Induce Defense Gene Expression Research Articles

Silencing Osa-miR827 via CRISPR/Cas9 protects rice against the blast fungus Magnaporthe oryzae

MicroRNAs (miRNAs) are short, non-coding RNAs that regulate gene expression at the post-transcriptional level. In plants, miRNAs participate in diverse developmental processes and adaptive responses to biotic and abiotic stress. MiR827 has long been recognized to be involved in plant responses to phosphate starvation. In rice, the miR827 regulates the expression of OsSPX-MFS1 and OsSPX-MFS2, these genes encoding vacuolar phosphate transporters. In this study, we demonstrated that miR827 plays a role in resistance to infection by the fungus Magnaporthe oryzae in rice. We show that MIR827 overexpression enhances susceptibility to infection by M. oryzae which is associated to a weaker induction of defense gene expression during pathogen infection. Conversely, CRISPR/Cas9-induced mutations in the MIR827 gene completely abolish miR827 production and confer resistance to M. oryzae infection. This resistance is accompanied by a reduction of leaf Pi content compared to wild-type plants, whereas Pi levels increase in leaves of the blast-susceptible miR827 overexpressor plants. In wild-type plants, miR827 accumulation in leaves decreases during the biotrophic phase of the infection process. Taken together, our data indicates that silencing MIR827 confers resistance to M. oryzae infection in rice while further supporting interconnections between Pi signaling and immune signaling in plants. Unravelling the role of miR827 during M. oryzae infection provides knowledge to improve blast resistance in rice by CRISPR/Cas9-editing of MIR827.

Small RNAs contribute to citrus Huanglongbing tolerance by manipulating methyl salicylate signaling and exogenous methyl salicylate primes citrus groves from emerging infection.

Citrus production is severely threatened by the devastating Huanglongbing (HLB) disease globally. By studying and analyzing the defensive behaviors of an HLB-tolerant citrus cultivar 'Shatangju', we discovered that citrus can sense Candidatus Liberibacter asiaticus (CLas) infection and induce immune responses against HLB, which can be further strengthened by both endogenously produced and exogenously applied methyl salicylate (MeSA). This immune circuit is turned on by an miR2977-SAMT (encoding a citrus Salicylate [SA] O-methyltransferase) cascade, by which CLas infection leads to more in planta MeSA production and aerial emission. We provided both transgenic and multi-year trail evidences that MeSA is an effective community immune signal. Ambient MeSA accumulation and foliage application can effectively induce defense gene expression and significantly boost citrus performance. We also found that miRNAs are battle fields between citrus and CLas, and about 30% of the differential gene expression upon CLas infection are regulated by miRNAs. Furthermore, CLas hijacks host key processes by manipulating key citrus miRNAs, and citrus employs miRNAs that coordinately regulate defense-related genes. Based on our results, we proposed that miRNAs and associated components are key targets for engineering or breeding resistant citrus varieties. We anticipate that MeSA-based management, either induced expression or external application, would be a promising tool for HLB control.

The R2R3-MYB-SG7 transcription factor CaMYB39 orchestrates surface phenylpropanoid metabolism and pathogen resistance in chickpea.

Flavonoids are important plant pigments and defense compounds; understanding the transcriptional regulation of flavonoid biosynthesis may enable engineering crops with improved nutrition and stress tolerance. Here, we characterize R2R3-MYB domain subgroup 7 transcription factor CaMYB39, which regulates flavonol biosynthesis primarily in chickpea trichomes. CaMYB39 overexpression in chickpea was accompanied by a change in flux availability for the phenylpropanoid pathway, particularly flavonol biosynthesis. Lines overexpressing CaMYB39 showed higher isoflavonoid levels, suggesting its role in regulating isoflavonoid pathway. CaMYB39 transactivates the transcription of early flavonoid biosynthetic genes (EBG). FLAVONOL SYNTHASE2, an EBG, encodes an enzyme with higher substrate specificity for dihydrokaempferol than other dihydroflavonols explaining the preferential accumulation of kaempferol derivatives as prominent flavonols in chickpea. Interestingly, CaMYB39 overexpression increased trichome density and enhanced the accumulation of diverse flavonol derivatives in trichome-rich tissues. Moreover, CaMYB39 overexpression reduced reactive oxygen species levels and induced defense gene expression which aids in partially blocking the penetration efficiency of the fungal pathogen, Ascochyta rabiei, resulting in lesser symptoms, thus establishing its role against deadly Ascochyta blight (AB) disease. Overall, our study reports an instance where R2R3-MYB-SG7 member, CaMYB39, besides regulating flavonol biosynthesis, modulates diverse pathways like general phenylpropanoid, isoflavonoid, trichome density, and defense against necrotrophic fungal infection in chickpea.

Nutrient Elements Promote Disease Resistance in Tomato by Differentially Activating Immune Pathways.

Nutrient elements play essential roles in plant growth, development, and reproduction. Balanced nutrition is critical for plant health and the ability to withstand biotic stress. Treatment with essential elements has been shown to induce disease resistance in certain cases. Understanding the functional mechanisms underlying plant immune responses to nutritional elements has the potential to provide new insights into crop improvement. In the present study, we investigated the effect of various elements-potassium (K), calcium (Ca), magnesium (Mg), and sodium (Na)-in promoting resistance against the necrotrophic fungus Botrytis cinerea and the hemibiotrophic bacterium Xanthomonas euvesicatoria in tomato. We demonstrate that spray treatment of essential elements was sufficient to activate immune responses, inducing defense gene expression, cellular leakage, reactive oxygen species, and ethylene production. We report that different defense signaling pathways are required for induction of immunity in response to different elements. Our results suggest that genetic mechanisms that are modulated by nutrient elements can be exploited in agricultural practices to promote disease resistance.

Nitrogen nutrition modifies the susceptibility of Arabidopsis thaliana to the necrotrophic fungus, Alternaria brassicicola.

The impact of the form of nitrogen (N) source (nitrate versus ammonium) on the susceptibility to Alternaria brassicicola, a necrotrophic fungus, has been examined in Arabidopsis thaliana at the rosette stage. Nitrate nutrition was found to increase fungal lesions considerably. There was a similar induction of defence gene expression following infection under both N nutritions, except for the phytoalexin deficient 3 gene, which was overexpressed with nitrate. Nitrate also led to a greater nitric oxide production occurring in planta during the saprophytic growth and lower nitrate reductase (NIA1) expression 7 days after inoculation. This suggests that nitrate reductase-dependent nitric oxide production had a dual role, whereby, despite its known role in the generic response to pathogens, it affected plant metabolism, and this facilitated fungal infection. In ammonium-grown plants, infection with A. brassicicola induced a stronger gene expression of ammonium transporters and significantly reduced the initially high ammonium content in the leaves. There was a significant interaction between N source and inoculation (presence versus absence of the fungus) on the total amino acid content, while N nutrition reconfigured the spectrum of major amino acids. Typically, a higher content of total amino acid, mainly due to a stronger increase in asparagine and glutamine, is observed under ammonium nutrition while, in nitrate-fed plants, glutamate was the only amino acid which content increased significantly after fungal inoculation. N nutrition thus appears to control fungal infection via a complex set of signalling and nutritional events, shedding light on how nitrate availability can modulate disease susceptibility.

A Small Cysteine-Rich Phytotoxic Protein of Phytophthora capsici Functions as Both Plant Defense Elicitor and Virulence Factor.

Small cysteine-rich (SCR) proteins, including fungal avirulence proteins, play important roles in pathogen-plant interactions. SCR protein-encoding genes have been discovered in the genomes of Phytophthora pathogens but their functions during pathogenesis remain obscure. Here, we report the characterization of one Phytophthora capsici SCR protein (namely, SCR82) with similarity to Phytophthora cactorum phytotoxic protein PcF. The scr82 gene has 10 allelic sequences in the P. capsici population. Homologs of SCR82 were not identified in fungi or other organisms but in Phytophthora relative species. Initially, scr82 was weakly expressed during the mycelium, sporangium, and zoospore stages but quickly upregulated when the infection initiated. Both ectopic expression of SCR82 and recombinant yeast-expressed protein (rSCR82) caused cell death on tomato leaves. Upon treatment, rSCR82 induced plant defense responses, including the induction of defense gene expression, reactive oxygen species burst, and callose deposition. Knockout of scr82 in P. capsici by CRISPR/Cas9 severely impaired its virulence on host plants and significantly reduced its resistance against oxidative stress. Inversely, its overexpression increased the pathogen's virulence and tolerance to oxidative stress. Our results collectively demonstrate that SCR82 functions as both an important virulence factor and plant defense elicitor, which is conserved across Phytophthora spp.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Surfactin and fengycin contribute to the protection of a Bacillus subtilis strain against grape downy mildew by both direct effect and defence stimulation.

Summary Bacillus subtilis GLB191 (hereafter GLB191) is an efficient biological control agent against the biotrophic oomycete Plasmopara viticola, the causal agent of grapevine downy mildew. In this study, we show that GLB191 supernatant is also highly active against downy mildew and that the activity results from both direct effect against the pathogen and stimulation of the plant defences (induction of defence gene expression and callose production). High‐performance thin‐layer chromatography analysis revealed the presence of the cyclic lipopeptides fengycin and surfactin in the supernatant. Mutants affected in the production of fengycin and/or surfactin were thus obtained and allowed us to show that both surfactin and fengycin contribute to the double activity of GLB191 supernatant against downy mildew. Altogether, this study suggests that GLB191 supernatant could be used as a new biocontrol product against grapevine downy mildew.

OsDCL1a activation impairs phytoalexin biosynthesis and compromises disease resistance in rice.

MicroRNAs (miRNAs) are small non-coding RNAs that act as post-transcriptional regulators of gene expression via sequence-specific cleavage or translational repression of target transcripts. They are transcribed as long single-stranded RNA precursors with unique stem-loop structures that are processed by a DICER-Like (DCL) ribonuclease, typically DCL1, to produce mature miRNAs. Although a plethora of miRNAs have been found to be regulated by pathogen infection in plants, the biological function of most miRNAs remains largely unknown. Here, the contribution of OsDCL1 to rice immunity was investigated. Activation-tagged Osdcl1a (Osdcl1a-Ac) rice mutants were examined for resistance to pathogen infection. mRNA and small RNA deep sequencing, quantitative real-time PCR (RT-qPCR) and stem-loop reverse tanscripion-PCR (RT-PCR) were used to examine DCL1a-mediated alterations in the rice transcriptome. Rice diterpene phytoalexins were quantified by liquid chromatography-tandem mass spectrometry (LC-MSMS). Accumulation of O2·- was determined by nitroblue tetrazolium (NBT) staining. dcl1a-Ac mutants exhibit enhanced susceptibility to infection by fungal pathogens which was associated with a weaker induction of defence gene expression. Comparison of the mRNA and miRNA transcriptomes of dcl1a-Ac and wild-type plants revealed misregulation of genes involved in detoxification of reactive oxygen species. Consequently, dcl1a-Ac plants accumulated O2·- in their leaves and were more sensitive to methyl viologen-induced oxidative stress. Furthermore, dcl1a-Ac plants showed downregulation of diterpenoid phytoalexin biosynthetic genes, these genes also being weakly induced during pathogen infection. Upon pathogen challenge, dcl1a-Ac plants failed to accumulate major diterpenoid phytoalexins. OsDCL1a activation resulted in marked alterations in the rice miRNAome, including both upregulation and downregulation of miRNAs. OsDCL1a activation enhances susceptibility to infection by fungal pathogens in rice. Activation of OsDCL1a represses the pathogen-inducible host defence response and negatively regulates diterpenoid phytoalexin production. These findings provide a basis to understand the molecular mechanisms through which OsDCL1a mediates rice immunity.

Differential Suppression of Nicotiana benthamiana Innate Immune Responses by Transiently Expressed Pseudomonas syringae Type III Effectors.

The plant pathogen Pseudomonas syringae injects about 30 different virulence proteins, so-called effectors, via a type III secretion system into plant cells to promote disease. Although some of these effectors are known to suppress either pattern-triggered immunity (PTI) or effector-triggered immunity (ETI), the mode of action of most of them remains unknown. Here, we used transient expression in Nicotiana benthamiana, to test the abilities of type III effectors of Pseudomonas syringae pv. tomato (Pto) DC3000 and Pseudomonas syringae pv. tabaci (Pta) 11528 to interfere with plant immunity. We monitored the sequential and rapid bursts of cytoplasmic Ca2+ and reactive oxygen species (ROS), the subsequent induction of defense gene expression, and promotion of cell death. We found that several effector proteins caused cell death, but independently of the known plant immune regulator NbSGT1, a gene essential for ETI. Furthermore, many effectors delayed or blocked the cell death-promoting activity of other effectors, thereby potentially contributing to pathogenesis. Secondly, a large number of effectors were able to suppress PAMP-induced defense responses. In the majority of cases, this resulted in suppression of all studied PAMP responses, suggesting that these effectors target common elements of PTI. However, effectors also targeted different steps within defense pathways and could be divided into three major groups based on their suppressive activities. Finally, the abilities of effectors of both Pto DC3000 and Pta 11528 to suppress plant immunity was conserved in most but not all cases. Overall, our data present a comprehensive picture of the mode of action of these effectors and indicate that most of them suppress plant defenses in various ways.

Soil applications of acibenzolar-S-methyl induce defense gene expression in tomato plants against bacterial spot

Acibenzolar-S-methyl (ASM), a plant activator known to induce plant resistance, has been used as foliar sprays to manage several plant diseases including bacterial spot on tomato caused by four distinct Xanthomonas species. This study aimed to investigate the effects of soil application rates of ASM on bacterial spot of tomato and the expression levels of the two pathogenesis-related (PR) genes, PR1a and PR1b, in leaf tissues. Tomato seedlings were leaf-applied with ASM at 18.8 mg/l corresponding to the labeled rate, soil-applied with ASM at 0.84 and 10 mg/l, and sprayed with water served as an untreated control. The soil application of ASM at 10 mg/l consistently reduced the final disease severity and disease progress compared to the untreated control in four growth chamber experiments, whereas the soil application of ASM at 0.84 mg/l and foliar spray of ASM significantly reduced the final disease severity and area under disease progress curve (AUDPC) in three out of the four experiments. The expression levels of PR1a and PR1b in the leaf tissues were significantly induced by both soil and foliar applications of ASM. In addition, field trial results suggested that the soil applications of ASM at 10 mg/l markedly reduced disease progress compared to the control and copper standard. Although the control efficiency of soil applications of ASM depends on rates used, this study suggests that ASM can be used as soil applications to induce tomato resistance against bacterial spot.

How Streptomyces anulatus Primes Grapevine Defenses to Cope with Gray Mold: A Study of the Early Responses of Cell Suspensions.

Gray mold, caused by Botrytis cinerea, is one of the most destructive diseases of grapevine and is controlled with an intense application of fungicides. As alternatives to chemicals, beneficial microbes may promote plant health by stimulating the plant’s immune system. An actinomycete, Streptomyces anulatus S37, has been screened from the rhizosphere microbiome of healthy Vitis vinifera on the basis of its ability to promote grapevine growth and to induce resistance against various phytopathogens, including B. cinerea. However, molecular mechanisms involved locally after direct perception of these bacteria by plant cells still remain unknown. This study focuses on local defense events induced in grapevine cells during interactions with S. anulatus S37 before and after pathogen challenge. We demonstrated that S. anulatus S37 induced early responses including oxidative burst, extracellular alkalinization, activation of protein kinases, induction of defense gene expression and phytoalexin accumulation, but not the programmed cell death. Interestingly, upon challenge with the B. cinerea, the S. anulatus S37 primed grapevine cells for enhanced defense reactions with a decline in cell death. In the presence of the EGTA, a calcium channel inhibitor, the induced oxidative burst, and the protein kinase activity were inhibited, but not the extracellular alkalinization, suggesting that Ca2+ may also contribute upstream to the induced defenses. Moreover, desensitization assays using extracellular pH showed that once increased by S. anulatus S37, cells became refractory to further stimulation by B. cinerea, suggesting that grapevine cells perceive distinctly beneficial and pathogenic microbes.

Root assays to study pattern-triggered immunity in plant-nematode interactions

Plants employ extracellular immune receptors to perceive conserved pathogen-associated molecular patterns (PAMPs), triggering the first layer of defense known as pattern-triggered immunity (PTI). The understanding of PTI is mainly based on studies focusing on leaves. Plants are vulnerable to attack by various root pathogens including plant-parasitic nematodes. Evidence is accumulating that phytonematodes utilize their secreted effectors to suppress PTI to enable infection. PTI assays used for characterizing nematode effectors are often conducted in a non-host plant or tissue, such as leaves, because of lacking of root assays. Thus, there is a need for PTI assays in roots of host plants. Here, we tested two bacterial PAMPs (flg22 and flgII-28) and two nonpathogenic bacteria (Pseudomonas fluorescens and P. syringae strain DC3000 ΔhrcQ-U) for their ability to induce PTI responses, including the induction of defense gene expression and callose deposition, in roots of tomato and potato. We found that flg22 and the two nonpathogenic bacteria are potent in inducing defense gene expression and callose deposition in tested roots, demonstrating for the first time induction of PTI in roots of solanaceous plants. Effectors GrCEP12 and Hs10A06 were previously indicated to be involved in PTI suppression. Consistently, upon elicitor treatment, roots of transgenic plants overexpressing GrCEP12 and Hs10A06, respectively, showed a reduced level of defense gene expression or no induction of callose deposition compared to control roots. Taken together, our established root PTI assays represent a valuable tool that will facilitate the study of phytonematodes and potentially other root pathogens in their manipulation of plant immunity.

Suppression of bacterial infection in rice by treatment with a sulfated peptide.

The rice XA21 receptor kinase confers robust resistance to bacterial blight disease caused by Xanthomonas oryzae pv. oryzae (Xoo). A tyrosine-sulfated peptide from Xoo, called RaxX, triggers XA21-mediated immune responses, including the production of ethylene and reactive oxygen species and the induction of defence gene expression. It has not been tested previously whether these responses confer effective resistance to Xoo. Here, we describe a newly established post-inoculation treatment assay that facilitates investigations into the effect of the sulfated RaxX peptide in planta. In this assay, rice plants were inoculated with a virulent strain of Xoo and then treated with the RaxX peptide 2 days after inoculation. We found that post-inoculation treatment of XA21 plants with the sulfated RaxX peptide suppresses the development of Xoo infection in XA21 rice plants. The treated plants display restricted lesion development and reduced bacterial growth. Our findings demonstrate that exogenous application of sulfated RaxX activates XA21-mediated immunity in planta, and provides a potential strategy for the control of bacterial disease in the field.

Editorial: "Agrobacterium biology and its application to transgenic plant production".

Editorial: "Agrobacterium biology and its application to transgenic plant production".

Arabidopsis serotonin N-acetyltransferase knockout mutant plants exhibit decreased melatonin and salicylic acid levels resulting in susceptibility to an avirulent pathogen.

Serotonin N-acetyltransferase (SNAT) is the penultimate enzyme in the melatonin biosynthesis pathway in plants. We examined the effects of SNAT gene inactivation in two Arabidopsis T-DNA insertion mutant lines. After inoculation with the avirulent pathogen Pseudomonas syringe pv. tomato DC3000 harboring the elicitor avrRpt2 (Pst-avrRpt2), melatonin levels in the snat knockout mutant lines were 50% less than in wild-type Arabidopsis Col-0 plants. The snat knockout mutant lines exhibited susceptibility to pathogen infection that coincided with decreased induction of defense genes including PR1, ICS1, and PDF1.2. Because melatonin acts upstream of salicylic acid (SA) synthesis, the reduced melatonin levels in the snat mutant lines led to decreased SA levels compared to wild-type, suggesting that the increased pathogen susceptibility of the snat mutant lines could be attributed to decreased SA levels and subsequent attenuation of defense gene induction. Exogenous melatonin treatment failed to induce defense gene expression in nahG Arabidopsis plants, but restored the induction of defense gene expression in the snat mutant lines. In addition, melatonin caused translocation of NPR1 (nonexpressor of PR1) protein from the cytoplasm into the nucleus indicating that melatonin-elicited pathogen resistance in response to avirulent pathogen attack is SA-dependent in Arabidopsis.

Non-proteinaceous yeast extract induces arabidopsis defense responses independently of salicylic acid

In spite of more severe effect on crop productivity, difficulties in artificial culturing and plant inoculation lead to less understanding of plant defense responses to fungal pathogens compared to bacteria. Therefore, it is of great importance to better understand detailed plant defense responses to fungal pathogens. We here show that the commercially available yeast extract includes at least a fungal pathogen-associated molecular pattern (PAMP) to induce defense gene expression via the mitogen-activated protein kinase activation in Arabidopsis plants. Yeast extract induced the expression of PR1 and EDS5 genes in the salicylic acid-deficient nahG plants. Induction of PR1 and EDS5 gene expression by proteinase K-treated yeast extract suggests that at least a non-proteinaceous fungal PAMP in yeast extract induces the PAMP-triggered immunity in Arabidopsis plants in a manner independent of salicylic acid.

Endophytic Bacillus spp. produce antifungal lipopeptides and induce host defence gene expression in maize

Endophytes are mutualistic symbionts within healthy plant tissues. In this study we isolated Bacillus spp. from seeds of several varieties of maize. Bacillus amyloliquifaciens or Bacillus subtilis were found to be present in all maize varieties examined in this study. To determine whether bacteria may produce antifungal compounds, generally lipopeptides in Bacillus spp., bacterial cultures were screened for production of lipopeptides. Lipopeptides were extracted by acid precipitation from liquid cultures of Bacillus spp. Lipopeptide extracts from Bacillus spp. isolated from Indian popcorn and yellow dent corn showed inhibitory activity against Fusarium moniliforme at 500μg per disk. Using MALDI-TOF mass spectrometry we detected the presence of antifungal iturin A, fengycin and bacillomycin in these isolates. PCR amplification also showed the presence of genes for iturin A and fengycin. B. subtilis (SG_JW.03) isolated from Indian popcorn showed strong inhibition of Arabidopsis seed mycoflora and enhanced seedling growth. We tested for the induction of defence gene expression in the host plant after treatment of plants with B. subtilis (SG_JW.03) and its lipopeptide extract using RT-qPCR. Roots of Indian popcorn seedlings treated with a suspension of B. subtilis (SG_JW.03) showed the induction of pathogenesis-related genes, including PR-1 and PR-4, which relate to plant defence against fungal pathogens. The lipopeptide extract alone did not increase the expression of these pathogenesis-related genes. Based on our study of maize endophytes, we hypothesize that, bacterial endophytes that naturally occur in many maize varieties may function to protect hosts by secreting antifungal lipopeptides that inhibit pathogens as well as inducing the up-regulation of pathogenesis-related genes of host plants (systemic acquired resistance).

Jasmonate-dependent induction of polyphenol oxidase activity in tomato foliage is important for defense against Spodoptera exigua but not against Manduca sexta

BackgroundJasmonates are involved in plant defense, participating in the timely induction of defense responses against insect herbivores from different feeding guilds and with different degrees of host specialization. It is less clear to what extent the induction of plant defense is controlled by different members of the jasmonate family and how specificity of the response is achieved. Using transgenic plants blocked in jasmonic acid (JA) biosynthesis, we previously showed that JA is required for the formation of glandular trichomes and trichome-borne metabolites as constitutive defense traits in tomato, affecting oviposition and feeding behavior of the specialist Manduca sexta. In contrast, JA was not required for the local induction of defense gene expression after wounding. In JA-deficient plants, the JA precursor oxophytodienoic acid (OPDA) substituted as a regulator of defense gene expression maintaining considerable resistance against M. sexta larvae. In this study, we investigate the contribution of JA and OPDA to defense against the generalist herbivore Spodoptera exigua.ResultsS. exigua preferred JA-deficient over wild-type tomato plants as a host for both oviposition and feeding. Feeding preference for JA-deficient plants was caused by constitutively reduced levels of repellent terpenes. Growth and development of the larvae, on the other hand, were controlled by additional JA-dependent defense traits, including the JA-mediated induction of foliar polyphenol oxidase (PPO) activity. PPO induction was more pronounced after S. exigua herbivory as compared to mechanical wounding or M. sexta feeding. The difference was attributed to an elicitor exclusively present in S. exigua oral secretions.ConclusionsThe behavior of M. sexta and S. exigua during oviposition and feeding is controlled by constitutive JA/JA-Ile-dependent defense traits involving mono- and sesquiterpenes in both species, and cis-3-hexenal as an additional chemical cue for M. sexta. The requirement of jasmonates for resistance of tomato plants against caterpillar feeding differs for the two species. While the OPDA-mediated induction of local defense is sufficient to restrict growth and development of M. sexta larvae in absence of JA/JA-Ile, defense against S. exigua relied on additional JA/JA-Ile dependent factors, including the induction of foliar polyphenol oxidase activity in response to S. exigua oral secretions.

Long-Term Induction of Defense Gene Expression in Potato by Pseudomonas sp. LBUM223 and Streptomyces scabies

Streptomyces scabies is a causal agent of common scab of potato, which generates necrotic tuber lesions. We have previously demonstrated that inoculation of potato plants with phenazine-1-carboxylic acid (PCA)- producing Pseudomonas sp. LBUM223 could significantly reduce common scab symptoms. In the present study, we investigated whether LBUM223 or an isogenic phzC- mutant not producing PCA could elicit an induced systemic resistance response in potato. The expression of eight defense-related genes (salicylic acid [SA]-related ChtA, PR-1b, PR-2, and PR-5; and jasmonic acid and ethylene-related LOX, PIN2, PAL-2, and ERF3) was quantified using newly developed TaqMan reverse-transcription quantitative polymerase chain reaction assays in 5- and 10-week-old potted potato plants. Although only wild-type LBUM223 was capable of significantly reducing common scab symptoms, the presence of both LBUM223 and its PCA-deficient mutant were equally able to upregulate the expression of LOX and PR-5. The presence of S. scabies overexpressed all SA-related genes. This indicates that (i) upregulation of potato defense-related genes by LBUM223 is unlikely to contribute to common scab's control and (ii) LBUM223's capacity to produce PCA is not involved in this upregulation. These results suggest that a direct interaction occurring between S. scabies and PCA-producing LBUM223 is more likely involved in controlling common scab development.

Water extracts from winery by-products as tobacco defense inducers.

Water extracts from winery by-products exhibited significant plant defense inducer properties. Experiments were conducted on three marc extracts containing various amounts of polyphenols and anthocyanins. Infiltration of red, white and seed grape marc extracts into tobacco leaves induced hypersensitive reaction-like lesions with cell death evidenced by Evans Blue staining. The infiltration zones and the surrounding areas revealed accumulation of autofluorescent compounds under UV light. Leaf infiltration of the three winery by-product extracts induced defense gene expression. The antimicrobial PR1, β-1,3-glucanase PR2, and chitinase PR3 target genes were upregulated locally in tobacco plants following grape marc extract treatments. The osmotin PR5 transcripts accumulated as well in red marc extract treated-tobacco leaves. Overall, the winery by-product extracts elicited an array of plant defense responses making the grape residues a potential use of high value compounds.