Induction Of Systemic Acquired Resistance Research Articles

New bifunctional ionic liquid-based plant systemic acquired resistance (SAR) inducers with an improved environmental hazard profile

SAR inducers with minimized environmental hazards and high biological activity are a step towards more sustainable agricultural practice.

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.

Derivatives of Isonicotinic Acid as New Efficient Systemic Acquired Resistance (SAR) Inducers

AbstractSystemic acquired resistance (SAR) is one of the most promising ways to support plants in fight against viruses, bacteria and fungi. This phenomenon is activated by pathogen attack or artificially, by using resistance inducer (elicitor) which imitates the plant‐pathogen interaction. Both, biological or chemical factors interact with plants and stimulate their immune system against infections before first symptoms of diseases occurs. SAR inducing properties of elicitors could be potentially utilized to develop a new plant protection strategies. This paper presents synthesis, phytotoxicity and SAR induction efficacy of four newly obtained amide derivatives of 2,6‐dichloroisonicotinic acid and four known amide derivatives of isonicotinic acid. Induction of plant resistance tests were performed on tobacco Nicotiana tabacum var. Xanthi infected by Tobacco mosaic virus (TMV). Six of the eight presented substances indicate resistance inducing properties, which were expressed by decrease of necrotic area on leaves caused by viral infection by 44 to 92 % in comparison to the control plants. Moreover, obtained results show that presented compounds exhibited better plant resistance inducing properties than salicylic acid, oxalic acid and β‐aminobutyric acid used as reference.

Reticine A, a new potent natural elicitor: isolation from the fruit peel of Citrus reticulate and induction of systemic resistance against tobacco mosaic virus and other plant fungal diseases.

Systemic acquired resistance (SAR) induced by elicitors is a highly satisfying form of resistance that protects plants against invading pathogens. Exploration and development of new elicitors is a promising alternative to conventional biocides in resistant pest management. In our previous broad screening, fruit peel extract of Citrus reticulata Blanco exhibited the ability to induce SAR in tobacco. A new potent elicitor reticine A was isolated from the fruit peel extract of industrial crop C. reticulate and its structure was well elucidated. In vivo assays showed that reticine A had considerable control efficacies at 100 and 500 μg mL-1 , being superior to commercial elicitor benzothiadiazole (BTH) (100 μg mL-1 ). Reticine A had no significant impact on the virulence of tobacco mosaic virus (TMV) particles under in vitro conditions. Application of reticine A induced a local hypersensitive reaction (HR), systemic accumulation of H2 O2 and salicylic acid (SA), systemic increase in defensive enzyme activities and systemic upregulated expression of pathogenesis-related (PR) proteins, suggesting its induction of SAR in tobacco. The expression of NPR1 and SA biosynthesis genes ICS and PAL were systemically upregulated. SAR induced by reticine A against TMV in tobacco was demonstrated and the mechanism might be attributed to activating the expression of several defensive genes mediated by an SA signal. This study highlights the potential of reticine A which is recommended to be applied directly or as an active ingredient in the crude extract formulation ahead of time in the field, as well as being a potential lead compound for further optimization.

Activation of defense mechanism in Lehsua (Cordia myxa) through systemic acquired resistance inducers against Alternaria leaf spot disease

Indian cherry or Lehsua (Cordia myxa Roxb.) is vernacularly known as Lasoda and Gunda and it is a major fruit of the family Boraginaceae which well flourished in arid conditions of Rajasthan and India. The immature fruits are used for preparing mix vegetable and pickle in Indian cuisine. Alternaria leaf spot of lehsua, caused by Alternaria alternata is an important disease of this crop and causes economic yield loss. In present investigation, four systemic acquired resistance (SAR) activators (salicylic acid, isonicotinic acid, hydrogen peroxide and β-amino butyric acid) were sprayed in orchard and this was the foremost case of study in perennial and arid fruit crops in Rajasthan. Among these, spray of salicylic acid (@200 ppm) proved most effective in reducing disease intensity (54.87%) followed by 2,6-dichloroisonicotinic acid (47.85%) possibly by activating the cellular genes and defense mechanism and thereby spreading throughout the plant body. These SAR inducers can be used as an alternative to fungicides as green chemicals by incorporating in integrated disease management system, because they do not cause bio-accumulation in eatables and environmental hazards.

Inducible biosynthesis and immune function of the systemic acquired resistance inducer N-hydroxypipecolic acid in monocotyledonous and dicotyledonous plants

Recent work has provided evidence for the occurrence of N-hydroxypipecolic acid (NHP) in Arabidopsis thaliana, characterized its pathogen-inducible biosynthesis by a three-step metabolic sequence from l-lysine, and established a central role for NHP in the regulation of systemic acquired resistance. Here, we show that NHP is biosynthesized in several other plant species in response to microbial attack, generally together with its direct metabolic precursor pipecolic acid and the phenolic immune signal salicylic acid. For example, NHP accumulates locally in inoculated leaves and systemically in distant leaves of cucumber in response to Pseudomonas syringae attack, in Pseudomonas-challenged tobacco and soybean leaves, in tomato inoculated with the oomycete Phytophthora infestans, in leaves of the monocot Brachypodium distachyon infected with bacterial (Xanthomonas translucens) and fungal (Magnaporthe oryzae) pathogens, and in M. oryzae-inoculated barley. Notably, resistance assays indicate that NHP acts as a potent inducer of acquired resistance to bacterial and fungal infection in distinct monocotyledonous and dicotyledonous species. Pronounced systemic accumulation of NHP in leaf phloem sap of locally inoculated cucumber supports a function for NHP as a phloem-mobile immune signal. Our study thus generalizes the existence and function of an NHP resistance pathway in plant systemic acquired resistance.

Sting nematodes modify metabolomic profiles of host plants

Plant-parasitic nematodes are devastating pathogens of many important agricultural crops. They have been successful in large part due to their ability to modify host plant metabolomes to their benefit. Both root-knot and cyst nematodes are endoparasites that have co-evolved to modify host plants to create sophisticated feeding cells and suppress plant defenses. In contrast, the ability of migratory ectoparasitic nematodes to modify host plants is unknown. Based on global metabolomic profiling of sting nematodes in African bermudagrass, ectoparasites can modify the global metabolome of host plants. Specifically, sting nematodes suppress amino acids in susceptible cultivars. Upregulation of compounds linked to plant defense have negative impacts on sting nematode population densities. Pipecolic acid, linked to systemic acquired resistance induction, seems to play a large role in protecting tolerant cultivars from sting nematode feeding and could be targeted in breeding programs.

Studies on the mechanism of agricultural chemicals focused on plant hormone signals.

In recent years, it has become clear that the crosstalk of various plant hormones controls plant growth and disease resistance. Plant hormone signals may also be involved in the actions of a variety of pesticides and disease control techniques used for crop protection. From this point of view, we have focused on plant hormones to analyze the mode of action of pesticides that function in plants. Disease resistance inducers are pesticides that induce systemic acquired resistance (SAR) by activating the salicylic acid (SA)-mediated signaling pathway. However, when under unfavorable climate conditions, such as cold and cloudy weather, the resistance inducers are not sufficiently effective. Since the environmental stress response mediated by abscisic acid (ABA) may affect disease resistance, extensive studies of tobacco and tomato plants were performed, which clarified that SAR induction was suppressed by ABA. On the other hand, it was shown that transient high temperature treatment enhanced disease resistance via SA biosynthesis. These results suggest that changes in temperature due to climate change have an impact on disease resistance. The mode of action of a plant-growth regulator was analyzed by focusing on plant hormones. Isoprothiolane (IPT), an active ingredient of Fuji-one, is used as a plant-growth regulator and a fungicide. In Arabidopsis thaliana, we demonstrated that jasmonic acid and ethylene are required for the root elongation-promoting effect of IPT. As shown above, mode-of-action studies on pesticides in relation to plant hormones will lead to the development of new techniques for the better cultivation and protection of crops.

Novel bacterial control agent tolprocarb enhances systemic acquired resistance in Arabidopsis and rice as a second mode of action

The fungicide tolprocarb (TPC) is a melanin biosynthesis inhibitor, but it may also have another mode of action. Here in tests of TPC for inducing plant systemic acquired resistance (SAR), TPC induced promoter activity of the tobacco pathogenesis-related gene PR-1a in Arabidopsis thaliana and genes for PBZ1, β-1,3-glucanase, and chitinase 1 in the defense-related salicylic acid (SA) signaling pathway in rice, but not genes for the jasmonate signaling pathway. Probenazole (PBZ), a commercially used plant defense activator, induced genes in both signaling pathways. The antibacterial activity of TPC was equivalent to that of PBZ. Irrigation with 200 μM TPC prevented growth by Pseudomonas syringae pv. maculicola in A. thaliana, and 30 μM TPC inhibited Xanthomonas oryzae pv. oryzae growth in rice. The results of this study suggest that TPC functions not only as a melanin biosynthesis inhibitor but also as an SAR inducer and is applicable as a novel bacterial control agent that induces SAR activity in both A. thaliana and rice.

Extracellular pyridine nucleotides trigger plant systemic immunity through a lectin receptor kinase/BAK1 complex

Systemic acquired resistance (SAR) is a long-lasting broad-spectrum plant immunity induced by mobile signals produced in the local leaves where the initial infection occurs. Although multiple structurally unrelated signals have been proposed, the mechanisms responsible for perception of these signals in the systemic leaves are unknown. Here, we show that exogenously applied nicotinamide adenine dinucleotide (NAD+) moves systemically and induces systemic immunity. We demonstrate that the lectin receptor kinase (LecRK), LecRK-VI.2, is a potential receptor for extracellular NAD+ (eNAD+) and NAD+ phosphate (eNADP+) and plays a central role in biological induction of SAR. LecRK-VI.2 constitutively associates with BRASSINOSTEROID INSENSITIVE1-ASSOCIATED KINASE1 (BAK1) in vivo. Furthermore, BAK1 and its homolog BAK1-LIKE1 are required for eNAD(P)+ signaling and SAR, and the kinase activities of LecR-VI.2 and BAK1 are indispensable to their function in SAR. Our results indicate that eNAD+ is a putative mobile signal, which triggers SAR through its receptor complex LecRK-VI.2/BAK1 in Arabidopsis thaliana.

Assessment of the Efficacy and Mode of Action of Benzo(1,2,3)-Thiadiazole-7-Carbothioic Acid S-Methyl Ester (BTH) and Its Derivatives in Plant Protection Against Viral Disease.

Systemic acquired resistance (SAR) induction is one of the primary defence mechanisms of plants against a broad range of pathogens. It can be induced by infectious agents or by synthetic molecules, such as benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH). SAR induction is associated with increases in salicylic acid (SA) accumulation and expression of defence marker genes (e.g., phenylalanine ammonia-lyase (PAL), the pathogenesis-related (PR) protein family, and non-expressor of PR genes (NPR1)). Various types of pathogens and pests induce plant responses by activating signalling pathways associated with SA, jasmonic acid (JA) and ethylene (ET). This work presents an analysis of the influence of BTH and its derivatives as resistance inducers in healthy and virus-infected plants by determining the expression levels of selected resistance markers associated with the SA, JA, and ET pathways. The phytotoxic effects of these compounds and their influence on the course of viral infection were also studied. Based on the results obtained, the best-performing BTH derivatives and their optimal concentration for plant performance were selected, and their mode of action was suggested. It was shown that application of BTH and its derivatives induces increased expression of marker genes of both the SA- and JA-mediated pathways.

Influence of different SAR elicitors on induction and expression of PR-proteins in Potato and Muskmelon against Oomycete pathogens

Two cultivars each of potato namely; Kufri Badshah, Kufri Jyoti and muskmelon namely; Kajri and Punjab Hybrid were treated with varying concentrations of four different elicitors for induction of systemic acquired resistance (SAR) against Phytophthora infestans causing late blight in potato and Pseudoperonospora cubensis causing downy mildew in muskmelon. Attempts were made to induce SAR with application of various SAR elicitors as spray i.e. Jasmonic acid (JA), Salicylic acid (SA), Benzothiadiazole (BTH) and Beta Amino butyric acid (BABA). These elicitors were sprayed on 3-week-old sprouts and effect on defence related proteins along with late blight and downy mildew severity was studied. Leaf samples were collected up to 7 days post treatment and used for estimation of total proteins. Sporangial solution (4.0 × 104 sporangia per ml) was sprayed after 1 week of elicitors spray. Per cent disease severity was recorded at 14 days post inoculation. SA, JA, BTH and BABA caused statistically significant (at 5% level) increase in total soluble protein in leaf samples. The best treatments for maximum induction of proteins were 500 μM each of SA, JA, and BTH and 50 mM for BABA. SDS–PAGE electrophoresis confirmed that exogenous application of JA, SA, BABA and BTH resulted in the induction of PR proteins of low molecular size along with some other proteins in both potato and muskmelon, which helped in induction of SAR against oomycete pathogens.

A MPK3/6-WRKY33-ALD1-Pipecolic Acid Regulatory Loop Contributes to Systemic Acquired Resistance.

Plants induce systemic acquired resistance (SAR) upon localized exposure to pathogens. Pipecolic acid (Pip) production via AGD2-LIKE DEFENSE RESPONSE PROTEIN1 (ALD1) is key for SAR establishment. Here, we report a positive feedback loop important for SAR induction in Arabidopsis thaliana We showed that local activation of the MAP kinases MPK3 and MPK6 is sufficient to trigger Pip production and mount SAR. Consistent with this, mutations in MPK3 or MPK6 led to compromised Pip accumulation upon inoculation with the bacterial pathogen Pseudomonas syringae pv tomato DC3000 (Pto) AvrRpt2, which triggers strong sustained MAPK activation. By contrast, P. syringae pv maculicola and Pto, which induce transient MAPK activation, trigger Pip biosynthesis and SAR independently of MPK3/6. ALD1 expression, Pip accumulation, and SAR were compromised in mutants defective in the MPK3/6-regulated transcription factor WRKY33. Chromatin immunoprecipitation showed that WRKY33 binds to the ALD1 promoter. We found that Pip triggers activation of MPK3 and MPK6 and that MAPK activation after Pto AvrRpt2 inoculation is compromised in wrky33 and ald1 mutants. Collectively, our results reveal a positive regulatory loop consisting of MPK3/MPK6, WRKY33, ALD1, and Pip in SAR induction and suggest the existence of distinct SAR activation pathways that converge at the level of Pip biosynthesis.

Salicylic acid and its derivatives elicit the production of diterpenes and sterols in corals and their algal symbionts: a metabolomics approach to elicitor SAR

The production of marine drugs in its normal habitats is often low and depends greatly on ecological conditions. Chemical synthesis of marine drugs is not economically feasible owing to their complex structures. Biotechnology application via elicitation represents a promising tool to enhance metabolites yield that has yet to be explored in soft corals. Study the elicitation impact of salicylic acid (SA) and six analogues in addition to a systemic acquired resistance inducer on secondary metabolites accumulation in the soft coral Sarcophyton ehrenbergi along with the symbiont zooxanthellae and if SA elicitation effect is extended to other coral species S. glaucum and Lobophyton pauciliforum. Post elicitation in the three corals and zooxanthella, metabolites were extracted and analyzed via UHPLC-MS coupled with chemometric tools. Multivariate data analysis of the UHPLC-MS data set revealed clear segregation of SA, amino-SA, and acetyl-SA elicited samples. An increased level ca. 6- and 8-fold of the diterpenes viz., sarcophytonolide I, sarcophine and a C28-sterol, was observed in SA and amino-SA groups, respectively. Post elicitation, the level of diepoxy-cembratriene increased 1.5-fold and 2.4-fold in 1mM SA, and acetyl-SA (aspirin) treatment groups, respectively. S. glaucum and Lobophyton pauciliforum showed a 2-fold increase of diepoxy-cembratriene levels. These results suggest that SA could function as a general and somewhat selective diterpene inducing signaling molecule in soft corals. Structural consideration reveals initial structure-activity relationship (SAR) in SA derivatives that seem important for efficient diterpene and sterol elicitation.

Direct antifungal activity of tiadinil, a systemic acquired resistance inducer, and thymol formulations on Stagonosporopsis citrulli and control of watermelon gummy stem blight

This study evaluated the direct antifungal activity of tiadinil [N-(3-chloro-4-methylphenyl)-4-methyl-1,2,3-thiadiazole-5-carboxamide], a systemic acquired resistance (SAR) inducer and two formulations of thymol (thymol I and thymol II) against Stagonosporopsis citrulli, the causal agent of gummy stem blight (GSB) disease of watermelon. Tiadinil, thymol I and thymol II completely inhibited the mycelial growth of S. citrulli in vitro at ≥ 100 ppm. Conidial germination and germ tube elongation were completely inhibited by tiadinil at ≥ 2000 ppm and by thymol-based formulations at ≥ 100 ppm. A single foliar application of tiadinil at ≥ 10 ppm or a single application of thymol I and II at ≥ 1 ppm, 48 h before or after pathogen inoculation, significantly reduced disease severity of watermelon seedlings inoculated with 105/ml conidial suspension of S. citrulli, compared to respective nontreated controls. Plants treated with foliar application of tiadinil at ≥ 1000 ppm before pathogen inoculation had significantly lower disease severity than plants that received an equivalent drench application. The efficacy of foliar application of tiadinil was affected by concentration and frequency of application. The study suggests direct antifungal activity of tiadinil, indicating a new mode of action of tiadinil against GSB disease of watermelon. The study also demonstrated direct antifungal action of thymol, a formulated active compound of essential oils, against S. citrulli and GSB disease of watermelon.

Plants Pack a Quiver Full of Arrows

Systemic acquired resistance (SAR) is a process wherein plants use chemical signals to communicate broad-spectrum systemic immunity to distant tissue. Two studies recently identified N-hydroxypipecolic acid as an additional essential SAR inducer. These findings assemble another piece in the SAR puzzle.

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.

Indispensable Role of Proteases in Plant Innate Immunity.

Plant defense is achieved mainly through the induction of microbe-associated molecular patterns (MAMP)-triggered immunity (MTI), effector-triggered immunity (ETI), systemic acquired resistance (SAR), induced systemic resistance (ISR), and RNA silencing. Plant immunity is a highly complex phenomenon with its own unique features that have emerged as a result of the arms race between plants and pathogens. However, the regulation of these processes is the same for all living organisms, including plants, and is controlled by proteases. Different families of plant proteases are involved in every type of immunity: some of the proteases that are covered in this review participate in MTI, affecting stomatal closure and callose deposition. A large number of proteases act in the apoplast, contributing to ETI by managing extracellular defense. A vast majority of the endogenous proteases discussed in this review are associated with the programmed cell death (PCD) of the infected cells and exhibit caspase-like activities. The synthesis of signal molecules, such as salicylic acid, jasmonic acid, and ethylene, and their signaling pathways, are regulated by endogenous proteases that affect the induction of pathogenesis-related genes and SAR or ISR establishment. A number of proteases are associated with herbivore defense. In this review, we summarize the data concerning identified plant endogenous proteases, their effect on plant-pathogen interactions, their subcellular localization, and their functional properties, if available, and we attribute a role in the different types and stages of innate immunity for each of the proteases covered.

Management of late leaf spot (Phaeoisariopsis personata) and root rot (Macrophomina phaseolina) diseases of groundnut (Arachis hypogaea L.) with plant growth-promoting rhizobacteria, systemic acquired resistance inducers and plant extracts

Greenhouse experiments were conducted to study the efficacy of plant growth-promoting rhizobacteria (PGPR) for control of root rot (RR) in groundnut (Arachis hypogaea L.) caused by Macrophomina phaseolina and to test the ability of plant systemic acquired resistance (SAR) inducers and plant extracts to protect groundnut plants from late leaf spot (LLS) caused by Phaeoisariopsis personata. Seed treatment and soil application of a talc-based formulation of B. subtilis strain G1 significantly reduced the incidence of root rot under greenhouse conditions. In experiments with SAR inducers, foliar application of salicylic acid (SA) (7 mM) on 45 days after sowing significantly reduced LLS incidence and increased the pod yield. Foliar application of aqueous extract (10%) from leaves of Adhatoda vasica and zimmu (Allium sativum x A. cepa) on 45 days after sowing significantly decreased the LLS incidence and increased the pod yield compared with the untreated control. Field experiments were conducted to develop an integrated method for the management of LLS and RR of groundnut using the best performing PGPR, SAR inducer and plant extract in combinations. Combined application of B. subtilis strain G1 through seed (10 g/kg) and soil (2.5 kg/ha) followed by foliar application of A. vasica extract (10%) on 30, 45 and 60 days after sowing significantly reduced LLS and RR diseases in groundnut and increased the pod yield under field conditions. The above treatment resulted in significant reductions in the area under the disease progress curve (AUDPC) for LLS compared with that of untreated control.

New ionic liquids based on systemic acquired resistance inducers combined with the phytotoxicity reducing cholinium cation

Systemic acquired resistance (SAR) is one of the most promising ways to support plants in the fight against viruses.