Expression Of Pathogenesis-related Genes Research Articles

Interference with developmental and pathogenesis related genes expression in 16Sr-IX associated phytoplasma infectedBrassica campestris

Interference with developmental and pathogenesis related genes expression in 16Sr-IX associated phytoplasma infected<i>Brassica campestris</i>

PpWRKY45 is involved in methyl jasmonate primed disease resistance by enhancing the expression of jasmonate acid biosynthetic and pathogenesis-related genes of peach fruit

• 10 μM of MeJA effectively inhibited soft rot caused by R. stolonifer. • MeJA elevated the expression of JA biosynthetic and PR genes. • MeJA induced disease resistance against R. stolonifer by priming of defense responses. • PpWRKY45 activated defense-related gene expression by bind to their promoters. • PpWRKY45 is involved in MeJA-primed defense by enhancing defense-related gene expression. Methyl jasmonate (MeJA) and WRKY transcription factors (TFs) are documented to exert vital defensive functions. Research concerning WRKY TFs together with MeJA-primed defense against Rhizopus stolonifer in peach fruit is still in its infancy. In the present study, the mode of MeJA on inducing resistance against soft rot caused by Rhizopus stolonifer in peaches during the postharvest storage and the involvement of WRKY TFs were investigated, and the results manifested that MeJA at 10 μM significantly reduced disease occurrence and lesion diameter after R. stolonifer infection during 60 h of storage at 20 °C. Moreover, the MeJA treatment promoted the activity of CHI and GLU and elevated the expression levels of PpLOX , PpAOS and PpOPR3 . More importantly, the expression of PpCHI , PpGLU , PpPR-like , PpLOX , PpAOS and PpOPR3 were substantially and rapidly elevated in the peach fruit that pretreated with MeJA and inoculated with R.stolonifer , indicating that MeJA stimulated a specific priming defense against Rhizopus rot in peaches. Furthermore, a MeJA-related transcription factor PpWRKY45 was identified and characterized as a nucleus-localized protein that could activate the expression of PpCHI, PpGLU, PpPR-like, PpLOX, PpAOS and PpOPR3 by binding to W-box elements in their promoters. These results indicate that PpWRKY45 is involved in MeJA-primed defense against R . stolonifer by activating JA biosynthetic and PR genes of peach fruit.

Overexpression of JcWRKY2 confers increased resistance towards Macrophomina phaseolina in transgenic tobacco.

WRKY proteins are plant-specific transcription factors (TFs), and form one of the largest families and are involved in plant development and responses to stress. The salicylic acid (SA) responsive WRKY family auto or cross-regulate the defence stress signalling pathways. In this study, we functionally validated the role of JcWRKY2 gene from biofuel crop Jatropha curcas towards improving resistance to tobacco transgenic against charcoal rot causing necrotrophic fungus, Macrophomina phaseolina. The microscopic studies revealed that JcWRKY2 participated in preventing the spread of infection in transgenic. The generation of H2O2 during M. phaseolina and combinatorial stress in transgenic induces the expression and activity of antioxidant enzymes. The transcript expression of SA biosynthetic (NtICS1) gene, pathogenesis-related (NtPR-10) gene and antioxidative enzymes (NtCAT1 and NtSOD) gene revealed that JcWRKY2 transgenic play a role in SA-mediated, antioxidative enzymes regulation during biotic challenges. The study highlights the potential of JcWRKY2 as an important regulator for plant biotic stress responses through the SA-dependent pathway.

Mediation of induced systemic resistance by the plant growth-promoting rhizobacteria Bacillus pumilus S2-3-2.

Plant-rhizobacteria interaction and co-evolution developed adaptive strategies which may help the plant survive in nature. Plant rhizosphere soil isolates were analyzed to investigated the effects of rhizobacteria for promoting plant growth and suppress plant disease.Bacterial strains which isolated from plant rhizosphere soil were screened for elicitation of induced systemic resistance (ISR) on tobacco. Strain S2-3-2 results in significant reduction of disease severity on tobacco, it was identified as Bacillus pumilus by multilocus sequence analysis (MLSA). Strain S2-3-2 was deeper studied for pepper plant growth promotion and biological control activity against pepper bacterial spot disease. It was found that the pepper disease severity was decreased when the roots were drenched with strain S2-3-2, and the pepper plants had a higher weight and chlorophyll content, as compared with the mock-treated plants. Transcriptional expression of pathogenesis-related (PR) protein genes in pepper was analyzed by real-time PCR, gene expressions of CaPR1, CaPR4, and CaPR10 were increased when the plants were treated with strain S2-3-2. Moreover, strain S2-3-2 was tested for the production of indole-3-acetic acid (IAA), and it was determined to emit volatiles that enhance the growth of the tobacco plants. Interesting, heat-killed S2-3-2 enhance the pepper root growth, increase the gene expressions of CaPR4 and CaPR10 after pathogen challenge for 6h, but limited to suppress the pepper bacterial spot disease as compare to the mock-treated plants.Strain S2-3-2 can be a potential biological control agent on the plant root for plant growth promoting and disease suppression.

Transcriptome Analyses of β-Thalassemia -28(A>G) Mutation Using Isogenic Cell Models Generated by CRISPR/Cas9 and Asymmetric Single-Stranded Oligodeoxynucleotides (assODNs).

β-thalassemia, caused by mutations in the human hemoglobin β (HBB) gene, is one of the most common genetic diseases in the world. The HBB −28(A>G) mutation is one of the five most common mutations in Chinese patients with β-thalassemia. However, few studies have been conducted to understand how this mutation affects the expression of pathogenesis-related genes, including globin genes, due to limited homozygote clinical materials. Therefore, we developed an efficient technique using CRISPR/Cas9 combined with asymmetric single-stranded oligodeoxynucleotides (assODNs) to generate a K562 cell model with HBB −28(A>G) named K562–28(A>G). Then, we systematically analyzed the differences between K562–28(A>G) and K562 at the transcriptome level by high-throughput RNA-seq before and after erythroid differentiation. We found that the HBB −28(A>G) mutation not only disturbed the transcription of HBB, but also decreased the expression of HBG, which may further aggravate the thalassemia phenotype and partially explain the more severe clinical outcome of β-thalassemia patients with the HBB −28(A>G) mutation. Moreover, we found that the K562–28(A>G) cell line is more sensitive to hypoxia and shows a defective erythrogenic program compared with K562 before differentiation. Importantly, all abovementioned abnormalities in K562–28(A>G) were reversed after correction of this mutation with CRISPR/Cas9 and assODNs, confirming the specificity of these phenotypes. Overall, this is the first time to analyze the effects of the HBB −28(A>G) mutation at the whole-transcriptome level based on isogenic cell lines, providing a landscape for further investigation of the mechanism of β-thalassemia with the HBB −28(A>G) mutation.

Constitutive Expression of Arabidopsis Senescence Associated Gene 101 in Brachypodium distachyon Enhances Resistance to Puccinia brachypodii and Magnaporthe oryzae.

Brachypodium distachyon, as an effective model of cereal grains, is susceptible to most destructive cereal pathogens. Senescence associated gene 101 (SAG101) has been studied extensively in Arabidopsis. SAG101 is one of the important regulators of plant immunity. However, no homologous genes of AtSAG101 were found in B. distachyon. In this study, the AtSAG101 gene was transformed into B. distachyon. Three transgenic plant lines containing the AtSAG101 gene were confirmed by PCR and GUS gene activity. There were fewer Puccinia brachypodii urediospores in the AtSAG101-overexpressing plants compared to wild type plants. P. brachypodii biomass was obviously decreased in AtSAG101 transgenic plants. The length of infection hyphae and infection unit areas of P. brachypodii were significantly limited in transgenic plants. Moreover, there were small lesions in AtSAG101 transgenic plants challenged by Magnaporthe oryzae. Salicylic acid accumulation was significantly increased, which led to elevated pathogenesis-related gene expression in transgenic B. distachyon inoculated by P. brachypodii or M. oryzae compared to wild type plants. These results were consistent with infected phenotypes. Overexpression of AtSAG101 in B. distachyon caused resistance to M. oryzae and P. brachypodii. These results suggest that AtSAG101 could regulate plant resistance in B. distachyon.

Overexpression of Pti4, Pti5, and Pti6 in tomato promote plant defense and fruit ripening

Pseudomonas syringae pv. tomato (Pst) is a pathogenic microorganism that causes bacterial speck disease and affects tomato yield and quality. Pto is a disease resistant gene for plant to recognize and defense against Pst. Pto interacts with Pti (Pto interacting) proteins, which include three transcription factors, Pti4, Pti5, Pti6, and they were thought to be downstream of Pto-mediated pathway to promote the expression of disease-related genes. In the present work, the overexpression plants of Pti4, Pti5 or Pti6 were obtained by Agrobacterium-mediated transformation in tomato. The Pti4/5/6-overexpressed lines indicated enhanced expression of pathogenesis-related genes and resistance to pathogenic bacteria Pst DC3000. Meanwhile, the transgenic plants showed that Pti4/5/6 function in ripening but performed no obvious adverse influence on flowering time, seed-setting rate, weight and soluble solids content of fruits. Furthermore, Pti-overexpressed fruits exhibited increased enzymatic activities of phenylalnine ammonialyase, catalase, peroxidase and decreased content of malondialdehyde. Additionally, cell-free and in vivo ubiquitination assay indicated that Pti4, Pti5 and Pti6 degraded by 26S proteasome which suggested that these Pti transcription regulators’ functions could be regulated by ubiquitin-mediated post translational regulation in tomato.

OsExo70B1 Positively Regulates Disease Resistance to Magnaporthe oryzae in Rice.

The exocyst, an evolutionarily conserved octameric protein complex, mediates tethering of vesicles to the plasma membrane in the early stage of exocytosis. Arabidopsis Exo70, a subunit of the exocyst complex, has been found to be involved in plant immunity. Here, we characterize the function of OsExo70B1 in rice. OsExo70B1 mainly expresses in leaf and shoot and its expression is induced by pathogen-associated molecular patterns (PAMPs) and rice blast fungus Magnaporthe oryzae (M. oryzae). Knocking out OsExo70B1 results in significantly decreased resistance and defense responses to M. oryzae compared to the wild type, including more disease lesions and enhanced fungal growth, downregulated expression of pathogenesis-related (PR) genes, and decreased reactive oxygen species accumulation. In contrast, the exo70B1 mutant does not show any defects in growth and development. Furthermore, OsExo70B1 can interact with the receptor-like kinase OsCERK1, an essential component for chitin reception in rice. Taken together, our data demonstrate that OsExo70B1 functions as an important regulator in rice immunity.

Molecular Cloning and Functional Analysis of the NPR1 Homolog in Kiwifruit (Actinidia eriantha)

Kiwifruit bacterial canker, caused by the bacterial pathogen Pseudomonas syringae pv. actinidiae (Psa), is a destructive disease in the kiwifruit industry globally. Consequently, understanding the mechanism of defense against pathogens in kiwifruit could facilitate the development of effective novel protection strategies. The Non-expressor of Pathogenesis-Related genes 1 (NPR1) is a critical component of the salicylic acid (SA)-dependent signaling pathway. Here, a novel kiwifruit NPR1-like gene, designated AeNPR1a, was isolated by using PCR and rapid amplification of cDNA ends techniques. The full-length cDNA consisted of 1952 base pairs with a 1,746-bp open-reading frame encoding a 582 amino acid protein. Homology analysis showed that the AeNPR1a protein is significantly similar to the VvNPR1 of grape. A 2.0 Kb 5′-flanking region of AeNPR1a was isolated, and sequence identification revealed the presence of several putative cis-regulatory elements, including basic elements, defense and stress response elements, and binding sites for WRKY transcription factors. Real-time quantitative PCR results demonstrated that AeNPR1a had different expression patterns in various tissues, and its transcription could be induced by phytohormone treatment and Psa inoculation. The yeast two-hybrid assay revealed that AeNPR1a interacts with AeTGA2. Constitutive expression of AeNPR1a induced the expression of pathogenesis-related gene in transgenic tobacco plants and enhanced tolerance to bacterial pathogens. In addition, AeNPR1a expression could restore basal resistance to Pseudomonas syringae pv. tomato DC3000 (Pst) in Arabidopsis npr1-1 mutant. Our data suggest that AeNPR1a gene is likely to play a pivotal role in defense responses in kiwifruit.

RIN13-mediated disease resistance depends on the SNC1-EDS1/PAD4 signaling pathway in Arabidopsis.

Plants have evolved an innate immune system to protect themselves from pathogen invasion with the help of intracellular nucleotide-binding leucine-rich repeat (NLR) receptors, though the mechanisms remain largely undefined. RIN13 (RPM1-interacting protein 13) was previously reported to enhance disease resistance, and suppress RPM1 (a CNL-type NLR)-mediated hypersensitive response in Arabidopsis via an as yet unknown mechanism. Here, we show that RIN13 is a nuclear-localized protein, and functions therein. Overexpression of RIN13 leads to autoimmunity with high accumulation of salicylic acid (SA), constitutive expression of pathogenesis-related genes, enhanced resistance to a virulent pathogen, and dwarfism. In addition, genetic and transcriptome analyses show that SA-dependent and SA-independent pathways are both required for RIN13-mediated disease resistance, with the EDS1/PAD4 complex as an integration point. RIN13-induced dwarfism was rescued completely by either the pad4-1 or the eds1-2 mutant but partially by snc1-r1, a mutant of the TNL gene SNC1, suggesting the involvement of EDS1/PAD4 and SNC1 in RIN13 functioning. Furthermore, transient expression assays indicated that RIN13 promotes the nuclear accumulation of PAD4. Collectively, our study uncovered a signaling pathway whereby SNC1 and EDS1/PAD4 act together to modulate RIN13-triggered plant defense responses.

Variation in several pathogenesis - Related (PR) protein genes in wheat (Triticum aestivum) involved in defense against Puccinia striiformis f. sp. tritici

Proteins participate in defense mechanisms after a pathogen attacks a host. Pathogenesis-related (PR) proteins in wheat plants play the important role of defense against Puccinia striiformis f. sp. Tritici. This study uses various analytic methods to describe the expression of five (PR) protein genes (PR1, PR2, PR4, PR9 and PR10) and genetic polymorphism among sixteen wheat genotypes to understand the mechanisms underlying of PR genes in Egyptian wheat compared with the Yr genotype. The PR protein genes were semi-quantitatively and quantitatively assessed in four selected wheat genotypes; two resistant (Misr-3 and Yr-29) and two susceptible (Sids-12 and Avocet-S). Reverse transcriptase PCR analysis results indicated that all PR protein genes exhibited different expression levels among the four studied genotypes and overexpression at the resistant genotypes in the different time points of Pst inoculation. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated that the expression of PR protein genes was elevated in adult plant resistant (APR) genotypes. Plant stage in combination with the infection process by stripe rust provided even greater upregulation in the expression of PR genes and enhanced induction of defense enzyme activities in the same sample at different time points for various wheat genotypes.

The Cysteine-Rich Repeat Protein TaCRR1 Participates in Defense against Both Rhizoctonia cerealis and Bipolaris sorokiniana in Wheat.

The domain of unknown function 26 (DUF26), harboring a conserved cysteine-rich motif (C-X8-C-X2-C), is unique to land plants. Several cysteine-rich repeat proteins (CRRs), belonging to DUF26-containing proteins, have been implicated in the defense against fungal pathogens in ginkgo, cotton, and maize. However, little is known about the functional roles of CRRs in the important staple crop wheat (Triticum aestivum). In this study, we identified a wheat CRR-encoding gene TaCRR1 through transcriptomic analysis, and dissected the defense role of TaCRR1 against the soil-borne fungi Rhizoctonia cerealis and Bipolaris sorokiniana, causal pathogens of destructive wheat diseases. TaCRR1 transcription was up-regulated in wheat towards B. Sorokiniana or R. cerealis infection. The deduced TaCRR1 protein contained a signal peptide and two DUF26 domains. Heterologously-expressed TaCRR1 protein markedly inhibited the mycelia growth of B. sorokiniana and R. cerealis. Furthermore, the silencing of TaCRR1 both impaired host resistance to B. sorokiniana and R. cerealis and repressed the expression of several pathogenesis-related genes in wheat. These results suggest that the TaCRR1 positively participated in wheat defense against both B. sorokiniana and R. cerealis through its antifungal activity and modulating expression of pathogenesis-related genes. Thus, TaCRR1 is a candidate gene for improving wheat resistance to B. sorokiniana and R. cerealis.

Translocation of PpNPR1 is required for β-aminobutyric acid-triggered resistance against Rhizopus stolonifer in peach fruit

This study attempted to determine the transcriptomic responses of peach fruit to β-aminobutyric acid (BABA) elicitation and Rhizopus stolonifer inoculation and to identify the key differentially expressed genes (DEGs) and significantly enriched pathways between BABA + inoculation-versus-control and inoculation-versus-control pairwise comparisons. Furthermore, the regulatory linkage between the critical DEGs and their interacting proteins was investigated based on the expression of pathogenesis-related (PR) genes in the BABA-treated samples. The results showed that the transcription factor PpTGA1 possessed the highest degree in the protein-protein interaction (PPI) network and was identified as a critical DEG. PpTGA1 could directly interact with the significantly expressed PpNPR1 gene in the nucleus through the salicylic acid (SA)-dependent signalling pathway, eventually leading to high transcript levels of PpPR1 and PpPR1-like. In addition, an increased generation of redox signalling molecules (nitric oxide (NO) and SA) was accompanied by an obvious effect on the suppression of Rhizopus rot in peaches pretreated with BABA and subsequent R. stolonifer challenge. Thus, BABA elicitation and subsequent R. stolonifer inoculation positively modulated the intercellular redox state and consequently induced the translocation of the product of the highly expressed PpNPR1 gene to the nucleus, where it interacts with PpTGA1 and eventually strengthens defence-related gene expression and disease resistance.

Species-Specific Impact of Fusarium Infection on the Root and Shoot Characteristics of Asparagus.

Soil-borne pathogens can have considerable detrimental effects on asparagus (Asparagus officinalis) growth and production, notably caused by the Fusarium species F. oxysporum f.sp. asparagi, F. proliferatum and F. redolens. In this study, their species-specific impact regarding disease severity and root morphological traits was analysed. Additionally, various isolates were characterised based on in vitro physiological activities and on protein extracts using matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS). The response of two asparagus cultivars to the different Fusarium species was evaluated by inoculating experiments. Differences in aggressiveness were observed between Fusarium species and their isolates on roots, while no clear disease symptoms became visible in ferns eight weeks after inoculation. F. redolens isolates Fred1 and Fred2 were the most aggressive strains followed by the moderate aggressive F. proliferatum and the less and almost non-aggressive F. oxysporum isolates, based on the severity of disease symptoms. Fungal DNA in stem bases and a significant induction of pathogenesis-related gene expression was detectable in both asparagus cultivars. A significant negative impact of the pathogens on the root characteristics total root length, volume, and surface area was detected for each isolate tested, with Fred1 causing the strongest effects. No significant differences between the tested asparagus cultivars were observed.

VdNPS, a Nonribosomal Peptide Synthetase, Is Involved in Regulating Virulence in Verticillium dahliae.

Nonribosomal peptide synthetases (NPS) are known for the biosynthesis of antibiotics, toxins, and siderophore production. They are also a virulence determinant in different phytopathogens. However, until now, the functional characterization of NPS in Verticillium dahliae has not been reported. Deletion of the NPS gene in V. dahliae led to the decrease of conidia, microsclerotia, and pathogenicity. ΔVdNPS strains were tolerant to H2O2, and the genes involved in H2O2 detoxification, iron/copper transport, and cytoskeleton were differentially expressed in ΔVdNPS. Interestingly, ΔVdNPS strains exhibited hypersensitivity to salicylic acid (SA), and the genes involved in SA hydroxylation were up-regulated in ΔVdNPS compared with wild-type V. dahliae under SA stress. Additionally, during infection, ΔVdNPS induced more pathogenesis-related gene expression, higher reactive oxygen species production, and stronger SA-mediated signaling transduction in host to overcome pathogen. Uncovering the function of VdNPS in pathogenicity could provide a reliable theoretical basis for the development of cultivars with durable resistance against V. dahliae-associated diseases.

A Novel ERF Transcription Factor, ZmERF105, Positively Regulates Maize Resistance to Exserohilum turcicum.

The ethylene response factor (ERF) plays a crucial role in plant innate immunity. However, the molecular function of ERF in response to Exserohilum turcicum (E. turcicum) remains unknown in maize. In this study, a novel ERF gene, designated as ZmERF105, was firstly isolated and characterized. The ZmERF105 protein contains an APETALA2/ETHYLENE RESPONSIVE FACTOR (AP2/ERF) domain and a conserved LSPLSPHP motif in its C-terminal region. ZmERF105 protein was exclusively localized to the nucleus. ZmERF105 expression responded to E. turcicum treatment. Yeast one-hybrid and transcription activity assays revealed that ZmERF105 is an activator of transcription and binds to GCC-box elements. Over-expression of ZmERF105 was shown to increase maize resistance against E. turcicum, and erf105 mutant lines displayed opposite phenotype. Moreover, the activities of superoxide dismutase (SOD) and peroxidase (POD) in the ZmERF105 over-expression lines were markedly higher than in the wild-type maize lines (WT) after infection with E. turcicum, and were compromised in the erf105 mutant lines. Simultaneously, ZmERF105 over-expression lines enhanced the expression of several pathogenesis-related (PR) genes, including ZmPR1a, ZmPR2, ZmPR5, ZmPR10.1, and ZmPR10.2 after infection with E. turcicum. In contrast, the expression of PR genes was reduced in erf105 mutant lines. Our work reveals that ZmERF105 as a novel player of the ERF network and positively regulates the maize resistance response to E. turcicum.

Unregulated Sphingolipid Biosynthesis in Gene-Edited Arabidopsis ORM Mutants Results in Nonviable Seeds with Strongly Reduced Oil Content.

Orosomucoid-like proteins (ORMs) interact with serine palmitoyltransferase (SPT) to negatively regulate sphingolipid biosynthesis, a reversible process critical for balancing the intracellular sphingolipid levels needed for growth and programmed cell death. Here, we show that ORM1 and ORM2 are essential for life cycle completion in Arabidopsis (Arabidopsis thaliana). Seeds from orm1 -/- orm2 -/- mutants, generated by crossing CRISPR/Cas9 knockout mutants for each gene, accumulated high levels of ceramide, indicative of unregulated sphingolipid biosynthesis. orm1 -/- orm2 -/- seeds were nonviable, displayed aberrant embryo development, and had >80% reduced oil content versus wild-type seeds. This phenotype was mimicked in Arabidopsis seeds expressing the SPT subunit LCB1 lacking its first transmembrane domain, which is critical for ORM-mediated regulation of SPT. We identified a mutant for ORM1 lacking one amino acid (Met-51) near its second transmembrane domain that retained its membrane topology. Expressing this allele in the orm2 background yielded plants that did not advance beyond the seedling stage, hyperaccumulated ceramides, and showed altered organellar structures and increased senescence- and pathogenesis-related gene expression. These seedlings also showed upregulated expression of genes for sphingolipid catabolic enzymes, pointing to additional mechanisms for maintaining sphingolipid homeostasis. ORM1 lacking Met-51 had strongly impaired interactions with LCB1 in a yeast (Saccharomyces cerevisiae) model, providing structural clues about regulatory interactions between ORM and SPT.

Characterization of the wheat cultivars against Tilletia controversa K\xfchn, causal agent of wheat dwarf bunt

Wheat is one of the most important staple crops. Tilletia controversa Kühn is the causal agent of wheat dwarf bunt. In this study, a resistant wheat cultivar displayed significantly higher expression of pathogenesis-related genes than a susceptible cultivar at 7 days post inoculation (DPI) with T. controversa. Similarly, the expression was high in the resistant cultivar after exogenous application of phytohormones, including salicylic acid. The expression of pathogenesis-related genes, especially chitinase 4, was high in the resistant cultivar, while LPT-1 was down regulated after T. controversa infection. Callose deposition was greater in the resistant cultivar than in the susceptible cultivar at 10 DPI. Confocal microscopy was used to track the fungal hyphae in both cultivars in anther and ovary cells. The anthers and ovaries of the susceptible cultivar were infected by T. controversa at 7 and 15 DPI. There were no fungal hyphae in anther and ovary cells in the resistant cultivar until 10 and 23 DPI, respectively. Moreover, anther length and width were negatively influenced by T. controversa at 16 DPI. The plant height was also affected by fungal infection. Ultimately, resistance to T. controversa was achieved in cultivars via the regulation of the expression of defense-related and pathogenesis-related genes.

SlARG2 contributes to MeJA‐induced defense responses to Botrytis cinerea in tomato fruit

Arginase plays key roles in methyl jasmonate (MeJA)-mediated quality maintenance in vegetables and fruits. MeJA treatment induced the Arginase 2 (SlARG2) expression, which is one of the most important encoding genes of arginase. In addition, the treatment with MeJA induced resistance to pathogenic infection in many plants. However, the functions of SlARG2 in MeJA-induced defense to Botrytis cinerea are unclear. In our work, control and SlARG2-silenced tomato fruits (Solanum lycopersicum) were treated with 0.05 mmoL L-1 MeJA before storage to assay the roles of SlARG2 in MeJA-induced defense responses to Botrytis cinerea. Our results indicated that MeJA treatment induced both pathogenesis-related gene expression (PR1, PR2a, PR2b and PR3b), and the activity of defense-related enzymes, as well as upregulated arginine metabolism. Compared to control fruits, the treatment with MeJA also induced the activity of arginase, arginine decarboxylase (ADC) and ornithine aminotransferase (OAT), and expression of SlARG2, SlADC, ornithine decarboxylase (SlODC) and SlOAT, and consequently increased the accumulation of arginine, proline, glutamate, putrescine and spermine. However, the induction effects by MeJA were significantly reduced in fruits in which SlARG2 was silenced and severe disease symptoms were observed. MeJA fumigation could inhibit disease development by inducing pathogenesis-related gene expression (PR1, PR2a, PR2b and PR3b) and defense-related enzymes activity, as well as upregulated arginine metabolism. In addition, SlARG2 silencing could inhibit the functions of MeJA in inducing the accumulation of the above substances. Overall, our study provided strong evidence that SlARG2 was essential for MeJA-induced tomato fruit defense responses to Botrytis cinerea. © 2020 Society of Chemical Industry.

Tomato T2 ribonuclease LE is involved in the response to pathogens

T2 ribonucleases (RNases) are RNA‐degrading enzymes that function in various cellular processes, mostly via RNA metabolism. T2 RNase‐encoding genes have been identified in various organisms, from bacteria to mammals, and are most diverse in plants. The existence of T2 RNase genes in almost every organism suggests an important biological function that has been conserved through evolution. In plants, T2 RNases are suggested to be involved in phosphate scavenging and recycling, and are implicated in defence responses to pathogens. We investigated the function of the tomato T2 RNase LE, known to be induced by phosphate deficiency and wounding. The possible involvement of LE in pathogen responses was examined. Expression analysis showed LE induction during fungal infection and by stimuli known to be associated with pathogen inoculation, including oxalic acid and hydrogen peroxide. Analysis of LE‐suppressed transgenic tomato lines revealed higher susceptibility to oxalic acid, a cell death‐inducing factor, compared to the wild type. This elevated sensitivity of LE‐suppressed lines was evidenced by visual signs of necrosis, and increased ion leakage and reactive oxygen species levels, indicating acceleration of cell death. Challenge of the LE‐suppressed lines with the necrotrophic pathogen Botrytis cinerea resulted in accelerated development of disease symptoms compared to the wild type, associated with suppressed expression of pathogenesis‐related marker genes. The results suggest a role for plant endogenous T2 RNases in antifungal activity.