Defense Priming Research Articles

VOZ‐dependent priming of salicylic acid‐dependent defense against Rhizopus stolonifer by β‐aminobutyric acid requires the TCP protein TCP2 in peach fruit

SUMMARYVascular plant one‐zinc finger (VOZ) transcription factors (TFs) play crucial roles in plant immunity. Nevertheless, how VOZs modulate defense signaling in response to elicitor‐induced resistance is not fully understood. Here, the defense elicitor β‐aminobutyric acid (BABA) resulted in the visible suppression of Rhizopus rot disease of peach fruit caused by Rhizopus stolonifer. Defense priming by BABA was notably associated with increased levels of salicylic acid (SA) and SA‐dependent gene expression. Data‐independent acquisition proteomic analysis revealed that two VOZ proteins (PpVOZ1 and PpVOZ2) were substantially upregulated in BABA‐induced resistance (BABA‐IR). Furthermore, the interaction of PpVOZ1 and PpVOZ2 and their potential target of the TEOSINTE‐BRANCHED1/CYCLOIDEA/PCF (TCP)‐family protein PpTCP2 screened from protein–protein interaction networks was confirmed by yeast two‐hybrid (Y2H), luciferase complementation imaging and glutathione S‐transferase pull‐down assays. Furthermore, subcellular localization, yeast one‐hybrid, electrophoretic mobility shift assay and dual‐luciferase reporter assays demonstrated that nuclear localization of both PpVOZ1 and PpVOZ2 was critical for their contribution to BABA‐IR, as these proteins potentiated the PpTCP2‐mediated transcriptional activation of isochorismate synthase genes (ICS1/2). The overexpression of both PpVOZ1 and PpVOZ2 could activate the transcription of SA‐dependent genes and provide disease resistance in transgenic Arabidopsis. In contrast, the ppvoz1cas9 and ppvoz2cas9 loss‐of‐function mutations and the voz1cas9 voz2cas9 double mutation attenuated BABA‐IR against R. stolonifer. Therefore, the three identified positive TFs, PpVOZ1, PpVOZ2, and PpTCP2, synergistically contribute to the BABA‐activated priming of systemic acquired resistance in postharvest peach fruit by a VOZ‐TCP‐ICS regulatory module.

Developmentally regulated generation of a systemic signal for long-lasting defence priming in tomato.

Tomato is a major global crop. However, its production is limited by Botrytis cinerea. Due to the toxicity of postharvest pesticide application, alternative control methods such as priming are being investigated. Plants were treated with β-aminobutyric acid (BABA) at two developmental stages and resistance against B. cinerea was tested in fruit tissue and in progenies. DNA methylation and RNA sequencing were conducted to characterise the (epi)genetic changes associated with long-lasting resistance. Grafting experiments were done to assess the systemic nature of this signal, which was further characterised by small RNA (sRNA) sequencing of scions. Only BABA-treated seedlings displayed induced resistance (IR). DNA methylation analysis revealed seedling-specific changes, which occurred in the context of lower basal methylation. BABA-IR was found to be transmissible from primed rootstock to grafted unprimed scions. In these scions, we identified a subset of mobile 24 nt sRNAs associated with genes showing primed expression during infection in fruit. Our results demonstrate the functional association of a systemic signal with long-lasting IR and priming. Through integrated omics approaches, we have identified markers of long-lasting priming in tomato fruit which could also serve as targets for durable resistance in other crops.

Potato β-aminobutyric acid receptor IBI1 manipulates VOZ1 and VOZ2 transcription factor activity to promote disease resistance.

Upon infection with non-pathogenic microorganisms or treatment with natural or synthetic compounds, plants exhibit a more rapid and potent response to both biotic and abiotic stresses. However, the molecular mechanisms behind this phenomenon, known as defense priming, are poorly understood. β-aminobutyric acid (BABA) is an endogenous stress metabolite that enhances plant tolerance to various abiotic stresses and primes plant defense responses, providing the ability to resist a variety of pathogens (broad-spectrum resistance). In this study, we identified an aspartyl-tRNA synthetase (AspRS), StIBI1 (named after Arabidopsis IMPAIRED IN BABA-INDUCED IMMUNITY 1; IBI1), as a BABA receptor in Solanum tuberosum. We elucidated the regulatory mechanisms by which StIBI1 interacts with two NAC (NAM, ATAF1, 2, and CUC2) transcription factors (TFs), StVOZ1 and StVOZ2 (VASCULAR PLANT ONE ZINC FINGER, VOZ), to activate BABA-induced resistance (BABA-IR). StVOZ1 represses, whereas StVOZ2 promotes, immunity to the late blight pathogen Phytophthora infestans. Interestingly, BABA and StIBI1 influence StVOZ1- and StVOZ2-mediated immunity. StIBI1 interacts with StVOZ1 and StVOZ2 in the cytoplasm, reducing the nuclear accumulation of StVOZ1 and promoting the nuclear accumulation of StVOZ2. Our findings indicate that StVOZ1 and StVOZ2 finely regulate potato resistance to late blight through distinct signaling pathways. In summary, our study provides insights into the interaction between the potato BABA receptor StIBI1 and the TFs StVOZ1 and StVOZ2, which affects StVOZ1 and StVOZ2stability and nuclear accumulation to regulate late blight resistance during BABA-IR. This research advances our understanding of the primary mechanisms of BABA-IR in potato and contributes to a theoretical basis for the prevention and control of potato late blight using BABA-IR.

Overexpressing CsSABP2 enhances tolerance to Huanglongbing and citrus canker in C. sinensis.

Huanglongbing (HLB) and citrus canker, arising from Candidatus Liberibacter asiaticus (CaLas) and Xanthomonas citri pv. Citri (Xcc), respectively, have been imposing tremendous losses to the global citrus industry. Systemic acquired resistance (SAR) has been shown to be crucial for priming defense against pathogen in citrus. Salicylic acid (SA) binding protein 2 (SABP2), which is responsible for converting methyl salicylate (MeSA) to SA, is essential for full SAR establishment. Here, we characterized the functions of four citrus SABP2 genes (CsSABP2-1, CsSABP2-1V18A , CsSABP2-2 and CsSABP2-3) against HLB and citrus canker. In vitro enzymatic assay revealed that all four proteins had MeSA esterase activities, and CsSABP2-1 and CsSABP2-1V18A has the strongest activity. Their activities were inhibited by SA except for CsSABP2-1V18A. Four genes controlled by a strong promoter 35S were induced into Wanjincheng orange (Citrus sinensis Osbeck) to generate transgenic plants overexpressing CsSABP2. Overexpressing CsSABP2 increased SA and MeSA content and CsSABP2-1V18A had the strongest action on SA. Resistance evaluation demonstrated that only CsSABP2-1V18A had significantly enhanced tolerance to HLB, although all four CsSABP2s had increased tolerance to citrus canker. The data suggested the amino acid Val-18 in the active site of CsSABP2 plays a key role in protein function. Our study emphasized that balancing the levels of SA and MeSA is crucial for regulating SAR and conferring broad-spectrum resistance to HLB and citrus canker. This finding offers valuable insights for enhancing resistance through SAR engineering.

Butterfly eggs prime anti-herbivore defense in an annual but not perennial Arabidopsis species

Main conclusionUnlike Arabidopsis thaliana, defenses of Arabidopsis lyrata against Pieris brassicae larval feeding are not primable by P. brassicae eggs. Thus, egg primability of plant anti-herbivore defenses is not phylogenetically conserved in the genus Arabidopsis.While plant anti-herbivore defenses of the annual species Arabidopsis thaliana were shown to be primable by Pieris brassicae eggs, the primability of the phylogenetically closely related perennial Arabidopsis lyrata has not yet been investigated. Previous studies revealed that closely related wild Brassicaceae plant species, the annual Brassica nigra and the perennial B. oleracea, exhibit an egg-primable defense trait, even though they have different life spans. Here, we tested whether P. brassicae eggs prime anti-herbivore defenses of the perennial A. lyrata. We exposed A. lyrata to P. brassicae eggs and larval feeding and assessed their primability by (i) determining the biomass of P. brassicae larvae after feeding on plants with and without prior P. brassicae egg deposition and (ii) investigating the plant transcriptomic response after egg deposition and/or larval feeding. For comparison, these studies were also conducted with A. thaliana. Consistent with previous findings, A. thaliana’s response to prior P. brassicae egg deposition negatively affected conspecific larvae feeding upon A. thaliana. However, this was not observed in A. lyrata. Arabidopsis thaliana responded to P. brassicae eggs with strong transcriptional reprogramming, whereas A. lyrata responses to eggs were negligible. In response to larval feeding, A. lyrata exhibited a greater transcriptome change compared to A. thaliana. Among the strongly feeding-induced A. lyrata genes were those that are egg-primed in feeding-induced A. thaliana, i.e., CAX3, PR1, PR5, and PDF1.4. These results suggest that A. lyrata has evolved a robust feeding response that is independent from prior egg exposure.

NAD+ deficiency primes defense metabolism via 1O2-escalated jasmonate biosynthesis in plants

Nicotinamide adenine dinucleotide (NAD+) is a redox cofactor and signal central to cell metabolisms. Disrupting NAD homeostasis in plant alters growth and stress resistance, yet the underlying mechanisms remain largely unknown. Here, by combining genetics with multi-omics, we discover that NAD+ deficiency in qs-2 caused by mutation in NAD+ biosynthesis gene-Quinolinate Synthase retards growth but induces biosynthesis of defense compounds, notably aliphatic glucosinolates that confer insect resistance. The elevated defense in qs-2 is resulted from activated jasmonate biosynthesis, critically hydroperoxidation of α-linolenic acid by the 13-lipoxygenase (namely LOX2), which is escalated via the burst of chloroplastic ROS-singlet oxygen (1O2). The NAD+ deficiency-mediated JA induction and defense priming sequence in plants is recapitulated upon insect infestation, suggesting such defense mechanism operates in plant stress response. Hence, NAD homeostasis is a pivotal metabolic checkpoint that may be manipulated to navigate plant growth and defense metabolism for stress acclimation.

Bolstering Wheat's immunity: BABA-mediated defense priming against Bipolaris sorokiniana amid competition

Wheat productivity is significantly limited by biotic stresses, notably Bipolaris sorokiniana, which are exacerbated by competition for nutrients and space among plants. This study addresses the critical problem of developing sustainable, pesticide-free strategies to enhance wheat defense against pathogens, particularly under competitive conditions. In the present study, we demonstrated BABA priming in wheat against Bipolaris sorokiniana under varying degrees of competition. Our findings indicate that growth parameters were reduced in both primed and non-primed wheat with an increase in the level of competition. However, overall, the primed wheat showed better morphological growth compared to the non-primed at each level of competition. Under disease pressure, BABA-primed performed better than non-primed wheat at each level of competition, and the level of protection was consistent across all competition levels. Biochemical parameters such as photosynthetic pigments, enzymatic and non-enzymatic antioxidants, and defence enzymes suggested that the defence mechanism was activated in primed wheat only after pathogen challenge, ensuring the establishment of the priming effect. Non-primed HD plants exhibited maximum susceptibility under high competition and disease pressure. BABA priming enabled the plants to outweigh the fitness cost and mitigate the effects of competition under pathogen pressure. Additionally, the yield performance of BABA-primed wheat was superior across all competition levels. These results suggest that BABA priming is a viable, pesticide-free approach to enhancing wheat resilience against Bipolaris sorokiniana, particularly in competitive environments. Our research highlights the potential of BABA priming to contribute to sustainable agricultural practices by improving crop protection and yield.

Companion basil plants prime the tomato wound response through volatile signaling in a mixed planting system

Key messageVolatile compounds released from basil prime the tomato wound response by promoting jasmonic acid, mitogen-activated protein kinase, and reactive oxygen species signaling.Within mixed planting systems, companion plants can promote growth or enhance stress responses in target plants. However, the mechanisms underlying these effects remain poorly understood. To gain insight into the molecular nature of the effects of companion plants, we investigated the effects of basil plants (Ocimum basilicum var. minimum) on the wound response in tomato plants (Solanum lycopersicum cv. ‘Micro-Tom’) within a mixed planting system under environmentally controlled chamber. The results showed that the expression of Pin2, which specifically responds to mechanical wounding, was induced more rapidly and more strongly in the leaves of tomato plants cultivated with companion basil plants. This wound response priming effect was replicated through the exposure of tomato plants to an essential oil (EO) prepared from basil leaves. Tomato leaves pre-exposed to basil EO showed enhanced expression of genes related to jasmonic acid, mitogen-activated protein kinase (MAPK), and reactive oxygen species (ROS) signaling after wounding stress. Basil EO also enhanced ROS accumulation in wounded tomato leaves. The wound response priming effect of basil EO was confirmed in wounded Arabidopsis plants. Loss-of-function analysis of target genes revealed that MAPK genes play pivotal roles in controlling the observed priming effects. Spodoptera litura larvae-fed tomato leaves pre-exposed to basil EO showed reduced growth compared with larvae-fed control leaves. Thus, mixed planting with basil may enhance defense priming in both tomato and Arabidopsis plants through the activation of volatile signaling.

Leveraging air-borne VOC-mediated plant defense priming to optimize Integrated Pest Management

Leveraging air-borne VOC-mediated plant defense priming to optimize Integrated Pest Management

Exogenous systemin peptide treatment in olive alters Bactrocera oleae oviposition preference

AbstractThe olive fruit fly, Bactrocera oleae (Rossi), is a key pest of the olive crop, whose control relies mostly on the use of insecticides. Plant peptides may represent a more environmentally-friendly tool to manage olive fly, due to their recognized role to activate and/or prime plant defence responses against pests. In this work, behavioural experiments (no-choice and two-choice) and analysis of volatile compounds were carried out to evaluate the impact of the exogenous application of the peptide systemin to olive tree on olive fly infestation, and to elucidate its mode of action to prime plant defence. The treatment of olive branches with 10 nM systemin showed to confer protection against olive fly, by reducing significantly the ovipositions (up to 3.0-fold) and the number of infested fruits (up to 2.9-fold) when compared to non-treated branches. This protective effect was even detected in neighbouring non-treated branches, suggesting the ability of systemin to trigger plant-to-plant communication. The deterrent activity of the primed olives was associated with the emission of the volatiles 2-ethyl-1-hexanol, 4-tert-butylcyclohexyl acetate and 1, 2, 3-trimethyl-benzene, which were negatively correlated with oviposition and fly infestation. Systemin has also showed to trigger the biosynthesis of specific volatiles (esters) in olives in response to fly attacks. Overall, the observed protection conferred by systemin against olive fly is likely due to the emission of specific volatiles that can act as a defence and/or as signalling molecules to upregulate the plant defence response. Thus, systemin represents a novel and useful tool to manage olive fruit fly.

Exploring Synergistic Effects of Levan and Levan-Metabolizing Bacillaceae in Promoting Growth and Enhancing Immunity of Tomato and Wheat.

Boosting plant immunity by priming agents can lower agrochemical dependency in plant production. Levan and levan-derived oligosaccharides (LOS) act as priming agents against biotic stress in several crops. Additionally, beneficial microbes can promote plant growth and protect against fungal diseases. This study assessed possible synergistic effects caused by levan, LOS and five levan- and LOS-metabolizing Bacillaceae (Bacillus and Priestia) strains in tomato and wheat. Leaf and seed defense priming assays were conducted in non-soil (semi-sterile substrate) and soil-based systems, focusing on tomato-Botrytis cinerea and wheat-Magnaporthe oryzae Triticum (MoT) pathosystems. In the non-soil system, seed defense priming with levan, the strains (especially Bacillus velezensis GA1), or their combination significantly promoted tomato growth and protection against B. cinerea. While no growth stimulatory effects were observed for wheat, disease protective effects were also observed in the wheat-MoT pathosystem. When grown in soil and subjected to leaf defense priming, tomato plants co-applied with levan and the bacterial strains showed increased resistance to B. cinerea compared with plants treated with levan or single strains, and these effects were synergistic in some cases. For seed defense priming in soil, more synergistic effects on disease tolerance were observed in a non-fertilized soil as compared to a fertilized soil, suggesting that potential prebiotic effects of levan are more prominent in poor soils. The potential of using combinations of Bacilliaceae and levan in sustainable agriculture is discussed.

Cytokinin-deficient Chlamydomonas reinhardtii CRISPR-Cas9 mutants show reduced ability to prime resistance of tobacco against bacterial infection.

Although microalgae have only recently been recognized as part of the plant and soil microbiome, their application as biofertilizers has a tradition in sustainable crop production. Under consideration of their ability to produce the plant growth-stimulating hormone cytokinin (CK), known to also induce pathogen resistance, we have assessed the biocontrol ability of CK-producing microalgae. All pro- and eukaryotic CK-producing microalgae tested were able to enhance the tolerance of tobacco against Pseudomonas syringae pv. tabaci (PsT) infection. Since Chlamydomonas reinhardtii (Cre) proved to be the most efficient, we functionally characterized its biocontrol ability. We employed the CRISPR-Cas9 system to generate the first knockouts of CK biosynthetic genes in microalgae. Specifically, we targeted Cre Lonely Guy (LOG) and isopentenyltransferase (IPT) genes, the key genes of CK biosynthesis. While Cre wild-type exhibits a strong protection, the CK-deficient mutants have a reduced ability to induce plant defence. The degree of protection correlates with the CK levels, with the IPT mutants showing less protection than the LOG mutants. Gene expression analyses showed that Cre strongly stimulates tobacco resistance through defence gene priming. This study functionally verifies that Cre primes defence responses with CK, which contributes to the robustness of the effect. This work contributes to elucidate microalgae-mediated plant defence priming and identifies the role of CKs. In addition, these results underscore the potential of CK-producing microalgae as biologicals in agriculture by combining biofertilizer and biocontrol ability for sustainable and environment-friendly crop management.

Guardians of wheat: Unleashing transgenerational immune priming with Trichoderma against spot blotch

Plants encounter various biotic stresses in their natural habitat, prompting the exploration of innovative protective measures to bolster their immune defences against pathogens. An effective strategy involves priming plant immunity, conferring enduring and comprehensive protection. Defence priming, characterized by heightened responsiveness to attackers following prior stress exposure, intriguingly manifests transgenerationally, transmitting the memory of priming to subsequent plant generations. In our investigation, we delved into the phenomenon of transgenerational immune priming (TGIP) in wheat facilitated by Trichoderma, spanning from the parental generation (G0) to the grand progeny (G2) generation. Successful establishment of transgenerational priming in the G2 generation was achieved by initially priming the G0 generation of wheat with Trichoderma. To assess priming inheritance, seeds from the G1 generation were collected, and both primed and non-primed wheat plants' grand progeny (G2 generation) were cultivated in soil. Subsequent evaluation encompassed the disease phenotype, biochemical parameters, and yield-related traits of the G2 generation wheat when confronted with Bipolaris sorokiniana, the causal agent of spot blotch disease. Our analysis revealed that the G2 generation of primed wheat displayed superior protection against spot blotch compared to non-primed wheat. Biochemical studies indicated no activation of defensive responses in the absence of disease pressure in G2 wheat plants. However, upon pathogen challenge, the grand progeny of primed wheat exhibited a robust defence response, surpassing that of non-primed wheat. This targeted defence response enabled efficient resource utilization and mitigated yield penalties. Furthermore, under biotic stress conditions, the grand progeny of primed wheat exhibited enhanced yield parameters compared to non-primed wheat. Our findings provide conclusive evidence that Trichoderma-mediated priming against B. sorokiniana can be inherited successfully from the parental (G0) generation to the grand progeny (G2) generation of wheat. This study represents the initial documentation of Trichoderma-mediated TGIP in wheat against B. sorokiniana, holding significant promise for sustainable agriculture by offering an environmentally friendly alternative to chemical pesticides.

Intergenerational priming by Trichoderma alleviates drought stress in barley

The escalating global climate anomalies and disruption of ecological equilibrium pose profound threats to agroecosystems and crop productivity. Drought stress, a prominent consequence of climate change, exert detrimental effects on plant physiology, thereby reducing crop yield. This study explores the implementation of Trichoderma-mediated defence priming in barley under drought stress. Through a comprehensive evaluation of encompassing morphological, stomatal, biochemical, and yield parameters, Trichoderma- primed barley is compared with non-primed barley under drought stress conditions. Our results revealed that Trichoderma-primed barley exhibits heightened tolerance to drought stress in comparison to non-primed barley. The improved stomatal characteristics indicate that Trichoderma priming enables the plant to better adapt to drought conditions. Biochemical analysis showed enhanced photosynthetic pigments and reduced MDA in primed barley under drought, suggesting that primed barley experiences less oxidative stress compared to non-primed barley. Additionally, Trichoderma-primed plants displayed heightened levels of both non-enzymatic and enzymatic antioxidant activities, aiding in ROS detoxification. Stress responses are activated upon encountering a triggering stimulus (drought stress), but not after Trichoderma priming. This means there was no wasteful diversion of resources towards resistance mechanisms in the absence of stressful situations, leading to enhanced yield for primed barley compared to non-primed barley under stressful conditions. Furthermore, our investigation into the heritability of priming effects in the succeeding generation demonstrates that the progeny of primed barley retains enhanced protection akin to their primed parents. In contrast the progeny of non-primed barley exhibits greater vulnerability under water-deficit conditions. The expression of the epigenetic regulator gene HvDME was more enhanced in primed barley and its subsequent generations than in non-primed barley. This research emphasizes the potential of Trichoderma priming as a cost-effective strategy to enhance barley production amidst climate change challenges. The priming defense approach offers an intelligent plant health management solution devoid of adverse environmental or yield-related implications. This discovery holds significant promise, particularly in regions with limited water resources where crops are highly susceptible to drought stress. The findings contribute to the advancement of sustainable and climate-resilient agriculture, offering insights for practitioners and policymakers aiming to address the adverse impacts of climate change on crop production.

Physiological mechanisms of cross-stress and memory in soybean plants subjected to water deficit and waterlogging

Defense priming is a strategy that enables plants to react faster, minimizing the damage suffered and allowing them to survive stress. Here, we investigated the physiological and morphological changes underpinning stress memory and adaptive capacity in soybean plants subjected to water deficit and/or waterlogging at two developmental stages. Our experiment was carried out in a greenhouse with seven treatments and four replicates: control (non-stressed plants), water deficit at R2; water deficit both at V4 and R2; water deficit in V4 and waterlogging in R2; waterlogging in R2; waterlogging in both V4 and R2; and waterlogging at V4 and water deficit at R2. Samples for biometric analyses, measurements of xylem water potential, leaf gas exchange, chlorophyll a fluorescence, and quantification of photosynthetic pigments were harvested on the last day of stress (R2) and four days after recovery. Primed and cross-primed plants showed improvements in leaf water potential and relative water content due to the greater water use efficiency. Together with the osmotic effects, these plants developed greater protection of the photosynthetic apparatus, showing a superior capacity to transfer energy between photosystems as well as to dissipate photon excess as heat. They also showed a greater ability to recover after periods of stress and maintained carotenoid levels. This study adds another piece of evidence that soybean plants have stress memory, important to maintain high productivity under a global climate change.

PGPR-Mediated Defense Priming: A Sustainable Paradigm for Combating Biotic and Abiotic Stresses in Agriculture

In the face of complex biotic and abiotic stresses, modern agriculture seeks innovative solutions to ensure sustainable crop production. Plant Growth-Promoting Rhizobacteria (PGPR) emerges as powerful allies, offering a sustainable approach to fortifying plant defense mechanisms. This review delves into harnessing PGPR-mediated defense priming to combat both biotic and abiotic stresses in agriculture. Defense priming, a sophisticated mechanism acquired through exposure to primary stimuli, empowers plants to mount quicker and more resilient defense responses against subsequent challenges. PGPR induce a pre-conditioned state of heightened alertness, enabling rapid and robust defense responses upon stress encounters. This paradigm not only enhances plant resilience to pathogens and environmental stressors but also promotes sustainable practices by reducing chemical inputs. The review critically evaluates the mechanisms underlying PGPR-mediated priming, emphasizing its potential to modulate plant physiology, metabolite production, increased antioxidants enzymes, defense related enzymes activities and enhance stress tolerance. It further explores how PGPR can improve plant responses to a spectrum of stressors. This review also highlights PGPR-mediated defense priming as a cost-effective, enduring, chemical-free, and sustainable method for managing abiotic and biotic stresses in agriculture. Implementing this strategy offers effective crop protection with minimal fitness and environmental costs, even in harsh conditions.

Integrative omics studies revealed synergistic link between sucrose metabolic isogenes and carbohydrates in poplar roots infected by Fusarium wilt.

Advances in carbohydrate metabolism prompted its essential role in defense priming and sweet immunity during plant-pathogen interactions. Nevertheless, upstream responding enzymes in the sucrose metabolic pathway and associated carbohydrate derivatives underlying fungal pathogen challenges remain to be deciphered in Populus, a model tree species. In silico deduction of genomic features, including phylogenies, exon/intron distributions, cis-regulatory elements, and chromosomal localization, identified 59 enzyme genes (11 families) in the Populus genome. Spatiotemporal expression of the transcriptome and the quantitative real-time PCR revealed a minuscule number of isogenes that were predominantly expressed in roots. Upon the pathogenic Fusarium solani (Fs) exposure, dynamic changes in the transcriptomics atlas and experimental evaluation verified Susy (PtSusy2 and 3), CWI (PtCWI3), VI (PtVI2), HK (PtHK6), FK (PtFK6), and UGPase (PtUGP2) families, displaying promotions in their expressions at 48 and 72h of post-inoculation (hpi). Using the gas chromatography-mass spectrometry (GC-MS)-based non-targeted metabolomics combined with a high-performance ion chromatography system (HPICS), approximately 307 metabolites (13 categories) were annotated that led to the quantification of 46 carbohydrates, showing marked changes between three compared groups. By contrast, some sugars (e.g., sorbitol, L-arabitol, trehalose, and galacturonic acid) exhibited a higher accumulation at 72 hpi than 0 hpi, while levels of α-lactose and glucose decreased, facilitating them as potential signaling molecules. The systematic overview of multi-omics approaches to dissect the effects of Fs infection provides theoretical cues for understanding defense immunity depending on fine-tuned Suc metabolic gene clusters and synergistically linked carbohydrate pools in trees.

Marker and readout genes for defense priming in Pseudomonas cannabina pv. alisalensis interaction aid understanding systemic immunity in Arabidopsis

Following localized infection, the entire plant foliage becomes primed for enhanced defense. However, specific genes induced during defense priming (priming-marker genes) and those showing increased expression in defense-primed plants upon rechallenge (priming-readout genes) remain largely unknown. In our Arabidopsis thaliana study, genes AT1G76960 (function unknown), CAX3 (encoding a vacuolar Ca2+/H+ antiporter), and CRK4 (encoding a cysteine-rich receptor-like protein kinase) were strongly expressed during Pseudomonas cannabina pv. alisalensis-induced defense priming, uniquely marking the primed state for enhanced defense. Conversely, PR1 (encoding a pathogenesis-related protein), RLP23 and RLP41 (both encoding receptor-like proteins) were similarly activated in defense-primed plants before and after rechallenge, suggesting they are additional marker genes for defense priming. In contrast, CASPL4D1 (encoding Casparian strip domain-like protein 4D1), FRK1 (encoding flg22-induced receptor-like kinase), and AT3G28510 (encoding a P loop-containing nucleoside triphosphate hydrolases superfamily protein) showed minimal activation in uninfected, defense-primed, or rechallenged plants, but intensified in defense-primed plants after rechallenge. Notably, mutation in only priming-readout gene NHL25 (encoding NDR1/HIN1-like protein 25) impaired both defense priming and systemic acquired resistance, highlighting its previously undiscovered pivotal role in systemic plant immunity.

Endogenous rhythmic growth and ectomycorrhizal fungi modulate priming of antiherbivore defences in subsequently formed new leaves of oak trees

Abstract Priming of plant defences provides increased plant protection against herbivores and reduces the allocation costs of defence. Defence priming in woody plants remains obscure, in particular, due to plant development traits such as endogenous rhythmic growth. By using bioassays with oak microcuttings (Quercus robur) and combining transcriptomic and metabolomic analyses, we investigated how leaf herbivory by Lymantria dispar and root inoculation with the ectomycorrhizal fungus Piloderma croceum prime oak defences. We further investigated how defence priming is modulated by the rhythmic growth of the oaks. A first herbivory challenge in oak leaves primed newly grown leaves for enhanced induction of jasmonic acid (JA)‐related direct defences, or enhanced emission of volatiles, depending on the specific growth stage at which the plants were first challenged. Root inoculation with Piloderma abolished the enhanced induction of JA‐related defences and volatile emission. Synthesis: Our results indicate that a first herbivore attack primes direct and indirect defences of newly formed oak leaves and that the specific display of defence priming is regulated by rhythmic growth, and modulated by the interaction Piloderma. Our results show that the priming memory in oaks can be transmitted to the next growth cycle, to the leaves of the new shoot unit.

Functional dissection of the PME and PMEI super families in the protection of Populus pectin during Fusarium wilt

Integration of the pectin homogalacturonan (HG) integrity and remodeling into defense immunity facilitates building blocks of stress resilience within the host and pathogen interactions in the forests. Recent advances in apoplastic pectin methylesterases (PMEs) and coordinated PME inhibitors (PMEIs) deployed immune signaling, highlighting their critical roles in regulating biotic stress adaptation. Nevertheless, the PME and PMEI multiple isoenzyme families involved in defense priming in the model tree species are largely unknown. The in silico demonstration of the molecular profile identified 70 PME- (50 type-I proPME and 20 type-II PME) and 49 PMEI-encoding genes in the Populus genome. Evaluation of the transcript abundance verified 20 PtproPMEs and 18 PtPMEIs that showed predominant expression in the root and genetic redundancy for type-II PtPMEs. Dynamic changes in the transcriptome and experimental evaluation revealed a minuscule number of type-I proPMEs and PMEIs that showed marked promotions during a time-course Fusarium solani (Fs) infection. Using GC/LC-mass-based non-targeted metabolomics to inspect structure-related carbohydrates and PME activities suggested a correlation between proPME-PMEI co-expression and deformed pectin HG biosynthesis. Further functional examination of the enzyme inhibitory affinities of recombinant PtPMEI1 and 39 in vitro and apoplastic patterns suggested a clustering of the type-I proPME in the regulation of the innate immunity by maintaining homeostasis of the pectin HG de-methylesterification (DME) in the cell wall. The coherent omics-wide survey of the fungi pathogen-induced PME-PMEIs and disturbed carbohydrate accumulation provide theoretical cues for in-depth mining of the biological significance of the underlying immune signaling networks within an apoplastic niche in trees.