrier and scaffold supporting plant cells but also influences cell fates and identities. Cell wall remodeling involves the selective degradation of certain cell wall components or the integration of new components. Recently, accumulating evidence has underscored the importance of cell wall remodeling in plant regeneration. Wounding signals, transmitted by transcription factors, trigger the expressions of genes responsible for cell wall loosening, which is essential for tissue repair. In de novo organ regeneration and somatic embryogenesis, phytohormones orchestrate a transcriptional regulatory network to induce cell wall remodeling, which promotes cell fate reprogramming and organ formation. This review summarizes the effects of cell wall remodeling on various regenerative processes and provides novel insights into the future research of uncharacterized roles of cell wall in plant regeneration.

Tomato (Solanum lycopersicum) is an important crop but frequently experiences saline–alkali stress. Our previous studies have shown that exogenous spermidine (Spd) could significantly enhance the saline–alkali resistance of tomato seedlings, in which a high concentration of Spd and jasmonic acid (JA) exerted important roles. However, the mechanism of Spd and JA accumulation remains unclear. Herein, SlWRKY42, a Group II WRKY transcription factor, was identified in response to saline–alkali stress. Overexpression of SlWRKY42 improved tomato saline–alkali tolerance. Meanwhile, SlWRKY42 knockout mutants, exhibited an opposite phenotype. RNA-sequencing data also indicated that SlWRKY42 regulated the expression of genes involved in JA signaling and Spd synthesis under saline–alkali stress. SlWRKY42 is directly bound to the promoters of SlSPDS2 and SlNHX4 to promote Spd accumulation and ionic balance, respectively. SlWRKY42 interacted with SlMYC2. Importantly, SlMYC2 is also bound to the promoter of SlSPDS2 to promote Spd accumulation and positively regulated saline–alkali tolerance. Furthermore, the interaction of SlMYC2 with SlWRKY42 boosted SlWRKY42's transcriptional activity on SlSPDS2, ultimately enhancing the tomato's saline–alkali tolerance. Overall, our findings indicated that SlWRKY42 and SlMYC2 promoted saline–alkali tolerance by the Spd biosynthesis pathway. Thus, this provides new insight into the mechanisms of plant saline–alkali tolerance responses triggered by polyamines (PAs).

Increasing plant density has been recognized as an effective strategy for boosting maize yields over the past few decades. However, dense planting significantly reduces the internal light intensity and the red to far-red (R:FR) light ratio in the canopy, which subsequently triggers shade avoidance responses (SAR) that limit further yield enhancements, particularly under high-density conditions. In this study, we identified double B-box containing protein DBB2, a member of the ZmBBX family that is rapidly induced by shade, as a crucial regulator of plant height and SAR. Disruption of DBB2 resulted in shorter internodes, reduced plant height, decreased cell elongation, and diminished sensitivity to shade in maize, effects that can be largely alleviated by external treatment with gibberellins (GA). Furthermore, we discovered that DBB2 physically interacted with the transcription factor HY5, inhibiting its transcriptional activation of ZmGA2ox4, a gene encoding a GA2 oxidase that can deactivate GA. This interaction positively influences maize plant height through the GA pathway. Additionally, we found that the induction of ZmDBB2 by shade is mediated by the transcription factor PIF4. Interestingly, DBB2 then interacted with PIF4 to enhance the transcriptional activation of cell elongation-related genes, such as ZmEXPA1, thereby establishing a positive feedback loop promoting cell elongation under canopy shade conditions. Our findings highlight the critical role of BBX proteins in modulating plant height and SAR, presenting them as key genetic targets for developing maize varieties suited to high-density planting conditions. This study also provides new insights into the molecular mechanisms underlying SAR and offers potential strategies for the genetic improvement of maize plant architecture and grain yield.

Poplar plantations are often established on nitrogen-poor land, and poplar growth and wood formation are constrained by low nitrogen (LN) availability. However, the molecular mechanisms by which specific genes regulate wood formation in acclimation to LN availability remain unclear. Here, we report a previously unrecognized module, basic region/leucine zipper 55 (PtobZIP55)–PtoMYB170, which regulates the wood formation of Populus tomentosa in acclimation to LN availability. PtobZIP55 was highly expressed in poplar wood and induced by LN. Altered wood anatomical properties and increased lignification were detected in PtobZIP55-overexpressing poplars, whereas the opposite results were detected in PtobZIP55-knockout poplars. Molecular and transgenic analyses revealed that PtobZIP55 directly binds to the promoter sequence of PtoMYB170 to activate its transcription. The phenotypes of PtoMYB170 transgenic poplars were similar to those of PtobZIP55 transgenic poplars under LN conditions. Further molecular analyses revealed that PtoMYB170 directly bound the promoter sequences of lignin biosynthetic genes to activate their transcription to increase lignin concentrations in LN-treated poplar wood. These results suggest that PtobZIP55 activates PtoMYB170 transcription, which in turn positively regulates lignin biosynthetic genes, increasing lignin deposition in the wood of P. tomentosa in the context of acclimation to LN availability.

One mechanism plants use to tolerate high salinity is the deposition of cutin and suberin to form apoplastic barriers that limit the influx of ions. However, the mechanism underlying barrier formation under salt stress is unclear. Here, we characterized the glycerol-3-phosphate acyltransferase (GPAT) family gene TaGPAT6, encoding a protein involved in cutin and suberin biosynthesis for apoplastic barrier formation in wheat (Triticum aestivum). TaGPAT6 has both acyltransferase and phosphatase activities, which are responsible for the synthesis of sn-2-monoacylglycerol (sn-2 MAG), the precursor of cutin and suberin. Overexpressing TaGPAT6 promoted the deposition of cutin and suberin in the seed coat and the outside layers of root tip cells and enhanced salt tolerance by reducing sodium ion accumulation within cells. By contrast, TaGPAT6 knockout mutants showed increased sensitivity to salt stress due to reduced cutin and suberin deposition and enhanced sodium ion accumulation. Yeast-one-hybrid and electrophoretic mobility shift assays identified TaABI5 as the upstream regulator of TaGPAT6. TaABI5 knockout mutants showed suppressed expression of TaGPAT6 and decreased barrier formation in the seed coat. These results indicate that TaGPAT6 is involved in cutin and suberin biosynthesis and the resulting formation of an apoplastic barrier that enhances salt tolerance in wheat.