Plant Defense Responses Research Articles

Molecular and metabolic insights into the mechanism of exogenous methyl jasmonate in enhancing the postharvest resistance of kiwifruit to Botrytis cinerea

Gray mold, caused by Botrytis cinerea infection, significantly impacts postharvest kiwifruit, leading to spoilage and food safety issues. Meanwhile, methyl jasmonate (MeJA) induces defense responses in plants. This study aimed to identify the optimal MeJA concentration to induce disease resistance in kiwifruits. Notably, various plant defense enzymes were detected in MeJA-treated kiwifruit. Moreover, 0.01 mol/L MeJA was found to enhance B. cinerea resistance in kiwifruit by increasing antioxidant enzyme activity. Widely targeted metabolomics analysis revealed 62–64 differentially expressed metabolites (DEMs) between the different treatment groups. Additionally, RNA-seq analysis revealed 930–2900 differentially expressed genes (DEGs), between these groups. Subsequently, combined DEG and DEM analysis highlighted phenylpropanoid biosynthesis and plant hormone signaling as key transduction pathway associated with MeJA-induced resistance. Overall, these findings identified the mechanism of MeJA-induced resistance in kiwifruit, providing a theoretical basis for the safe and effective control of postharvest diseases in kiwifruit.

Expansion and functional divergence of terpene synthase genes in angiosperms: a driving force of terpene diversity

Abstract Angiosperms are prolific producers of structurally diverse terpenes, which are essential for plant defense responses, as well as the formation of floral scents, fruit flavors, and medicinal constituents. Terpene synthase genes (TPSs) play crucial roles in the biosynthesis of terpenes. This study specifically focuses on the catalytic products of 222 functionally characterized TPSs in 24 angiosperms, which mainly comprise monoterpenes, sesquiterpenes, diterpenes, and sesterterpene. Our systematic analysis of these TPSs uncovered a significant expansion of the angiosperm-specific TPS-a, b, and g subfamilies in comparison to the TPS-e/f and c subfamilies. The expanded subfamilies can be further partitioned into distinct branches, within which considerable functional innovation and diversification have been observed. Numerous TPSs exhibit bifunctional or even trifunctional activities in vitro, yet they exhibit only a single activity in vivo, which may be largely determined by their inherent properties, subcellular localization, and the availabilities of endogenous substrates. Additionally, we explored the biological functions of terpenes in various organs and tissues of angiosperms. We propose that the expansion and functional divergence of TPSs contribute to the adaptability and diversity of angiosperms, facilitating the production of a broad spectrum of terpenes that enable diverse interactions with the environment and other organisms. Our findings provide a foundation for comprehending the correlation between the evolutionary features of TPSs and the diversity of terpenes in angiosperms, which is significant for terpene biosynthesis research.

MiR472 deficiency enhances Arabidopsis thaliana defense without reducing seed production.

After having co-existed in plant genomes for at least 200 million years, the products of microRNA (miRNA) and Nucleotide-Binding Leucine Rich Repeat protein (NLR) genes formed a regulatory relationship in the common ancestor of modern gymnosperms and angiosperms. From then on, DNA polymorphisms occurring at miRNA target sequences within NLR transcripts must have been compensated by mutations in the corresponding mature miRNA sequence. The potential evolutionary advantage of such regulation remains largely unknown and might be related to two non-exclusive scenarios: miRNA-dependent regulation of NLR levels might prevent defense mis-activation with negative effects on plant growth and reproduction; or reduction of active miRNA levels in response to pathogen derived molecules (PAMPS and silencing suppressors) might rapidly release otherwise silent NLR transcripts for rapid translation and thereby enhance defense. Here, we used Arabidopsis thaliana plants deficient for miR472 function to study the impact of releasing its NLR targets on plant growth and reproduction and on defense against the fungal pathogen Plectospharaella cucumerina. We show that miR472 regulation has a dual role, participating both in the tight regulation of plant defense and growth. MIM472 lines, with reduced active miR472, are more resistant to pathogens and, correlatively, have reduced relative growth compared to wild-type plants although the end of their reproductive phase is delayed, exhibiting higher adult biomass and similar seed yield as the wild-type. Our study highlights how negative consequences of defense activation might be compensated by changes in phenology and that miR472 reduction is an integral part of plant defense responses.

Identifying the key motif essential for enhancing plant defense against Botrytis cinerea by β-glucanase from Bacillus velezensis LJ02

The glycosyl hydrolase β-glucanase elicits plant immune responses against pathogens and enhances plant immunity by activating signaling pathways. The specific functional domains responsible for disease prevention remain unclear. In this study, transient expression of β-glucanase significantly increased leaves resistance to Botrytis cinerea in Nicotiana benthamiana systemic leaves. Through sequence alignment and similarity analysis, five conserved motifs in the amino acid sequence of β-glucanase were identified, and five deletion mutants were generated to investigate its essential regions further. Notably, the N-terminal amino acid sites 5-54 deletion mutation of β-glucanase decreased resistance to B. cinerea infection. These results indicate that N-terminal amino acids 5-54 (N54) are crucial for β-glucanase induced N. benthamiana defense response and for enhancing resistance to B. cinerea. Further analysis using real-time quantitative fluorescent PCR (qRT-PCR) revealed a significant reduction in gene expression within the N54 region compared to that of unmutated β-glucanase. Additionally, there was a notable reduction in the relative expression levels of FRK, CYP71D20, WRKY7, WRKY8, ACRE31 and PTI genes. Therefore, the first 50 amino acids at positions 5-54 within the N-terminal domain were essential for triggering plant defense responses and enhancing resistance against B. cinerea infection. This study provides an important theoretical foundation for systematic investigation into key functional domains within β-glucanase that trigger defensive responses in plants against B. cinerea.

Regional propolis extracts suppress Fusarium fujikuroi and boost rice seedling growth and response against Bakanae disease

Bakanae, caused by Fusarium fujikuroi, is a serious seed-borne disease affecting rice production worldwide. Herein, we investigated the potential of propolis, a natural honeybee product, collected from three geographic locations in Egypt: North (Menofia), Middle (Cairo), and South (Sohag), and extracted with four different solvents (ethanol, olive oil, hexane, and water) as a sustainable eco-friendly alternative to control the Bakanae disease in rice. In vitro experiments showed that propolis extracts exhibited antifungal activity against F. fujikuroi and significantly suppressed its mycelial growth and sporulation. The antifungal activity of propolis extracts was associated with its chemical composition which varied depending on geographical origin, and the extraction solvents. Moreover, SEM-based analysis revealed that ethanolic extract of northern propolis prominently altered the microconidia morphology of F fujikuroi, which shrunken and seemed to lose their viability. These findings were further confirmed in vivo under greenhouse conditions. Soaking F fujikuroi-inoculated rice seeds in 1000 ppm of propolis extract significantly reduced the bakanae disease incidence, and disease severity index compared to mock-treated controls. Although infection with F fujikuroi induced the accumulation of H2O2, the application of propolis extracts alleviated oxidative stress and significantly reduced the H2O2 levels within infected rice seedlings. Moreover, propolis extracts enhanced the profile of enzymatic antioxidants (guaiacol-dependent peroxidase [POX] and polyphenol oxidase [PPO]) in F. fujikuroi-infected rice seedlings. Finally, Propolis extracts-treated rice seedlings exhibited higher transcript levels of three PTI-marker genes including nonexpressor of pathogenesis-related genes 1 (OsNPR1), WRKY transcription factor 21(OsWRKY21), and phenylalanine ammonia-lyase (OsPAL1), which are associated with systemic acquired resistance (SAR) and crucial for the plant's defense response against pathogens. Collectively, these findings suggest that propolis might be a promising sustainable, eco-friendly alternative to control bakanae disease and other fungal seed-borne phytopathogens due to its antifungal properties and ability to induce a complex multilayered defense system within infected plants.

The Vacuolar H+-ATPase subunit C is involved in oligogalacturonide (OG) internalization and OG-triggered immunity

In plants, the perception of cell wall fragments initiates signal transduction cascades that activate the immune response. Previous research on early protein dynamics induced by oligogalacturonides (OGs), pectin fragments acting as damage-associated molecular patterns (DAMPs), revealed significant phosphorylation changes in several proteins. Among them, the subunit C of the vacuolar H+-ATPase, known as DE-ETIOLATED 3 (DET3), was selected to elucidate its role in the OG-triggered immune response. The Arabidopsis det3 knockdown mutant exhibited defects in H2O2 accumulation, mitogen-activated protein kinases (MAPKs) activation, and induction of defense marker genes in response to OG treatment. Interestingly, the det3 mutant showed a higher basal resistance to the fungal pathogen Botrytis cinerea that, in turn, was completely reversed by the pre-treatment with OGs. Our results suggest a compromised ability of the det3 mutant to maintain a primed state over time, leading to a weaker defense response when the plant is later exposed to the fungal pathogen. Using fluorescently labelled OGs, we demonstrated that endocytosis of OGs was less efficient in the det3 mutant, implicating DET3 in the internalization process of OGs. This impairment aligns with the observed defect in the priming response in the det3 mutant, underscoring that proper internalization and signaling of OGs are crucial for initiating and maintaining a primed state in plant defense responses.

Salivary Protein Sfapyrase of Spodoptera frugiperda Stimulates Plant Defence Response.

Plants have developed various resistance mechanisms against herbivorous insects through prolonged coevolution. Plant defence responses can be triggered by specific compounds present in insect saliva. Apyrase, a known enzyme that catalyzes the hydrolysis of adenosine triphosphate (ATP) and adenosine diphosphate (ADP) into adenosine monophosphate (AMP) and inorganic phosphorus, has recently been identified in some herbivorous insects. However, whether insect salivary apyrase induces or inhibits plant responses remains poorly understood. In this study, we identified an apyrase-like protein in the salivary proteome of the fall armyworm, Spodoptera frugiperda, named Sfapyrase. Sfapyrase was primarily expressed in the salivary gland and secreted into plants during insect feeding. Transient expression of Sfapyrase in tobacco and maize enhanced plant resistance and resulted in decreased insect feeding. Knockdown of Sfapyrase through RNA interference led to increased growth and feeding of S. frugiperda. Furthermore, we showed that Sfapyrase activates the jasmonic acid signalling pathway and promotes the synthesis of secondary metabolites, especially benzoxazinoids, thereby enhancing resistance to S. frugiperda. In summary, our findings demonstrated that Sfapyrase acts as a salivary elicitor, inducing maize jasmonic acid defence responses and the production of insect-resistant benzoxazinoids. This study provides valuable insights into plant-insect interactions and offers potential targets for developing innovative insect pest management strategies.

Plant Immunity Modulation in Arbuscular Mycorrhizal Symbiosis and Its Impact on Pathogens and Pests

Arbuscular mycorrhizal (AM) symbiosis is the oldest and most widespread mutualistic association on Earth and involves plants and soil fungi belonging to Glomeromycotina. A complex molecular, cellular, and genetic developmental program enables partner recognition, fungal accommodation in plant tissues, and activation of symbiotic functions such as transfer of phosphorus in exchange for carbohydrates and lipids. AM fungi, as ancient obligate biotrophs, have evolved strategies to circumvent plant defense responses to guarantee an intimate and long-lasting mutualism. They are among those root-associated microorganisms able to boost plants’ ability to cope with biotic stresses leading to mycorrhiza-induced resistance (MIR), which can be effective across diverse hosts and against different attackers. Here, we examine the molecular mechanisms underlying the modulation of plant immunity during colonization by AM fungi and at the onset and display of MIR against belowground and aboveground pests and pathogens. Understanding the MIR efficiency spectrum and its regulation is of great importance to optimizing the biotechnological application of these beneficial microbes for sustainable crop protection.

Suprathreshold Water Spray Stimulus Enhances Plant Defenses against Biotic Stresses in Tomato.

Mechanical stimuli can affect plant growth, development, and defenses. The role of water spray stimulation, as a prevalent mechanical stimulus in the environment, in crop growth and defense cannot be overlooked. In this study, the effects of water spray on tomato plant growth and defense against the chewing herbivore Helicoverpa armigera and necrotrophic fungus Botrytis cinerea were investigated. Suprathreshold water spray stimulus (LS) was found to enhance tomato plant defenses against pests and pathogens while concurrently modifying plant architecture. The results of the phytohormone and chemical metabolite analysis revealed that LS improved the plant defense response via jasmonic acid (JA) signaling. LS significantly elevated the level of a pivotal defensive metabolite, chlorogenic acid, and reduced the emissions of volatile organic compounds (VOCs) from tomato plants, thereby defending against pest and pathogen attacks. The most obvious finding to emerge from this study is that LS enhances tomato plant defenses against biotic stresses, which will pave the way for further work on the application of mechanical stimuli for pest management.

Improving transient expression in N. benthamiana by suppression of the Nb-SABP2 and Nb-COI1 plant defence response related genes.

Currently transient expression is one of the preferred plant-based technologies for recombinant protein manufacturing, particularly in respect to pharmaceutically relevant products. Modern hybrid transient expression systems combine the features of Agrobacterium tumefaciens and viral vectors. However, host plant reaction to Agrobacterium-mediated delivery of gene of interest can negatively affect foreign protein accumulation. In this study, we investigated whether the modulation of plant immune response through knockdown of the Nb-SABP2 and Nb-COI1 N. benthamiana genes could improve recombinant protein yield. In plants, the SABP2 and COI1 proteins are involved in the salicylic acid and jasmonic acid metabolism, respectively. We exemplified the utility of this approach with the green fluorescence (GFP) and β nerve growth factor (βNGF) proteins: compared to the tobacco mosaic virus (TMV)-based vector the Nb-SABP2 and Nb-COI1-suppressed plants provided an increased recombinant protein accumulation. We also show that this strategy is extendable to the expression systems utilizing potato virus X (PVX) as the vector backbone: the enhanced amounts of βNGF were detected in the Nb-SABP2 and Nb-COI1-depleted leaves co-infiltrated with the PVX-βNGF. These findings suggest that modulating host plant reaction to agrodelivery of expression vectors could be useful for improving transient foreign protein production in N. benthamiana.

Integration of Transcriptomics and WGCNA to Characterize Trichoderma harzianum-Induced Systemic Resistance in Astragalus mongholicus for Defense against Fusarium solani.

Beneficial fungi of the genus Trichoderma are among the most widespread biocontrol agents that induce a plant's defense response against pathogens. Fusarium solani is one of the main pathogens that can negatively affect Astragalus mongholicus production and quality. To investigate the impact of Trichoderma harzianum on Astragalus mongholicus defense responses to Fusarium solani, A. mongholicus roots under T. harzianum + F. solani (T + F) treatment and F. solani (F) treatment were sampled and subjected to transcriptomic analysis. A differential expression analysis revealed that 6361 differentially expressed genes (DEGs) responded to T. harzianum induction. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the 6361 DEGs revealed that the genes significantly clustered into resistance-related pathways, such as the plant-pathogen interaction pathway, phenylpropanoid biosynthesis pathway, flavonoid biosynthesis pathway, isoflavonoid biosynthesis pathway, mitogen-activated protein kinase (MAPK) signaling pathway, and plant hormone signal transduction pathway. Pathway analysis revealed that the PR1, formononetin biosynthesis, biochanin A biosynthesis, and CHIB, ROS production, and HSP90 may be upregulated by T. harzianum and play important roles in disease resistance. Our study further revealed that the H2O2 content was significantly increased by T. harzianum induction. Formononetin and biochanin A had the potential to suppress F. solani. Weighted gene coexpression network analysis (WGCNA) revealed one module, including 58 DEGs associated with T. harzianum induction. One core hub gene, RPS25, was found to be upregulated by T. harzianum, SA (salicylic acid) and ETH (ethephon). Overall, our data indicate that T. harzianum can induce induced systemic resistance (ISR) and systemic acquired resistance (SAR) in A. mongholicus. The results of this study lay a foundation for a further understanding of the molecular mechanism by which T. harzianum induces resistance in A. mongholicus.

Arbuscular mycorrhizal fungi improve the disease resistance of Lycium barbarum to root rot by activating phenylpropane metabolism.

Root rot is one of the common diseases of Lycium barbarum. Pathogens can cause devastating disasters to plants after infecting host plants. This study investigated the effect of arbuscular mycorrhizal fungi (AMF) Rhizophagus intraradices inoculation on phenylpropane metabolism in L. barbarum and evaluated its resistance to root rot. The experiment was set up with AMF inoculation treatments (inoculated or not) and root rot pathogen-Fusarium solani inoculation treatments (inoculated or not). The results showed that AMF was able to form a symbiosis with the root system of L. barbarum, thereby promoting plant growth significantly and increasing plants' resistance to disease stress. The plant height of AMF-colonized L. barbarum increased by 24.83% compared to non-inoculated diseased plants. After inoculation with AMF, the plant defense response induced by pathogen infection was stronger. When the enzyme activity of the leaves reached the maximum after the onset of mycorrhizal L. barbarum, phenylalanine ammonia-lyase, cinnamic acid-4-hydroxylase, and 4-coumaric acid-CoA ligase increased by 3.67%, 31.47%, and 13.61%, respectively, compared with the non-inoculated diseased plants. The products related to the lignin pathway and flavonoid pathway downstream of phenylpropane metabolism such as lignin and flavonoids were also significantly increased by 141.65% and 44.61% compared to nonmycorrhizal diseased plants. The activities of chitinase and β-1,3-glucanase increased by 36.00% and 57.96%, respectively. The contents of salicylic acid and jasmonic acid were also 17.7% and 31.63% higher than those of nonmycorrhizal plants in the early stage of plant growth, respectively. The results indicated that AMF significantly promoted plant growth and enhanced disease resistance by increasing enzyme activities and the production of lignin and flavonoids.

Linear β-1,2-glucans trigger immune hallmarks and enhance disease resistance in plants.

Immune responses in plants are triggered by molecular patterns or elicitors, recognized by plant pattern recognition receptors. Such molecular patterns are consequence of host-pathogen interactions and the response cascade activated after their perception is known as pattern-triggered immunity (PTI). Glucans have emerged as key players in PTI, but the ability of certain glucans to stimulate defensive responses in plants remains understudied. This work focused on identifying novel glucan oligosaccharides as molecular patterns. The ability of various microorganism-derived glucans to prompt PTI responses was tested, revealing that specific microbial-derived molecules, such as short linear β-1,2-glucans, trigger this response in plants by increasing the production of reactive oxygen species (ROS), MAP kinase phosphorylation, and differential expression of defence-related genes in Arabidopsis thaliana. Pretreatments with β-1,2-glucan trisaccharide (B2G3) improved Arabidopsis defence against bacterial and fungal infections in a hypersusceptible genotype. The knowledge generated was then transferred to the monocotyledonous model species maize and wheat, confirming that these plants also respond to β-1,2-glucans, with increased ROS production and improved protection against fungal infections following B2G3 pretreatments. In summary, as with other β-glucans, plants perceive β-1,2-glucans as warning signals and stimulate defence responses against phytopathogens.

The module consisting of transcription factor WRKY14 and thaumatin-like protein TLP25 is involved in winter adaptation in Ammopiptanthus mongolicus.

Thaumatin-like proteins (TLPs) are conserved proteins involved in the defense and stress responses of plants. Previous studies showed that several TLPs were accumulated in leaf apoplast in Ammopiptanthus mongolicus in winter, indicating that TLPs might be related to the adaptation to winter climate in A. mongolicus. To investigate the roles of TLPs in winter adaptation, we first analyzed the expression pattern of TLP genes in A. mongolicus and then focused on the biological function and regulation pathway of AmTLP25 gene. Several TLP genes, including AmTLP25, were upregulated during winter and in response to both cold and osmotic stress. Overexpression of the AmTLP25 gene led to an increased tolerance of transgenic Arabidopsis to freezing and osmotic stress. Furthermore, the elevated AmWRKY14 transcription factor during winter activated AmTLP25 gene expression by specifically binding to its promoter. It is speculated that the AmWRKY14 - AmTLP25 module contributes to the adaptation to temperate winter climate in A. mongolicus. Our research advances the current understanding of the biological function and regulatory pathway of TLP genes and provides valuable information for understanding the molecular mechanism of temperate evergreen broad-leaved plants adapting to winter climate.

Rice sheath blight disease control by native endophytic Bacillus subtilis from Kuttanad, a Globally Important Agricultural Heritage System

ABSTRACT Kuttanad in India is known as ‘the rice bowl of Kerala’ and renowned for its unique rice cultivation system that is below sea level. However, rice production in this wetland ecosystem is seriously threatened by sheath blight disease caused by the fungus Rhizoctonia solani, which has resulted in a significant decline in grain quality and a 50% loss in yield. In the present study, we isolated two endophytic Bacillus subtilis strains, NIIST B616 and NIIST B627, from the roots of rice plants growing in the Kuttanad region and tested their efficacy in controlling sheath blight disease. We found that rice plants inoculated with a combination of both Bacillus strains and R. solani had a 64.71% lower disease incidence and a 31.02% higher harvest index than plants treated with R. solani alone. An analysis of defence-related enzymes and chemicals revealed that treatment with both isolates together greatly enhanced levels of l-phenylalanine ammonia-lyase (PAL; 2.8 fold), peroxidase (POX; 7.4 fold), polyphenol oxidase (PPO; 4.5 fold), and total phenol (8.5 fold) compared with plants treated solely with R. solani, indicating that these isolates significantly increased plant defence responses through induced systemic resistance (ISR), and this finding was validated through a split-root experiment. More detailed analysis using gas chromatography–tandem mass spectrometry (GC-MS/MS) and liquid chromatography–tandem mass spectrometry (LC-MS/MS) techniques revealed that cyclo-(Pro-Leu) and cyclo-(Pro-Phe) were the major components responsible for the antifungal properties of the two isolates and their capacity to trigger ISR in plants. Thus, the Bacillus strains and their metabolites may be deemed viable alternatives for controlling the prevalence of sheath blight disease in rice plants and augmenting rice production at unique locations such as Kuttanad.

LrHSP17.2 Plays an Important Role in Abiotic Stress Responses by Regulating ROS Scavenging and Stress-Related Genes in Lilium regale.

As an important part of heat shock response module, heat shock proteins (HSP) play an important role in plant defense response against heat stress; however, the involvement of the majority of the HSP family members against other abiotic stresses remains poorly understood. In the present study, LrHSP17.2 was identified and its function against abiotic stress was analyzed. The expression level of LrHSP17.2 was significantly induced by heat. Heterologous transgenes of LrHSP17.2 showed that LrHSP17.2 can increase the activity of catalase, peroxidase, superoxide dismutase to removes excess reactive oxygen species (ROS), maintain the stability of the membrane structure, and regulate genes related to antioxidant enzymes and defense under abiotic stress. In addition, LrHSP17.2 could be regulated by exogenous abscisic acid and melatonin, and the related hormone synthesis genes of transgenic plants were significantly up-regulated under heat stress. Taken together, our results revealed that LrHSP17.2 is involved in regulating abiotic stress responses by regulating ROS scavenging and stress-related genes in Lilium regale.

Effect of exogenous melatonin on growth and antioxidant system of pumpkin seedlings under waterlogging stress.

Melatonin regulates defense responses in plants under environmental stress. This study aimed to explore the impact of exogenous melatonin on the phenotype and physiology of 'BM1' pumpkin seedlings subjected to waterlogging stress. Waterlogging stress was induced following foliar spraying of melatonin at various concentrations (CK, 0, 10, 100, 200, and 300 μmol·L-1). The growth parameters, malondialdehyde (MDA) content, antioxidant enzyme activity, osmoregulatory substance levels, and other physiological indicators were assessed to elucidate the physiological mechanisms underlying the role of exogenous melatonin in mitigating waterlogging stress in pumpkin seedlings. The results indicate that pumpkin seedlings exhibit waterlogging symptoms, such as leaf wilting, water loss, edge chlorosis, and fading, under waterlogging stress conditions. Various growth indicators of the seedlings, including plant height, stem diameter, root length, fresh and dry weight, and leaf chlorophyll content, were significantly reduced. Moreover, the MDA content in leaves and roots increased significantly, along with elevated activities of superoxide dismutase, catalase, peroxidase, and soluble protein contents. When different concentrations of melatonin were sprayed on the leaves post waterlogging stress treatment, pumpkin seedlings showed varying degrees of recovery, with the 100 μmol·L-1 treatment displaying the best growth status and plant morphological phenotypes. There were no significant differences compared to the control group. Seedling growth indicators, chlorophyll content, root activity, antioxidant enzyme activities, soluble protein content, and osmotic adjustment substance content all increased to varying degrees with increasing melatonin concentration, peaking at 100 μmol·L-1. Melatonin also reduced membrane damage caused by oxidative stress and alleviated osmotic imbalance. Exogenous melatonin enhanced the activities of antioxidant enzymes and systems involved in scavenging reactive oxygen species, with 100 μmol·L-1 as the optimal concentration. These findings underscore the crucial role of exogenous melatonin in alleviating waterlogging stress in pumpkins. The findings of this study offer a theoretical framework and technical assistance for cultivating waterlogging-resistant pumpkins in practical settings. Additionally, it establishes a theoretical groundwork for the molecular breeding of pumpkins with increased tolerance to waterlogging.

A novel antifungal peptide, SP1.2, from Rhodopseudomonas palustris against the rice blast pathogen.

Rice blast has a significant detrimental impact on rice yields, so developing efficient biological control technologies is an effective means for rice blast prevention and control. The GroEL protein has proven to be effective at preventing and managing the pathogenicity of rice blast. Here, we analyzed the amino acid sequence of the GroEL protein and synthesized the '60 kDa chaperonin signature' (350-373 amino acids) peptide SP1.2, which has potent antifungal activity. Notably, the SP1.2 peptide exhibited potent fungicidal activity against Magnaporthe oryzae, effectively inhibiting appressorium germination. Electron microscopy revealed that SP1.2 disrupted the fungal plasma membrane and bound to multiple bioactive phosphoinositides in vitro, triggering the production of reactive oxygen species. Furthermore, it also caused an increase in the acetylation of M. oryzae and induced autophagy in cells. The spray application of SP1.2 significantly reduced the number of disease spots caused by the fungal pathogen M. oryzae in rice, enhancing the defense response of rice plants. Field trials showed that the control effect was 64.59% after spraying SP1.2. Our study illustrates the antifungal activity of the structurally unique SP1.2 peptide against plant fungal pathogens and paves the way for the future development of this class of peptides as antifungal agents. © 2024 Society of Chemical Industry.

Alternaria solani core effector Aex59 is a new member of the Alt a 1 protein family and is recognized as a PAMP

Early blight caused by Alternaria solani is a destructive disease in potato production. Here, through systematically screening of an effector protein pool consisting of 115 small cysteine-containing candidate Aex (Alternariaextracellular proteins) in A. solani, we identified a core effector protein named Aex59, a pathogen-associated molecular pattern (PAMP) molecule. Aex59 is uniquely present in the Ascomycota of fungi and can activate defense responses in multiple plants. Targeted gene disruption showed that Aex59 is a virulence factor and participates in spore development. Perception of Aex59 in Nicotiana benthamiana does not depend on the receptor-like kinases Brassinosteroid-associated kinase1 (BAK1) and Suppressor of BIR1-1 (SOBIR1), which are required for multiple pattern recognition receptors (PRR) pathways. Sequence analysis revealed that Aex59 is a new member of the Alt a 1 protein family and is a potential molecular marker capable of aiding in the classification of the fungi Alternaria spp.

Bioinformatics deciphers the thebaine biosynthesis pathway in opium poppy: Hub genes, network analysis, and miRNA regulation

Thebaine, a vital precursor in the codeine and morphine pathway, shows promise in addiction treatment. We conducted a comprehensive study on the thebaine biosynthesis pathway in opium poppy, utilizing bioinformatics tools. The dataset comprising the thirteen genes associated with the thebaine biosynthesis pathway was compiled from an extensive review of published literature and validated using the NCBI BLAST tool. Utilizing STRING and Cytoscape, we analyzed gene interactions and visualized the molecular interaction network, respectively. To identify hub proteins, CytoHubba was administered. The Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) at STRING were used for the enrichment analysis of the hub genes. CytoCluster was used to analyze the network in clusters. Promoter regions of hub genes and potential miRNAs were explored using MEME and the psRNATarget database. Hub genes crucial to thebaine biosynthesis were identified, contributing to essential cellular functions like growth, development, stress response, and signal transduction. Metabolic processes emerged as pivotal for thebaine production, indicating a broader role for the thebaine pathway gene network beyond primary metabolite production. Cell component subnetwork genes demonstrated associations with anatomical units, indicating involvement in plant defense responses. Dominant molecular functions drove plant defense responses. KEGG pathway analysis highlighted the significance of metabolic pathways and biosynthesis of secondary metabolites. Cluster analysis emphasized the relevance of the biosynthesis of amino acids, confirming the link between primary and secondary metabolites. Promoter analysis suggested the potential involvement of signal transduction in thebaine production. Hub genes were targeted by 40 miRNAs, suggesting potential novel biomarkers or target genes within the thebaine biosynthesis pathway. Based on the role of miRNAs identified in connection with the hub genes of the thebaine production process, the secondary metabolite pathway of thebaine appears to be associated with several key plant pathways, e.g. growth, development and stress response. However, these findings, based on bioinformatics analysis, warrant further experimental validation and promise to advance our understanding of the biosynthesis of thebaine and its interactions with other genes and metabolic pathways that influence the production of metabolites.