Defense Responses In Roots Research Articles

A genome-wide association study reveals cytokinin as a major component in the root defense responses against Ralstonia solanacearum.

Bacterial wilt caused by the soil-borne pathogen Ralstonia solancearum is economically devastating, with no effective methods to fight the disease. This pathogen invades plants through their roots and colonizes their xylem, clogging the vasculature and causing rapid wilting. Key to preventing colonization are the early defense responses triggered in the host’s root upon infection, which remain mostly unknown. Here, we have taken advantage of a high-throughput in vitro infection system to screen natural variability associated with the root growth inhibition phenotype caused by R. solanacearum in Arabidopsis during the first hours of infection. To analyze the genetic determinants of this trait, we have performed a genome-wide association study, identifying allelic variation at several loci related to cytokinin metabolism, including genes responsible for biosynthesis and degradation of cytokinin. Further, our data clearly demonstrate that cytokinin signaling is induced early during the infection process and cytokinin contributes to immunity against R. solanacearum. This study highlights a new role for cytokinin in root immunity, paving the way for future research that will help in understanding the mechanisms underpinning root defenses.

Effects of Salicylic Acid and Indole Acetic Acid Exogenous Applications on Induction of Faba Bean Resistance against Orobanche crenata.

The parasitic weed, Orobanche crenata, is one of the most devastating constraint for faba bean production in Mediterranean regions. Plant host defense induction was reported as one of the most appropriate control methods in many crops. The aim of this study was to elucidate the effect of salicylic acid (SA) and indole acetic acid (IAA) on the induction of faba bean resistance to O. crenata under the field and controlled experimental conditions. Both hormones were tested on two contrasting faba bean genotypes: Giza 843 (partially resistant to O. crenata) and Lobab (susceptible) at three different application methods (seed soaking, foliar spray, and the combination of both seed soaking and foliar spray). Soaking seeds in SA or IAA provided the highest protection levels reaching ~75% compared to the untreated control plants. Both elicitors limited the chlorophyll content decrease caused by O. crenata infestation and increased phenolic compound production in host plants. Phenylalanine ammonia lyase, peroxidase, and polyphenol oxidase activities were stimulated in the host plant roots especially in the susceptible genotype Lobab. The magnitude of induction was more obvious in infested than in non-infested plants. Histological study revealed that both SA and IAA decreased the number of attached O. crenata spikes which could be related to specific defense responses in the host plant roots.

Proteomic and Transcriptomic Analyses Indicate Metabolic Changes and Reduced Defense Responses in Mycorrhizal Roots of Oeceoclades maculata (Orchidaceae) Collected in Nature.

Orchids form endomycorrhizal associations with fungi mainly belonging to basidiomycetes. The molecular events taking place in orchid mycorrhiza are poorly understood, although the cellular changes necessary to accommodate the fungus and to control nutrient exchanges imply a modulation of gene expression. Here, we used proteomics and transcriptomics to identify changes in the steady-state levels of proteins and transcripts in the roots of the green terrestrial orchid Oeceoclades maculata. When mycorrhizal and non-mycorrhizal roots from the same individuals were compared, 94 proteins showed differential accumulation using the label-free protein quantitation approach, 86 using isobaric tagging and 60 using 2D-differential electrophoresis. After de novo assembly of transcriptomic data, 11,179 plant transcripts were found to be differentially expressed, and 2175 were successfully annotated. The annotated plant transcripts allowed the identification of up- and down-regulated metabolic pathways. Overall, proteomics and transcriptomics revealed, in mycorrhizal roots, increased levels of transcription factors and nutrient transporters, as well as ethylene-related proteins. The expression pattern of proteins and transcripts involved in plant defense responses suggested that plant defense was reduced in O. maculata mycorrhizal roots sampled in nature. These results expand our current knowledge towards a better understanding of the orchid mycorrhizal symbiosis in adult plants under natural conditions.

Plant microbiota modified by plant domestication

Human life became largely dependent on agricultural products after distinct crop-domestication events occurred around 10,000 years ago in different geographical sites. Domestication selected suitable plants for human agricultural practices with unexpected consequences on plant microbiota, which has notable effects on plant growth and health. Among other traits, domestication has changed root architecture, exudation, or defense responses that could have modified plant microbiota. Here we present the comparison of reported data on the microbiota from widely consumed cereals and legumes and their ancestors showing that different bacteria were found in domesticated and wild plant microbiomes in some cases. Considering the large variability in plant microbiota, adequate sampling efforts and function-based approaches are needed to further support differences between the microbiota from wild and domesticated plants. The study of wild plant microbiomes could provide a valuable resource of unexploited beneficial bacteria for crops.

Mild cadmium stress induces auxin synthesis and accumulation, while severe cadmium stress causes its rapid depletion in barley root tip

The aim of this study was to investigate the effect of 4-phenoxyphenylboronic acid (PPBo), an effective inhibitor of YUCCA enzyme on mild, moderate and severe Cd stress-induced responses in the root tips of young barley seedlings. While mild and moderate Cd stresses increased, severe Cd stress decreased the root IAA level. PPBo post-treatment strongly inhibited the Cd-induced IAA accumulation without affecting the Cd level in roots and mitigated the mild Cd stress-induced root growth inhibition, root swelling and lipoxygenase and glutathione peroxidase activity. In addition, PPBo reduced the H2O2 and NO generation under mild but had no effect under severe Cd stress, indicating that both H2O2 and NO act downstream of IAA in root responses to mild Cd stress. In turn, moderate or severe Cd stress strongly reduced the expression of IAA synthesis and cell cycle genes in the barley root tip. Therefore, we can conclude that IAA accumulation in the barley root tips is a distinctive feature of mild and moderate Cd stresses, which is involved in both root growth reorientation and defense response activation. PPBo completely reversed this Cd-induced IAA accumulation and stress responses, indicating that Cd induces IAA synthesis via indole-3-pyruvic acid biosynthesis pathway. On the contrary, severe Cd stress reduced IAA level was coupled with high ROS in the root tips and root growth arrest.

A nucleoside diphosphate kinase gene OsNDPK4 is involved in root development and defense responses in rice (Oryza sativa L.)

Dysfunctional mutation of OsNDPK4 resulted in severe defects in root development of rice. However, the resistance of Osndpk4 against bacterial blight was significantly enhanced. Nucleoside diphosphate kinases (NDPKs) are an evolutionarily conserved family of important enzymes balancing the energy currency nucleoside triphosphates by catalyzing the transfer of their phosphate groups. The aim of this study was to elucidate the function of OsNDPK4 in rice. A dysfunctional rice mutant was employed to characterize the function of OsNDPK4. Its expression and subcellular localization were examined. The transcriptomic change in roots of Osndpk4 was analyzed by RNA-seq. The rice mutant Osndpk4 showed severe defects in root development from the early seedling stage. Further analysis revealed that meristematic activity and cell elongation were significantly inhibited in primary roots of Osndpk4, together with reduced accumulation of reactive oxygen species (ROS). Map-based cloning identified that the mutation occurred in the OsNDPK4 gene. OsNDPK4 was found to be expressed in a variety of tissues throughout the plant and OsNDPK4 was located in the cytosol. Osndpk4 showed enhanced resistance to the bacterial pathogen Xanthomonas oryzae pv. oryzae (Xoo) and up-regulation of pathogenesis-related marker genes. In addition, transcriptomic analysis showed that OsNDPK4 was significantly associated with a number of biological processes, including translation, protein modification, metabolism, biotic stress response, etc. Detailed analysis revealed that the dysfunction of OsNDPK4 might reorchestrate energy homeostasis and hormone metabolism and signalling, resulting in repression of translation, DNA replication and cell cycle progression, and priming of biotic stress defense. Our results demonstrate that OsNDPK4 plays important roles in energy homeostasis, development process, and defense responses in rice.

Expression of the Legume-Specific Nod Factor Receptor Proteins Alters Developmental and Immune Responses in Rice

Legumes form symbiosis with rhizobia, which fix nitrogen for the benefit of host plant in return for carbon resources. Development of this unique symbiosis in legumes is triggered by rhizobia-secreted nodulation (Nod) factors (NFs). NFs, upon perception, activate Nod signaling cascade, leading to reprogramming of host cell (root) developmental networks to pave way for accommodating rhizobial symbionts. A long-cherished goal of legume-rhizobia symbiosis research is to extend this symbiotic nitrogen-fixing capacity to cereal plants such as rice. As a part of achieving this ultimate goal, in this work, initially we expressed legume-specific Nod factor receptor protein (NFRP) genes, MtNFP, MtLYK3, and LjLNP, in rice and assessed their impact on NF perception and consequently triggered biological responses in roots. RNA-seq analysis revealed that roots of both control and NFRP-expressing plants perceive NFs, but NFs elicited contrasting impacts on gene expression patterns in roots of these plants. In contrast to suppressive role of NFs on expression of several genes involved in innate immune response in roots of control plants, in NFRP-expressing plants, NFs triggered massive upregulation of a vast array of genes associated with signaling, defense response, and secondary metabolism networks in roots. Expression of NFRPs in rice also conferred root hairs the ability to respond to NFs in terms of exhibiting deformations, albeit at low levels. Together, results of the study demonstrated that rice plants have inherent ability to perceive NFs, but the expression of legume NFRPs rendered rice roots hypersensitive to NFs.

Effect of Aluminum Stress on Onion (Allium cepa L.)

This study analyzed the effect of Aluminium (Al) on onion (Allium cepa) root growth and its defense response. The experiment was carried out by exposing onion to 10, 100, 500 and 1000 µM AlCl3 for 24, 48 and 72 hour. Parameters observed were root length and diameter, root tip mitotic index, cortical parenchyma cell length and cortical cell layer thickness. Onion defense response observed by measuring mallic, citric and oxalic acid quantitatively, and the accumulation of Al and callose in root qualitatively. Results showed that an increase in the concentration of Al caused a decrease in root length. At concentrations up to 100 µM, Al increased root diameter cell length whilst at higher concentrations, Al decreased both parameters. Compared to control, all Al-treated root had lower index mitosis with 10 uM Al-treated root displayed highest IM. Since root treated with Al higher than 10 µM showed none or very low mitotic activity, chromosome aberration can only be observed in 10 uM Al–treated roots. Root defense response via Al and callose accumulation was increased with the increasing of Al concentrations. Organic acids were present in higher amounts in root than in medium with a ratio of 5000:1. Oxalic acid appeared to be the major organic acid that played a role in defense mechanisms against Al stress. Based on the results, it can be concluded that (1) Al3+ were toxic to Allium cepa because it caused root growth inhibition and chromosomal aberrations; (2) defense response for Al3+ stress were via the increase of callose as well as Al accumulation and organic acids content in root cells.

Jasmonate Signal Receptor Gene Family ZmCOIs Restore Male Fertility and Defense Response of Arabidopsis mutant coi1-1

Jasmonates (JAs) play an important role in many developmental processes, such as root growth, leaf senescence, male fertility, and defense responses against insects and pathogens. The F-box protein COI1, which plays a central role in JA signal transduction, perceives the JA signal and is required for all the JA-mediated defense responses against biotic and abiotic stresses. JA signaling elements including COI1 have been extensively investigated in Arabidopsise. However, the elements of the JA signaling pathway in maize are largely unknown. In this study, we identified four F-box protein genes from the maize genome, which share high homology with AtCOI1, designated as ZmCOI1a, ZmCOI1b, ZmCOI1c, and ZmCOI2, collectively ZmCOIs. To test whether or not the homologous genes of maize are functionally conservative in JA signaling, we over-expressed ZmCOIs in the Arabidopsis coi1-1 mutant. The results showed that over-expression of ZmCOI1a, ZmCOI1b or ZmCOI1c in the coi1-1 mutant resulted in the restoration of male fertility, indicating successful complementation of coi1-1 sterility by ZmCOI1a, ZmCOI1b, and ZmCOI1c. However, ZmCOI2 was not able to restore male fertility of the mutant, indicating that ZmCOI2 has a function diverged from JA signaling. Furthermore, over-expression of the ZmCOI1a, ZmCOI1b, and ZmCOI1c genes, except ZmCOI2, which, in the coi1-1 mutant, caused restoration of resistance to the leaf pathogen Botrytis cinerea and the soil-borne pathogen Pythium aristosporum. In addition, a set of JA-dependent genes are highly induced by wounding in the transformants of ZmCOIa, ZmCOI1b, and ZmCOI1c, but not inducible in transformants of ZmCOI2 or in the coi1-1 mutant, indicating that ZmCOIa, ZmCOI1b, and ZmCOI1c, except ZmCOI2, which can compensate coi1-1 mutation of Arabidopsis for the stress defense response. Putting all the data together, our results suggested that ZmCOIa, ZmCOI1b, and ZmCOI1c, but not ZmCOI2, act as AtCOI1 orthologues in maize for JA signal transduction.

Wheat Resistances to Fusarium Root Rot and Head Blight Are Both Associated with Deoxynivalenol- and Jasmonate-Related Gene Expression.

Fusarium graminearum is a major pathogen of wheat causing Fusarium head blight (FHB). Its ability to colonize wheat via seedling root infection has been reported recently. Our previous study on Fusarium root rot (FRR) has disclosed histological characteristics of pathogenesis and pathogen defense that mirror processes of spike infection. Therefore, it would be interesting to understand whether genes relevant for FHB resistance are induced in roots. The concept of similar-acting defense mechanisms provides a basis for research at broad Fusarium resistance in crop plants. However, molecular defense responses involved in FRR as well as their relation to spike resistance are unknown. To test the hypothesis of a conserved defense response, a candidate gene expression study was conducted to test the activity of selected prominent FHB defense-related genes in seedling roots, adult plant roots, spikes, and shoots. FRR was examined at seedling and adult plant stages to assess age-related pattern of disease and pathogen resistance. This study offers first evidence for a significant genetic overlap in root and spike defense responses, both in local and distant tissues. The results point to plant development-specific rather than organ-specific determinants of resistance, and suggest roots as an interesting model for studies on wheat-Fusarium interactions.

Identification of major quantitative trait loci for root diameter in synthetic hexaploid wheat under phosphorus-deficient conditions.

Synthetic hexaploid wheat (SHW) possesses numerous genes for resistance to stress, including phosphorus (P) deficiency. Root diameter (RDM) plays an important role in P-deficiency tolerance, but information related to SHW is still limited. Thus, the objective of this study was to investigate the genetic architecture of RDM in SHW under P-deficient conditions. To this end, we measured the RDM of 138F9 recombinant inbred lines derived from an F2 population of a synthetic hexaploid wheat line (SHW-L1) and a common wheat line (Chuanmai32) under two P conditions, P sufficiency (PS) and P deficiency (PD), and mapped quantitative trait loci (QTL) for RDM using an enriched high-density genetic map, containing 120,370 single nucleotide polymorphisms, 733 diversity arrays technology markers, and 119 simple sequence repeats. We identified seven RDM QTL for P-deficiency tolerance that individually explained 11-14.7% of the phenotypic variation. Five putative candidate genes involved in root composition, energy supply, and defense response were predicted. Overall, our results provided essential information for cloning genes related to P-deficiency tolerance in common wheat that might help in breeding P-deficiency-tolerant wheat cultivars.

Plant endophytic Pseudomonas putida BP25 induces expression of defense genes in black pepper roots: Deciphering through suppression subtractive hybridization analysis

Black pepper associated endophytic Pseudomonas putida BP25 displayed broad spectrum antagonistic activity against a wide range of plant pathogens. The molecular responses of black pepper upon endophytic colonization by P. putida BP25 was studied using a PCR based suppression subtractive hybridization (SSH) method. The transcripts induced by endophytic P. putida BP25 in black pepper included pathogenesis related proteins such as PR-4 and PR-1; reactive oxygen species (ROS) scavenging catalase and metallothionein; stress induced glutathione S-transferase; 5-enolpyruvylshikimate-3-phosphate synthase involved in phenylpropanoid metabolism. Differential expression of transcripts was validated by quantitative real time PCR (qPCR) for selected ESTs. This information helps in understanding the ability of an endophytic strain P. putida BP25 in inducing a wide array of defence responses in black pepper roots.

The effect of gibberellin and abscisic acid on plant defense responses and on disease severity caused by Meloidogyne javanica on tomato plants

Stimulating plant innate resistance is a choice method for developing safe alternatives to harmful chemical nematicides to control root-knot nematodes. Plant hormones have a key role in inducing resistance. Here, the ability of gibberellin and abscisic acid to induce biochemical defense responses in tomato roots was investigated by monitoring the activity of cytoplasmic enzymes phenylalanine ammonia lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO) and catalase (CAT) every other day for the first week after inoculation with Meloidogyne javanica, then weekly for 8 weeks. Two second-stage juveniles were added to 1 g of sandy loam soil in each pot. Tomato seedlings were sprayed with gibberellin (GA) or abscisic acid (ABA) when free of nematodes and in pots with M. javanica (GA + N and ABA + N). There were also healthy plants (C) as well as plants that were inoculated only with M. javanica (N). The highest PAL, POX and PPO activity was found in gibberellin-sprayed plants during the first week, demonstrating that gibberellin induced a plant defense response. The amount of enzymes then decreased during weeks 2 and 3 and rose at the fourth week, then decreased weekly till the end of the experiment. On the basis of plant growth and nematode reproduction, gibberellin stimulated plant innate defense responses and increase plant resistance, whereas abscisic acid decreased plant resistance. Therefore, spraying tomato plants with gibberellin is recommended to control M. javanica.

Exogenous glutamate rapidly induces the expression of genes involved in metabolism and defense responses in rice roots

BackgroundGlutamate is an active amino acid. In addition to protein synthesis and metabolism, increasing evidence indicates that glutamate may also function as a signaling molecule in plants. Still, little is known about the nutritional role of glutamate and genes that are directly regulated by glutamate in rice.ResultsExogenous glutamate could serve as a nitrogen nutrient to support the growth of rice seedlings, but it was not as effective as ammonium nitrate or glutamine. In nitrogen-starved rice seedlings, glutamate was the most abundant free amino acid and feeding of glutamate rapidly and significantly increased the endogenous levels of glutamine, but not glutamate. These results indicated that glutamate was quickly metabolized and converted to the other nitrogen-containing compounds in rice. Transcriptome analysis revealed that at least 122 genes involved in metabolism, transport, signal transduction, and stress responses in the roots were rapidly induced by 2.5 mM glutamate within 30 min. Many of these genes were also up-regulated by glutamine and ammonium nitrate. Still, we were able to identify some transcription factor, kinase/phosphatase, and elicitor-responsive genes that were specifically or preferentially induced by glutamate.ConclusionsGlutamate is a functional amino acid that plays important roles in plant nutrition, metabolism, and signal transduction. The rapid and specific induction of transcription factor, kinase/phosphatase and elicitor-responsive genes suggests that glutamate may efficiently amplify its signal and interact with other signaling pathways to regulate metabolism, growth and defense responses in rice.

NADP-dependent enzymes are involved in response to salt and hypoosmotic stress in cucumber plants.

Salt stress is one of the most damaging plant stressors, whereas hypoosmotic stress is not considered to be a dangerous type of stress in plants and has been less extensively studied. This study was performed to compare the metabolism of cucumber plants grown in soil with plants transferred to distilled water and to a 100 mM NaCl solution. Even though hypoosmotic stress caused by distilled water did not cause such significant changes in the relative water content, Na+/K+ ratio and Rubisco content as those caused by salt stress, it was accompanied by more pronounced changes in the specific activities of NADP-dependent enzymes. After 3 days, the specific activities of NADP-isocitrate dehydrogenase, glucose-6-phosphate dehydrogenase, NADP-malic enzyme and non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase in leaves were highest under hypoosmotic stress, and lowest in plants grown in soil. In roots, salt stress caused a decrease in the specific activities of major NADP-enzymes. However, at the beginning of salt stress, NADP-galactose-1-dehydrogenase and ribose-1-dehydrogenase were involved in a plant defense response in both roots and leaves. Therefore, the enhanced demands of NADPH in stress can be replenished by a wide range of NADP-dependent enzymes.

AtMYB44 Positively Regulates the Enhanced Elongation of Primary Roots Induced by N-3-Oxo-Hexanoyl-Homoserine Lactone in Arabidopsis thaliana.

N-acyl-homoserine lactones (AHL) are the quorum-sensing (QS) signal molecules used by many gram-negative bacteria to coordinate their collective behavior in a population. Recent evidence demonstrates their roles in plant root growth and defense responses. AtMYB44 is a multifaceted transcriptional factor that functions in many physiological processes in plants but whether AtMYB44 modulates the plant response to AHL with aspects of primary root elongation remains unknown. Here, we show that the expression of AtMYB44 was upregulated upon treatment with N-3-oxo-hexanoyl-homoserine lactone (3OC6-HSL). The stimulatory effect of 3OC6-HSL on primary root elongation was abolished in the AtMYB44 functional-deficiency mutant atmby44. In contrast, an enhanced promoting-impact of 3OC6-HSL on primary root growth was observed in AtMYB44-overexpressing plant MYB44OTA. Cellular analysis indicated that the prolonged primary root elicited by 3OC6-HSL is the consequence of increased cell division in the meristem zone and enhanced cell elongation in the elongation zone, and AtMYB44 may act as a positive regulator in this process. Furthermore, we demonstrated that AtMYB44 might participate in the 3OC6-HSL-mediated primary root growth via regulating the expression of cytokinin- and auxin-related genes. The data establish a genetic connection between the regulatory role of AtMYB44 in phytohormones-related gene expression and plant response to the bacterial QS signal.

Effects of silicon treatment and inoculation with Fusarium oxysporum f. sp. vasinfectum on cellular defences in root tissues of two cotton cultivars.

Silicon has been shown to enhance the resistance of plants to fungal and bacterial pathogens. Here, the effect of potassium silicate was assessed on two cotton (Gossypium hirsutum) cultivars subsequently inoculated with Fusarium oxysporum f. sp. vasinfectum (Fov). Sicot 189 is moderately resistant whilst Sicot F-1 is the second most resistant commercial cultivar presently available in Australia. Transmission and light microscopy were used to compare cellular modifications in root cells after these different treatments. The accumulation of phenolic compounds and lignin was measured. Cellular alterations including the deposition of electron-dense material, degradation of fungal hyphae and occlusion of endodermal cells were more rapidly induced and more intense in endodermal and vascular regions of Sicot F-1 plants supplied with potassium silicate followed by inoculation with Fov than in similarly treated Sicot 189 plants or in silicate-treated plants of either cultivar not inoculated with Fov. Significantly more phenolic compounds were present at 7 d post-infection (dpi) in root extracts of Sicot F-1 plants treated with potassium silicate followed by inoculation with Fov compared with plants from all other treatments. The lignin concentration at 3 dpi in root material from Sicot F-1 treated with potassium silicate and inoculated with Fov was significantly higher than that from water-treated and inoculated plants. This study demonstrates that silicon treatment can affect cellular defence responses in cotton roots subsequently inoculated with Fov, particularly in Sicot F-1, a cultivar with greater inherent resistance to this pathogen. This suggests that silicon may interact with or initiate defence pathways faster in this cultivar than in the less resistant cultivar.

Transcriptome changes specifically associated with apple (Malus domestica) root defense response during Pythium ultimum infection

RNA-seq technology was applied to identify the transcriptomic changes associated with apple root defense response to Pythium ultimum infection. Genes encoding homolog proteins with functions of pathogen detection such as chitin elicitor receptor kinase (CERK) and wall-associated receptor kinase (WAK) were among the differentially expressed apple genes. The biosynthesis and signaling of several plant hormones including ethylene, jasmonate and cytokinin were specifically induced in response to P. ultimum inoculation. Genes encoding enzymes of secondary metabolisms, cell wall fortification and pathogenesis related (PR) protein, laccase, mandelonitrile lyase and cyanogenic beta-glucosidase were consistently up-regulated in the later stages of infection.

Tracing Plant Defense Responses in Roots upon MAMP/DAMP Treatment.

This chapter describes how to apply microbe-associated molecular pattern (MAMP) or damage-associated molecular pattern (DAMP) solutions to Arabidopsis roots to trace defense responses in the root. Plants sense the presence of microbes via the perception of MAMPs or DAMPs by surface-localized pattern recognition receptors. The mechanisms governing plant root immunity are poorly characterized compared with those underlying plant immunity in the leaf, despite the fact that plant roots constantly interact with countless microbes living in soils that carry potential MAMPs and could stimulate the production of plant-derived DAMPs during colonization. To understand how a plant root immune system detects and reacts to the potential sources of a stimulus, we describe a simple method to monitor activation of root immunity upon MAMP/DAMP treatment by measuring relative expression of defense-related genes by RT-qPCR.

Salicylic acid alleviates cadmium-induced stress responses through the inhibition of Cd-induced auxin-mediated reactive oxygen species production in barley root tips

Auxin is a master regulator of root growth by modulating its development under the constantly changing environment. Recently, an antagonistic interaction was suggested between SA and IAA signaling. Therefore, the purpose of this work was to analyze and compare the effect of the indole-3-acetic acid (IAA) signaling inhibitor p-chlorophenoxyisobutyric acid (PCIB) and salicylic acid (SA) as a potential IAA signaling inhibitor on the root growth, enzyme activity and reactive oxygen species (ROS) production in Cd- and IAA-treated barley root tips. Exposure of plants to Cd resulted in a more than threefold increase of IAA content in the root apex even 3h after the treatment. In addition, exogenously applied IAA evoked root responses such as root growth inhibition and swelling, ROS generation and activation of lipoxygenase or glutathione peroxidase identical to those induced by Cd. Furthermore, both Cd- and IAA-induced stress responses were markedly reversed by PCIB or SA post-treatment. Similarly to PCIB, SA did not affect the IAA content of root tips, suggesting the action of SA on the IAA signaling pathway in barley roots. SA probably does not alleviate the Cd toxicity in roots, but rather prevents or partially inhibits the root defense response to the presence of Cd through the inhibition of Cd-induced IAA-mediated ROS generation in roots.