Systemic Acquired Resistance Response Research Articles

Hollow Mesoporous Silica Nanoparticles as a New Nanoscale Resistance Inducer for Fusarium Wilt Control: Size Effects and Mechanism of Action.

In agriculture, soil-borne fungal pathogens, especially Fusarium oxysporum strains, are posing a serious threat to efforts to achieve global food security. In the search for safer agrochemicals, silica nanoparticles (SiO2NPs) have recently been proposed as a new tool to alleviate pathogen damage including Fusarium wilt. Hollow mesoporous silica nanoparticles (HMSNs), a unique class of SiO2NPs, have been widely accepted as desirable carriers for pesticides. However, their roles in enhancing disease resistance in plants and the specific mechanism remain unknown. In this study, three sizes of HMSNs (19, 96, and 406 nm as HMSNs-19, HMSNs-96, and HMSNs-406, respectively) were synthesized and characterized to determine their effects on seed germination, seedling growth, and Fusarium oxysporum f. sp. phaseoli (FOP) suppression. The three HMSNs exhibited no side effects on cowpea seed germination and seedling growth at concentrations ranging from 100 to 1500 mg/L. The inhibitory effects of the three HMSNs on FOP mycelial growth were very weak, showing inhibition ratios of less than 20% even at 2000 mg/L. Foliar application of HMSNs, however, was demonstrated to reduce the FOP severity in cowpea roots in a size- and concentration-dependent manner. The three HMSNs at a low concentration of 100 mg/L, as well as HMSNs-19 at a high concentration of 1000 mg/L, were observed to have little effect on alleviating the disease incidence. HMSNs-406 were most effective at a concentration of 1000 mg/L, showing an up to 40.00% decline in the disease severity with significant growth-promoting effects on cowpea plants. Moreover, foliar application of HMSNs-406 (1000 mg/L) increased the salicylic acid (SA) content in cowpea roots by 4.3-fold, as well as the expression levels of SA marker genes of PR-1 (by 1.97-fold) and PR-5 (by 9.38-fold), and its receptor gene of NPR-1 (by 1.62-fold), as compared with the FOP infected control plants. Meanwhile, another resistance-related gene of PAL was also upregulated by 8.54-fold. Three defense-responsive enzymes of POD, PAL, and PPO were also involved in the HMSNs-enhanced disease resistance in cowpea roots, with varying degrees of reduction in activity. These results provide substantial evidence that HMSNs exert their Fusarium wilt suppression in cowpea plants by activating SA-dependent SAR (systemic acquired resistance) responses rather than directly suppressing FOP growth. Overall, for the first time, our results indicate a new role of HMSNs as a potent resistance inducer to serve as a low-cost, highly efficient, safe and sustainable alternative for plant disease protection.

Exogenous expression of barley HvWRKY6 in wheat improves broad-spectrum resistance to leaf rust, Fusarium crown rot, and sharp eyespot

Systemic acquired resistance (SAR) is a broad-spectrum plant defense phenomena controlled by the salicylic acid receptor NPR1. Key regulators of the SAR signaling pathway showed great potentials to improve crop resistance to various diseases. In our previous investigation, a barley transcription factor gene HvWRKY6 was identified as downstream of NPR1 during SAR. However, the broad-spectrum resistance features and molecular mechanisms of HvWRKY6 remain to be explored. In this study, a transgenic wheat line exogenously expressing HvWRKY6 showed improved resistance to leaf rust, Fusarium crown rot (FCR), and sharp eyespot. The model pathogen Pseudomonas syringae pv. tomato DC3000 was employed to induce the SAR response in wheat plants' leaf region adjacent to the infiltration area. Transcriptome sequencing revealed activation of broad-spectrum defense responses by expressing HvWRKY6 in a pathogen-independent manner. Based on the differentially expressed genes in plant hormone signal transduction, we speculated that the enhanced resistance in HvWRKY6-OE wheat transgenic line was associated with activation of the salicylic acid pathway and suppression of the abscisic acid and jasmonic acid pathways. These findings suggest that the transgenic line HvWRKY6-OE might be applied for the genetic improvement of wheat to several fungal diseases; the underlying resistance mechanism was clarified.

Insight into melatonin-mediated response and signaling in the regulation of plant defense under biotic stress.

Melatonin plays a crucial role in the mitigation of plant biotic stress through induced defense responses and pathogen attenuation. Utilizing the current knowledge of signaling and associated mechanism of this phytoprotectant will be invaluable in sustainable plant disease management. Biotic stress in plants involves complex regulatory networks of various sensory and signaling molecules. In this context, the polyfunctional, ubiquitous-signaling molecule melatonin has shown a regulatory role in biotic stress mitigation in plants. The present review conceptualized the current knowledge concerning the melatonin-mediated activation of the defense signaling network that leads to the resistant or tolerant phenotype of the infected plants. Fundamentals of signaling networks involved in melatonin-induced reactive oxygen species (ROS) or reactive nitrogen species (RNS) scavenging through enzymatic and non-enzymatic antioxidants have also been discussed. Increasing evidence has suggested that melatonin acts upstream of mitogen-activated proteinase kinases in activation of defense-related genes and heat shock proteins that provide immunity against pathogen attack. Besides, the direct application of melatonin on virulent fungi and bacteria showed disrupted spore morphology, destabilization of cell ultrastructure, reduced biofilm formation, and enhanced mortality that led to attenuate disease symptoms on melatonin-treated plants. The transcriptome analysis has revealed the down-regulation of pathogenicity genes, metabolism-related genes, and up-regulation of fungicide susceptibility genes in melatonin-treated pathogens. The activation of melatonin-mediated systemic acquired resistance (SAR) through cross-talk with salicylic acid (SA), jasmonic acid (JA) has been essential for viral disease management. The high endogenous melatonin concentration has also been correlated with the up-regulation of genes involved in pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI). The present review highlights the versatile functions of melatonin towards direct inhibition of pathogen propagule along with active participation in mediating oxidative burst and simulating PTI, ETI and SAR responses. The hormonal cross-talk involving melatonin mediated biotic stress tolerance through defense signaling network suggests its suitability in a sustainable plant protection system.

The role of salicylic acid and benzothiadiazole in decreasing phytoplasma titer of sugarcane white leaf disease

The objective of this research was to study the effect of Benzothiadiazole (BTH) and Salicylic acid (SA) on the systemic acquired resistance (SAR) of sugarcane the phytoplasma associated with the sugarcane white leaf (SCWL) disease. The experiment was conducted on plants of the sugarcane variety Khon Kaen 3 (KK3) infected with SCWL phytoplasma using insect vectors. Biochemical changes related to the SAR such as SA and total phenolic compounds were followed according to 4 different timepoints: 7, 14, 21 and 28 days after inoculation. Together, phytoplasma were quantified by RT-qPCR using the secA gene of phytoplasma. According to our results, the spraying of BTH and SA tended to increase the amounts of SA, total phenolic compounds and a lower presence of phytoplasma in the plants in comparison with the inoculated control. Spraying BTH at a concentration of 2.4 mM and SA at a concentration of 2.4 mM exhibited the best efficiency to reduce the concentration of phytoplasma. According to RT-qPCR results, the inoculated plants sprayed with BTH displayed a significantly lower concentration of phytoplasma compared to the inoculated controls. Overall, our results indicated that the spray of BTH and SA could induce an efficient SAR response to the phytoplasma associated with the SCWL disease. We expect these results will give support to the development of new products for controlling white leaf disease in sugarcane.

Enriched Agriculture Horticulture Science Technology Socio-EconomyCommunication-Issue by Biomedicines Suppressing Tomato-Disease and Coronavirus 2-Like-Pandemic-Diseases!

To avoid traditional control of root-knot disease, there remains a need for developing effective biomedicines of animal origins. The animal biomedicines; Nematode Extract (NE) and Gall Protein (GP) when applied by foliar spray, are highly effective at 1.3mg/plant and 2.01mg/plant, respectively in ameliorating root-knot disease of tomato (Lycopersicon lycopersicum, Cv. Pusa Ruby) caused by Meloidogyne incognita (Kofoid & White) Chitwood, promoting plant growth and fruit production. The nematode extract (NE) is more effective than the gall protein (GP) in this respect. Both the extract is thought to induce systemic acquired resistance response of the treated plants through the expression of pathogenesis-related proteins, and may prevent 21st –century pandemics COVID-19 like virus diseases by boosting immunity, resisting toxic effects on the environment, and improved agriculture green-socio-economy aquatic-science-technology-communication application issues, and it may also give a good scope for new advanced-development and research in the field “Agricultural, Horticulture, Earth-Environmental-Clinical-Toxicology-Health-Research, Science-Technology-Communication-Socio-Economy and May be Controlled 21st-Century Pandemic Diseases in Future New Normal Situation Also”.

Potentiators of Disease During Barley Infection by Pyrenophora teres f. teres in a Susceptible Interaction.

Pyrenophora teres f. teres is a necrotrophic fungal pathogen and causal agent of net form net blotch (NFNB), a significant disease of barley. RNA-seq data encompassing asymptomatic and subsequent necrotrophic phases of the pathogen was obtained for P. teres f. teres isolate W1-1 in NFNB-sensitive cultivar Baudin. Host genes notably regulated during infection included concerted induction of over half the repertoire of disease resistance genes, together with genes involved in oxidation-reduction processes, characteristic of a hypersensitive response. Several systemic acquired resistance response genes were suppressed and there was a complete absence of defense-related thionin gene expression. In P. teres f. teres, genes involved in hydrolase activities and cell-wall catabolic processes were induced during infection, while nitrate assimilation and response to oxidative stress processes were suppressed. Timecourse data allowed a number of predicted P. teres f. teres effector genes with differing expression profiles to be identified that may underlie barley sensitivity to NFNB. Candidate genes involved in the host-pathogen interaction provide a basis for functional characterization and control strategies based on fungicide or mutation targets, which will facilitate further research aimed at controlling NFNB disease.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

UGT76B1 controls the growth-immunity trade-off during systemic acquired resistance

UGT76B1 controls the growth-immunity trade-off during systemic acquired resistance

Genetic Effects of Ultra-High-Diluted-Biomedicines Gall 30C, Gall 200C, and Gall 1000C may be Vaccines against Plant and COVID-19 Diseases: Improved Agriculture-Health-Medical-Pharmaceutical-ScienceTechnology-Communication-Issues!

The current coronavirus 2 (SARS-CoV-2) outbreaks of Coronavirus disease 2019 (COVID-19) pandemic have emphasized the vulnerability of human populations to novel viral pressures, causing an emergent global pandemic and badly impacts on agriculture-environment-health-socio-economy medical-pharmaceutical-science-technology-communication-issues. And India emphasis on okra, Abelmoschus esculentus (L.) Moench Cv. Ankur-40, the ‘Nature’s-Gift to Human-Disease-Free-Healthy-Life’ and the most ‘Economically-Important Number-One Consumption-Vegetable-Crops,’ is damaged by different pathogens like nematodes, causing the root-knot (RK) disease which is easily controlled by different chemical-pesticides, and so it is an urgent need to develop potential epidemiological and biomedical vaccines. The Ultra-High-Diluted Biomedicines (UHDBM) Gall 30C, Gall 200C, and Gall 100C, prepared from Okra Root-Galls (ORG) or Gall MT, applied by foliar spray @ 20 mL/plant each group respectively, are highly effective against the root-knot disease of okra, with increasing fresh-plant growth and fruit production. The UHDBM; Gall 30C, Gall 200C, and Gall 1000-pretreatments are more effective than the untreated ones, and Gall 200C shows the most potential result in all respects. The genetic-effects of UHDBM thought to induce systemic acquired resistance response of the treated plants through the expression of pathogenesis-related (PR)- proteins-genes (22 to 4 numbers), which are more or less similar molecular range (295 kD to 11 kD) of many coronaviruses, and it will responsible for preventing RK and COVID-19 like variant-virus diseases by inducing defense-resistance or increasing innate-immunity, and enriching agricultureenvironment-health-socio-economy-medical-pharmaceutical-science-technology-communication-issues (AEHSEMPSTCI) with the toxic-free world, and it may help to develop best potential new preventive treatments methods or drug or vaccines, in the field of ‘21st Century COVID-19 like Pandemic in the new normal situation in future.

Glycosylation of N-hydroxy-pipecolic acid equilibrates between systemic acquired resistance response and plant growth

N-hydroxy-pipecolic acid (NHP) activates plant systemic acquired resistance (SAR). Enhanced defense responses are typically accompanied by deficiency in plant development and reproduction. Despite of extensive studies on SAR induction, the effects of NHP metabolism on plant growth remain largely unclear. In this study, we discovered that NHP glycosylation is a critical factor that fine-tunes the tradeoff between SAR defense and plant growth. We demonstrated that a UDP-glycosyltransferase (UGT76B1) forming NHP glycoside (NHPG) controls the NHP to NHPG ratio. Consistently, the ugt76b1 mutant exhibits enhanced SAR response and an inhibitory effect on plant growth, while UGT76B1 overexpression attenuates SAR response, promotes growth, and delays senescence, indicating that NHP levels are dependent on UGT76B1 function in the course of SAR. Furthermore, our results suggested that, upon pathogen attack, UGT76B1-mediated NHP glycosylation forms a “hand brake” on NHP accumulation by attenuating the positive regulation of NHP biosynthetic pathway genes, highlighting the complexity of SAR-associated networks. In addition, we showed that UGT76B1-mediated NHP glycosylation in the local site is important for fine-tuning SAR response. Our results implicate that engineering plant immunity through manipulating the NHP/NHPG ratio is a promising method to balance growth and defense response in crops.

Multi-Omics Revealed Molecular Mechanisms Underlying Guard Cell Systemic Acquired Resistance.

Systemic Acquired Resistance (SAR) improves immunity of plant systemic tissue after local exposure to a pathogen. Guard cells that form stomatal pores on leaf surfaces recognize bacterial pathogens via pattern recognition receptors, such as Flagellin Sensitive 2 (FLS2). However, how SAR affects stomatal immunity is not known. In this study, we aim to reveal molecular mechanisms underlying the guard cell response to SAR using multi-omics of proteins, metabolites and lipids. Arabidopsis plants previously exposed to pathogenic bacteria Pseudomonas syringae pv. tomato DC3000 (Pst) exhibit an altered stomatal response compared to control plants when they are later exposed to the bacteria. Reduced stomatal apertures of SAR primed plants lead to decreased number of bacteria in leaves. Multi-omics has revealed molecular components of SAR response specific to guard cells functions, including potential roles of reactive oxygen species (ROS) and fatty acid signaling. Our results show an increase in palmitic acid and its derivative in the primed guard cells. Palmitic acid may play a role as an activator of FLS2, which initiates stomatal immune response. Improved understanding of how SAR signals affect stomatal immunity can aid biotechnology and marker-based breeding of crops for enhanced disease resistance.

Methyl Salicylate Glucosylation Regulates Plant Defense Signaling and Systemic Acquired Resistance.

Plant systemic acquired resistance (SAR) provides an efficient broad-spectrum immune response to pathogens. SAR involves mobile signal molecules that are generated by infected tissues and transported to systemic tissues. Methyl salicylate (MeSA), a molecule that can be converted to salicylic acid (SA), is an essential signal for establishing SAR, particularly under a short period of exposure to light after pathogen infection. Thus, the control of MeSA homeostasis is important for an optimal SAR response. Here, we characterized a uridine diphosphate-glycosyltransferase, UGT71C3, in Arabidopsis (Arabidopsis thaliana), which was induced mainly in leaf tissue by pathogens including Pst DC3000/avrRpt2 (Pseudomonas syringae pv tomato strain DC3000 expressing avrRpt2). Biochemical analysis indicated that UGT71C3 exhibited strong enzymatic activity toward MeSA to form MeSA glucosides in vitro and in vivo. After primary pathogen infection by Pst DC3000/avrRpt2, ugt71c3 knockout mutants exhibited more powerful systemic resistance to secondary pathogen infection than that of wild-type plants, whereas systemic resistance in UGT71C3 overexpression lines was compromised. In agreement, after primary infection of local leaves, ugt71c3 knockout mutants accumulated significantly more systemic MeSA and SA than that in wild-type plants. whereas UGT71C3 overexpression lines accumulated less. Our results suggest that MeSA glucosylation by UGT71C3 facilitates negative regulation of the SAR response by modulating homeostasis of MeSA and SA. This study unveils further SAR regulation mechanisms and highlights the role of glucosylation of MeSA and potentially other systemic signals in negatively modulating plant systemic defense.

Isochorismate-based salicylic acid biosynthesis confers basal resistance to Fusarium graminearum in barley.

Salicylic acid (SA) plays an important role in signal transduction and disease resistance. In Arabidopsis, SA can be made by either of two biosynthetic branches, one involving isochorismate synthase (ICS) and the other involving phenylalanine ammonia-lyase (PAL). However, the biosynthetic pathway and the importance of SA remain largely unknown in Triticeae. Here, we cloned one ICS and seven PAL genes from barley, and studied their functions by their overexpression and suppression in that plant. Suppression of the ICS gene significantly delayed plant growth, whereas PAL genes, both overexpressed and suppressed, had no significant effect on plant growth. Similarly, suppression of ICS compromised plant resistance to Fusarium graminearum, whereas similar suppression of PAL genes had no significant effect. We then focused on transgenic plants with ICS. In a leaf-based test with F.graminearum, transgenic plants with an up-regulated ICS were comparable with wild-type control plants. By contrast, transgenic plants with a suppressed ICS lost the ability to accumulate SA during pathogen infection and were also more susceptible to Fusarium than the wild-type controls. This suggests that ICS plays a unique role in SA biosynthesis in barley, which, in turn, confers a basal resistance to F.graminearum by modulating the accumulation of H2 O2 , O2- and reactive oxygen-associated enzymatic activities. Although SA mediates systemic acquired resistance (SAR) in dicots, there was no comparable SAR response to F.graminearum in barley. This study expands our knowledge about SA biosynthesis in barley and proves that SA confers basal resistance to fungal pathogens.

Signals of Systemic Immunity in Plants: Progress and Open Questions.

Systemic acquired resistance (SAR) is a defence mechanism that induces protection against a wide range of pathogens in distant, pathogen-free parts of plants after a primary inoculation. Multiple mobile compounds were identified as putative SAR signals or important factors for influencing movement of SAR signalling elements in Arabidopsis and tobacco. These include compounds with very different chemical structures like lipid transfer protein DIR1 (DEFECTIVE IN INDUCED RESISTANCE1), methyl salicylate (MeSA), dehydroabietinal (DA), azelaic acid (AzA), glycerol-3-phosphate dependent factor (G3P) and the lysine catabolite pipecolic acid (Pip). Genetic studies with different SAR-deficient mutants and silenced lines support the idea that some of these compounds (MeSA, DIR1 and G3P) are activated only when SAR is induced in darkness. In addition, although AzA doubled in phloem exudate of tobacco mosaic virus (TMV) infected tobacco leaves, external AzA treatment could not induce resistance neither to viral nor bacterial pathogens, independent of light conditions. Besides light intensity and timing of light exposition after primary inoculation, spectral distribution of light could also influence the SAR induction capacity. Recent data indicated that TMV and CMV (cucumber mosaic virus) infection in tobacco, like bacteria in Arabidopsis, caused massive accumulation of Pip. Treatment of tobacco leaves with Pip in the light, caused a drastic and significant local and systemic decrease in lesion size of TMV infection. Moreover, two very recent papers, added in proof, demonstrated the role of FMO1 (FLAVIN-DEPENDENT-MONOOXYGENASE1) in conversion of Pip to N-hydroxypipecolic acid (NHP). NHP systemically accumulates after microbial attack and acts as a potent inducer of plant immunity to bacterial and oomycete pathogens in Arabidopsis. These results argue for the pivotal role of Pip and NHP as an important signal compound of SAR response in different plants against different pathogens.

Pathogenesis Related Protein 5 Expression is Induced by 1,4‐dimethylnaphthalene treatment of Potato Meristems

The agricultural compound 1,4‐dimethyl naphthalene (DMN) can be used to inhibit sprouting and prolong the storage of Solanum tuberosum. Potatoes treated with DMN exhibit characteristics that enable greater crop yields, including pathogen response, water retention, and greater shelf life. However, the molecular mode of action as to how DMN brings about these changes within the potato is not well understood. Two years of RNA‐seq data demonstrate extensive changes in gene expression within potato samples treated with DMN at different times throughout the year. Significant changes have been highlighted among a particular group of genes located on chromosome one. This group of genes codes for a pathogenesis‐related‐protein, PR5. As an extracellular protein, PR5 has an extensive signal transduction that responds to pathogens and abiotic stress. Among the PR5 family is a group of cytosine‐rich proteins called osmotins. Osmotin is a ubiquitous protein among several plant species and serves as a useful molecular marker for the systemic acquired resistance response, which is analogous to the innate immune system in plants. Gene expression measurements were taken over a six month period, during which dormancy was naturally breached. RNA‐seq data reveals 2‐3 fold increases between measurements taken before and as dormancy was breaking. After dormancy was breached, PR5 gene expression continued to rise significantly in comparison to previous measurements. These trends in gene regulation imply that the PR5 expression proceeds concurrently with sprout growth in DMN treated potatoes, and are related to the systemic changes seen from the outside of the potato. Phylogenetic analysis reveals the PR5 genes with the greatest changes in expression (7865 and 7870) demonstrate the greatest homology among their transcripts. Current analysis is being done using qPCR primers directed towards genes 7865 and 7870. Expression levels from qPCR should mirror the RNA‐seq data, in addition to demonstrate more immediate expression following DMN treatment. It has been hypothesized that gene expression spikes in prominent PR5 genes (7865 and 7870) will signify a broad molecular mode of action DMN plays inside S.tuberosum.Support or Funding InformationSupported in part by a grant from the 1,4‐Group, Meridian, ID.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato.

To combat pathogen infection, plants employ local defenses in infected sites and elicit systemic acquired resistance (SAR) in distant tissues. MicroRNAs have been shown to play a significant role in local defense, but their association with SAR is unknown. In addition, no such studies of the interaction between potato and Phytophthora infestans have been reported. We investigated the role of miR160 in local and SAR responses to P. infestans infection in potato. Expression analysis revealed induced levels of miR160 in both local and systemic leaves of infected wild-type plants. miR160 overexpression and knockdown plants exhibited increased susceptibility to infection, suggesting that miR160 levels equivalent to those of wild-type plants may be necessary for mounting local defense responses. Additionally, miR160 knockdown lines failed to elicit SAR, and grafting assays indicated that miR160 is required in both local and systemic leaves to trigger SAR. Consistently, SAR-associated signals and genes were dysregulated in miR160 knockdown lines. Furthermore, analysis of the expression of defense and auxin pathway genes and direct regulation of StGH3.6, a mediator of salicylic acid-auxin cross-talk, by the miR160 target StARF10 revealed the involvement of miR160 in antagonistic cross-talk between salicylic acid-mediated defense and auxin-mediated growth pathways. Overall, our study demonstrates that miR160 plays a crucial role in local defense and SAR responses during the interaction between potato and P. infestans.

In Vitro Induced Resistance of Fusarium Wilt Disease (Fusarium oxysporum f.sp. cepae) by Salicylic Acid in Shallot CV ‘Bima Brebes’

Fusarium wilt caused by Fusarium oxysporum f.sp.cepae (Foc) is one of the most important diseases of onion in Indonesia. Induced resistance was one of the effective techniques to acquire resistance to fusarium wilt in shallot. Salicylic acid (SA) had been inferred to be in endogenous signal in the systemic acquired resistance response of plants. This research aimed to study the effect of exogenously added SA to resistance in shallot callus cv ‘Bima Brebes’ to fusarium wilt disease in vitro; and to determine the effective concentration of SA to induce resistance. A group of shallot callus was grown in MS medium containing varying SA concentration (0, 15, 20 and 25 ppm) in vitro for 2 weeks. Then, the callus were treated with toxin of Fusarium oxysporum, namely fusaric acid, to have observe the resistance response. In vitro selection was done twice in different fusaric acid concentration. Application of exogenous SA at all concentration did not suppres time of appearance of disease symptom yet. Toxic symptom in the callus was shown by the browning or blackening (off) of callus. Salicylic acid at concentration of 20 ppm effectively reduced the toxic symptom up to 16.66% and supported callus regeneration better than the concentration of 15 ppm and 25 ppm. The number of resistant callus regenerated was 66.67% at pretreatment of 20 ppm of SA.

The Elicitor Protein AsES Induces a Systemic Acquired Resistance Response Accompanied by Systemic Microbursts and Micro-Hypersensitive Responses in Fragaria ananassa.

The elicitor AsES (Acremonium strictum elicitor subtilisin) is a 34-kDa subtilisin-like protein secreted by the opportunistic fungus Acremonium strictum. AsES activates innate immunity and confers resistance against anthracnose and gray mold diseases in strawberry plants (Fragaria × ananassa Duch.) and the last disease also in Arabidopsis. In the present work, we show that, upon AsES recognition, a cascade of defense responses is activated, including: calcium influx, biphasic oxidative burst (O2⋅- and H2O2), hypersensitive cell-death response (HR), accumulation of autofluorescent compounds, cell-wall reinforcement with callose and lignin deposition, salicylic acid accumulation, and expression of defense-related genes, such as FaPR1, FaPG1, FaMYB30, FaRBOH-D, FaRBOH-F, FaCHI23, and FaFLS. All these responses occurred following a spatial and temporal program, first induced in infiltrated leaflets (local acquired resistance), spreading out to untreated lateral leaflets, and later, to distal leaves (systemic acquired resistance). After AsES treatment, macro-HR and macro-oxidative bursts were localized in infiltrated leaflets, while micro-HRs and microbursts occurred later in untreated leaves, being confined to a single cell or a cluster of a few epidermal cells that differentiated from the surrounding ones. The differentiated cells initiated a time-dependent series of physiological and anatomical changes, evolving to idioblasts accumulating H2O2 and autofluorescent compounds that blast, delivering its content into surrounding cells. This kind of systemic cell-death process in plants is described for the first time in response to a single elicitor. All data presented in this study suggest that AsES has the potential to activate a wide spectrum of biochemical and molecular defense responses in F. ananassa that may explain the induced protection toward pathogens of opposite lifestyle, like hemibiotrophic and necrotrophic fungi.

Biochemical Principles and Functional Aspects of Pipecolic Acid Biosynthesis in Plant Immunity.

The nonprotein amino acid pipecolic acid (Pip) regulates plant systemic acquired resistance and basal immunity to bacterial pathogen infection. In Arabidopsis (Arabidopsis thaliana), the lysine (Lys) aminotransferase AGD2-LIKE DEFENSE RESPONSE PROTEIN1 (ALD1) mediates the pathogen-induced accumulation of Pip in inoculated and distal leaf tissue. Here, we show that ALD1 transfers the α-amino group of l-Lys to acceptor oxoacids. Combined mass spectrometric and infrared spectroscopic analyses of in vitro assays and plant extracts indicate that the final product of the ALD1-catalyzed reaction is enaminic 2,3-dehydropipecolic acid (DP), whose formation involves consecutive transamination, cyclization, and isomerization steps. Besides l-Lys, recombinant ALD1 transaminates l-methionine, l-leucine, diaminopimelate, and several other amino acids to generate oxoacids or derived products in vitro. However, detailed in planta analyses suggest that the biosynthesis of 2,3-DP from l-Lys is the major in vivo function of ALD1. Since ald1 mutant plants are able to convert exogenous 2,3-DP into Pip, their Pip deficiency relies on the inability to form the 2,3-DP intermediate. The Arabidopsis reductase ornithine cyclodeaminase/μ-crystallin, alias SYSTEMIC ACQUIRED RESISTANCE-DEFICIENT4 (SARD4), converts ALD1-generated 2,3-DP into Pip in vitro. SARD4 significantly contributes to the production of Pip in pathogen-inoculated leaves but is not the exclusive reducing enzyme involved in Pip biosynthesis. Functional SARD4 is required for proper basal immunity to the bacterial pathogen Pseudomonas syringae Although SARD4 knockout plants show greatly reduced accumulation of Pip in leaves distal to P. syringae inoculation, they display a considerable systemic acquired resistance response. This suggests a triggering function of locally accumulating Pip for systemic resistance induction.

Characterization of a Pipecolic Acid Biosynthesis Pathway Required for Systemic Acquired Resistance

Systemic acquired resistance (SAR) is an immune response induced in the distal parts of plants following defense activation in local tissue. Pipecolic acid (Pip) accumulation orchestrates SAR and local resistance responses. Here, we report the identification and characterization of SAR-DEFICIENT4 (SARD4), which encodes a critical enzyme for Pip biosynthesis in Arabidopsis thaliana Loss of function of SARD4 leads to reduced Pip levels and accumulation of a Pip precursor, Δ1-piperideine-2-carboxylic acid (P2C). In Escherichia coli, expression of the aminotransferase ALD1 leads to production of P2C and addition of SARD4 results in Pip production, suggesting that a Pip biosynthesis pathway can be reconstituted in bacteria by coexpression of ALD1 and SARD4. In vitro experiments showed that ALD1 can use l-lysine as a substrate to produce P2C and P2C is converted to Pip by SARD4. Analysis of sard4 mutant plants showed that SARD4 is required for SAR as well as enhanced pathogen resistance conditioned by overexpression of the SAR regulator FLAVIN-DEPENDENT MONOOXYGENASE1. Compared with the wild type, pathogen-induced Pip accumulation is only modestly reduced in the local tissue of sard4 mutant plants, but it is below detection in distal leaves, suggesting that Pip is synthesized in systemic tissue by SARD4-mediated reduction of P2C and biosynthesis of Pip in systemic tissue contributes to SAR establishment.

Diversity analysis, pathogen load and gene expression studies of mango cultivars infected by mango malformation disease

Mango (Mangifera indica L.) is known as “king of fruits” in India. More than 1000 mango varieties are currently cultivated in Indian Sub-continent. However most of the orchards of mango are infected with mango malformation disease (MMD), which every year leads to huge losses in yield of mango in range of 40 to 80 % in India. Till date there is no effective control measure against MMD. Floral Malformation, in contrast to vegetative one, is very virulent and can cause the loss of the entire crop. In the present study, six mango cultivars commonly grown in Gujarat, and all infected with various degrees of MMD were taken for studying their molecular relatedness, pathogen load and defense responsiveness via gene expression to rate whether hybrids or landrace among mango cultivars are better equipped to fight MMD. Genetic diversity analysis was performed using 30 SSR markers in order to bring out clustering pattern among the six cultivars belonging to orchards of Balisana and Prantij, Gujarat. The diversity analysis gave clues to the existence of wide genetic base among the six cultivars. Fungal load studies using Real Time PCR lead to the ranking of cultivars based on maximum and minimum infection load of pathogen. Absolute quantitation studies found that cultivars like Totapuri, Neelam and Amrapali were more resistant to MMD than highly popular cultivars like Kesar. The six mango cultivars were further quantified for pathogen responsiveness with 21 defense responsive genes using Real Time PCR. Among the 21 genes selected for the study, 11 genes were directly part of defense responsive pathways like Phenyl propanoid pathway and jasmonic acid pathway. Gene expression studies aided in ranking mango hybrid like Amrapali having better systemic acquired resistance response as 11 defense responsive genes were found upregulated in this cultivar followed by landrace Neelam which is in fact a parental line of Amrapali. If MMD remains unchecked it may lead to evolution of more virulent strains of Fusarium; propelling devastating consequences in mango cultivation. Hence mango hybrids developed via molecular and expressional screening will fasten process of establishment of resistant mango cultivars.