Elicitor-induced Defense Responses Research Articles

Advancements in Breeding Strategies for Enhancing Wilt and Root Rot Resistance in Castor (Ricinus communis L.): A Comprehensive Review

Castor (Ricinus communis L.) is an economically important oilseed crop cultivated worldwide for its versatile uses in various industries. However, the cultivation of castor is challenged by wilt and root rot diseases caused by fungal pathogens, leading to significant yield losses. Enhancing wilt and root rot resistance in castor is crucial for sustainable and productive cultivation. This review explores advancements in breeding strategies for enhancing resistance against these diseases through various approaches along with introduction to castor cultivation and the challenges posed by wilt and root rot diseases in castor. It emphasizes the importance of breeding for disease resistance and discusses the significance of genetic diversity in castor germplasm. Traditional breeding approaches including phenotypic selection, recurrent selection, hybridization, mutation breeding and polyploidy induction are examined for their effectiveness in introducing and accumulating resistance genes and traits. Biotechnological approaches such as marker-assisted selection (MAS), genetic transformation and RNA interference (RNAi) are highlighted as powerful tools for targeted selection and manipulation of specific genes associated with disease resistance. The utilization of physiological and biochemical approaches including understanding host-pathogen interactions, elicitor-induced defense responses and metabolic engineering are discussed for enhancing resistance mechanisms in castor. These advancements offer promising avenues for accelerating the development of wilt and root rot-resistant castor varieties.

Efficacy evaluation of some fumigants against Fusarium oxysporum and enhancement of tomato growth as elicitor-induced defense responses

Fusarium wilt, the most serious soil-borne pathogen, is a serious problem for tomato production worldwide. The presented study evaluated the antifungal activity against Fusarium oxysporum f. sp. lycopersici in vitro and in vivo for nine fumigants. In addition, the research examined the possibility of enhancing the growth of tomato plants in order to increase resistance against this disease by using four chemical inducers. The results indicated that at 20 mg/L, the radial growth of the pathogen was inhibited 100% by formaldehyde and > 80% by phosphine. Among the essential oils investigated, neem oil was the most effective, however, it only achieved 40.54% at 500 mg/L. The values of EC50 for all fumigants, except dimethyl disulfide (DMDS) and carbon disulfide (CS2), were lower than those for thiophanate-methyl. Phosphine was the highest efficient. The elicitors can be arranged based on their effectiveness, gibberellic acid (GA3) > sorbic acid > cytokinin (6-benzylaminopurine) > indole-3-butyric acid. The change in root length, fresh weight, and dry weight was greater with soil drench than with foliar application. The fumigant generators formaldehyde, phosphine and 1,4-dichlorobenzene and bio-fumigants citrus and neem oils as well as elicitors gibberellic and sorbic acid could be one of the promising alternatives to methyl bromide against Fusarium oxysporum as an important component of integrated management of Fusarium wilt.

Transcriptome dynamics underlying elicitor-induced defense responses against Septoria leaf spot disease of tomato (Solanum lycopersicum L.).

The online version contains supplementary material available at 10.1007/s12298-021-00970-y.

Elicitor-Induced Defense Response in Soybean Plants Challenged by Bemisia tabaci

The whitefly, Bemisia tabaci Biotype B (Hemiptera: Aleyrodidae), is considered one of the world’s major agricultural pest groups, attacking a wide range of crop hosts and causing considerable crop loss. Understanding the interactions between whiteflies and host plants promotes the development of novel strategies for controlling whiteflies. This study aimed to evaluate the biochemical alterations caused by induced resistance in soybean plants, challenged by B. tabaci. The experiment was performed at the Federal University of Technology, Parana. Soybean seeds (cv. BRS 284) were sown in polyethylene pots in the greenhouse. The elicitors sprayed on the soybean leaves were: acibenzolar-S-Methyl (ASM-0.005%); Salicylic Acid (SA-2Mm); foliar phosphite (ULTRA K®-0.004%); Chitosan (CH-1%); and silicon (SI-0.25%); whilst the control group was sprayed with distilled water. Plants were infested with 50 adult whiteflies in the cages that were released after applying the treatments. At 0, 24, 48 and 96 hours after the beginning of the experiment, biochemical analyses of total proteins and the activity of the phenylalanine ammonia-lyase (PAL), peroxidases (PO), phenolic compounds, chitinase, and β-1,3-glucanase were made. The results showed that the application of elicitors increased the route of the phenylpropanoids with the activation of PAL and formation of phenolic compounds. It was also verified the activation of pathogenicity-related enzymes such as peroxidases and chitinase.

Synthetic Elicitor-Induced Defense Responses in Tomato (Solanum lycopersicum) Cultivated in Côte d'Ivoire against Bacterial Wilt Caused by Ralstonia solanacearum

Synthetic Elicitor-Induced Defense Responses in Tomato (Solanum lycopersicum) Cultivated in Côte d'Ivoire against Bacterial Wilt Caused by Ralstonia solanacearum

Elicitor-Induced Cellular and Molecular Events Are Responsible for Productivity Enhancement in Hairy Root Cultures: An Insight Study

A wide range of external stress stimuli triggers a plant cell to undergo a complex network of reactions that ultimately lead to the synthesis and accumulation of secondary metabolites. These secondary metabolites help the plant to survive under stress challenge. The potential of biotic and abiotic elicitors for the induction and enhancement of secondary metabolite production in various culture systems including hairy root (HR) cultures is well-known. The elicitor-induced defense responses involves signal perception of elicitor by a cell surface receptor followed by its transduction involving some major cellular and molecular events including activation of major secondary message signaling pathways. This result in induction of gene expressions escorting to the synthesis of various proteins mainly associated with plant defense responses and secondary metabolite synthesis and accumulation. The review discusses the elicitor-induced various cellular and molecular events and correlates them with enhanced secondary metabolite synthesis in HR systems. Further, this review also concludes that combining elicitation with in-silico approaches enhances the usefulness of this practice in better understanding and identifying the rate-limiting steps of biosynthetic pathways existing in HRs which in turn can contribute towards better productivity by utilizing metabolic engineering aspects.

Arabidopsis MPK3 and MPK6 Play Different Roles in Basal and Oligogalacturonide- or Flagellin-Induced Resistance against Botrytis cinerea

Mitogen-activated protein kinases (MAPKs) are fundamental components of the plant innate immune system. MPK3 and MPK6 are Arabidopsis (Arabidopsis thaliana) MAPKs activated by pathogens and elicitors such as oligogalacturonides (OGs), which function as damage-associated molecular patterns, and flg22, a well-known microbe-associated molecular pattern. However, the specific contribution of MPK3 and MPK6 to the regulation of elicitor-induced defense responses is not completely defined. In this work we have investigated the roles played by these MAPKs in elicitor-induced resistance against the fungal pathogen Botrytis cinerea. Analysis of single mapk mutants revealed that lack of MPK3 increases basal susceptibility to the fungus, as previously reported, but does not significantly affect elicitor-induced resistance. Instead, lack of MPK6 has no effect on basal resistance but suppresses OG- and flg22-induced resistance to B. cinerea. Overexpression of the AP2C1 phosphatase leads to impaired OG- and flg22-induced phosphorylation of both MPK3 and MPK6, and to phenotypes that recapitulate those of the single mapk mutants. These data indicate that OG- and flg22-induced defense responses effective against B. cinerea are mainly dependent on MAPKs, with a greater contribution of MPK6.

C-Terminal Region of Plant Ferredoxin-Like Protein Is Required to Enhance Resistance to Bacterial Disease in Arabidopsis thaliana

Protein phosphorylation is an important biological process associated with elicitor-induced defense responses in plants. In a previous report, we described how plant ferredoxin-like protein (PFLP) in transgenic plants enhances resistance to bacterial pathogens associated with the hypersensitive response (HR). PFLP possesses a putative casein kinase II phosphorylation (CK2P) site at the C-terminal in which phosphorylation occurs rapidly during defense response. However, the contribution of this site to the enhancement of disease resistance and the intensity of HR has not been clearly demonstrated. In this study, we generated two versions of truncated PFLP, PEC (extant CK2P site) and PDC (deleted CK2P site), and assessed their ability to trigger HR through harpin (HrpZ) derived from Pseudomonas syringae as well as their resistance to Ralstonia solanacearum. In an infiltration assay of HrpZ, PEC intensified harpin-mediated HR; however, PDC negated this effect. Transgenic plants expressing these versions indicate that nonphosphorylated PFLP loses its ability to induce HR or enhance disease resistance against R. solanacearum. Interestingly, the CK2P site of PFLP is required to induce the expression of the NADPH oxidase gene, AtrbohD, which is a reactive oxygen species producing enzyme. This was further confirmed by evaluating the HR on NADPH oxidase in mutants of Arabidopsis. As a result, we have concluded that the CK2P site is required for the phosphorylation of PFLP to enhance disease resistance.

Effect of sodium nitroprusside on growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root cultures

Nitric oxide (NO) is known as a signaling molecule involved in elicitor-induced defense responses of plants. Sodium nitroprusside (SNP), a donor of NO, stimulates catharanthine formation in Catharanthus roseus cells.1 Two important terpenoid indole alkaloids produced in small quantities within C. roseus are vinblastine and vincristine which are being used clinically as anticancer drugs. We are interested in engineering C. roseus hairy roots to increase the production of the TIAs. The present work investigates the effects of treating different concentrations of SNP to the hairy root cultures from line LBE-6-1. The alkaloid concentrations were analyzed 9, 14, 17, 20, 23, 26, and 30 days after treatment of SNP on day 0. We also studied the transient effects of SNP treatment during the exponential phase in C. roseus hairy roots. Analysis of the results showed that treatment of 0.1-mM SNP did not affect the growth of hairy roots, whereas 1-mM SNP suppressed the growth significantly, and 10-mM SNP almost completely inhibited the growth of hairy roots. 0.1-mM SNP treatment on day 0 caused a significant increase in the concentration of serpentine, catharanthine, ajmalicine, lochnericine and tabersonine production. SNP treatment on day 12 stimulated the formation of serpentine, catharanthine, ajmalicine, hörhammericine, lochnericine and tabersonine by day 21. After the initial stimulation, serpentine, horhammericine and lochnericine concentrations returned to the basal level by day 28. Treatment of 0.1-mM SNP on day 0 caused significant decrease in the mRNA levels for TDC, ASA, STR, ORCA3, ZCT1, and Crgbf1 on day 23. Treating 0.1-mM SNP on day 12 caused decreases in the expression levels of STR, ORCA3, ZCT1, and Crgbf1 on day 21 and day 28. Compared with day 28, the mRNA transcript of ZCT1 on day 21 is about twofold higher. Expression levels of G10H increased significantly.

A novel regulatory system in plants involving medium-chain fatty acids

Polyethylene glycol sorbitan monoacylates (Tween) are detergents of widespread use in plant sciences. However, little is known about the plant response to these compounds. Interestingly, the structure of Tweens' detergents (especially from Tween 20) resembles the lipid A structure from gram-negative bacteria polysaccharides (a backbone with short saturated fatty acids). Thus, different assays (microarray, GC-MS, RT-PCR, Northern blots, alkalinization and mutant analyses) were conducted in order to elucidate physiological changes in the plant response to Tween 20 detergent. Tween 20 causes a rapid and complex change in transcript abundance which bears all characteristics of a pathogenesis-associated molecular pattern (PAMP)/elicitor-induced defense response, and they do so at concentrations which cause no detectable deleterious effects on plant cellular integrity. In the present work, it is shown that the PAMP/elicitor-induced defense responses are caused by medium-chain fatty acids which are efficiently released from the Tween backbone by the plant, notably lauric acid (12:0) and methyl lauric acid. These compounds induce the production of ethylene, medium alkalinization and gene activation in a jasmonate-independent manner. Medium-chain fatty acids are thus novel elicitors/regulators of plant pathogen defense as they have being proved in animals.

Quantitative iTRAQ proteome and comparative transcriptome analysis of elicitor‐induced Norway spruce (Picea abies) cells reveals elements of calcium signaling in the early conifer defense response

Long-lived conifer trees depend on both constitutive and induced defenses for resistance against a myriad of potential pathogens and herbivores. In species of spruce (Picea spp.), several of the late events of pathogen-, insect-, or elicitor-induced defense responses have previously been characterized at the anatomical, biochemical, transcriptome, and proteome levels in stems and needles. However, accurately measuring the early events of induced cellular responses in a conifer is technically challenging due to limitations in the precise timing of induction and tissue sampling from intact trees following insect or fungal treatment. In the present study, we used the advantages of Norway spruce (Picea abies) cell suspensions combined with chitosan elicitation to investigate the early proteome response in a conifer. A combination of iTRAQ labeling and a new design of iterative sample analysis employing data-dependent exclusion lists were used for proteome analysis. This approach improved the coverage of the spruce proteome beyond that achieved in any prior study in a conifer system. Comparison of elicitor-induced proteome and transcriptome responses in Norway spruce cells consistently identified features associated with calcium-mediated signaling and response to oxidative stress that have not previously been observed in the response of intact trees to fungal attack.

Spent growth medium of Pantoea agglomerans primes wheat suspension cells for augmented accumulation of hydrogen peroxide and enhanced peroxidase activity upon elicitation

Induced disease resistance in plants is based on multiple mechanisms, including cell "priming", i.e. an enhancement of the capacity to mobilize cellular defense responses upon pathogen attack. Potent inducers of priming are, for example, salicylic acid, synthetic compounds such as a benzothiadiazole, and certain rhizosphere bacteria. While priming is well characterized for a number of dicot plants, only few cases of priming are documented in monocots. Here, we report that the spent growth medium of the Gram negative bacterium Pantoea agglomerans is capable of priming wheat cells (Triticum aestivum L. cv Prelude-Sr5) for elicitor-induced defense responses. Pre-incubation of suspension-cultured wheat cells with growth medium of P. agglomerans led to a strong enhancement of an oxidative burst that has been induced by chitin or chitosan and to an increase in extracellular peroxidase activity. Moreover, exopolysaccharides (EPS) were isolated from the spent growth medium and demonstrated to be sufficient for the induction of H2O2 priming. The EPS-induced priming was shown to be time- and concentration-dependent. We conclude that EPS are the or one of several priming-active component(s) in the spent growth medium of P. agglomerans. The present work is the first report of priming in a monocot plant by a specific component of bacterial origin. A comparison with known chemical inducers of resistance revealed that a benzothiadiazole was able to enhance the oxidative burst similar to the spent growth medium or the EPS of P. agglomerans, while salicylic acid was not.

Proteomics of Rac GTPase signaling reveals its predominant role in elicitor-induced defense response of cultured rice cells.

We have previously shown that a human small GTPase Rac homolog, OsRac1, from rice (Oryza sativa) induces cascades of defense responses in rice plants and cultured cells. Sphingolipid elicitors (SEs) have been similarly shown to activate defense responses in rice. Therefore, to systematically analyze proteins whose expression levels are altered by OsRac1 and/or SE treatment, we performed a differential display analysis of proteins by the use of two-dimensional gel electrophoresis and mass spectrometry. A total of 271 proteins whose expression levels were altered by constitutively active (CA)-OsRac1 or SE were identified. Interestingly, of 100 proteins that were up-regulated by a SE, 87 were also induced by CA-OsRac1, suggesting that OsRac1 plays a pivotal role in defense responses induced by SE in cultured rice cells. In addition, CA-OsRac1 induces the expression of 119 proteins. Many proteins, such as pathogenesis-related proteins, SGT1, and prohibitin, which are known to be involved in the defense response, were found among these proteins. Proteins involved in redox regulation, chaperones such as heat shock proteins, BiP, and chaperonin 60, proteases and protease inhibitors, cytoskeletal proteins, subunits of proteasomes, and enzymes involved in the phenylpropanoid and ethylene biosynthesis pathways were found to be induced by CA-OsRac1 or SE. Results of our proteomic analysis revealed that OsRac1 is able to induce many proteins in various signaling and metabolic pathways and plays a predominant role in the defense response in cultured rice cells.

Identification of a putative voltage‐gated Ca2+ channel as a key regulator of elicitor‐induced hypersensitive cell death and mitogen‐activated protein kinase activation in rice

Elicitor-triggered transient membrane potential changes and Ca2+ influx through the plasma membrane are thought to be important during defense signaling in plants. However, the molecular bases for the Ca2+ influx and its regulation remain largely unknown. Here we tested effects of overexpression as well as retrotransposon (Tos17)-insertional mutagenesis of the rice two-pore channel 1 (OsTPC1), a putative voltage-gated Ca(2+)-permeable channel, on a proteinaceous fungal elicitor-induced defense responses in rice cells. The overexpressor showed enhanced sensitivity to the elicitor to induce oxidative burst, activation of a mitogen-activated protein kinase (MAPK), OsMPK2, as well as hypersensitive cell death. On the contrary, a series of defense responses including the cell death and activation of the MAPK were severely suppressed in the insertional mutant, which was complemented by overexpression of the wild-type gene. These results suggest that the putative Ca(2+)-permeable channel determines sensitivity to the elicitor and plays a role as a key regulator of elicitor-induced defense responses, activation of MAPK cascade and hypersensitive cell death.

Cell Cycle Dependence of Elicitor-induced Signal Transduction in Tobacco BY-2 Cells

The molecular links between the cell cycle and defense responses in plants are largely unknown. Using synchronized tobacco BY-2 cells, we analyzed the cell cycle dependence of elicitor-induced defense responses. In synchronized cultured apoaequorin-expressing cells, the increase in cytosolic free Ca2+ induced by a proteinaceous elicitor, cryptogein, was greatly suppressed during the G2 and M phases in comparison with G1 or S phases. Treatment with cryptogein during the G1 or S phases also induced biphasic (rapid/transient and slow/prolonged) responses in activation of mitogen-activated protein kinases (MAPKs) and production of reactive oxygen species (ROS). In contrast, elicitor treatment during the G2 or M phases induced only a rapid and transient phase of MAPK activation and ROS production. Their slow and prolonged phases as well as expression of defense-related genes, cell cycle arrest and cell death were induced only after the cell cycle progressed to the G1 phase; removal of the elicitor before the start of the G1 phase inhibited these responses. These results suggest that although cryptogein recognition occurred at all phases of the cell cycle, the recognition during the S or G1 phases, but not at the G2 or M phases, induces the prolonged activation of MAPKs and the prolonged production of ROS, followed by cell cycle arrest, accumulation of defense-related gene transcripts and cell death. Elicitor signal transduction depends on the cell cycle and is regulated differently at each phase.

Involvement of nitric oxide in elicitor-induced defense responses and secondary metabolism of Taxus chinensis cells

This work was to characterize the generation of nitric oxide (NO) in Taxus chinensis cells induced by a fungal elicitor extracted from Fusarium oxysporum mycelium and the signal role of NO in the elicitation of plant defense responses and secondary metabolite accumulation. The fungal elicitor at 10-100 microg/ml (carbohydrate equivalent) induced a rapid and dose-dependent NO production in the Taxus cell culture, which exhibited a biphasic time course, reaching the first plateau within 1 h and the second within 12 h of elicitor treatment. The NO donor sodium nitroprusside potentiated elicitor-induced H2O2 production and cell death but had little influence on elicitor-induced membrane K+ efflux and H+ influx (medium alkalinization). NO inhibitors Nomega-nitro-L-arginine and 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide partially blocked the elicitor-induced H2O2 production and membrane ion fluxes. Moreover, the NO inhibitors suppressed elicitor-induced activation of phenylalanine ammonium-lyase and accumulation of diterpenoid taxanes (paclitaxel and baccatin III). These results suggest that NO plays a signal role in the elicitor-induced responses and secondary metabolism activities in the Taxus cells.

Differential regulation of MBP kinases by a glycoproptein elicitor and a polypeptide suppressor from Mycosphaerella pinodes in pea

A polypeptide fungal suppressor from a pea pathogen Mycosphaerella pinodes plays a key role in pathogenesis by suppressing elicitor-induced defense response(s) in pea ( Pisum sativum L). In this study, we show that treatment of pea tissues with the polysaccharide elicitor secreted by M. pinodes results in rapid increased activation of two myelin basic protein (MBP)-dependent kinases p44 (≈44 kDa) and p48 (≈48 kDa) within 15–30 min upon elicitation. Interestingly, the suppressor inhibited the elicitor-induced activation of only p44 kinase. While the defense-inducing signalling molecules, chitosan and salicylic acid (SA) activated the p44 and p48 kinases, methyl jasmonate (MeJA) did not. The abiotic stress signals, abscisic acid (ABA), NaCl and wounding activated the p48 kinase alone. These results demonstrate that MAPKs are differentially activated in response to pathogen invasion and abiotic stress in pea. Furthermore, specific inhibition of elicitor-induced p44 kinase activation by a MAPKK inhibitor, PD098059 and protein kinase inhibitor, K252a correlated with the suppression of elicitor-induced phenylalanine ammonia lyase (PAL) gene expression, supporting a role for p44 in the elicitor-induced defense response(s) in pea. Inhibition of p44 by the phosphoinositide (PI) turnover inhibitor, neomycin (a fungal suppressor mimic), and potentiation of p44 by the diacylglycerol (DAG) kinase inhibitor, R59022 indicated that p44 may be acting downstream of (PI) metabolism. Taken together, our results indicate that suppressor of defense elicitation from M. pinodes acts through inhibition of a MAPK (p44), possibly through a PI signaling pathway, facilitating the establishment of basic compatibility during infection of pea.

Identification and characterization of two new members of the GRAS gene family in rice responsive to N-acetylchitooligosaccharide elicitor

We identified two new members of the GRAS gene family from rice, CIGR1 and CIGR2, which are rapidly induced upon N-acetylchitooligosaccharide elicitor perception. The predicated proteins encoded by CIGR1 and CIGR2 possess significant sequence similarity with previously identified members of the GRAS family, such as Arabidopsis SCARECROW, GAI, RGA, tomato Lateral suppressor, and rice SLR1, all of which have VHIID regions, likely to play a role in cellular signaling. Fusions of CIGR1 and CIGR2 with Green Fluorescent Protein were detected exclusively in the nuclei of onion epidermal cells. The expression of CIGR1 and CIGR2 was dependent on the structure of N-acetylchitooligosaccharides, which parallels the structural specificity for chitin binding to the plasma membrane-localized chitin-binding protein, and independent of de novo protein synthesis. Co-cultivation of rice cells with rice blast fungus strongly induced the expression of CIGR1 and CIGR2, whereas inoculation of suspension cells with phytopathogenic bacteria did not. We hypothesize that CIGR1 and CIGR2 act as transcriptional regulators in the early events of the elicitor-induced defense response in rice.

A functional homolog of mammalian protein kinase C participates in the elicitor-induced defense response in potato.

The elicitor-induced activation of the potato pathogenesis-related gene PR-10a is positively controlled by a protein kinase(s) that affects the binding of the nuclear factors PBF-1 (for PR-10a binding factor-1) and PBR-2 to an elicitor response element (ERE). In this study, we have identified a kinase that has properties similar to the conventional isoenzymes of the mammalian protein kinase C (PKC) family. the treatment of potato tuber discs with specific inhibitors of PKC abolished the elicitor-induced binding of the nuclear factor PBF-2 to the ERE. This correlated with a reduction in the accumulation of the PR-10a protein. In contrast, treatment of the tuber discs with 12-O-tetradecanoylphorbol 13-acetate (TPA), an activator of PKC, led to an increase in binding of PBF-2 to the ERE and the corresponding increase in the level of the PR-10a protein, mimicking the effect seen with the elicitor arachidonic acid. Biochemical characterization of proteins extracted from the particulate fraction of potato tubers demonstrated that a kinase belonging to the conventional isoforms of PKC is present. This was confirmed by immunoprecipitation with antibodies specific to the conventional isoforms of human PKC and in-gel kinase assays. The ability of the immunoprecipitates to phosphorylate the alpha-peptide (a PKC specific substrate) in the presence of the coactivators calcium, phosphatidylserine, and TPA strongly suggested that the immunoprecipitated kinase is similar to the kinase characterized biochemically. Finally, the similar effects of the various modulators of PKC activity on the elicitor-induced resistance against a compatible race of Phytophthora infestans implicate this kinase in the overall defense response in potato.

A Functional Homolog of Mammalian Protein Kinase C Participates in the Elicitor-Induced Defense Response in Potato

A Functional Homolog of Mammalian Protein Kinase C Participates in the Elicitor-Induced Defense Response in Potato