Systemic Acquired Resistance Pathway Research Articles

Cadmium toxicity promotes hormonal imbalance and induces the expression of genes involved in systemic resistances in barley.

Cadmium (Cd) is a widely distributed pollutant that adversely affects plants' metabolism and productivity. Phytohormones play a vital role in the acclimation of plants to metal stress. On the other hand, phytohormones trigger systemic resistances, including systemic acquired resistance (SAR) and induced systemic resistance (ISR), in plants in response to biotic interactions. The present study aimed to investigate the possible induction of SAR and ISR pathways in relation to the hormonal alteration of barley seedlings in response to Cd stress. Barley seedlings were exposed to 1.5mgg-1 Cd in the soil for three days. The nutrient content, oxidative status, phytohormones profile, and expression of genes involved in SAR and ISR pathways of barley seedlings were examined. Cd accumulation resulted in a reduction in the nutrient content of barley seedlings. The specific activity of superoxide dismutase and the hydrogen peroxide content significantly increased in response to Cd toxicity. Abscisic acid, jasmonic acid, and ethylene content increased under Cd exposure. Cd treatment resulted in the upregulation of NPR1, PR3, and PR13 genes in SAR pathways. The transcripts of PAL1 and LOX2.2 genes in the ISR pathway were also significantly increased in response to Cd treatment. These findings suggest that hormonal-activated systemic resistances are involved in the response of barley to Cd stress.

Drought Shifts Sorghum Root Metabolite and Microbiome Profiles and Enriches for Pipecolic Acid

Plant-associated microbial communities shift in composition as a result of environmental perturbations, such as drought. It has been shown that Actinobacteria are enriched in plant roots and rhizospheres during drought stress. However, the correlations between microbiome dynamics and plant response to drought are poorly understood. Here we apply a combination of bacterial community composition analysis and plant metabolite profiling in Sorghum bicolor roots, rhizospheres, and soil during drought and drought recovery to investigate potential contributions of host metabolism to shifts in bacterial composition. Our results provide a detailed view of metabolic shifts across the plant root during drought and show that the response to rewatering differs between root and soil; additionally, we identify drought-responsive metabolites that are highly correlated with the observed changes in Actinobacteria abundance. Furthermore, we find that pipecolic acid is a drought-enriched metabolite in sorghum roots, and that exogenous application of pipecolic acid inhibits root growth. Finally, we show that this activity functions independent of the systemic acquired resistance pathway and has the potential to impact Actinobacterial taxa within the root microbiome.

Pseudomonas fluorescens SP007S Formulations in Controlling Soft Rot Disease and Promoting Growth in Kale

Kale has gained popularity as a healthy food choice due to its rich nutrient profile. However, kale production systems are also affected by various pests and diseases, especially plant pathogenic bacteria, such as Pectobacterium carotovorum. The purpose of this research was to investigate the biocontrol activity of the Pseudomonas fluorescens strain SP007s and develop a formulation that provides stability, long shelf life, and disease control, as well as promoting growth in kale; we expect this formulation to have further commercial applications in the cultivation of kale. The results indicated that a unique mixture of paper sludge, sugar sludge, and glycerol at 40% relative humidity promoted SP007 survival for 6 months at room temperature (30 °C) with measures of 5.92 × 106 CFU/g. This treatment maintained the control efficacy of P. carotovorum in infected soil within 28 days of soil drenching. We evaluated the SP007s formula for controlling soft rot disease in greenhouse conditions, and our results revealed that it can reduce the disease in kale by 65.2% and promotes growth in terms of canopy width, shoot height, number of leaves per plant, fresh weight, and dry weight, which were superior to the control. In addition, the formula can induce the accumulation of endogenous salicylic acid and phenolic compounds, as well as a systemic acquired resistance pathway in the plant defense system. Therefore, the formula of SP007s can be a promising novel biocontrol formula for use in kale production, as it promotes plant growth and acts against P. carotovorum causal soft rot disease.

Aureobasidium pullulans from the Fire Blight Biocontrol Product, Blossom Protect, Induces Host Resistance in Apple Flowers.

Fire blight, caused by Erwinia amylovora, is a devastating disease of apple. Blossom Protect, a product that contains Aureobasidium pullulans as the active ingredient, is one of the most effective biological controls of fire blight. It has been postulated that the mode of action of A. pullulans is to compete against and antagonize epiphytic growth of E. amylovora on flowers, but recent studies have found that flowers treated with Blossom Protect harbored similar to or only slightly reduced E. amylovora populations compared with nontreated flowers. In this study, we tested the hypothesis that A. pullulans-mediated biocontrol of fire blight is the result of induced host resistance. We found that PR genes in the systemic acquired resistance pathway, but not genes in the induced systemic resistance pathway, were induced in hypanthial tissue of apple flowers after the Blossom Protect treatment. Additionally, the induction of PR gene expression was coupled with an increase of plant-derived salicylic acid in this tissue. After inoculation with E. amylovora, PR gene expression was suppressed in nontreated flowers, but in flowers pretreated with Blossom Protect, the heightened PR expression offset the immune repression caused by E. amylovora, and prevented infection. Temporal and spatial analysis of PR gene induction showed that induction of PR genes occurred 2days after the Blossom Protect treatment, and required direct flower-yeast contact. Finally, we observed deterioration of the epidermal layer of the hypanthium in some of the Blossom Protect-treated flowers, suggesting that PR gene induction in flowers may be a result of pathogenesis by A. pullulans.

Cross-Talk between Iron Deficiency Response and Defense Establishment in Plants

Plants are at risk of attack by various pathogenic organisms. During pathogenesis, microorganisms produce molecules with conserved structures that are recognized by plants that then initiate a defense response. Plants also experience iron deficiency. To address problems caused by iron deficiency, plants use two strategies focused on iron absorption from the rhizosphere. Strategy I is based on rhizosphere acidification and iron reduction, whereas Strategy II is based on iron chelation. Pathogenic defense and iron uptake are not isolated phenomena: the antimicrobial phenols are produced by the plant during defense, chelate and solubilize iron; therefore, the production and secretion of these molecules also increase in response to iron deficiency. In contrast, phytohormone jasmonic acid and salicylic acid that induce pathogen-resistant genes also modulate the expression of genes related to iron uptake. Iron deficiency also induces the expression of defense-related genes. Therefore, in the present review, we address the cross-talk that exists between the defense mechanisms of both Systemic Resistance and Systemic Acquired Resistance pathways and the response to iron deficiency in plants, with particular emphasis on the regulation genetic expression.

Articulating beneficial rhizobacteria-mediated plant defenses through induced systemic resistance: A review

Induced systemic resistance (ISR) is a mechanism by which certain plant beneficial rhizobacteria (PBR) and fungi produce immunity, which can stimulate crop growth and resilience against various phytopathogens, insects, and parasites. These beneficial rhizobacteria and fungi improve plant performance by regulating hormone signaling, including salicylic acid (SA), jasmonic acid (JA), prosystemin, pathogenesis-related gene 1 (PR1), and ethylene (ET) pathways which activate the gene expression of ISR, the synthesis of secondary metabolites, various enzymes, and volatile compounds that ultimately induce the defense mechanism of the plant. To protect themselves from disease, plants have various advanced defense mechanisms in which local acquired resistance (LAR), systemic gene silencing (SGS), systemic wounding response (SWR), systemic acquired resistance (SAR), and ISR are involved. Several rhizobacteria activate the SA- dependent SAR pathway by producing SA at the root's surface. In contrast, other rhizobacteria can activate different signaling pathways independent of SA (SA-independent ISR pathways) such as those dependent on JA and ET signaling. The main objective of this review is to provide insight into the types of induced resistance utilized for plant defense. Further to this, the genetic approaches used to suppress disease-causing genes i.e., RNA interference and antisense RNA, which are still underutilized in sustainable agriculture along with the current vision for VIGS (a virus-based tool) are also discussed.

Overexpression of the salicylic acid binding protein 2 (SABP2) from tobacco enhances tolerance against Huanglongbing in transgenic citrus.

Overexpression of the salicylic acid binding protein 2 (SABP2) gene from Tobacco results in enhanced tolerance to Huanglongbing (HLB; citrus greening disease) in transgenic sweet oranges. Huanglongbing (HLB), the most destructive citrus disease, is caused by Candidatus Liberibacter asiaticus (CaLas). Currently, no cure for this disease exists, and all commercially planted cultivars are highly susceptible. Salicylic Acid Binding Protein 2 (SABP2) is a well-characterized protein essential for establishing systemic acquired resistance (SAR) in tobacco.The constitutive over expression of SABP2 from tobacco (NtSABP2) in 'Hamlin' sweet orangeresulted in the production of several transgenic lines with variable transcript levels. Transient expression ofthe NtSABP2-EGFP fusion protein in Nicotiana benthamiana plants demonstrated thatNtSABP2was cytosolicin its subcellular localization. In a long-term fieldstudy, we identified a SABP2 transgenic linewithsignificantly reduced HLB symptoms that maintained aconsistently low CaLas titer. Transcriptome analysis of this selectedtransgenic line demonstrated upregulation of several genes related to plant defense and SAR pathways. Genes, such as NPR family genes and those coding for monooxygenases and lipoxygenases, were upregulated in the35S-NtSABP2 overexpressing line and might be candidates for incorporation into our citrus improvement program.

Transcriptome Analysis of Tryptophan-Induced Resistance against Potato Common Scab.

Potato common scab (CS) is a worldwide soil-borne disease that severely reduces tuber quality and market value. We observed that foliar application of tryptophan (Trp) could induce resistance against CS. However, the mechanism of Trp as an inducer to trigger host immune responses is still unclear. To facilitate dissecting the molecular mechanisms, the transcriptome of foliar application of Trp and water (control, C) was compared under Streptomyces scabies (S) inoculation and uninoculation. Results showed that 4867 differentially expressed genes (DEGs) were identified under S. scabies uninoculation (C-vs-Trp) and 2069 DEGs were identified under S. scabies inoculation (S-vs-S+Trp). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that Trp induced resistance related to the metabolic process, response to stimulus, and biological regulation. As phytohormone metabolic pathways related to inducing resistance, the expression patterns of candidate genes involved in salicylic acid (SA) and jasmonic acid/ethylene (JA/ET) pathways were analyzed using qRT-PCR. Their expression patterns showed that the systemic acquired resistance (SAR) and induced systemic resistance (ISR) pathways could be co-induced by Trp under S. scabies uninoculation. However, the SAR pathway was induced by Trp under S. scabies inoculation. This study will provide insights into Trp-induced resistance mechanisms of potato for controlling CS, and extend the application methods of Trp as a plant resistance inducer in a way that is cheap, safe, and environmentally friendly.

Elemental Sulfur Nanoparticles Enhance Disease Resistance in Tomatoes.

In agriculture, loss of crop yield to pathogen damage seriously threatens efforts to achieve global food security. In the present work, "organic" elemental sulfur nanoparticles (SNPs) were investigated for management of the fungal pathogen Fusarium oxysporum f. sp. lycopersici on tomatoes. Foliar application and seed treatment with SNPs (30-100 mg/L, 30 and 100 nm) suppressed pathogen infection in tomatoes, in a concentration- and size-dependent fashion in a greenhouse experiment. Foliar application with 1 mg/plant of 30 nm SNPs (30-SNPs) exhibited the best performance for disease suppression, significantly decreasing disease incidence by 47.6% and increasing tomato shoot biomass by 55.6% after 10 weeks application. Importantly, the disease control efficacy with 30-SNPs was 1.43-fold greater than the commercially available fungicide hymexazol. Mechanistically, 30-SNPs activated the salicylic acid-dependent systemic acquired resistance pathway in tomato shoots and roots, with subsequent upregulation of the expression of pathogenesis-related and antioxidase-related genes (upregulated by 11-352%) and enhancement of the activity and content of disease-related biomolecules (enhanced by 5-49%). In addition, transmission electron microscopy imaging shows that SNPs were distributed in the tomato stem and directly inactivated in vivo pathogens. The oxidative stress in tomato shoots and roots, the root plasma membrane damage, and the growth of the pathogen in stem were all significantly decreased by SNPs. The findings highlight the significant potential of SNPs as an eco-friendly and sustainable crop protection strategy.

Floro-pyrazolo[3,4-d]pyrimidine derivative as a novel plant activator induces two-pathway immune system

Plants are known to develop a multi-faceted innate immune system for pathogenic defense. Systemic acquired resistance (SAR) and induced systemic resistance (ISR) are the two main pathways. Many chemical inducers known as plant activators that activate innate immunity to defend against pathogens have been discovered. Currently, the exploitation of new plant activators is mainly done to develop analogs of salicylic acid as SAR-signaling molecules; however, the ISR pathway is hardly investigated for new plant activators. Based on recent studies on 1-methyl pyrazolo [3,4-d]pyrimidine bioactivity and ATP-induced resistance to biotrophic and necrotrophic pathogens, a new lead compound, 1-methyl-4-amino-pyrazolo [3,4-d]pyrimidine, was obtained as a new scaffold of plant activators for possible inducing ISR immunity system. Additionally, fluorine atom plays an important role in the design and development of new pesticides due to the unique physical chemistry effect, a series of pyrazolo [3,4-d]pyrimidine derivatives were designed and synthesized. Several compounds showed good broad-spectrum induced resistance in vivo, but there was no direct antibacterial activity in vitro. Notably, the introduction of fluorine atom at the para-position of the benzene ring greatly enhanced the induction activity of P1d both involved in SAR and ISR pathways, which implied the inducing resistance both in defending pathogens and insects.

Systemic propagation of immunity in plants.

Systemic immunity triggered by local plant-microbe interactions is studied as systemic acquired resistance (SAR) or induced systemic resistance (ISR) depending on the site of induction and the lifestyle of the inducing microorganism. SAR is induced by pathogens interacting with leaves, whereas ISR is induced by beneficial microbes interacting with roots. Although salicylic acid (SA) is a central component of SAR, additional signals exclusively promote systemic and not local immunity. These signals cooperate in SAR- and possibly also ISR-associated signaling networks that regulate systemic immunity. The non-SA SAR pathway is driven by pipecolic acid or its presumed bioactive derivative N-hydroxy-pipecolic acid. This pathway further regulates inter-plant defense propagation through volatile organic compounds that are emitted by SAR-induced plants and recognized as defense cues by neighboring plants. Both SAR and ISR influence phytohormone crosstalk towards enhanced defense against pathogens, which at the same time affects the composition of the plant microbiome. This potentially leads to further changes in plant defense, plant-microbe, and plant-plant interactions. Therefore, we propose that such inter-organismic interactions could be combined in potentially highly effective plant protection strategies.

Exogenous application of the plant signalers methyl jasmonate and salicylic acid induces changes in volatile emissions from citrus foliage and influences the aggregation behavior of Asian citrus psyllid (Diaphorina citri), vector of Huanglongbing.

Huanglongbing, also known as citrus greening, is a destructive disease that threatens citrus production worldwide. It is putatively caused by the phloem-limited bacterium Candidatus Liberibacter asiaticus (Las). Currently, the disease is untreatable and efforts focus on intensive insecticide use to control the vector, Asian citrus psyllid (Diaphorina citri). Emerging psyllid resistance to multiple insecticides has generated investigations into the use of exogenously applied signaling compounds to enhance citrus resistance to D. citri and Las. In the present study, we examined whether foliar applications of methyl jasmonate (MJ), a volatile signaling compound associated with the induced systemic resistance pathway, and salicylic acid, a constituent of the systemic acquired resistance pathway, would elicit the emission of defense-related volatiles in citrus foliage, and what effect this might have on the host-plant searching behavior of D. citri. Comparisons were made of volatiles emitted from growing shoots of uninfected and Las-infected ‘Valencia’ sweet orange (Citrus sinensis) trees over two consecutive sampling days. A settling behavioral assay was used to compare psyllid attraction to MJ-treated vs. Tween-treated citrus sprigs. All three main effects, Las infection status, plant signaler application, and sampling day, influenced the proportions of individual volatile compounds emitted in different treatment groups. MJ- and SA-treated trees had higher emission rates than Tween-treated trees. Methyl salicylate (MeSA) and β-caryophyllene were present in higher proportions in the volatiles collected from Las-infected + trees. On the other hand, Las-infected + MJ-treated trees emitted lower proportions of MeSA than did Las-infected + Tween-treated trees. Because MeSA is a key D. citri attractant, this result suggests that MJ application could suppress MeSA emission from Las-infected trees, an approach that could be used to discourage psyllid colonization during shoot growth. MJ application enhanced emission of E-β-ocimene, indole, volatiles attractive to many of the psyllid’s natural enemies, indicating that MJ application could be used in an ‘attract and reward’ conservation biological control strategy. Volatile emissions in SA-treated trees were dominated by MeSA. MJ application elicited aggregation behavior in D. citri. Similar numbers of psyllids settled on MJ-treated versus Tween-treated sprigs, but a significantly greater percentage of the MJ-treated sprigs had aggregations of nine or more psyllids on them. Taken together, the results of this study indicate that exogenous applications of MJ or SA could be used to influence Asian citrus psyllid settling behavior and attract its natural enemies.

A Risk Assessment Model for Bacterial Leaf Spot of Pepper (Capsicum annuum), Caused by Xanthomonas euvesicatoria, Based on Concentrations of Macronutrients, Micronutrients, and Micronutrient Ratios.

The phytopathogenic bacterium Xanthomonas euvesicatoria causes bacterial leaf spot (BLS) of pepper and has a worldwide distribution. BLS is difficult to control and an integrated management strategy that incorporates crop rotation, use of clean seed and clean plants, weed control, resistant varieties, applications of bactericides, biocontrol agents, and systemic acquired resistance (SAR) inducers is generally recommended. However, even with that arsenal of weapons, BLS can still be responsible for severe losses under favorable environmental conditions. Thus, additional tools need to be added to an overall integrated management strategy to combat BLS. In this article, we developed several models from 2012 to 2014 that were based on how macronutrients, micronutrients, and micronutrient ratios affect BLS severity. Factors used to select a model for validation included highly significant P values, high adjusted R2 values, low variance inflation factor values (<5), root mean square error, Mallow's Cp, and high Akaike's information criterion correction values. In addition, salicylic acid (SA) concentrations and relative expression of nonexpresser pathogenesis-related gene1 (NPR1) and pathogenesis-related protein 1 (PR1) in pepper tissues were also considered in model selection. A model (ECGA1) consisting of concentrations of copper, manganese, potassium, and the iron/zinc ratio as independent variables was used for validation in three different commercial pepper fields in Georgia: Colquitt County and Worth County in 2015 and Tift County in 2016. When area under the disease progress curve (AUDPC) values for two field sites (Colquitt and Worth Counties) in 2015 were pulled together and plotted against ECGA1-predicted values for both sites, the resulting relationship was highly significant (P = 0.0001) with an R2 value of 0.92. A significant relationship between observed AUDPC versus predicted values was also observed in Tift County in 2016 (P < 0.001; adjusted R2 = 0.98). Relative gene expression of both NPR1 and PR1 genes was significantly (P < 0.01) higher in pepper grown in predicted low-risk sites compared with pepper from high-risk sites in Colquitt, Worth, and Tift Counties. Although BLS severity will fluctuate depending on environmental conditions, the data indicate that the level of risk at a particular location may be influenced by how macronutrient and micronutrient concentrations affect plant disease resistance genes in the SAR pathway.

Salicylic Acid Is Involved in the Basal Resistance of Tomato Plants to Citrus Exocortis Viroid and Tomato Spotted Wilt Virus.

Tomato plants expressing the NahG transgene, which prevents accumulation of endogenous salicylic acid (SA), were used to study the importance of the SA signalling pathway in basal defence against Citrus Exocortis Viroid (CEVd) or Tomato Spotted Wilt Virus (TSWV). The lack of SA accumulation in the CEVd- or TSWV-infected NahG tomato plants led to an early and dramatic disease phenotype, as compared to that observed in the corresponding parental Money Maker. Addition of acibenzolar-S-methyl, a benzothiadiazole (BTH), which activates the systemic acquired resistance pathway downstream of SA signalling, improves resistance of NahG tomato plants to CEVd and TSWV. CEVd and TSWV inoculation induced the accumulation of the hydroxycinnamic amides p-coumaroyltyramine, feruloyltyramine, caffeoylputrescine, and feruloylputrescine, and the defence related proteins PR1 and P23 in NahG plants earlier and with more intensity than in Money Maker plants, indicating that SA is not essential for the induction of these plant defence metabolites and proteins. In addition, NahG plants produced very high levels of ethylene upon CEVd or TSWV infection when compared with infected Money Maker plants, indicating that the absence of SA produced additional effects on other metabolic pathways. This is the first report to show that SA is an important component of basal resistance of tomato plants to both CEVd and TSWV, indicating that SA-dependent defence mechanisms play a key role in limiting the severity of symptoms in CEVd- and TSWV-infected NahG tomato plants.

Induction of pathogenesis-related gene 1 (PR-1) by acibenzolar-s-methyl application in pineapple and its effect on reniform nematodes (Rotylenchulus reniformis)

The induction of systemic acquired resistance (SAR) in pineapples (Ananas comosus) was studied as shown by the up-regulation of the PR-1 gene (the SAR marker) and examination of the SAR effect on the reniform nematode, Rotylenchulus reniformis. Real-time polymerase chain reaction assay was performed using degenerate primers designed from the PR-1 genes of several monocotyledonous (monocots) and dicotyledonous (dicots) plants. A 266 bp cDNA band was evident only in plants treated with the SAR inducer, acibenzolar-s-methyl. This 266 bp cDNA was sequenced and found to be highly homologous to a number of PR-1 genes from monocots. In addition, the amino acid sequence deduced from the 266 bp cDNA showed a high identity to PR-1 proteins from both monocots and dicots. Therefore, it was highly likely that this cloned fragment was part of the A. comosus PR-1 gene, indicating that A. comosus has an SAR pathway. The time course of PR-1 expression was studied. The results showed that PR-1 induction was initiated as early as 1 d after acibenzolar application and continued through 3 wk thereafter. The effect of SAR on the nematodes, R. reniformis, in pineapples was also elucidated. The results showed that the reproduction of nematodes on the pineapples treated with 100 mg/L or 200 mg/L was 55% lower than that on pineapples treated with 0 mg/L or 50 mg/L. Nematode reproduction on pineapples treated with the same concentration but inoculated at different times was not significantly different (p > 0.05).

Response of phytohormones and correlation of SAR signal pathway genes to the different resistance levels of grapevine against Plasmopara viticola infection

Phytohormones play an important role in the process of disease resistance in plants. Here, we investigated which among salicylic acid, jasmonic acid, and abscisic acid performs a key role in plant defense after Plasmopara viticola infection in grapevine. We used grapevines possessing different resistance levels against P. viticola infection to study the relationship between the expression of key genes in the related resistance signaling pathways and the level of resistance. We performed high-performance liquid chromatography–mass spectrometry to estimate the phytohormone contents in grape leaves at different time points after the infection. Furthermore, we performed quantitative analyses of key genes such as EDS1, PAD4, ICS2, PAL, NPR1, TGA1, and PR1 in the systemic acquired resistance pathway by quantitative reverse transcription-polymerase chain reaction. The results showed an increased variation in the SA content, which was maintained at high levels, after P. viticola infection in plant species exhibiting stronger resistance to the pathogen; this finding highlights the importance of SA in plant defense mechanisms. Moreover, EDS1 and PAD4 expression did not show a positive correlation with disease resistance in grape; however, higher expression of other genes that were analyzed was observed in highly resistant grape varieties. Our results provide insights into the role of phytohormone regulation in the induction and maintenance of plant defense response to pathogens.

Nitric oxide and reactive oxygen species are required for systemic acquired resistance in plants

Systemic acquired resistance (SAR) is a form of broad-spectrum disease resistance that is induced in response to primary infection and that protects uninfected portions of the plant against secondary infections by related or unrelated pathogens. SAR is associated with an increase in chemical signals that operate in a collective manner to confer protection against secondary infections. These include, the phytohormone salicylic acid (SA), glycerol-3-phosphate (G3P), azelaic acid (AzA) and more recently identified signals nitric oxide (NO) and reactive oxygen species (ROS). NO, ROS, AzA and G3P function in the same branch of the SAR pathway, and in parallel to the SA-regulated branch. NO and ROS function upstream of AzA/G3P and different reactive oxygen species functions in an additive manner to mediate chemical cleavage of the C9 double bond on C18 unsaturated fatty acids to generate AzA. The parallel and additive functioning of various chemical signals provides important new insights in the overlapping pathways leading to SAR.

Ethylene Signaling Modulates Herbivore-Induced Defense Responses in the Model Legume Medicago truncatula

One or more effectors in the labial saliva (LS) of generalist Noctuid caterpillars activate plant signaling pathways to modulate jasmonate (JA)-dependent defense responses; however, the exact mechanisms involved have yet to be elucidated. A potential candidate in this phytohormone interplay is the ethylene (ET) signaling pathway. We compared the biochemical and molecular responses of the model legume Medicago truncatula and the ET-insensitive skl mutant to herbivory by fourth instar Spodoptera exigua (Hübner) caterpillars with intact or impaired LS secretions. Cellular oxidative stress increases rapidly after herbivory, as evidenced by changes in oxidized-to-reduced ascorbate (ASC) and glutathione (GSH) ratios. The caterpillar-specific increase in GSH ratios and the LS-specific increase in ASC ratios are alleviated in the skl mutant, indicating that ET signaling is required. Ten hours postherbivory, markers of the JA and JA/ET pathways are differentially expressed; MtVSP is induced and MtHEL is repressed in a caterpillar LS- and ET-independent manner. In contrast, expression of the classic marker of the systemic acquired resistance pathway, MtPR1, is caterpillar LS-dependent and requires ET signaling. Caterpillar LS further suppresses the induction of JA-related trypsin inhibitor activity in an ET-dependent manner. Findings suggest that ET is involved in the caterpillar LS-dependent, salicylic acid/NPR1-mediated attenuation of JA-dependent induced responses.

Transgenic pearl millet male fertility restorer line (ICMP451) and hybrid (ICMH451) expressing Brassica juncea Nonexpressor of pathogenesis related genes 1 (BjNPR1) exhibit resistance to downy mildew disease.

Brassica juncea Nonexpressor of pathogenesis-related genes 1 (BjNPR1) has been introduced into pearl millet male fertility restorer line ICMP451 by Agrobacterium tumefaciens-mediated genetic transformation. Transgenic pearl millet plants were regenerated from the phosphinothricin-resistant calli obtained after co-cultivation with A. tumefaciens strain LBA4404 harbouring Ti plasmid pSB111-bar-BjNPR1. Molecular analyses confirmed the stable integration and expression of BjNPR1 in transgenic pearl millet lines. Transgenes BjNPR1 and bar were stably inherited and disclosed co-segregation in subsequent generations in a Mendelian fashion. Transgenic pearl millet hybrid ICMH451-BjNPR1 was developed by crossing male-sterile line 81A X homozygous transgenic line ICMP451-BjNPR1. T3 and T4 homozygous lines of ICMP451-BjNPR1 and hybrid ICMH451-BjNPR1 exhibited resistance to three strains of downy mildew pathogen, while the untransformed ICMP451 and the isogenic hybrid ICMH451 plants were found susceptible. Following infection with S. graminicola, differential expression of systemic acquired resistance pathway genes, UDP-glucose salicylic acid glucosyl transferase and pathogenesis related gene 1 was observed in transgenic ICMP451-BjNPR1 and untransformed plants indicating the activation of systemic acquired resistance pathway contributing to the transgene-mediated resistance against downy mildew. The transgenic pearl millet expressing BjNPR1 showed resistance to multiple strains of S. graminicola and, as such, seems promising for the development of durable downy mildew resistant hybrids.

Benzothiadiazole (BTH) induces resistance to Pepper golden mosaic virus (PepGMV) in pepper (Capsicum annuum L.)

Pepper is an economically important crop in many countries around the world but it is susceptible to many diseases. In Mexico, diseases caused by bipartite begomoviruses have emerged as important problems in pepper. Several control strategies have been explored wiht little success; most of them are based on the avoidance of virus transmission and the breeding for resistance. Abiotic inducers can act at various points in the signaling pathways involved in disease resistance, providing long-lasting, wide-spectrum resistance. Benzothiadiazole (BTH) shares the property of activating the systemic acquired resistance pathway downstream from the SA signaling. In this work, resistance to PepGMV infection was induced in pepper plants by activating the SA pathway using BTH treatment. The resistance was characterized by evaluating symptom appearance, virus accumulation and viral movement. Our results showed that BTH could be an attractive alternative to induce geminivirus resistance in pepper plants without a significant damage of the fruit quality and productivity.