Defense Gene Expression Research Articles

Enhanced fungicidal activity and mechanism of pyraclostrobin nanoparticle with reactive oxygen species responsiveness against Rhizoctonia solani

The frequent application of pesticides has caused adverse effects on humans and the environment. Stimuli-responsive nanoparticles can enhance the control efficacy of pesticides to pest insects or fungus and reduce impacts on non-target organisms. In the present work, 4-(hydroxymethyl) phenylboronic acid pinacol ester (PBAP) groups were conjugated onto β-cyclodextrin (β-CD) to form a H2O2-eliminating material (Ox-bCD), and then the nanoparticiles (PYR@OxbCD) loaded pyraclostrobin (PYR) were successfully fabricated by a nanoprecipitation. The prepared PYR@OxbCD had excellent reactive oxygen species (ROS) release dependency, and significantly enhanced fungicidal activity against Rhizoctonia solani in vitro. PYR@OxbCD could accurately release PYR at infected rice plants, providing a better control efficacy than PYR suspension concentrate (SC). The contact angle of PYR@OxbCD on leaves was reduced, and the spreading and antiphotolysis properties were increased, which may lead to an increase of deposition and a lower dissipation rate. Furthermore, the released PYR from PYR@OxbCD enhanced the activity of peroxidase (POD) and catalase (CAT), and up-regulated the expression of defense genes including PAL, PR10, CHI, CAT, GluPOD and AsPOD. PYR@OxbCD exhibited a much weaker effect on microbial abundance in rice cropping systems. The study provides an idea for smart delivery of fungicides to prevent plant pathogens and offers a more comprehensive view about the resistance of R. solani-infected rice triggered by PYR@OxbCD in levels of phenotype, enzymatic activity, gene expression, and microbiome.

A prophage tail-like protein facilitates the endophytic growth of Burkholderia gladioli and mounting immunity in tomato.

A prophage tail-like protein (Bg_9562) of Burkholderia gladioli strain NGJ1 possesses broad-spectrum antifungal activity, and it is required for the bacterial ability to forage over fungi. Here, we analyzed whether heterologous overexpression of Bg_9562 or exogenous treatment with purified protein can impart disease tolerance in tomato. The physiological relevance of Bg_9562 during endophytic growth of NGJ1 was also investigated. Bg_9562 overexpressing lines demonstrate fungal and bacterial disease tolerance. They exhibit enhanced expression of defense genes and activation of mitogen-activated protein kinases. Treatment with Bg_9562 protein induces defense responses and imparts immunity in wild-type tomato. The defense-inducing ability lies within 18-51 aa region of Bg_9562 and is due to sequence homology with the bacterial flagellin epitope. Interaction studies suggest that Bg_9562 is perceived by FLAGELLIN-SENSING 2 homologs in tomato. The silencing of SlSERK3s (BAK1 homologs) prevents Bg_9562-triggered immunity. Moreover, type III secretion system-dependent translocation of Bg_9562 into host apoplast is important for elicitation of immune responses during colonization of NGJ1. Our study emphasizes that Bg_9562 is important for the endophytic growth of B. gladioli, while the plant perceives it as an indirect indicator of the presence of bacteria to mount immune responses. The findings have practical implications for controlling plant diseases.

Plant Elicitor Peptide (Pep) Signaling and Pathogen Defense in Tomato.

Endogenous signaling compounds are intermediaries in signaling pathways that plants use to respond to the perception of harmful and beneficial organisms. The plant elicitor peptides (Peps) of plants are important endogenous signaling molecules that induce elements of defense responses such as hormone production, increased expression of defensive genes, the activation of phosphorelays, and the induction of cell secondary messenger synthesis. The processes by which Peps confer resistance to pathogenic microorganisms have been extensively studied in Arabidopsis but are less known in crop plants. Tomato and many other solanaceous plants have an endogenous signaling polypeptide, systemin, that is involved in the defense against herbivorous insects and necrotrophic pathogens. This paper explores the similarity of the effects and chemical properties of Pep and systemin in tomato. Additionally, the relationship of the Pep receptor and systemin receptors is explored, and the identification of a second tomato Pep receptor in the literature is called into question. We suggest future directions for research on Pep signaling in solanaceous crops during interactions with microbes.

Is localized acquired resistance the mechanism for effector-triggered disease resistance in plants?

Plant nucleotide-binding leucine-rich repeat receptors (NLRs) are intracellular immune receptors that are activated by their direct or indirect interactions with virulence effectors. NLR activation triggers a strong immune response and consequent disease resistance. However, the NLR-driven immune response can be targeted by virulence effectors. It is thus unclear how immune activation can occur concomitantly with virulence effector suppression of immunity. Recent observations suggest that the activation of effector-triggered immunity does not sustain defence gene expression in tissues in contact with the hemi-biotrophic pathogen Pseudomonas syringae pv. tomato. Instead, strong defence was observed on the border of the infection area. This response is reminiscent of localized acquired resistance (LAR). LAR is a strong defence response occurring in a ~2 mm area around cells in contact with the pathogen and probably serves to prevent the spread of pathogens. Here we propose that effector-triggered immunity is essentially a quarantining mechanism to prevent systemic pathogen spread and disease, and that the induction of LAR is a key component of this mechanism.

Colonization of Serendipita indica promotes resistance against Spodoptera exigua in onion (Allium cepa L.).

Plant-endophyte symbiosis influences plant defense and growth. Serendipita indica is a root endophyte that promotes growth and induces tolerance against biotic and abiotic stress in plants. In this study, we examined the effect of S. indica colonization on herbivore (Spodoptera exigua) resistance of onion (Allium cepa L.). We found that colonization of S. indica in the roots of onion significantly reduced the feeding damage of leaves by S. exigua larvae, and also resulted in a reduction in weight gain of the larvae when fed on S. indica plants. This enhanced resistance is a result of modulation of antioxidant and defense enzymes/genes in the host by S. indica mutualism. Specifically, the activities of enzymes such as Superoxide dismutase, peroxidase, polyphenol oxidase, phenylalanine ammonia-lyase, and H2O2 content were significantly higher in the early stages of S. exigua feeding in the S. indica colonized plants compared to the non-colonized counterparts. Similarly, defense genes also showed modulation in response to this tripartite interaction of onion -S. indica mutualism and S. exigua herbivory. The hierarchical cluster analysis and principal component analysis indicated a clear difference in the onion biochemical responses, which is due to the S. indica symbiosis. Our investigation demonstrates that onion-S. indica symbiosis significantly decreases chewing injury by efficiently modulating antioxidant and defense enzyme activities and gene expression in response to S. exigua herbivory. Therefore, S. indica can be used as a potential biocontrol agent for sustainable management of this important pest of Alliums.

A chitin deacetylase PsCDA2 from Puccinia striiformis f. sp. tritici confers disease pathogenicity by suppressing chitin-triggered immunity in wheat.

Plants have the ability to recognize the essential chitin molecule present in the fungal cell wall, which stimulates the immune response. Phytopathogenic fungi have developed various strategies to inhibit the chitin-triggered immune response. Here, we identified a chitin deacetylase of Puccinia striiformis f. sp. tritici (Pst), known as PsCDA2, that was induced during the initial invasion of wheat and acted as an inhibitor of plant cell death. Knockdown of PsCDA2 in wheat enhanced its resistance against Pst, highlighting the significance of PsCDA2 in the host-pathogen interaction. Moreover, PsCDA2 can protect Pst urediniospores from being damaged by host chitinase in vitro. PsCDA2 also suppressed the basal chitin-induced plant immune response, including the accumulation of callose and the expression of defence genes. Overall, our results demonstrate that Pst secretes PsCDA2 as a chitin deacetylase involved in establishing infection and modifying the acetyl group to prevent the breakdown of chitin in the cell wall by host endogenous chitinases. Our research unveils a mechanism by which the fungus suppresses plant immunity, further contributing to the understanding of wheat stripe rust control. This information could have significant implications for the development of suitable strategies for protecting crops against the devastating effects of this disease.

PIF3 is phosphorylated by MAPK to modulate plant immunity.

Surface-localized pattern recognition receptors perceive pathogen-associated molecular patterns (PAMPs) to activate pattern-triggered immunity (PTI). Activation of mitogen-activated protein kinases (MAPKs) represents a major PTI response. Here, we report that Arabidopsis thaliana PIF3 negatively regulates plant defense gene expression and resistance to Pseudomonas syringae DC3000. PAMPs trigger phosphorylation of PIF3. Further study reveals that PIF3 interacts with and is phosphorylated by MPK3/6. By mass spectrometry and site-directed mutagenesis, we identified the corresponding phosphorylation sites which fit for SP motif. We further show that a phospho-mimicking PIF3 variant (PIF36D /pifq) conferred increased susceptibility to P. syringae DC3000 and caused lower levels of defense gene expression in plants. Together, this study reveals that PIF3 is phosphorylated by MPK3/6 and phosphorylation of the SP motif residues is required for its negative regulation on plant immunity.

Expression comparisons of defense-related genes in resistant and susceptible chickpea cultivars in response to Ascochyta rabiei

Ascochyta blight caused by Ascochyta rabiei is a destructive disease affecting chickpea crops worldwide. Numerous studies have sought to identify resistance genes involved in the defense response against Ascochyta blight, but the function of potential resistant genes across different chickpea genetic backgrounds remains poorly understood. This study evaluated the potential role of 16 genes involved in defense responses in partially resistant (ILC482) and susceptible (Sarı98) chickpea cultivars. The leaf samples were collected from 2-week-old seedlings at 6, 12, 24, 48, and 72 h post-inoculation (hpi) with A. rabiei and analyzed for differential expression of defense genes using qRT-PCR. Twelve out of 16 genes were differentially regulated between chickpea cultivars. The qRT-PCR analysis indicated that the expression of defense genes was significantly higher in the partially resistant cultivar than that in the susceptible cultivar. The time point of the highest upregulation ratio for defense genes was variable between the cultivars. The expression of glutathione S-transferase, serine/threonine-protein kinase, WRKY gene (CaWRKY16), and wall associated-receptor kinase genes reached maximum levels in the resistant cultivar at 6 hpi, while antimicrobial peptide precursor (SNAKIN2), polymorphic antigen membrane protein, ethylene receptor-like sequences (CaETR1), and two nucleotide-binding site-leucine-rich repeat genes showed the highest activity at 24 hpi. In particular, CaWRKY16, CaWRKY50, glutathione S-transferase, CaETR1, and wall-associated-receptor kinase genes provided valuable information for assessing chickpea cultivars with different resistance levels. The results indicated that the expression changes of these genes involved in defense pathways play a crucial role in enhanced resistance of chickpea plants against Ascochyta blight and, therefore could be potential candidate genes for future breeding studies.

Biogenic silicon nanoparticles mitigate cadmium (Cd) toxicity in rapeseed (Brassica napus L.) by modulating the cellular oxidative stress metabolism and reducing Cd translocation

Nano-enabled strategies have emerged as promising alternatives to resolve heavy metals (HMs) related harms in an eco-friendly manner. Here, we explored the potential of biogenic silicon nanoparticles (SiNPs) in alleviating cadmium (Cd) stress in rapeseed (Brassica napus L.) plants by modulating cellular oxidative repair mechanisms. Biogenic SiNPs of spherical shapes with size ranging between 14 nm and 35 nm were synthesized using rice straw extract and characterized through advanced characterization techniques. A greenhouse experiment results showed that SiNPs treatment at 250 mg kg−1 significantly improved growth parameters, including fresh weight (33.3 %) and dry weight (32.6 %) of rapeseed plants than Cd-treated control group. Photosynthesis and leaf gas exchange parameters were also positively influenced by SiNPs treatment, indicating enhanced photosynthetic efficiency. Additionally, SiNPs treatment at 250 mg kg−1 increased the activities of antioxidant enzymes such as superoxide dismutase (19.1 %), peroxidase (33.4 %), catalase (14.4 %), and ascorbate peroxidase (33.8 %), which may play a crucial role in ROS scavenging and reduction in Cd-induced oxidative stress. TEM analysis revealed that SiNPs treatment effectively mitigated Cd-induced damage to leaf ultrastructure, while qPCR analysis showed that SiNPs treatment changed the expressions of the antioxidant defense and stress related genes. Moreover, SiNPs treatment significantly influenced the Cd accumulation and Si contents in plants. Overall, our findings revealed that biogenic SiNPs have great potential to serve as a sustainable, eco-friendly, and non-toxic alternative for the remediation of Cd toxicity in rapeseed plants.

Entomopathogenic Fungus-Related Priming Defense Mechanisms in Cucurbits Impact Spodoptera littoralis (Boisduval) Fitness.

Entomopathogenic fungi (EPF) exhibit direct and indirect mechanisms to increase plant resistance against biotic and abiotic stresses. Plant responses to these stresses are interconnected by common regulators such as ethylene (ET), which is involved in both iron (Fe) deficiency and induced systemic resistance responses. In this work, the roots of cucurbit seedlings were primed with Metarhizium brunneum (EAMa 01/58-Su strain), and relative expression levels of 18 genes related to ethylene (ET), jasmonic acid (JA), and salicylic acid (SA) synthesis, as well as pathogen-related (PR) protein genes, were studied by reverse transcription-quantitative PCR (qRT-PCR). Effects of priming on Spodoptera littoralis were studied by feeding larvae for 15 days with primed and control plants. Genes showed upregulation in studied species; however, the highest relative expression was observed in roots and shoots of plants with Fe deficiency, demonstrating the complexity and the overlapping degree of the regulatory network. EIN2 and EIN3 should be highlighted; both are key genes of the ET transduction pathway that enhanced their expression levels up to eight and four times, respectively, in shoots of primed cucumber. Also, JA and SA synthesis and PR genes showed significant upregulation during the observation period (e.g., the JA gene LOX1 increased 506 times). Survival and fitness of S. littoralis were affected with significant effects on mortality of larvae fed on primed plants versus controls, length of the larval stage, pupal weight, and the percentage of abnormal pupae. These results highlight the role of the EAMa 01/58-Su strain in the induction of resistance, which could be translated into direct benefits for plant development. IMPORTANCE Entomopathogenic fungi are multipurpose microorganisms with direct and indirect effects on insect pests. Also, EPF provide multiple benefits to plants by solubilizing minerals and facilitating nutrient acquisition. A very interesting and novel effect of these fungi is the enhancement of plant defense systems by inducing systematic and acquired resistance. However, little is known about this function. This study sheds light on the molecular mechanisms involved in cucurbits plants' defense activation after being primed by the EPF M. brunneum. Furthermore, the subsequent effects on the fitness of the lepidopteran pest S. littoralis are shown. In this regard, a significant upregulation was recorded for the genes that regulate JA, SA, and ET pathways. This increased expression of defense genes caused lethal and sublethal effects on S. littoralis. This could be considered an added value for the implementation of EPF in integrated pest management programs.

G-type receptor-like kinase AsNIP43 interacts with rhizobia effector nodulation outer protein P and is required for symbiosis.

In the Rhizobium-Legume symbiosis, the nodulation outer protein P (NopP) effector is one of the key regulators for rhizobial infection and nodule organogenesis. However, the molecular mechanism through which host legume plants sense NopP remains largely unknown. Here, we constructed an nopP deletion mutant of Mesorhizobium huakuii and found that nopP negatively regulates nodulation on Chinese milk vetch (Astragalus sinicus). Screening for NopP interacting proteins in host plants using the yeast 2-hybrid system identified NopP interacting protein 43 (AsNIP43), which encodes a G-type receptor-like kinase (LecRLK). The B-lectin domain at the N terminus of AsNIP43 was essential in mediating its interaction with NopP, which was confirmed in vitro and in vivo. Subcellular localization, co-localization, and gene expression analyses showed that AsNIP43 and NopP function tightly associated with earlier infection events. RNA interference (RNAi) knockdown of AsNIP43 expression by hairy root transformation led to decreased nodule formation. AsNIP43 plays a positive role in symbiosis, which was further verified in the model legume Medicago truncatula. Transcriptome analysis indicated that MtRLK (a homolog of AsNIP43 in M. truncatula) may function to affect defense gene expression and thus to regulate early nodulation. Taken together, we show that LecRLK AsNIP43 is a legume host target that interacts with rhizobia effector NopP is essential for rhizobial infection and nodulation.

9,10-KODA, an α-ketol produced by the tonoplast-localized 9-lipoxygenase ZmLOX5, plays a signaling role in maize defense against insect herbivory

13-Lipoxygenases (LOXs) initiate the synthesis of jasmonic acid (JA), the best-understood oxylipin hormone in herbivory defense. However, the roles of 9-LOX-derived oxylipins in insect resistance remain unclear. Here, we report a novel anti-herbivory mechanism mediated by a tonoplast-localized 9-LOX, ZmLOX5, and its linolenic acid-derived product, 9-hydroxy-10-oxo-12(Z),15(Z)-octadecadienoic acid (9,10-KODA). Transposon-insertional disruption of ZmLOX5 resulted in the loss of resistance to insect herbivory. lox5 knockout mutants displayed greatly reduced wound-induced accumulation of multiple oxylipins and defense metabolites, including benzoxazinoids, abscisic acid (ABA), and JA-isoleucine (JA-Ile). However, exogenous JA-Ile failed to rescue insect defense in lox5 mutants, while applications of 1 μM 9,10-KODA or the JA precursor, 12-oxo-phytodienoic acid (12-OPDA), restored wild-type resistance levels. Metabolite profiling revealed that exogenous 9,10-KODA primed the plants for increased production of ABA and 12-OPDA, but not JA-Ile. While none of the 9-oxylipins were able to rescue JA-Ile induction, the lox5 mutant accumulated lower wound-induced levels of Ca2+, suggesting this as a potential explanation for lower wound-induced JA. Seedlings pretreated with 9,10-KODA exhibited rapid or more robust wound-induced defense gene expression. In addition, an artificial diet supplemented with 9,10-KODA arrested fall armyworm larvae growth. Finally, analysis of single and double lox5 and lox10 mutants showed that ZmLOX5 also contributed to insect defense by modulating ZmLOX10-mediated green leaf volatile signaling. Collectively, our study uncovered a previously unknown anti-herbivore defense and hormone-like signaling activity for a major 9-oxylipin α-ketol.

Overexpression of Sly-miR398b Compromises Disease Resistance against Botrytis cinerea through Regulating ROS Homeostasis and JA-Related Defense Genes in Tomato.

MicroRNAs (miRNAs) have been shown to be critical components in plant immunity. MicroRNA398 (miR398) is a highly conserved miRNA in all land plants and plays crucial roles in diverse biotic stress responses. However, the role of miR398 has not yet been characterized in tomato resistance against Botrytis cinerea. In this report, the transcript levels of sly-miR398b were strongly decreased in B. cinerea-infected leaves and the overexpression of sly-miR398b resulted in enhanced susceptibility. The attenuated expression of cytosol Cu/Zn-SOD (CSD1), chloroplast Cu/Zn-SOD (CSD2), and guaiacol peroxidase (GPOD), as well as the decreased activities of superoxide dismutase (SOD) and GPOD, collectively led to increased hydrogen peroxide (H2O2) accumulation in sly-miR398b overexpressing plants. Furthermore, sly-miR398b was induced by methyl jasmonate (MeJA) treatment. The overexpression of sly-miR398b suppressed the expression of TomLoxD, LapA, and PR-STH2 in response to B. cinerea and MeJA treatment. Our data demonstrate that sly-miR398b overexpression negatively regulates the resistance to B. cinerea in tomato by inducing the accumulation of reactive oxygen species (ROS) and downregulating the expression of MeJA-responsive defense genes.

The Slow Progression of Diabetic Retinopathy Is Associated with Transient Protection of Retinal Vessels from Death.

The purpose of this study was to investigate the reason that diabetic retinopathy (DR) is delayed from the onset of diabetes (DM) in diabetic mice. To this end, we tested the hypothesis that the deleterious effects of DM are initially tolerated because endogenous antioxidative defense is elevated and thereby confers resistance to oxidative stress-induced death. We found that this was indeed the case in both type 1 DM (T1D) and type 2 DM (T2D) mouse models. The retinal expression of antioxidant defense genes was increased soon after the onset of DM. In addition, ischemia/oxidative stress caused less death in the retinal vasculature of DM versus non-DM mice. Further investigation with T1D mice revealed that protection was transient; it waned as the duration of DM was prolonged. Finally, a loss of protection was associated with the manifestation of both neural and vascular abnormalities that are diagnostic of DR in mice. These observations demonstrate that DM can transiently activate protection from oxidative stress, which is a plausible explanation for the delay in the development of DR from the onset of DM.

The Medicago truncatula nodule-specific cysteine-rich peptides, NCR343 and NCR-new35 are required for the maintenance of rhizobia in nitrogen-fixing nodules.

In the nodules of IRLC legumes, including Medicago truncatula, nitrogen-fixing rhizobia undergo terminal differentiation resulting in elongated and endoreduplicated bacteroids specialized for nitrogen fixation. This irreversible transition of rhizobia is mediated by host produced nodule-specific cysteine-rich (NCR) peptides, of which c. 700 are encoded in the M.truncatula genome but only few of them have been proved to be essential for nitrogen fixation. We carried out the characterization of the nodulation phenotype of three ineffective nitrogen-fixing M. truncatula mutants using confocal and electron microscopy, monitored the expression of defence and senescence-related marker genes, and analysed the bacteroid differentiation with flow cytometry. Genetic mapping combined with microarray- or transcriptome-based cloning was used to identify the impaired genes. Mtsym19 and Mtsym20 mutants are defective in the same peptide NCR-new35 and the lack of NCR343 is responsible for the ineffective symbiosis of NF-FN9363. We found that theexpression of NCR-new35 is significantly lower and limited to the transition zone of the nodule compared with other crucial NCRs. The fluorescent protein-tagged version of NCR343 and NCR-new35 localized to the symbiotic compartment. Our discovery added two additional members to the group of NCR genes essential for nitrogen-fixing symbiosis in M. truncatula.

Physiological and Gene Expression Response of Interspecific Hybrids of Fraxinus mandshurica × Fraxinus americana to MJ or SNP under Drought

Drought affects the growth and production of Fraxinus tree species, such as the precious woody plant Fraxinus mandshurica. Based on interspecific hybrid F1 combinations, D110 plants of F. mandshurica × F. americana with strong drought resistance were selected for this study. To further reveal their heterosis mechanism under drought, in this study, an analysis was conducted pertaining to the physiological indexes and gene expression of related key gene changes in materials of 5 yr D110 seedlings and their female and male parental controls (D113 and 4–3) in response to drought, as well as to the addition of sodium nitrate (SNP, a donor of nitric oxide) and methyl jasmonate (MJ, a donor of jasmonate) signal molecules after drought. The results showed that under drought stress, hybrid D110 plants performed significantly better than their parents, especially compared to D113, in plant growth (the plant height growth was 29.48% higher), photosynthesis (the net photosynthetic rate was 38.4% higher), peroxidation (the increase in MDA content was 71.77% lower), defense enzyme activity (SOD and POD activities were 36.63% and 65.58% higher), hormone contents (ABA, IAA and GA were 33.9%~51.2% higher) and gene expression (the LHY and TOC1 rhythmic genes were 131.97%~165.81% higher). When an exogenous additive agent (SNP or MJ) was applied after drought, the negative effects of drought on growth were effectively alleviated (the tree height growth of D110 increased from 22.76% to 22.32% in comparison to drought conditions); meanwhile, the height growth of D110 plants was significantly higher than that of their parents. Further results of physiological indexes and the expression of related key gene changes in response to SNP or MJ also indicated that D110 plants can recover faster from drought than their parents after application of SNP or MJ. This article provides new ideas for revealing the heterosis mechanism of the drought resistance of interspecific F1 hybrids and supplies effective measures for improving drought resistance in F. mandshurica.

The RLCK subfamily VII-4 controls pattern-triggered immunity and basal resistance to bacterial and fungal pathogens in rice.

Receptor-like cytoplasmic kinases (RLCKs) mediate the intracellular signaling downstream of pattern-recognition receptors (PRRs). Several RLCKs from subfamily VII of rice (Oryza sativa) have important roles in plant immunity, but the role of RLCKVII-4 in pattern-triggered immune (PTI) signaling and resistance to pathogens has not yet been investigated. Here, we generated by multiplex CRISPR/Cas9-mediated genome editing rice sextuple mutant lines where the entire RLCKVII-4 subfamily is inactivated and, analyzed the resulting lines for their response to chitin and flg22 and for their immunity to Xanthomonas oryzae pv. oryzae (Xoo)and Magnaporthe oryzae. Analysis of the rlckvii-4 mutants revealed that they have an impaired ROS burst and reduced defense gene expression in response to flg22 and chitin. This indicates that the members of the rice RLCKVII-4 subfamily are required for immune signaling downstream of multiple PRRs. Furthermore, we found that the rice RLCKVII-4 subfamily is important for chitin-induced callose deposition and MAPK activation and that it is crucial for basal resistance against Xoo and M. oryzae pathogens. This establishes that the RLCKVII-4 subfamily has critical functions in the regulation of multiple PTI pathways in rice and opens the way for deciphering the precise role of its members in the control of rice PTI.

The flavour of grape colour: anthocyanin content tunes aroma precursor composition by altering the berry microenvironment.

Anthocyanin-less (white) instead of black/red (coloured) fruits develop in grapevine cultivars without functional VviMYBA1 and VviMYBA2 genes, which conditions the colour of wines that can be produced. To evaluate whether this genetic variation has additional consequences on fruit ripening and composition, we performed microenvironment, transcriptomics and metabolomics comparisons of developing grapes between near-isogenic white- and black-berried somatic variants of Garnacha and Tempranillo cultivars. Berry temperature was up to 3.5ºC lower in white- compared to black-berried Tempranillo. An RNA-seq study combined with targeted and untargeted metabolomics revealed that ripening fruits of white-berried variants were characterized by the upregulation of photosynthesis and other light-responsive genes and their higher accumulation of specific terpene aroma precursors, fatty acid-derived aldehyde volatiles and phenylpropanoid precursor amino acids. MYBA1-MYBA2 function proved essential for flavonol trihydroxylation in black-berried somatic variants, which were also characterized by enhanced pathogen defence gene expression in berry skin and increased accumulation of C6-derived alcohol and ester volatiles and GABA. Collectively, our results indicate that anthocyanin depletion entails side effects on grape composition by altering the berry internal microenvironment and the partition of the phenylpropanoid pathway. These findings show how fruit colour can condition other fruit features, such as flavour potential and stress homeostasis.

Antibacterial mechanism of the novel antimicrobial peptide Jelleine-Ic and its efficacy in controlling Pseudomonas syringae pv. actinidiae in kiwifruit.

Bacterial canker caused by Pseudomonas syringae pv. actinidiae (Psa) poses a severe threat to kiwifruit production. Because of the insufficient efficacy and environmental safety of available treatments, novel antibacterial agents should be urgently developed. Antimicrobial peptides (AMPs) can be used as antimicrobials for disease control. In this study, we designed a novel AMP, Jelleine-Ic, and evaluated its antibacterial activity and mechanism of action against Psa. Jelleine-Ic with a half maximal effective concentration of 1.67 μg/mL exhibited stronger antibacterial activity than did parent Jelleine-I. Jelleine-Ic targeted the Psa membrane, increased membrane permeabilization, and dissipated membrane potential, resulting in calcium leakage. Electron microscopy revealed that Jelleine-Ic disrupted cell morphology and caused intracellular alterations. Moreover, this AMP penetrated the cell membrane, bound to DNA, and reduced the expression of genes related to DNA replication and repair. Jelleine-Ic also reduced esterase activity and induced intracellular reactive oxygen species generation. This peptide inhibited the development of Psa canker. The control efficiency of Jelleine-Ic against Psa in the leaf discs and leaves of kiwifruit was 81.83% and 70.53%, respectively, which was superior to that of the commercial agricultural streptomycin. Furthermore, Jelleine-Ic upregulated the expression of kiwifruit defense genes (PR-10 and WRKY70a). Jelleine-Ic effectively controls Psa in vitro and in vivo, and may be developed as a bactericide for plant disease control. This article is protected by copyright. All rights reserved.

Engineering Climate-Resilient Rice Using a Nanobiostimulant-Based "Stress Training" Strategy.

Under a changing climate, cultivating climate-resilient crops will be critical to maintaining food security. Here, we propose the application of reactive oxygen species (ROS)-generating nanoparticles as nanobiostimulants to trigger stress/immune responses and subsequently increase the stress resilience of plants. We established three regimens of silver nanoparticles (AgNPs)-based "stress training": seed training (ST), leaf training (LT), and combined seed and leaf training (SLT). Trained rice seedlings were then exposed to either rice blast fungus (Magnaporthe oryzae) or chilling stress (10 °C). The results show that all "stress training" regimes, particularly SLT, significantly enhanced the resistance of rice against the fungal pathogen (lesion size reduced by 82% relative to untrained control). SLT also significantly enhanced rice tolerance to cold stress. The mechanisms for the enhanced resilience were investigated with metabolomics and transcriptomics, which show that "stress training" induced considerable metabolic and transcriptional reprogramming in rice leaves. AgNPs boosted ROS-activated stress signaling pathways by oxidative post-translational modifications of stress-related kinases, hormones, and transcriptional factors (TFs). These signaling pathways subsequently modulated the expression of defense genes, including specialized metabolites (SMs) biosynthesis genes, cell membrane lipid metabolism genes, and pathogen-plant interaction genes. Importantly, results showed that the "stress memory" can be transferred transgenerationally, conferring offspring seeds with improved seed germination and seedling vigor. This may provide an epigenetic breeding strategy to fortify stress resilience of crops. This nanobiostimulant-based stress training strategy will increase yield vigor against a changing climate and will contribute to sustainable agriculture by reducing agrochemical use.