Defense Hormone Research Articles

Species- and organ-specific contribution of peroxisomal cinnamate:CoA ligases to benzoic and salicylic acid biosynthesis.

Salicylic acid (SA) is a prominent defense hormone whose basal level, organ-specific accumulation, and physiological role vary widely among plant species. Of the two known pathways of plant SA biosynthesis, the phenylalanine ammonia lyase (PAL) pathway is more ancient and universal but its biosynthetic and physiological roles in diverse plant species remain unclear. Studies in which the PAL pathway is specifically or completely inhibited, as well as a direct comparison of diverse species and different organs within the same species, are needed. To this end, we analyzed the PAL pathway in rice (Oryza sativa) and Arabidopsis (Arabidopsis thaliana), two distantly related model plants whose basal SA levels and distributions differ tremendously at the organism and tissue levels. Based on our recent identification of the rice peroxisomal cinnamate:CoA ligases (CNLs), we identified two peroxisomal CNLs from Arabidopsis and showed CNL as the most functionally specific enzyme among the known enzymes of the PAL pathway. We then revealed the species- and organ-specific contribution of the PAL pathway to benzoic and salicylic acid biosynthesis and clarified its physiological importance in rice and Arabidopsis. Our findings highlight the necessity to consider species and organ types in future SA-related studies and may help to breed new disease-resistant crops.

Physiological characteristics during the formation of aromatic components in xylem of Aquilaria sinensis induced by exogenous substances.

An inductive combination of plant growth regulators, inorganic salts, and fungi is essential for the formation of aromatic components in the xylem of Aquilaria sinensis. However, the dynamics of xylem physiology and the relationships between physiological properties and aromatic components after artificial induction remain unclear. In this study, the changes in physiological properties of A. sinensis xylem during induction were determined and analyzed under four induction treatments and a control group. The defense hormone contents of jasmonic acid, salicylic acid, aminocyclopropane-1-carboxylic acid, and abscisic acid obtained from the four induction treatments increased significantly. However, the concentrations of gibberellin and indoleacetic acid were decreased compared to the control group. An initially upward and then downward trend was observed in the main antioxidant enzyme activities. Additionally, malonaldehyde content decreased obviously, while proline content tended to increase and then decrease as induction continued. The total and soluble sugar content was evidently reduced after treatment, and the soluble sugar content recovered more rapidly with time. Thirty-three aromatic components were identified in all treatments, and the primary aromatic components were terpenes, aromatics and chromones, the relative contents of which varied among treatments. These results provide new insights for optimization and innovation of agarwood induction techniques by exploring the formation of aromatics in the xylem of A. sinensis and its physiological responses following induction with exogenous substances (ethephon, NaCl, CaCl2 and fungal mixed solution).

Deciphering Whether Illite, a Natural Clay Mineral, Alleviates Cadmium Stress in Glycine max Plants via Modulation of Phytohormones and Endogenous Antioxidant Defense System

Globally, cadmium (Cd) stress dramatically reduces agricultural yield. Illite, a natural clay mineral, is a low-cost, environmentally acceptable, new promising method of reducing the heavy metal (HM) stress of cereal crops. In research statistics, there is little research on stress tolerance behavior of Illite (IL) on an experimental soybean plant. In the present study, we took IL and examined it for tolerance to Cd, as well as for other plant-growth-promoting (PGP) characteristics in Glycine max (soybean). The results showed that applying clay minerals in different concentrations enhanced the level of SA (defense hormone) and reduced the level of ABA (stress hormone). Cd 1 mM significantly reduces plant growth by altering their morphological characteristics. However, the application of IL significantly enhanced the seedling characteristics, such as root length (RL), 29.6%, shoot length (SL), 14.5%, shoot fresh biomass (SFW), 10.8%, and root fresh biomass (RFB), 6.4%, in comparison with the negative control group. Interestingly, IL 1% also enhanced the chlorophyll content (C.C), 15.5%, and relative water content (RWC), 12.5%, in all treated plants. Moreover, it resulted in an increase in the amount of superoxide dismutase (SOD), phenolics, and flavonoids in soybean plants, while lowering the levels of peroxidase (POD) and H2O2. Furthermore, compared to control plants, soybean plants treated with the Illite exhibited increased Si absorption and lower Cd levels, according to inductively coupled plasma mass spectrometry (ICP-MS). Thus, the IL can operate as an environmentally beneficial biofertilizer and sustainable approach under Cd stress by promoting plant development by activating signaling events.

Antagonistic manipulation of ER-protein quality control between biotrophic pathogenic fungi and host induced defense

The interaction of plants with pathogens during infection is a multifaceted process involving various molecules deriving from both partners. A current goal in combating pathogen virulence is to induce plant resistance using environmentally friendly compounds. Here, we show that chitosan-based nanoparticles loaded with the defense hormone salicylic acid, can efficiently activate defense responses and reactive oxygen species (ROS) production and PATHOGENESIS RELATED-1 (PR1) expression in Arabidopsis thaliana leaves, and reduce conidial germination of the biotrophic pathogenic fungus Podosphaera xanthii. Transcriptomic and proteomic analyses identified immune response-related upregulated transcripts and proteins after nanoparticle application, highlighting the Leucine Rich Repeat (LRR)-, Systemic Acquired Resistance (SAR)-, and glutathione-related protein groups. Examination of P. xanthii during infection at control conditions, identified ribosomal, hydrolase-related, putative secreted and effector proteins, while nanoparticle application significantly downregulated their expression. An in-depth investigation of the highly expressed proteins in P. xanthii and Arabidopsis revealed the involvement of components of endoplasmic reticulum protein quality control (ERQC) in the pathogen-host interaction. The RPS27A effector protein was identified in fungal virulence, while endoplasmic reticulum (ER) protein processing- and glycosyltransferase-related proteins were implicated in plant's induced defense response following nanoparticle application. Overall, these findings demonstrate that the ERQC is dynamically manipulated by both the pathogen for efficient virulence and by elicitors for plant induced defense.

Mechanism of Exogenous Jasmonic Acid-Induced Resistance to Thrips palmi in Hemerocallis citrina Baroni Revealed by Combined Physiological, Biochemical and Transcriptomic Analyses

Jasmonic acid (JA) is a regulator of plant resistance to phytophagous insects, and exogenous JA treatment induces plant insect resistance. This study investigated the mechanism of exogenous JA-induced resistance of Hemerocallis citrina Baroni (daylily) to Thrips palmi at the biochemical and molecular levels. Daylily leaves sprayed with JA showed significantly higher levels of secondary metabolites—tannins, flavonoids, and total phenols, and activity of defense enzymes—peroxidase, phenylalanine ammonia lyase, polyphenol oxidase, and protease inhibitor (PI) than control leaves; the most significant effects were observed with 1 mmol L−1 JA. Owing to an improved defense system, significantly fewer T. palmi were present on the JA-treated plants than control plants. The JA-treated leaves had a smoother wax layer and fewer stomata, which was unfavorable for insect egg attachment. The differentially expressed genes (DEGs) were significantly enriched in insect resistance pathways such as lignin and wax biosynthesis, cell wall thickening, antioxidant enzyme synthesis, PI synthesis, secondary metabolite synthesis, and defense hormone signaling. A total of 466 DEGs were predicted to be transcription factors, mainly bHLH and WRKY family members. Weighted gene co-expression network analysis identified 13 key genes; TRINITY_DN16412_c0_g1 and TRINITY_DN6953_c0_g1 are associated with stomatal regulation and lipid barrier polymer synthesis, TRINITY_DN7582_c0_g1 and TRINITY_DN11770_c0_g1 regulate alkaloid synthesis, and TRINITY_DN7597_c1_g3 and TRINITY_DN1899_c0_g1 regulate salicylic acid and ethylene biosynthesis. These results indicate that JA treatment of daylily improved its resistance to T. palmi. These findings provide a scientific basis for the utilization of JA as an antagonist to control T. palmi in daylily.

The early hormone signaling network underlying wound-induced de novo root regeneration.

Plants possess a remarkable capability to regenerate new organs after wounding. De novo root regeneration (DNRR) from aboveground tissues after physical wounding is observed in a wide range of plant species. Here, we provide an overview of recent progress in the elucidation of the molecular mechanisms that govern DNRR, with a particular emphasis on the early signaling components. Wound-inducible chemicals and hormones such as jasmonic acid, ethylene, and salicylic acid, which were originally identified as defense hormones, influence DNRR. Overall, the ongoing elucidation of the molecular network underlying DNRR provides insight into the plant strategy of coactivating regeneration and defense responses at the early stages of the wound response.

Influences of Ipomoea batatas Anti-Cancer Peptide on Tomato Defense Genes.

This study investigates the impact of IbACP (Ipomoea batatas anti-cancer peptide) on defense-related gene expression in tomato leaves, focusing on its role in plant defense mechanisms. Previously, IbACP was isolated from sweet potato leaves, and it was identified as a peptide capable of inducing an alkalinization response in tomato suspension culture media. Additionally, IbACP was found to regulate the proliferation of human pancreatic adenocarcinoma cells. Elucidate IbACP's molecular influence on defense-related gene expression in tomato leaves using next-generation sequencing analysis. To assess the impact of IbACP on defense-related gene expression, transcriptome data were analyzed, encompassing various functional categories such as photosynthesis, metabolic processes, and plant defense. Semi-quantitative reverse-transcription polymerase chain reaction analysis was employed to verify transcription levels of defense-related genes in tomato leaves treated with IbACP for durations ranging from 0 h (control) to 24 h. IbACP induced jasmonic acid-related genes (LoxD and AOS) at 2 h, with a significant up-regulation of salicylic acid-dependent gene NPR1 at 24 h. This suggested a temporal antagonistic effect between jasmonic acid and salicylic acid during the early hours of IbACP treatment. Downstream ethylene-responsive regulator genes (ACO1, ETR4, and ERF1) were consistently down-regulated by IbACP at all times. Additionally, IbACP significantly up-regulated the gene expressions of suberization-associated anionic peroxidases (TMP1 and TAP2) at all time points, indicating enhanced suberization of the plant cell wall to prevent pathogen invasion. IbACP enhances the synthesis of defense hormones and up-regulates downstream defense genes, improving the plant's resistance to biotic stresses.

Distinct profiles of plant immune resilience revealed by natural variation in warm temperature-modulated disease resistance among Arabidopsis accessions.

Elevated temperature suppresses the plant defence hormone salicylic acid (SA) by downregulating the expression of master immune regulatory genes CALMODULIN BINDING PROTEIN 60-LIKE G (CBP60g) and SYSTEMIC ACQUIRED RESISTANCE DEFICIENT1 (SARD1). However, previous studies in Arabidopsis thaliana plants have primarily focused on the accession Columbia-0 (Col-0), while the genetic determinants of intraspecific variation in Arabidopsis immunity under elevated temperature remain unknown. Here we show that BASIC HELIX LOOP HELIX 059 (bHLH059), a thermosensitive SA regulator at nonstress temperatures, does not regulate immune suppression under warmer temperatures. In agreement, temperature-resilient and -sensitive Arabidopsis accessions based on disease resistance to the bacterial pathogen Pseudomonas syringae pv. tomato (Pst) DC3000 did not correlate with bHLH059 polymorphisms. Instead, we found that temperature-resilient accessions exhibit varying CBP60g and SARD1 expression profiles, potentially revealing CBP60g/SARD1-dependent and independent mechanisms of immune resilience to warming temperature. We identified thermoresilient accessions that exhibited either temperature-sensitive or -insensitive induction of the SA biosynthetic gene ICS1 (direct target gene of CBP60g and SARD1) and SA hormone levels. Collectively, this study has unveiled the intraspecific diversity of Arabidopsis immune responses under warm temperatures, which could aid in predicting plant responses to climate change and provide foundational knowledge for climate-resilient crop engineering.

Citrus Flavanone Effects on the Nrf2-Keap1/GSK3/NF-κB/NLRP3 Regulation and Corticotroph-Stress Hormone Loop in the Old Pituitary.

Oxidative stress and inflammation are significant causes of aging. At the same time, citrus flavanones, naringenin (NAR), and hesperetin (HES) are bioactives with proven antioxidant and anti-inflammatory properties. Nevertheless, there are still no data about flavanone's influence and its potential effects on the healthy aging process and improving pituitary functioning. Thus, using qPCR, immunoblot, histological techniques, and biochemical assays, our study aimed to elucidate how citrus flavanones (15 mg/kg b.m. per os) affect antioxidant defense, inflammation, and stress hormone output in the old rat model. Our results showed that HES restores the redox environment in the pituitary by down-regulating the nuclear factor erythroid 2-related factor 2 (Nrf2) protein while increasing kelch-like ECH-associated protein 1 (Keap1), thioredoxin reductase (TrxR1), and superoxide dismutase 2 (SOD2) protein expression. Immunofluorescent analysis confirmed Nrf2 and Keap1 down- and up-regulation, respectively. Supplementation with NAR increased Keap1, Trxr1, glutathione peroxidase (Gpx), and glutathione reductase (Gr) mRNA expression. Decreased oxidative stress aligned with NLRP3 decrement after both flavanones and glycogen synthase kinase-3 (GSK3) only after HES. The signal intensity of adrenocorticotropic hormone (ACTH) cells did not change, while corticosterone levels in serum decreased after both flavanones. HES showed higher potential than NAR in affecting a redox environment without increasing the inflammatory response, while a decrease in corticosterone level has a solid link to longevity. Our findings suggest that HES could improve and facilitate redox and inflammatory dysregulation in the rat's old pituitary.

In-depth analysis of isochorismate synthase-derived metabolism in plant immunity: Identification of meta-substituted benzoates and salicyloyl-malate

Isochorismate-derived metabolism enables biosynthesis of the plant defense hormone salicylic acid (SA) and its derivatives. In Arabidopsis thaliana, the stress-induced accumulation of SA depends on ISOCHORISMATE SYNTHASE1 (ICS1) and also requires the presumed isochorismate transporter ENHANCED DISEASE SUSCEPTIBILITY5 (EDS5) and the GH3 enzyme avrPphB SUSCEPTIBLE3 (PBS3). By comparative metabolite and structural analyses, we identified several hitherto unreported ICS1- and EDS5-dependent, biotic stress-inducible Arabidopsis metabolites. These involve meta-substituted SA derivatives (5-formyl-SA, 5-carboxy-SA, 5-carboxymethyl-SA), their benzoic acid (BA) analogs (3-formyl-BA, 3-carboxy-BA, 3-carboxymethyl-BA), and besides the previously detected salicyloyl-aspartate (SA-Asp), the ester conjugate salicyloyl-malate (SA-Mal). SA functions as a biosynthetic precursor for SA-Mal and SA-Asp, but not for the meta-substituted SA- and BA-derivatives, which accumulate to moderate levels at later stages of bacterial infection. Interestingly, Arabidopsis leaves possess oxidizing activity to effectively convert meta-formyl- into meta-carboxy-SA/BAs. In contrast to SA, exogenously applied meta-substituted SA/BA-derivatives and SA-Mal exert a moderate impact on plant immunity and defence-related gene expression. While the isochorismate-derived metabolites are negatively regulated by the SA receptor NON-EXPRESSOR OF PR GENES1, SA conjugates (SA-Mal, SA-Asp, SA-glucose conjugates) and meta-substituted SA/BA-derivatives are oppositely affected by PBS3. Notably, our data indicate a PBS3-independent path to isochorismate-derived SA at later stages of bacterial infection, which does not considerably impact immune-related characteristics. Moreover, our results argue against a previously proposed role of EDS5 in the biosynthesis of the immune signal N-hydroxypipecolic acid and associated transport processes. We propose a significantly extended biochemical scheme of plant isochorismate metabolism that involves an alternative generation mode for benzoate- and salicylate-derivatives.

Plant Extracts for Production of Functionalized Selenium Nanoparticles.

In recent years, selenium nanoparticles (SeNPs) have attracted expanding consideration, particularly in the nanotechnology field. This element participates in important biological processes, such as antioxidant defense, immune function, and thyroid hormone regulation, protecting cells from oxidative damage. Selenium in the form of nanoscale particles has drawn attention for its biocompatibility, bioavailability, and low toxicity; thus, it has found several biomedical applications in diagnosis, treatment, and monitoring. Green methods for SeNP synthesis using plant extracts are considered to be single-step, inexpensive, and eco-friendly processes. Besides acting as natural reductants, compounds from plant extracts can also serve as natural capping agents, stabilizing the size of nanoparticles and contributing to the enhanced biological properties of SeNPs. This brief overview presents the recent developments in this area, focusing on the synthesis conditions and the characteristics of the obtained SeNPs.

Decoding nature's defense dance: Mechanistic insights into biochemical and metabolic shifts in Cajanus cajan and Cajanus platycarpus during combat with the lepidopteran pest Helicoverpa armigera provide evidence for non-host plant immunity

This study provides intriguing mechanistic insights into the innate immunity of the pigeonpea wild relative, Cajanus platycarpus, in comparison to the cultivated Cajanus cajan, based on biochemical and metabolic changes during Helicoverpa armigera infestation. Over a 96 h herbivory challenge, both species exhibited 164 metabolites, with 54 undergoing significant alterations. In C. platycarpus, primary carbohydrates such as sucrose, glucose, galactose, and mannose decreased, while structural carbohydrates like xylose, talose, and tagatose accumulated, indicating cell wall damage in C. cajan. Notably, C. platycarpus down-regulated most amino acids except for phenylalanine, which is crucial for the synthesis of defense compounds. The linoleic/linolenic acid pathway, which produces jasmonic acid, was exclusively identified in C. platycarpus, highlighting active defense. As herbivory continued, levels of galactinol, mannitol, and pinitol significantly increased in C. platycarpus, while sorbitol and xylitol levels rose in C. cajan. Essentially, C. platycarpus reduced nutrient availability to insects, enhanced specific defense hormones and pathways, and maintained reactive oxygen species (ROS) equilibrium. These findings offer significant potential for understanding the biology of the wild relative and developing innovative strategies for herbivore control and food sustainability.

Melon CmRLCK VII-8 kinase genes CmRLCK27, CmRLCK30 and CmRLCK34 modulate resistance against bacterial and fungal diseases in Arabidopsis.

Receptor-like cytoplasmic kinases (RLCKs) represent a distinct class of receptor-like kinases crucial for various aspects of plant biology, including growth, development, and stress responses. This study delves into the characterization of RLCK VII-8 members within cucurbits, particularly in melon, examining both structural features and the phylogenetic relationships of these genes/proteins. The investigation extends to their potential involvement in disease resistance by employing ectopic overexpression in Arabidopsis. The promoters of CmRLCK VII-8 genes harbor multiple phytohormone- and stress-responsive cis-acting elements, with the majority (excluding CmRLCK39) displaying upregulated expression in response to defense hormones and fungal infection. Subcellular localization studies reveal that CmRLCK VII-8 proteins predominantly reside on the plasma membrane, with CmRLCK29 and CmRLCK30 exhibiting additional nuclear distribution. Notably, Arabidopsis plants overexpressing CmRLCK30 manifest dwarfing and delayed flowering phenotypes. Overexpression of CmRLCK27, CmRLCK30, and CmRLCK34 in Arabidopsis imparts enhanced resistance against Botrytis cinerea and Pseudomonas syringae pv. tomato DC3000, concomitant with the strengthened expression of defense genes and reactive oxygen species accumulation. The CmRLCK VII-8 members actively participate in chitin- and flg22-triggered immune responses. Furthermore, CmRLCK30 interacts with CmMAPKKK1 and CmARFGAP, adding a layer of complexity to the regulatory network. In summary, this functional characterization underscores the regulatory roles of CmRLCK27, CmRLCK30, and CmRLCK34 in immune responses by influencing pathogen-induced defense gene expression and ROS accumulation.

Genome-wide Characterization of Small Secreted Peptides in Nicotiana tabacum and Functional Assessment of NtLTP25 in Plant Immunity.

Small secreted peptides (SSPs), serving as signaling molecules for intercellular communication, play significant regulatory roles in plant growth, development, pathogen immunity, and responses to abiotic stress. Despite several SSPs, such as PIP, PSK, and PSY having been identified to participate in plant immunity, the majority of SSPs remain understudied, necessitating the exploration and identification of SSPs regulating plant immunity from vast genomic resources. Here we systematically characterized 756 putative SSPs across the genome of Nicotiana tabacum. 173 SSPs were further annotated as established SSPs, such as nsLTP, CAPE, and CEP. Furthermore, we detected the expression of 484 putative SSP genes in five tissues, with 83 SSPs displaying tissue-specific expression. Transcriptomic analysis of tobacco roots under plant defense hormones revealed that 46 SSPs exhibited specific responsiveness to salicylic acid (SA), and such response was antagonistically regulated by methyl jasmonate. It's worth noting that among these 46 SSPs, 16 members belong to nsLTP family, and one of them, NtLTP25, was discovered to enhance tobacco's resistance against Phytophthora nicotianae. Overexpression of NtLTP25 in tobacco enhanced the expression of ICS1, subsequently stimulating the biosynthesis of SA and the expression of NPR1 and pathogenesis-related genes. Concurrently, NtLTP25 overexpression activated genes associated with ROS scavenging, consequently mitigating the accumulation of ROS during the subsequent phases of pathogenesis. These discoveries indicate that these 46 SSPs, especially the 16 nsLTPs, might have a vital role in governing plant immunity that relies on SA signaling. This offers a valuable source for pinpointing SSPs involved in regulating plant immunity.

The Impact of Antimicrobial Proteins on Plant Defense Strategies

Antimicrobial proteins (APs) are the short polypeptides, and have an important piece of the innate defense network present in all lifeforms, including microbes, arthropods, animals, and plants. Our literature survey revealed that APs have an inherent immunological reaction in several creatures including humans, but there has a little knowledge on how they act as defense mechanisms in plants. As a result, the goal of this study is to advancement of our understanding that how these plant peptides function as an immunological defense at the molecular level in plants. Thus, our research revealed that how the anxious message created by the biological risk is recognized by plant cells and then eventually transformed into an effective defensive action. Moreover, APs might kill pathogenic microbes, fungi, and viruses, and have capacity to promptly eradicate antibiotic-mediated multidrug-resistant infections. Further, our finding with these tiny peptides gives quick, regulated, and long-lasting immunity to a wide range of pathogenic organisms via stimulation of Ca2+ influx, reactive oxygen and nitrogen species generation, and activation of MAPK cascade for the transcription of APs genes and also plant defense hormones, assures plant survival in nature. Therefore, it may conclude that APs might be used as an admirable weapon for protecting plants from different infections, and recommends a bright future in agricultural and therapeutic studies.

The jasmonate pathway promotes nodule symbiosis and suppresses host plant defense in Medicago truncatula

Root nodule symbiosis (RNS) between legumes and rhizobia is a major source of nitrogen in agricultural systems. Effective symbiosis requires precise regulation of plant defense responses. The role of the defense hormone jasmonic acid (JA) in the immune response has been extensively studied. Current research shows that JA can play either a positive or negative regulatory role in RNS depending on its concentration, but the molecular mechanisms remain to be elucidated. In this study, we found that inoculation with the rhizobia Sm1021 induces the JA pathway in Medicago truncatula, and blocking the JA pathway significantly reduces the number of infection threads. Mutations in the MtMYC2 gene, which encodes a JA signaling master transcription factor, significantly inhibited rhizobia infection, terminal differentiation, and symbiotic cell formation. Combining RNA sequencing and chromatin immunoprecipitation sequencing, we discovered that MtMYC2 regulates the expression of nodule-specific MtDNF2, MtNAD1, and MtSymCRK to suppress host defense, while it activates MtDNF1 expression to regulate the maturation of MtNCRs, which in turn promotes bacteroid formation. More importantly, MtMYC2 participates in symbiotic signal transduction by promoting the expression of MtIPD3. Notably, the MtMYC2-MtIPD3 transcriptional regulatory module is specifically present in legumes, and the Mtmyc2 mutants are susceptible to the infection by the pathogen Rhizoctonia solani. Collectively, these findings reveal the molecular mechanisms of how the JA pathway regulates RNS, broadening our understanding of the roles of JA in plant-microbe interactions.

Fidelity in plant hormone modifications catalyzed by Arabidopsis GH3 acyl acid amido synthetases

GRETCHEN HAGEN 3 (GH3) acyl acid amido synthetases conjugate amino acids to acyl acid hormones to either activate or inactivate the hormone molecule. The largest subgroup of GH3 proteins modify the growth-promoting hormone auxin (indole-3-acetic acid; IAA) with the second largest class activating the defense hormone jasmonic acid (JA). The two-step reaction mechanism of GH3 proteins provides a potential proofreading mechanism to ensure fidelity of hormone modification. Examining pyrophosphate release in the first-half reaction of Arabidopsis GH3 proteins that modify IAA (AtGH3.2/YDK2, AtGH3.5/WES1, AtGH3.17/VAS2), JA (AtGH3.11/JAR1), and other acyl acids (AtGH3.7, AtGH3.12/PBS3) indicates that acyl acid-AMP intermediates are hydrolyzed into acyl acid and AMP in the absence of the amino acid, a typical feature of pre-transfer editing mechanisms. Single-turnover kinetic analysis of AtGH3.2/YDK2 and AtGH3.5/WES1 shows that non-cognate acyl acid-adenylate intermediates are more rapidly hydrolyzed than the cognate IAA-adenylate. In contrast, AtGH3.11/JAR1 only adenylates JA, not IAA. While some of the auxin-conjugating GH3 proteins in Arabidopsis (i.e., AtGH3.5/WES1) accept multiple acyl acid substrates, others, like AtGH3.2/YDK2, are specific for IAA; however, both these proteins share similar active site residues. Biochemical analysis of chimeric variants of AtGH3.2/YDK2 and AtGH3.5/WES1 indicates that the C-terminal domain contributes to selection of cognate acyl acid substrates. These findings suggest that the hydrolysis of non-cognate acyl acid-adenylate intermediates, or proofreading, proceeds via a slowed structural switch that provides a checkpoint for fidelity before the full reaction proceeds.

Unraveling Microbial Strategies: Targeting the Plant Ubiquitin - Proteasome Pathway

The Ubiquitin-Proteasome System (UPS) stands as a central regulator in the intricate web of plant immune responses, orchestrating the turnover of key immune components. Through the UPS, plants finely tune their defenses, from the initial recognition of pathogens to the activation of defense strategies. Immune receptors, signaling components associated with defense hormones, and transcription factors are among the targets modulated by the UPS, shaping both PAMP-triggered immunity (PTI) and effector-triggered immunity (ETI). However, pathogens have evolved sophisticated strategies to manipulate the UPS for their benefit. Fungi, bacteria, and viruses deploy effector proteins to exploit the UPS, either by promoting the degradation of host defense elements or by interfering with UPS-mediated processes. This intricate interplay underscores the pivotal role of the UPS in plant-pathogen dynamics, presenting both challenges and opportunities for understanding and manipulating plant immunity. Unraveling the complexities of UPS regulation and pathogen exploitation thereof is crucial for advancing our knowledge of plant-microbe interactions and developing strategies for sustainable disease management in agriculture.

Physiological responses and transcriptome analysis of Hemerocallis citrina Baroni exposed to Thrips palmi feeding stress.

Thrips are serious pests of Hemerocallis citrina Baroni (daylily), affecting crop yield and quality. To defend against pests, daylily has evolved a set of sophisticated defense mechanisms. In the present study, induction of systemic resistance in Hemerocallis citrina 'Datong Huanghua' by Thrips palmi feeding was investigated at both biochemical and molecular levels. The soluble sugar content of daylily leaves was significantly lower than that in control check (CK) at all time points of feeding by T. palmi, whereas the amino acid and free fatty acid contents started to be significantly lower than those in CK after 7 days. Secondary metabolites such as tannins, flavonoids, and total phenols, which are harmful to the growth and reproduction of T. palmi, were increased significantly. The activities of defense enzymes such as peroxidase (POD), phenylalanine ammonia lyase (PAL), and polyphenol oxidase (PPO) were significantly increased, and the degree of damage to plants was reduced. The significant increase in protease inhibitor (PI) activity may lead to disrupted digestion and slower growth in T. palmi. Using RNA sequencing, 1,894 differentially expressed genes (DEGs) were identified between control and treatment groups at five timepoints. DEGs were mainly enriched in secondary metabolite synthesis, jasmonic acid (JA), salicylic acid (SA), and other defense hormone signal transduction pathways, defense enzyme synthesis, MAPK signaling, cell wall thickening, carbohydrate metabolism, photosynthesis, and other insect resistance pathways. Subsequently, 698 DEGs were predicted to be transcription factors, including bHLH and WRKY members related to biotic stress. WGCNA identified 18 hub genes in four key modules (Purple, Midnight blue, Blue, and Red) including MYB-like DNA-binding domain (TRINITY_DN2391_c0_g1, TRINITY_DN3285_c0_g1), zinc-finger of the FCS-type, C2-C2 (TRINITY_DN21050_c0_g2), and NPR1 (TRINITY_DN13045_c0_g1, TRINITY_DN855_c0_g2). The results indicate that biosynthesis of secondary metabolites, phenylalanine metabolism, PIs, and defense hormones pathways are involved in the induced resistance to T. palmi in daylily.

The chloroplast singlet oxygen-triggered biosynthesis of salicylic acid and jasmonic acid is mediated by EX1 and GUN1 in Arabidopsis.

Reactive oxygen species (ROS) and defence hormones like salicylic acid (SA) and jasmonic acid (JA) play pivotal roles in triggering cell death. However, the precise mechanism governing the interaction between ROS and SA/JA remains elusive. Recently, our research revealed that RNAi mutants with suppressed expression of PROGRAMMED CELL DEATH8 (PCD8) exhibit an overabundance of tetrapyrrole intermediates, particularly uroporphyrinogen III (Uro III), leading to the accumulation of singlet oxygen (1O2) during the transition from darkness to light, thereby instigating leaf necrosis. In this investigation, we uncovered that 1O2 stimulates biosynthesis of SA and JA, activating SA/JA signalling and the expression of responsive genes in PCD8 RNAi (pcd8) mutants. Introducing NahG or knocking out PAD4 or NPR1 significantly alleviates the cell death phenotype of pcd8 mutants, while coi1 partially mitigates the pcd8 phenotype. Further exploration revealed that EX1 and GUN1 can partially rescue the pcd8 phenotype by reducing the levels of Uro III and 1O2. Notably, mutations in EX1 mutations but not GUN1, substantially diminish SA content in pcd8 mutants compared to the wild type, implying that EX1 acts as the primary mediator of 1O2 signalling-mediated SA biosynthesis. Moreover, the triple ex1 gun1 pcd8 displays a phenotype similar to ex1. Overall, our findings underscore that the 1O2-induced cell death phenotype requires EX1/GUN1-mediated retrograde signalling in pcd8 mutants, providing novel insights into the interplay between ROS and SA/JA.