Biochemical defence responses of resistant and susceptible rice genotypes against blast pathogen Magnaporthe oryzae

Abstract

Rice blast is the leading fungal disease which is caused by Magnaporthe oryzae that contributes for the significant decline in the rice yield throughout the globe. There is a need for the understanding of biochemical changes in rice plant during blast infection for the development of novel disease control strategies. In the present study, we isolated M. oryzae from the local paddy fields and the fungal isolates (VCF and PON) were identified by ITS-PCR using genomic DNA samples. Further, we inoculated resistant (BR2655 and TUNGA) and susceptible (INTAN and HR12) rice cultivars with PON and VCF isolates. PON isolate showed relatively high virulence compared to VCF and standard MTCC fungal strains. Therefore, we evaluated the effect of PON on the total protein content and plant defence-related key enzymes (peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, β-glucosidase, chitinase and lipoxygenase) activities between 24- and 120-hour post-inoculation (hpi). The results demonstrated the decrease in total protein content in all the inoculated cultivars. In addition, we observed the variation in the activity of peroxidase, polyphenol oxidase, β-glucosidase, chitinase and lipoxygenase at different time points in all the tested rice plants compared to respective controls. However, no significant difference was observed in the phenylalanine ammonia lyase activity relative to its control. Taken together, this study emphasizes on the variation in the activities of plant defence enzymes in different plant cultivars against the tested fungal pathogen and also implementation of defence enzymes as biochemical markers for resistant breeding.     

Expression of phenolics and defence-related enzymes in relation to red root rot disease of tea plants

Red root rot caused by Poria hypolateritia is a dreadful disease in tea plant due to sudden death of bushes. In response to fungal pathogen, variation in the defence-related enzymes was investigated. The infected tea root was undertaken to study about various defence-related and pathogen-related enzymes. The infected root, as a prime response to disease attack, was subjected to the analysis of phenolics, phenylalanine ammonia lyase, tyrosine ammonia lyase, peroxidase, polyphenol oxidase, catalase, chitinase, β-1,3-glucanase and protease were assayed. The results on assay of defence-related enzymes revealed that the activity was significantly higher in infected roots when compared with healthy roots. Phenolics were accumulated more in infected roots. The sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis further confirmed the presence of induced pathogenesis-related proteins in the infected root tissues. The activity of all enzymes was increased up to threefold amount when compared with normal ones. The accumulation of defence enzymes in plants revealed the virulence of root pathogen in stimulating induced systemic resistance of tea plants and phytopathogenicity causing pathogenesis. This study exemplify to recognise underlying processes in causing infection and to identify the existence of host–pathogen relationship.     

Control of root-knot nematode (Meloidogyne javanica) with combination of Arthrobotrys oligospora and salicylic acid and study of some plant defense responses

Root-knot nematodes (RKN) (Meloidogyne spp.) are economically the most important pathogens of agricultural products. The aim of the present study was to control Meloidogyne javanica by using Arthrobotrys oligospora and salicylic acid (SA) and to analyse the kinetics of enzymes, phenylalanine ammonia lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO) and phenolic compounds accumulation in the root system of tomato after inoculation with M. javanica, A. oligospora and SA. The ability of A. oligospora to produce extracellular proteases was also examined. In greenhouse studies, we used soil drenching of A. oligospora (10⁶ spores/ml) and soil drenching or leaf spraying of SA (5 mM) in six-leaf stage, separately and in combination. Experiments were performed in a completely randomised design. The efficiencies of treatments were appraised by using diameter of galls, number of galls per plant, number of egg masses per plant, number of eggs per egg mass, root and foliage fresh weight. The results showed that the combined application of A. oligospora and SA provided the best nematode control. The activity of the enzymes and phenolic compounds increased in comparison with the control. The nematophagous fungus A. oligospora produced extracellular proteases in the broth culture. Using A. oligospora and SA could be effective in control of M. javanica in tomato.     

Concomitant Induction of Systemic Resistance to Pseudomonas syringae pv. lachrymans in Cucumber by Trichoderma asperellum (T-203) and Accumulation of Phytoalexins

Most studies on the reduction of disease incidence in soil treated with Trichoderma asperellum have focused on microbial interactions rather than on plant responses. This study presents conclusive evidence for the induction of a systemic response against angular leaf spot of cucumber (Pseudomonas syringae pv. lachrymans) following application of T. asperellum to the root system. To ascertain that T. asperellum was the only microorganism present in the root milieu, plants were grown in an aseptic hydroponic growth system. Disease symptoms were reduced by as much as 80%, corresponding to a reduction of 2 orders of magnitude in bacterial cell densities in leaves of plants pretreated with T. asperellum. As revealed by electron microscopy, bacterial cell proliferation in these plants was halted. The protection afforded by the biocontrol agent was associated with the accumulation of mRNA of two defense genes: the phenylpropanoid pathway gene encoding phenylalanine ammonia lyase (PAL) and the lipoxygenase pathway gene encoding hydroxyperoxide lyase (HPL). This was further supported by the accumulation of secondary metabolites of a phenolic nature that showed an increase of up to sixfold in inhibition capacity of bacterial growth in vitro. The bulk of the antimicrobial activity was found in the acid-hydrolyzed extract containing the phenolics in their aglycone form. High-performance liquid chromatography analysis of phenolic compounds showed a marked change in their profile in the challenged, preelicited plants relative to that in challenged controls. The results suggest that similar to beneficial rhizobacteria, T. asperellum may activate separate metabolic pathways in cucumber that are involved in plant signaling and biosynthesis, eventually leading to the systemic accumulation of phytoalexins.     

Exogenous systemin has a contrasting effect on disease resistance in mycorrhizal tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis>) plants infected with necrotrophic or hemibiotrophic pathogens

A study was performed to determine the effect of the systemin polypeptide on the bio-protective effect of arbuscular mycorrhizal fungi (AMF) in tomato plants infected with Alternaria solani, Phytophthora infestans or P. parasitica. Before infection, tomato plants were colonized with two different AMF, Glomus fasciculatum or G. clarum. In addition, a group of inoculated plants was treated with systemin, just after emergence. The exogenous application of systemin marginally suppressed the resistance against A. solani leaf blight observed in G. fasciculatum mycorrhizal plants but significantly enhanced it in plants colonized with G. clarum. Systemin induced resistance to P. parasitica in leaves of G. fasciculatum mycorrhizal plants, in which AMF colonization alone was shown to have no protective effect. Conversely, none of the treatments led to resistance to root or stem rots caused by P. infestans or P. parasitica. The above effects did not correlate with changes in the activity levels of β-1,3-glucanase (BG), chitinase (CHI), peroxidase (PRX), and phenylalanine ammonium lyase (PAL) in leaves of infected plants. However, they corroborated previous reports showing that colonization by AMF can lead to a systemic resistance response against A. solani. Systemic resistance to A. solani was similarly observed in non-mycorrhizal systemin-treated plants, which, in contrast, showed increased susceptibility to P. infestans and P. parasitica. The results indicated that the pattern of systemic disease resistance conferred by mycorrhizal colonization was dependent on the AMF employed and could be altered by the exogenous application of systemin, by means of a still undefined mechanism.     

Salicylic acid induced systemic resistant on peanut against Alternaria alternata

Abstract

The effect of Salicylic Acid (SA) in inducing resistance in groundnut plants against Alternaria alternata was investigated. Foliar application of SA at the concentration of 1 mM significantly reduced the leaf blight disease intensity and increased the pod yield under glasshouse conditions. Changes in the activities of phenylalanine ammonium lyase, chitinase β-1,3 glucanase and in phenolic content on groundnut after application of SA and inoculation with A. alternate were studied. In SA-treated leaves (plants) an increase in phenolic content was observed five days after challenge inoculation with A. alternata in groundnut plants pretreated with SA. There was a marked increase in chitinase and pathogen inoculation in SA-treated leaves. In chitinase, β-1,3 glucanase activities were observed in response to plants with an increase in SA treated leaves. Foliar applications of SA-induced in peroxidase and polyphenol oxidase activities were observed upon challenge inoculation with pathogen.     

Evaluation of Ochrobactrum anthropi TRS-2 and its talc based formulation for enhancement of growth of tea plants and management of brown root rot disease

Aim: To evaluate Ochrobactrum anthropi TRS‐2 isolated from tea rhizosphere and its talc based formulation for growth promotion and management of brown root rot disease of tea. Methods and Results: Ochrobactrum anthropi TRS‐2, isolated from tea rhizosphere could solubilize phosphate, produce siderophore and IAA in vitro and also exhibited antifungal activity against six test pathogens. Application of an aqueous suspension of O. anthropi to the rhizosphere of nursery grown tea seedlings of five varieties of tea (TV‐18, T‐17, HV‐39, S‐449, UP‐3 and) led to enhanced growth of the treated plants, as evidenced by increase in height, in the number of shoots and number of leaves per shoot. Treatment with O. anthropi also decreased brown root rot of tea, caused by Phellinus noxius. Multifold increase in activities of chitinase, β‐1,3‐glucanase, peroxidase and phenylalanine ammonia lyase in tea plants was observed on application of O. anthropi to soil followed by inoculation with P. noxius. A concomitant increase in accumulation of phenolics was also obtained. Further, talc based formulation of O. anthropi was prepared and its survival determined every month up to a period of 12 months. Ochrobactrum anthropi could survive in the formulation up to a period of 9 months with a concentration of 7·0 log₁₀ CFU g^?1, after which there was a decline. Talc formulation was as effective as aqueous suspensions in both plant growth promotion and disease suppression. Conclusion: Ochrobactrum anthropi, either in aqueous suspension or as talc formulation induced growth of tea plants and suppressed brown root rot disease. It induced defense responses in tea plants. Significance and Impact of the Study: Ochrobactrum anthropi and its talc based formulation can be considered as an addition to available plant growth promoting rhizobacteria (PGPR) currently being used for field application. The present study offers a scope of utilizing this bacterium for growth promotion and disease management which would help in reduction of the use of chemicals in tea plantations.     

Concomitant induction of systemic resistance to Pseudomonas syringae pv. lachrymans in cucumber by Trichoderma asperellum (T-203) and accumulation of phytoalexins

Most studies on the reduction of disease incidence in soil treated with Trichoderma asperellum have focused on microbial interactions rather than on plant responses. This study presents conclusive evidence for the induction of a systemic response against angular leaf spot of cucumber (Pseudomonas syringae pv. lachrymans) following application of T. asperellum to the root system. To ascertain that T. asperellum was the only microorganism present in the root milieu, plants were grown in an aseptic hydroponic growth system. Disease symptoms were reduced by as much as 80%, corresponding to a reduction of 2 orders of magnitude in bacterial cell densities in leaves of plants pretreated with T. asperellum. As revealed by electron microscopy, bacterial cell proliferation in these plants was halted. The protection afforded by the biocontrol agent was associated with the accumulation of mRNA of two defense genes: the phenylpropanoid pathway gene encoding phenylalanine ammonia lyase (PAL) and the lipoxygenase pathway gene encoding hydroxyperoxide lyase (HPL). This was further supported by the accumulation of secondary metabolites of a phenolic nature that showed an increase of up to sixfold in inhibition capacity of bacterial growth in vitro. The bulk of the antimicrobial activity was found in the acid-hydrolyzed extract containing the phenolics in their aglycone form. High-performance liquid chromatography analysis of phenolic compounds showed a marked change in their profile in the challenged, preelicited plants relative to that in challenged controls. The results suggest that similar to beneficial rhizobacteria, T. asperellum may activate separate metabolic pathways in cucumber that are involved in plant signaling and biosynthesis, eventually leading to the systemic accumulation of phytoalexins.     

The Pathway and Regulation of Salicylic Acid Biosynthesis in Probenazole-Treated <Emphasis Type="Italic">Arabidopsis</Emphasis>

Probenazole (PBZ; 3-allyloxy-1,2-benzisothiazole-1,1-dioxide) is a highly effective chemical inducer of systemic-acquired resistance (SAR). It has been used widely to protect rice plants against the rice blast fungus Magnaporthe grisea. Previous studies have shown that PBZ induces SAR through enhanced accumulation of salicylic acid (SA). Plants synthesize SA by either a pathway that uses phenylalanine as substrate or another that involves isochorismate. To clarify how SA is produced in PBZ-treated Arabidopsis, we examined the expression patterns and enzyme activities of phenylalanine ammonia lyase (PAL) and isochorismate synthase (ICS), which are the main components of the phenylalanine and isochorismate pathways, respectively. PBZ exposure significantly improved the accumulation of SA and increased ICS activity. In the sid2–2 mutant, which has a defect in ICS1, PBZ had no effect on the level of endogenous SA or activity of ICS. In contrast, PAL activity and the expression of most PAL genes were down-regulated by such treatment in wild-type plants. These results suggest that SA is mainly synthesized via the ICS-mediated pathway in Arabidopsis.     

Comparative studies on the activities of chitinase, β-1,3-glucanase, peroxidase and phenylalanine ammonia lyase in the leaves of triticale and wheat infected with Stagonospora nodorum

The activities of chitinase, β-1,3-glucanase, peroxidase and phenylalanine ammonia lyase, constitutive and induced by Stagonospora nodorum were examined in the 10 – 14 day old seedlings of three triticale and two wheat cultivars under controlled environmental conditions and in flag leaves of two triticale cultivars in the field. Two S. nodorum isolates of different virulence were used. Both the constitutive and induced activities in triticale and wheat depended on genotype and in triticale the effect of growth conditions was also evidenced. The constitutive activities of chitinase, β-1,3-glucanase and peroxidase were several fold lower in flag triticale leaves in plants from the field than in the seedlings, growing under controlled conditions, but induction in the infected flag leaves was significantly more pronounced. In triticale genotypic differences in the response to infection were revealed only upon inoculation by S. nodorum isolate of higher virulence. The enzymatic activities increased several fold during successive days after the infection except for phenylalanine ammonia lyase. Induction of this enzyme was only transient and the activity decreased 48 or 96 h after infection when the activities of other enzymes were rising. In flag leaves in the field this activity was differentiated only after infection with more a virulent strain. A tendency appeared in triticale seedlings for association of the resistance to the pathogen with lower enzymatic constitutive activities. This relationship became more evident in triticale infected by S. nodorum and may imply that although the investigated enzymes are certainly involved in general, non-specific defense mechanism, they do not decide on the resistance to pathogen at least in the early stages of infection and cooperate with other factors in the complex pathogen-plant interaction. One can also assume that the enzymatic activities are associated with severity of infection rather than resistance to pathogen.     

Rhizobacterial bioformulation for the effective management of Macrophomina root rot in mungbean

Abstract

Fluorescent pseudomonads based bioformulation was evaluated for their ability to control Macrophomina root rot disease in mungbean (Vigna mungo). P. fluorescens isolate Pf1 showed the maximum inhibition in mycelial growth of Macrophomina phaseolina under in vitro conditions. Bioformulation of Pf1 with chitin was effective in reducing the root rot incidence in green gram both under glasshouse and field conditions. The rhizosphere colonization of P. fluorescens was observed appreciable with the green gram plants. However, Pf1 amended with chitin colonized effectively. Furthermore, the induction of defence-related enzymes and chemicals in plants by Pf1 amended with or without chitin and neem were tested. Increased accumulation of defence enzymes viz., phenylalanine ammonia lyase (PAL), peroxidase (PO), polyphenol oxidase (PPO), chitinase, β-1,3-glucanse and phenolics were observed in Pf1 bioformulation amended with chitin, pre-treated plants challenge inoculated with M. phaseolina under glasshouse conditions. The present study reveals that in addition to direct antagonism and plant-growth promotion, PGPR strains amended with chitin bioformulation induced defence-related enzymes and pathogenesis related (PR) proteins which collectively enhance the resistance in green gram against the infection of M. phaseolina.     

Induction of Defence Responses in Roots and Mesocotyls of Sorghum Seedlings by Inoculation with Fusarium thapsinum and F. proliferatum, Wounding and Light

The defence reactions of sorghum seedlings 7 days after inoculation with Fusarium thapsinum and F. proliferatum, and interactions with wounding and exposure to light were studied to determine whether responses to these fungi differed from those to abiotic stresses. In non‐wounded plants, inoculation with both fungi increased concentrations of anthocyanins and soluble phenolics and activities of peroxidase (POX), chitinase and β‐1,3‐glucanase in the roots, and increased β‐1,3‐glucanase activity in the mesocotyls. There was no effect of inoculation on phenylalanine ammonia‐lyase (PAL) activity. Wounding by itself increased anthocyanin content of mesocotyls. Wounding also had a variety of interactions with inoculation. Exposure to light had very little effect on any defence response measured. A time course experiment showed that induction of chitinase and β‐1,3‐glucanase occurred in less than 24 h after inoculation. POX activity increased 2 days after inoculation, followed by a transient increase in PAL activity. The content of anthocyanins and soluble phenolics in roots of inoculated seedlings increased gradually compared with controls over 6 days. The responses of sorghum seedlings to inoculation with F. thapsinum and F. proliferatum were similar to those found by other workers following challenge by necrotrophic pathogens and were different from those induced by wounding and exposure to light.     

Chitin-supplemented Foliar Application of Serratia marcescens GPS 5 Improves Control of Late Leaf Spot Disease of Groundnut by Activating Defence-related Enzymes

Chitinolytic Serratia marcescens GPS 5 and non‐chitinolytic Pseudomonas aeruginosa GSE 18, with and without supplementation of chitin, were tested for their ability to activate defence‐related enzymes in groundnut leaves. Thirty‐day‐old groundnut (cv. TMV 2) plants pretreated with GPS 5 and GSE 18 (with and without supplementation of 1% colloidal chitin) were challenge inoculated after 24 h with Phaeoisariopsis personata, the causal agent of late leaf spot (LLS) disease of groundnut. GPS 5 and GSE 18, applied as a prophylactic spray, reduced the lesion frequency by 23% and 67%, respectively, compared with control. Chitin supplementation had no effect on the control of LLS by GSE 18, unlike GPS 5, which upon chitin supplementation reduced the lesion frequency by 64%, compared with chitin alone. In a time course study the activities of chitinase, β‐1,3‐glucanase, peroxidase and phenylalanine ammonia lyase were determined for the different treatments. There was an enhanced activity of the four defence‐related enzymes with all the bacterial treatments when compared with phosphate buffer and colloidal chitin‐treated controls. In correlation to disease severity in bacterial treatments, chitin‐supplemented GSE 18 was similar to GSE 18, whereas chitin‐supplemented GPS 5 was much more effective than GPS 5, in activation of the defence‐related enzymes. The high levels of enzyme activities following chitin‐supplemented GPS 5 application continued up to the measured 13 days after pathogen inoculation.     

Biocontrol of aerial plant diseases in agriculture and horticulture: current approaches and future prospects

Until recently, the majority of research on the biological control of aerial plant diseases was focused on control of bacterial pathogens. Such research led to the commercialization of the biocontrol agent Pseudomonas fluorescens A506, as BlightBan A506™, for control of fire blight of pear. In contrast, chemical fungicides typically have provided adequate control of most foliar fungal pathogens. However, fungicide resistance problems, concerns regarding pesticide residues and revocation of registration of certain widely used fungicides have led to increased activity in the development of biocontrol agents of foliar fungal pathogens. Much of this activity has centered around the use of Trichoderma spp and Gliocladium spp to control Botrytis cinerea on grape and strawberry. The biocontrol agent Trichoderma harzianum T39 is commercially available in Israel, as Trichodex ™, for control of grey mold in grapes and may soon be registered for use in the US. Also targeted primarily against a foliar disease of grapes, in this case powdery mildew caused by Uncinula necator, is the biocontrol agent Ampelomyces quisqualis AQ10, marketed as AQ^{10  TM} biofungicide. Another promising development in the area of foliar disease control, though one which is not yet commercialized, is the use of rhizobacteria as seed treatments to induce systemic resistance in the host plant, a strategy which can protect the plant against a range of bacterial and fungal pathogens. Received 06 February 1997/ Accepted in revised form 05 June 1997     

Hydrogen Sulfide Prolongs Postharvest Storage of Fresh-Cut Pears (Pyrus pyrifolia) by Alleviation of Oxidative Damage and Inhibition of Fungal Growth

Hydrogen sulfide (H₂S) has proved to be a multifunctional signaling molecule in plants and animals. Here, we investigated the role of H₂S in the decay of fresh-cut pears (Pyrus pyrifolia). H₂S gas released by sodium hydrosulfide (NaHS) prolonged the shelf life of fresh-cut pear slices in a dose-dependent manner. Moreover, H₂S maintained higher levels of reducing sugar and soluble protein in pear slices. H₂S significantly reduced the accumulation of hydrogen peroxide (H₂O₂), superoxide radicals (•O₂ ⁻) and malondialdehyde (MDA). Further investigation showed that H₂S fumigation up-regulated the activities of antioxidant enzymes ascorbate peroxidase (APX), catalase (CAT), and guaiacol peroxidase (POD), while it down-regulated those of lipoxygenase (LOX), phenylalanine ammonia lyase (PAL) and polyphenol oxidase (PPO). Furthermore, H₂S fumigation effectively inhibited the growth of two fungal pathogens of pear, Aspergillus niger and Penicillium expansum, suggesting that H₂S can be developed as an effective fungicide for postharvest storage. The present study implies that H₂S is involved in prolonging postharvest storage of pears by acting as an antioxidant and fungicide.     

Change in phenylalanine ammonia lyase activity and isozyme patterns of polyphenol oxidase and peroxidase by salicylic acid leading to enhance resistance in cowpea against <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

In this paper, we describe a study concerning the determination of phenylalanine ammonia lyase (PAL) activities leading to decline in disease progress caused by Rhizoctonia solani after application of salicylic acid (SA). Two applications of 1.4 mM SA (pH 6.5) followed by inoculation with Rhizoctonia solani resulted in a quantitative change in polyphenol oxidase (PPO), peroxidase (PRX) isoforms and increase in PAL activities from 4.38 to 19.48 unit g⁻¹ (FM) h⁻¹ in Bundel-1, UPC-4200 and IFC-902 cowpea genotypes. Increase in PAL activities was further observed specifically in UPC-4200 when plants were exposed with Rhizoctonia solani spores. Total soluble proteins did not change after SA treatment, however they were significantly increased in SA sprayed-inoculated UPC-4200 genotype. Of the ten detected isoforms of polyphenol oxidase, isoforms 7 and 10, and isoform 4 of peroxidase showed increased activities by SA application. The disease symptoms measured as areas under progress curve (AUDPC) indicated less A value in SA sprayed Bundel-1 and UPC-4200 genotypes over their controls.     

Hyaluronic acid of Streptococcus sp. as a potent elicitor for induction of systemic resistance against plant diseases

The potential of hyaluronic acid (HA) in inducing systemic resistance to cucumber, tomato and pepper was tested in planta. In the study, HA was found to be a potent agent for suppressing disease caused by Cucumber Mosaic Virus (CMV) (in pepper), Pseudomonas syringae pv. tomato (tomato speck disease), Xanthomonas axonopodis pv. vesicatoria (tomato spot disease), Pseudomonas syringae pv. lachrymans (cucumber angular leaf spot), and Colletotrichum orbiculare (cucumber anthracnose). Disease control was obtained with spraying, injection and drenching of plants with HA. HA did not exhibit direct antimicrobial action against the pathogens tested. Studies carried out in transgenic tobacco indicated that defense genes PR 1a and PDF 1.2 were activated upon treatment with HA, demonstrating salicylic acid (SA) and jasmonic acid (JA) pathways getting activated during defense. Further work is warranted to evaluate the use of HA-mediated disease suppression in crop plants.     

Influence of beneficial microorganisms during in vivo acclimatization of in vitro-derived tea (<Emphasis Type="Italic">Camellia sinensis</Emphasis>) plants

The effects of native isolates of Pseudomonas fluorescens, Azospirillum brasilense, and Trichoderma harzianum on rooting and acclimatization of in vitro-grown shoots and plantlets of tea were evaluated. In vitro bacterization of P. fluorescens failed to establish, while both T. harzianum and A. brasilense retarded shoot growth, eventually overtaking shoot cultures in in vitro rooting. Acclimatization of rooted plantlets in soil amended with bioinoculants, either individually or in various combinations, promoted plantlet survival. Moreover, efficiency of nutrient uptake of plantlets was higher in the presence of microorganisms. Root rot or wilting of tissue culture-derived plants was not observed in bioinoculant-treated plants, as they possessed relatively higher activities of defense enzymes, including peroxidase and phenylalanine ammonia lyase.     

Induced defense-related proteins in soybean (Glycine max L. Merrill) plants by Carnobacterium sp. SJ-5 upon challenge inoculation of Fusarium oxysporum

The aim of the present study was to analyze induced expression of defense-related proteins in the soybean plants by rhizobacterial stain Carnobacterium sp. SJ-5 upon challenge inoculation with Fusarium oxysporum. Determination of the enzymatic activity of the different defense-related enzymes, phenylalanine ammonia lyase (PAL), lipoxygenase (LOX), peroxidase (POD) and polyphenol oxidase (PPO) was performed in the major parts of Glycine max L. Merrill using spectrophotometric method. Native-polyacrylamide gel electrophoresis analysis of the POD and PPO was employed followed by activity staining to find out the isoforms of respective enzymes. Activities of the PAL, LOX, POD and PPO were found to be highest in the bacterized root tissue of the soybean plants challenged with F. oxysporum. Isoform analysis revealed that PPO1, PPO4 and POD2 isoforms were expressed at higher levels in bacterized soybean root tissues challenge inoculated with the pathogen. Conclusively it was found that bacterial strain Carnobacterium sp. SJ-5 protect soybean plants from wilt disease caused by F. oxysporum by elicitation of the defense-related enzymes.