Induction of Defense Responses in Cucumber Plants by Using the Cell-free Filtrate of the Plant Growth-Promoting Fungus Penicillium simplicissimum GP17-2 (original) (raw)
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Plant growth-promoting fungi (PGPF) such as Phoma sp. (isolates GS8-1, GS8-2 and GS8-3) and nonsporulating fungus (isolate GU21-2) were tested for their ability to induce systemic resistance against Colletotrichum orbiculare in cucumber. These isolates, used as colonized barley kernels to natural soils, induced systemic resistance in the greenhouse as well as in the field. Different elicitors from PGPF such as cell walls retaining or lacking protein and lipids, cell walls lipid fraction, and three fractions of culture filtrate (CF) with different molecular weight ranges were tested for their ability to elicit the defense response of cucumber plants under controlled conditions. Cell wall fraction lacking protein and lipids and the cell wall lipid fraction from root colonizing isolates GS8-1, GS8-2, and GS8-3 protected plants against C. orbiculare. While, only cell wall lipid fraction and CF fractions of different molecular weight ranges of the isolate GU21-2 protected plants against C. orbiculare infection. Methanol-soluble substances from CF fractions with molecular weight greater than 12,000 and less than 8000 consistently protected plants against the pathogen. Germination of C. orbiculare spores decreased significantly at 72 h of incubation on leaves of PGPF-protected plants. Isolate GU21-2 was the most effective in inhibiting spore germination. On the other hand, isolates GS8-1 and GU21-2 induced lignifications in the hypocotyls of seven-day-old cucumber seedlings after challenge inoculation with C. orbiculare. Three-week-old cucumber plants treated with isolate GU21-2 and challenged with C. orbiculare showed increased activities of exo-and endo-forms of glucanase and chitinase, as well as peroxidase and polyphenol oxidase in the second true leaves. Induction treatment with isolate GS8-1 also increased the activities of these enzymes with the exception of exo-glucanase. This study shows that the inoculation of PGPF or its CF resulted in additive effect on the suppression of anthracnose disease in cucumber.
Physiological and molecular plant pathology, 2006
Expression profiles for major cell wall strengthening genes, lignin peroxidase (LPO), callose synthase (CALS), caffeoyl-CoA 3-O-methyl transferase (CCOAMT) and cinnamyl alcohol dehydrogenase (CAD) as well as reactive oxygen species forming and scavenging genes namely, Cu/Zn superoxide dismutase (Cu/Zn SOD) and catalase (CAT) genes were examined in cucumber during acibenzolar-S-methyl (ASM) induced systemic resistance against anthraconse disease caused by Colletotrichum orbiculare. The time course study revealed early increased expressions of LPO, CALS and Cu/Zn SOD genes, while transcripts of CCOAMT, CAD and CAT remained unchanged. The early gene expression of LPO and CALS coincided with subsequent deposition of lignin and callose which generally occurs within 24h after inoculation of the fungal pathogen on ASM treated plants. SOD enzyme activities were increased after 24h of treatment and enhanced CAT activity was noticed at 12 and 48h after treatment in inoculated plants. These results reveal that only certain cell wall strengthening genes (LPO and CALS) and the production and scavenging of ROS processes are involved in systemic acquired resistance induced by ASM in cucumber.
Susceptible and resistant cultivars of cucumber (cv. Suyo and cv. Shogoin fushinari, respectively) were evaluated for induction of systemic resistance by plant growth-promoting fungal isolates Phoma sp. GS8-1 and nonsporulating fungus GU21-2 against Colletotrichum orbiculare. Reduction in anthracnose disease was assessed at regular intervals of 0, 3, 6, and 9 days after challenge inoculation. Activities of endochitinase, β-1, 3-endoglucanase, peroxidase (PO), polyphenol oxidase, and phenylalanine ammonia lyase (PAL) were assayed at the above intervals, while the activities of exochitinase and β-1,3-exogluconase were assayed at 9 days after challenge inoculation. The data showed that all the enzymes activities increased in plants treated with plant growth-promoting fungi (PGPF) compared with nontreated plants of both cultivars. The activities of endochitinase and β-1,3-endoglucanase were highly increased in PGPF-induced plants than in nontreated plants of both cultivars at different time points. Activities of PO, polyphenol oxidase, and PAL also increased between 3 and 6 days due to inoculation with PGPF and decreased thereafter. The activity of PAL at 9 days in PGPF-induced plants was at the same level as that of nontreated plants of both cultivars. Results of this investigation suggest that PGPF-induced systemic resistance is associated with not only high increases in the activities of chitinase and β-1,3-gluconases but also other plant defense-related enzymes such as PO, PPO, and PAL. Roles of other mechanisms rather than accumulation of defense-related enzymes in PGPF-mediated induced resistance in susceptible cultivars were demonstrated.
How do plants defend themselves against pathogens-Biochemical mechanisms and genetic interventions
Physiology and Molecular Biology of Plants, 2022
In agro-ecosystem, plant pathogens hamper food quality, crop yield, and global food security. Manipulation of naturally occurring defense mechanisms in host plants is an effective and sustainable approach for plant disease management. Various natural compounds, ranging from cell wall components to metabolic enzymes have been reported to protect plants from infection by pathogens and hence provide specific resistance to hosts against pathogens, termed as induced resistance. It involves various biochemical components, that play an important role in molecular and cellular signaling events occurring either before (elicitation) or after pathogen infection. The induction of reactive oxygen species, activation of defensive machinery of plants comprising of enzymatic and non-enzymatic antioxidative components, secondary metabolites, pathogenesis-related protein expression (e.g. chitinases and glucanases), phytoalexin production, modification in cell wall composition, melatonin production, c...
Activation of defense-related genes in parsley leaves by infection withErwinia chrysanthemi
European Journal of Plant Pathology, 1995
Parsley leaves exhibited incomplete resistance following stomatal inoculation with the broad host range pathogen, Erwinia chrysanthemi, resulting in the development of local lesions in 58% of inoculated leaves. The responses of the same plant material to five mutants generated from the wild type bacterial strain varied from highly susceptible to completely resistant. While the susceptible phenotype was characterized by spreading maceration, hypersensitivelike necrotic lesions were typical for resistant leaves, with both types of symptom becoming visible two days after inoculation. Accumulation of transcripts of an entire set of defense-related genes and of furanocoumarin phytoalexins preceded visible onset of lesion development or maceration in the respective interactions. Escherichia coli did not elicit symptoms, gene activation or phytoalexin accumulation in parsley leaves. No correlation existed between the final extent of plant resistance on the one hand, and the timing or degree gene activation and phytoalexin accumulation on the other hand. Rapid activation of defense-related genes therefore appears to be insufficient, although it may be necessary, to protect parsley from attack by Erwinia chrysanthemi.
Suppression of Induced Resistance in Cucumber Through Disruption of the Flavonoid Pathway
Phytopathology, 2005
In this study, cucumber plants (Cucumis sativus) expressing induced resistance against powdery mildew (caused by Podosphaera xanthii) were infiltrated with inhibitors of cinnamate 4-hydroxylase, 4-coumarate:CoA ligase (4CL), and chalcone synthase (CHS) to evaluate the role of flavonoid phytoalexin production in induced disease resistance. Light and transmission electron microscopy demonstrated ultrastructural changes in inhibited plants, and biochemical analyses determined levels of CHS and β-glucosidase enzyme activity and 4CL protein accumulation. Our results showed that elicited plants displayed a high level of induced resistance. In contrast, down regulation of CHS, a key enzyme of the flavonoid pathway, resulted in nearly complete suppression of induced resistance, and microscopy confirmed the development of healthy fungal haustoria within these plants. Inhibition of 4CL ligase, an enzyme largely responsible for channeling phenylpropanoid metabolites into the lignin pathway, ha...
Plant defense-related enzymes against pathogens: A Review
Plant disease control is mainly based on the use of fungicides, bactericides, and insecticides-chemical compounds toxic to plant invaders, causative agents, or vectors of plant diseases. However, the detrimental effect of these chemicals or their degradation products on the environment and human health strongly imposes the search for novel, harmless means of disease control. Therefore, it is essential to introduce environmentally-friendly alternative measures for management of plant diseases. Induced plant resistance is one of the promising non-chemical strategies for the effective management of diseases. The host plant mediated resistance is governed by defense response genes encoding for production of various pathogenesis-related (PR) proteins. This review chiefly explains the biochemical response of plant defense mechanism pertaining to defense-related enzymes which have been identified as PR proteins.
The role of plant defence proteins in fungal pathogenesis
Molecular Plant Pathology, 2007
It is becoming increasingly evident that a plant-pathogen interaction may be compared to an open warfare, whose major weapons are proteins synthesized by both organisms. These weapons were gradually developed in what must have been a multimillionyear evolutionary game of ping-pong. The outcome of each battle results in the establishment of resistance or pathogenesis. The plethora of resistance mechanisms exhibited by plants may be grouped into constitutive and inducible, and range from morphological to structural and chemical defences. Most of these mechanisms are defensive, exhibiting a passive role, but some are highly active against pathogens, using as major targets the fungal cell wall, the plasma membrane or intracellular targets. A considerable overlap exists between pathogenesis-related (PR) proteins and antifungal proteins. However, many of the now considered 17 families of PR proteins do not present any known role as antipathogen activity, whereas among the 13 classes of antifungal proteins, most are not PR proteins. Discovery of novel antifungal proteins and peptides continues at a rapid pace. In their long coevolution with plants, phytopathogens have evolved ways to avoid or circumvent the plant defence weaponry. These include protection of fungal structures from plant defence reactions, inhibition of elicitor-induced plant defence responses and suppression of plant defences. A detailed understanding of the molecular events that take place during a plant-pathogen interaction is an essential goal for disease control in the future. Comparison of three pathogenesis-related proteins from plants of two cultivars of tobacco infected with TMV. J. Gen. Virol. 47, 79-87. Argos, P., Narayana, S.V. and Nielsen, N.C. (1985) Structural similarity between legumin and vicilin storage proteins from legumes. EMBO J. 4, 1111-1117. Arie, M., Hikichi, K., Takahashi, K. and Esaka, M. (2000) Characterization of a basic chitinase which is secreted by cultured pumpkin cells. Physiologia Plantarum, 110, 232-239. Ary, M.B., Richardson, M. and Shewry, P.R. (1989) Purification and characterization of an insect alpha-amylase inhibitor/endochitinase from seeds of Job's Tears (Coix lachryma-jobi ). Biochim. Biophys. Acta, 999, 260-266. Localized changes in peroxidase activity accompany hydrogen peroxide generation during the development of a nonhost hypersensitive reaction in Lettuce. Plant Physiol. 118, 1067-1078. Blackwell, J. (1988) Physical methods for the determination of chitin. structure and conformation. Methods Enzymol. 161, 435-442. Blilou, I., Ocampo, J.A. and Garcia-Garrido, J.M. (2000) Induction of Ltp (lipid transfer protein) and Pal (phenylalanine ammonia-lyase) gene expression in rice roots colonized by the arbuscular mycorrhizal fungus Glomus mosseae. HPLC analysis of grapevine phytoalexins coupling photodiode array detection and fluorometry. Anal. Chem. 69, 5172-5177. Jeandet, P., Douillet-Breuil, A.C., Bessis, R., Debord, S., Sbaghi, M. and Adrian, M. (2002) Phytoalexins from the Vitaceae: biosynthesis, phytoalexin gene expression in transgenic plants, antifungal activity, and metabolism. Constitutive expression of a celery mannitol dehydrogenase in tobacco enhances resistance to the mannitol-secreting fungal pathogen Alternaria alternata. Plant J. 32, 41-49. Jones, D.A. and Takemoto, D. (2004) Plant innate immunity-direct and indirect recognition of general and specific pathogen-associated molecules. Curr. Opin. Immunol. 16, 48-62. Jongedijk, E., Tigelaar, H., Roekel, J.S.C., Bres-Vloemans, S.A., Dekker, I., Elzen, P.J.M., Cornelissen, B.J.C. and Melchers, L.S. (1955) Synergistic activity of chitinases and beta-1,3-glucanases enhances fungal resistance in transgenic tomato plants. Euphytica, 85, 173-180. Jordá, L., Conejero, V. and Vera, P. (2000) Characterization of P69E and P69F, two differentially regulated genes encoding new members of the subtilisin-like proteinase family from tomato plants. Plant Physiol. 122, 67-74. Joshi, B.N., Sainani, M.N., Bastawade, K.B., Gupta, V.S. and Ranjekar, P.K. (1998) Cysteine protease inhibitor from pearl millet: a new class of antifungal protein. A novel yeast gene, RHK1, is involved in the synthesis of the cell wall receptor for the HM-1 killer toxin that inhibits beta-1,3-glucan synthesis. N-glycosylation is involved in the sensitivity of Saccharomyces cerevisiae to HM-1 killer toxin secreted from Hansenula mrakii IFO 0895. Appl. Microbiol. Biotechnol. 51, 176-184. Kitajima, S. and Sato, F. (1999) Plant pathogenesis-related proteins: molecular mechanisms of gene expression and protein function.