Role Of Chitinases Research Articles

Statistical optimization of medium components for chitinase production by Pseudomonas fluorescens strain HN1205: role of chitinase on egg hatching inhibition of root-knot nematode

In the present research, statistical Plackett–Burman design (PBD) and central composite design (CCD) were used to optimize the medium's components in order to enhance the chitinase activity of Pseudomonas fluorescens strain HN1205. Among the seven nutritional elements that were studied, crab shell powder, CaCl2·2H2O and yeast extract were proved using PBD to have significant effect on chitinase activity. An optimal medium was obtained by applying a three-factor central composite design, which consisted of crab shell powder (1.0 g/L), CaCl2 (0.5 g/L) and yeast extract (0.5 g/L), with the highest chitinase activity of 1.03 U/mL. This value was 2.87-fold higher than the activity obtained in the lowest productive medium in the design matrix. The chitinase produced by the HN1205 strain was partially purified with 80% ammonium sulphate and anion-exchange chromatography and assessed for inhibition of hatching of nematode eggs. The partially purified chitinase significantly reduced the hatch of Meloidogyne incognita eggs (8.1%), as compared to the effect of 10 mmol/L Tris-HCl buffer (pH 8.0) (49.4%) and boiled chitinase (54.7%). This study demonstrated the role of chitinolytic enzymes produced by P. fluorescens strain HN1205. The obtained optimal activity proved that the enzymes could be potential biological control agents.

Role of chitinase 3-like-1 and semaphorin 7a in pulmonary melanoma metastasis.

The prototypic chitinase-like protein Chi3l1 is induced in cancers and portends a poor prognosis, but whether it contributes to cancer progression is unknown. To address this gap in knowledge, we investigated the production of Chi3l1 in melanoma lung metastases. We found that Chi3l1 was induced during pulmonary melanoma metastasis and that this induction was regulated by the semaphorin Sema7a, interacting in stimulatory or inhibitory ways with its β1 integrin or Plexin C1 receptors, respectively. In mouse strains with genetic deletions of Chi3l1 or Sema7a, there was a significant reduction in pulmonary metastasis. Notably, antiserum raised against Chi3l1 or Sema7a phenocopied the reduction produced by genetic deletions. Melanoma lung metastasis was also decreased in the absence of IL13Rα2, a recently identified receptor for Chi3l1, consistent with a key role for Chi3l1 in melanoma spread. We confirmed roles for Sema7a and Chi3l1 in pulmonary metastasis of EMT6 breast cancer cells. Taken together, our studies establish a novel pathway through which Sem7a and its receptors regulate Chi3l1, revealing a host axis involving IL13Rα2 that plays a critical role in generating a pulmonary microenvironment that is critical to license metastasis.

Potential use and mode of action of the new strain Bacillus thuringiensis UM96 for the biological control of the grey mould phytopathogen Botrytis cinerea

The potential use of Bacillus thuringiensis UM96 as a biocontrol agent for the grey mould phytopathogen Botrytis cinerea was evaluated. In order to dissect the mode of action of this UM96 strain, we also examined the role of lytic activities in the antagonism. First, B. thuringiensis UM96 was characterised based on 16S rRNA and gyrA gene sequencing and phenotypic traits. Petri dish biocontrol assays demonstrated that when strain UM96 was inoculated 24 h previous to B. cinerea, the mycelial growth was inhibited by up to 70%. Test for lytic enzymes activities of cellulase and glucanase was negative. Chitinase was the only positive enzyme activity in two different culture media. PCR detection of the chiB gene was also positive. Chitinolytic supernatants, obtained from rich and minimal media supplemented with colloidal chitin as the sole carbon source, from B. thuringiensis UM96 showed a strong inhibitory effect of B. cinerea that was not observed with heat-treated supernatant. Interestingly, when the supernatant was supplemented with 100 µM allosamidin, a chitinase specific inhibitor, the antagonistic activity was suppressed significantly. A lack of chitinase activity was also observed in allosamidin-treated supernatants. Our pathogenic B. cinerea strain also exhibited susceptibility to pure Streptomyces griseus chitinase. Finally, the chitinolytic strain B. thuringiensis UM96 was able to protect Medicago truncatula plants in vitro from B. cinerea infection and significantly reduced the necrotic zones and root browning of the plants. Together, these results suggest a potential use of B. thuringiensis UM96 for the biological control of B. cinerea and a role for chitinases during the antagonism displayed.

Increased expression and possible role of chitinase 3-like-1 in a colitis-associated carcinoma model.

To investigate the possible role of chitinase 3-like-1 (CHI3L1) in the progression of colitis-associated carcinoma (CAC). Thirty-four Balb/c mice were randomly assigned to five groups, including the control, CAC control, CAC + caffeine, colitis control and colitis + caffeine. Three animals were sacrificed every two weeks for blinded macroscopic inspection, histological analysis, and total RNA extraction. An immunofluorescent assay was performed using specimens from the colitis control and colitis + caffeine groups to investigate whether the protective effect of caffeine was associated with less oxidative DNA damage. In vitro, HT29 cells pre-stimulated with different concentrations of recombinant CHI3L1 protein and H2O2 were loaded with the DCFH-DA fluorescent probe to determine the effect of CHI3L1 on intracellular reactive oxygen species production. CHI3L1 mRNA was increased during the progression of colon carcinogenesis. Tumors were mostly located in the distal end of the colon where the expression of CHI3L1 was higher than in the proximal colon. Caffeine-treated mice developed fewer tumors and milder inflammation than untreated mice. CHI3L1 protein increased reactive oxygen species in HT29 cells when exposed to H2O2. Caffeine reduces tumor incidence by decreasing oxidative DNA damage. CHI3L1 may contribute to CAC by increasing reactive oxygen species production.

Defense gene expression in Sorghum bicolor against Macrophomina phaseolina in leaves and roots of susceptible and resistant cultivars

The fungal disease, charcoal root rot, caused by Macrophomina phaseolina is a foremost yield restraining factor of Sorghum bicolor L. around the world. The expression analysis of genes induced in general defense response can endow with clues to reveal major defense mechanisms against pathogen infection in sorghum plant. The role of chitinase and Stilbene synthase in response to M. phaseolina in sorghum was studied under control growth conditions using a real-time polymerase chain reaction. Here, we report the expression analysis of antifungal genes in two cultivars viz. PJ-1430 (resistant) and SU-1080 (susceptible) at different hours after inoculation with M. phaseolina isolate MTCC 2165. Chitinase and stilbene synthase were induced in PJ-1430 within 0 h, 24 h and in SU-1080 in 48 h, 24 h, respectively, after inoculation. However, the expression levels of chitinase and stilbene synthase in resistant cultivar were significantly higher. The results showed that chitinase and stilbene synthase can be effective to enhance resistance to M. phaseolina.

The Role of Chitinases in Atopic Dermatitis

Amac: Bu calismada atopik dermatitli hastalardan alinan deri biyopsilerinde asidik memeli kitinaz (AMCase) gen ekspresyonu incelendi. Hastalar ve yontemler: Mayis 2005 Mayis 2007 tarihleri arasinda atopik dermatitli bes yetiskin hasta calismaya dahil edildi. Lezyonlu ve lezyonsuz bolgelerden deri biyopsileri alindi. Gercek zamanli polimeraz zincir reaksiyonu ile AMCase ekspresyonu incelendi. AMCase gen urunleri, beta-aktin ile karsilastirildi. Bulgular: Atopik lezyondan alinan numunelere kiyasla, AMCase yapimi derinin saglikli bolgelerinde daha yuksek oranda idi. Saglikli deri ve atopik deri dokusunda AMCase gen ekspresyonuna rastlandi. Sonuc: AMCase gen ekspresyonu her iki deri bolgesinde izole edildi. Bulgularimiz bu genin sadece mide ve akcigerde sinirli olmadigini ayni zamanda deride de bulundugunu gostermektedir. Anahtar sozcukler: Atopik dermatit; kitin mikropartikulu; kitin; kitinaz. Objectives: In this study, we investigated the acidic mammalian chitinase (AMCase) gene expression in skin biopsy samples taken from patients with atopic dermatitis. Patients and methods: Five adult patients with atopic dermatitis were enrolled in this study between May 2005 and May 2007. Skin biopsy samples were taken from lesional and unaffected areas. AMCase gene expression was tested by real-time polymerase chain reaction. The AMCase gene products were compared with beta-actin. Results: The AMCase production was higher in healthy regions of skin compared to the samples taken from atopic lesion. The AMCase gene expression was isolated in healthy and atopic skin regions. Conclusion: The AMCase gene expression was isolated both skin regions. Our study shown that this gene is not only expressed stomach and lung and also expressed in skin.

The role of chitinases and glucanases in somatic embryogenesis of black pine and hybrid firs

AbstractGlucanase and chitinase enzymes play an important role in different plant processes including defense against pathogens and morphogenesis. Moreover, their role in the processes of somatic embryogenesis has been demonstrated. It has been suggested, that the presence of this type of proteins might be a marker for embryogenic potential of callus cultures. In this work we screened for the presence of glucanases and chitinases in liquid growth media of a set of conifer embryogenic cell lines in order to find correlation with their embryogenic potential. We have found that none of the 12 chitinase isoforms detected in culture media of Pinus nigra Arn. or the nine chitinases detected in media with Abies alba × A. cephalonica and Abies alba × A. numidica embryogenic tissues could be linked to their embryogenic capacity. Similarly, none of the six glucanase isoforms detected in the extracellular fluid of Pinus nigra Arn. cultures can be assigned as a marker of embryogenic potential. Thus, our data indicate the large variability and doubtless importance of glucanases and chitinases for cell growth and development of somatic embryos, however, do not support the premise that they are markers of embryogenesis.

Invitro Screening of Antifungal Activity of Rhizospheric Bacteria and Possible Role of Chitinase in Antifungal Activity

The use of biocontrol agents is becoming an increasingly important alternative to chemical crop protection against weeds, insects and plant diseases in the field of agriculture. The success of biocontrol and yield increase depends on the nature of antagonistic properties and mechanisms of action of the biocontrol agent against the phytopathogens. In this study, 103 macroscopically different bacterial isolates (62 from Kirtipur and 41 from Khumaltar) from 21 different rhizosphere soil samples (11 from Kirtipur and 10 from Khumaltar) were screened for antagonism against five fungal phytopathogens, viz, Fusarium oxysporum, Alternaria solani, Sclerotium rolfsii, Exserohilum turcicum and Phytophthora infestans by dual culture technique on Potato dextrose agar. Out of 18 different active isolates two of them showed the chitinolytic potential and the most active fungal antagonist was identified as Bacillus subtilis on the basis of colonial, morphological, physiological and biochemical characteristics based on Bergey’s Manual of systemic bacteriology. The isolate produced maximum chitinase in colloidal chitin broth at pH7 and temperature 37oC after four days of inoculation. The corresponding culture filtrate supposed to contain chitinase showed maximum % inhibition of 53.29% with Fusarium oxysporum and no inhibition to Phytophthora infestans in agar well diffusion assay. Furthermore, chitinase was best fractionated at 40% ammonium sulphate salt fractionation which has almost similar inhibition potential as the crude culture filtrate. The 40% salt fraction of the enzyme showed the maximum chitinolytic potential at pH8 and temperature 40oC. Among the phytopathogens tested, sensitivity of Bacillus subtilis to fungi containing chitin on their cell wall demonstrates the possible role of chitinase in the antifungal activity.DOI: http://dx.doi.org/10.3126/njst.v12i0.6517 Nepal Journal of Science and Technology 12 (2011) 304-311

Self versus non-self: fungal cell wall degradation in Trichoderma

Lysis of the prey's cell wall is one of the key steps during mycoparasitism. Genome analysis of two mycoparasitic Trichoderma species, T. atroviride and T. virens, revealed an expanded arsenal of genes encoding enzymes potentially involved in cell wall hydrolysis. Glycoside hydrolase family 18, which contains all fungal chitinases, is the largest family of carbohydrate-active enzymes in mycoparasitic Trichoderma species. However, in addition to their aggressive functions during mycoparasitism, the roles of chitinases and other cell wall degrading enzymes also include remodelling and recycling of the fungus's own cell wall. In this review we discuss current knowledge about fungal cell wall degrading enzymes in Trichoderma and how the fungus distinguishes between self- and non-self fungal cell wall degradation. In the past few years, the chitinolytic enzyme machinery of Trichoderma has been used as a model system to address this question. Gene expression profiles of most investigated chitinases indicate an overlap of functions of the respective enzymes and an involvement in both self- and non-self fungal cell wall degradation. Similar sets of enzymes appear to be involved in mycoparasitism, exogenous chitin decomposition and recycling of the fungus's own cell wall. Thus, we hypothesize that the regulation of self and non-self fungal cell wall degradation is not due to a speciation of individual chitinases. Rather, we hypothesize that it is regulated by substrate accessibility due to cell wall protection in healthy hyphae vs deprotection during mycoparasitic attack, hyphal ageing and autolysis.

Role of Chitinase in Plant Defense

Role of Chitinase in Plant Defense

Bacillus pumilus SG2 chitinases induced and regulated by chitin, show inhibitory activity against Fusarium graminearum and Bipolaris sorokiniana

The possible role of chitinase in in vitro growth inhibition of the wheat pathogens Fusarium graminearum and Bipolaris sorokiniana by Bacillus pumilus SG2 was investigated. B. pumilus SG2, a chitinolytic bacterium producing two different chitinases, was previously isolated from the saline deserts of Iran. When grown in Spizizen salts medium with colloidal chitin, B. pumilus SG2 secreted two chitinases into the medium, resulting in growth inhibition of F. graminearum and abortion of hyphal elongation of B. sorokiniana. In contrast, when glucose was used as the carbon source, the chitinases were not expressed and antifungal activity of the B. pumilus SG2 was completely abolished. These results confirmed that expression of the B. pumilus SG2 chitinases is under the control of two types of regulation, special regulation by chitin and global regulation by glucose. We demonstrated that chitinases are the main components that caused hyphal inhibition activity of B. pumilus SG2. Hyphal inhibition of F. graminearum and B. sorokiniana was stable in agar for a minimum of 14 days.

Anti-fungal activity of maize silk proteins and role of chitinases in Aspergillus flavus resistance

Studies were conducted to identify proteins in maize silks that may be contributing to Aspergillus flavus resistance. We first performed bioassays using silk extracts collected from two A. flavus-resistant inbred lines and two susceptible inbred lines. Fungal biomass was quantified by measuring fluorescence of a green fluorescent protein (GFP)-tagged A. flavus and by measuring ergosterol levels. The silk extracts from resistant inbreds had greater anti-fungal activity compared to susceptible inbreds. Comparative proteomic analysis of the two resistant and susceptible inbreds led to the identification of several anti-fungal proteins. One of the anti-fungal proteins that we further investigated was chitinase. There were three chitinases that were differentially expressed in the resistant lines (PRm3 chitinase, chitinase I, and chitinase A). We conducted chitinase assays on silk proteins from extracts of resistant and susceptible inbred lines. Silk extracts from resistant inbred lines showed significantly higher activity in the resistant maize inbreds compared to the susceptible inbreds (P < 0.01). The differential expression of chitinases in maize resistant and susceptible inbred silks suggests that these proteins may contribute to A. flavus resistance.

Diverse mechanisms adopted by fluorescent Pseudomonas PGC2 during the inhibition of Rhizoctonia solani and Phytophthora capsici

Rhizoctonia solani and Phytophthora capsici are two of the most destructive phytopathogens occurring worldwide and are only partly being managed by traditional control strategies. Fluorescent Pseudomonas isolates PGC1 and PGC2 were checked for the antifungal potential against R. solani and P. capsici. Both the isolates were screened for the ability to produce a range of antifungal compounds. The results of this study indicated the role of chitinase and β-1,3-glucanase in the inhibition of R. solani, however, antifungal metabolites of a non-enzymatic nature were responsible for inhibition of P. capsici. The study confirmed that multiple and diverse mechanisms are adopted by the same antagonist to suppress different phytopathogens, as evidenced in case of R. solani and P. capsici.

Role of chitinase and β-1,3-glucanase activities produced by a fluorescent pseudomonad and in vitro inhibition ofPhytophthora capsiciandRhizoctonia solani

A study was conducted to investigate the possibility of involvement of chitinase and beta-1,3-glucanase of an antagonistic fluorescent Pseudomonas in growth suppression of phytopathogenic fungi, Phytophthora capsici and Rhizoctonia solani. Fluorescent Pseudomonas isolates GRC(3) and GRC(4) were screened for their antifungal potential against phytopathogenic fungi by using dual culture technique both on solid and liquid media. The percent inhibition was calculated. Various parameters were monitored for optimization of enzyme activities by fluorescent Pseudomonas GRC(3). The involvement of chitinases, beta-1,3-glucanases, and antifungal metabolites of nonenzymatic nature was correlated with the inhibition of P. capsici and R. solani. The results provide evidence for antibiosis as a mechanism for antagonism. The study also confirms that multiple mechanisms are involved in suppressing phytopathogens as evidenced by the involvement of chitinase and beta-1,3-glucanase in inhibition of R. solani but not P. capsici by isolate GRC3.

Control of green and blue mould on orange fruit by Serratia plymuthica strains IC14 and IC1270 and putative modes of action

Two biological control agents of Serratia plymuthica, strains IC1270 and IC14 applied separately and in combination were evaluated for suppressing Penicillium digitatum (green mould) or Penicillium italicum (blue mould) on orange. These bacteria were effective and controlled both pathogens at 1 × 10 8 cells/mL. Disease suppression was increased when both bacterial strains were combined. Possible modes of action studied in this work were antibiosis, chitinolytic activity and competition for nutrients. Two mutants of strain IC1270, one deficient in chitinolytic activity and the second deficient in pyrrolnitrin production were obtained by the gene replacement technique. On orange fruit, mutant IC1270-C7 deficient in chitinase production and mutant IC1270-P1 deficient in pyrrolnitrin production showed a similar efficiency against P. digitatum to the parental strain. However, in vitro IC1270-P1 lost its antifungal activity and no inhibition zone was observed when it was tested against P. digitatum or P. italicum. In similar experiments, a chitinase-deficient mutant of strain IC14 was as effective as the parental strain IC14, suggesting no evidence for a possible role of chitinases in controlling green mould caused by P. digitatum. Interactions between strains IC1270 or IC14 and P. digitatum were studied in tissue culture plates with diluted orange peel extract as the nutrient source. Strain IC1270 decreased germination of P. digitatum conidia when it was physically separated from the pathogen by a membrane filter, which permits nutrient and metabolite interchange, while strain IC14 did not affect germination. Significant inhibition of conidial germination of P. digitatum was achieved, however, when the pathogen and IC14 were in physical contact. Competition for nutrients appears to be the main mode of action of strain IC1270, while a direct cell-to-cell interaction between IC14 and the pathogen is needed for antagonism.

Exploiting worm and allergy models to understand Th2 cytokine biology

Helminthic parasites and many allergens trigger highly polarized Th2-type immune responses. In most helminth infections, the Th2 response often leads to parasite expulsion or sequestration. During murine Schistosoma mansoni infection, however, the parasites persist and the chronic Th2 response induces severe pathological changes in the gut and liver. Thus, the study of schistosome infections in mice has become a popular model to study the pathogenesis of Th2 cytokine-mediated disease. This review will discuss recent findings from the schistosomiasis model that may be relevant to the understanding of allergic inflammation, asthma and Th2 cytokine biology in general. Evidence is accumulating that the Th2 pathway is not a 'default pathway' but one that is actively instructed by mechanisms that are only beginning to be understood. Other areas of intensive investigation include studies on alternatively activated macrophages, the role of dendritic cells in Th2 response development, the inhibitory function of IL-10, regulatory T-cells and decoy receptors on chronic Th2-mediated inflammation, and the role of chitinases in mediating Th2 disease. Finally, the development of novel eosinophil-deficient mice has also accelerated our understanding of the contribution of this important cell type to Th2 immunity. Many findings from the schistosomiasis model have been subsequently demonstrated in models of allergic disease, illustrating the utility of this model to dissect basic mechanisms of Th2-mediated inflammation. Further study of helminth-induced Th2 responses may expedite the discovery of new therapeutics for a wide range of Th2-associated diseases.

Role of Chitinase and Sormatin Accumulation in the Resistance of Sorghum Cultivars to Grain Mold

Experiments were conducted to determine the association between resistance to grain mold and the accumulations of chitinase and sormatin. Eight sorghum lines were treated at 50% bloom with Fusarium thapsinum, Curvularia lunata, a mixture of the two fungi, and a water-sprayed control. At maturity, percent disease severity, seed germination rates, and kernel weight were recorded. Chitinase and sormatin content (mg/g of dry weight) were measured in seed samples taken at 30 and 50 days after treatment (DAT). Seed chitinase content was moderately affected by sorghum line (P = 0.10) and significantly affected by the developmental stage of the kernels (P = 0.05). Cultivars Sureno, 98LB650, and 98LB723 exhibited larger negative changes in chitinase content at 50 DAT over water-sprayed control treatment at 30 DAT than the susceptible cultivars Dorado, RTx2536, and RTx430. In 2000, significant negative correlations were observed for percent disease severity and chitinase content at 30 DAT, seed germination and sormatin content at 50 DAT, and between seed germination and kernel weight. There also was a significant positive correlation between germination and chitinase content at 30 DAT. No association between disease severity and changes in chitinase content at 50 DAT was observed. Sormatin content also was significantly affected by the stage of kernel development. Sorghum cultivars inoculated with fungal pathogens responded differently as indicated by the significant sorghum line x treatment interaction for sormatin content in 2000. In both years, larger increases in sormatin content over the water-sprayed control treatments were observed on moderately susceptible to susceptible cultivars such as 98LB650, 98LB723, 98LB789, RTx430, and RTx2536 than on Sureno. Except for percent disease severity and germination, there was no significant association among all of the other parameters measured in 2001. The results of this study did not clearly demonstrate a strong association between resistance to grain mold and the accumulation of sormatin and chitinase. Thus, there is the possibility that certain moderately resistant to resistant sorghum cultivars, such as Sureno, may employ other strategies to eschew or restrict fungal invasion either before or after physiological maturity.

Chitinase is crucial for parasite invasion of mosquito midgut

A set of two new papers provides compelling evidence supporting the crucial role of chitinase secreted by Plasmodium ookinetes for invasion of the mosquito midgut. Y-L. Tsai et al. [(2001) Infect. Immun. 69, 4048–4054,] disrupted the only identified P. falciparum chitinase gene PfCHT1 – this led to a significant impairment in the ability of affected parasites to form oocysts in the midguts of Anopheles freeborni. J. Dessens et al. [(2001) Infect. Immun. 69, 4041–4047] identified and characterized PbCHT1, the first chitinase gene from the rodent malaria parasite P. berghei. Targeted disruption of PbCHT1 also dramatically reduced parasite infectivity in A. stephensi mosquitoes. Interestingly, PbCHT1 null mutants had no residual ookinete-derived chitinase activity, suggesting that P. berghei ookinetes express only one chitinase gene. PbCHT1 is a structural orthologue of PfCHT1 and of the avian Plasmodium chitinase gene, PgCHT1. SHK

β‐1,3‐Glucanase and Chitinase Activities and the Resistance Response of Wheat to Leaf Rust

AbstractTo investigate the role of β‐1,3‐glucanases (EC 3.2.1.39) and chitinases (EC 3.2.1.14) in the resistance response of wheat (Triticum aestivum L.) to leaf rust (Puccinia recondita f. sp. tritici), their intercellular activities were studied in resistant (Karee/Lr35) and susceptible (Karee) near‐isogenic wheat lines under conditions of infection and non‐infection. Resistance was associated with high constitutively expressed chitinase activity and induced β‐1,3‐glucanase activity. At 168 hours post‐inoculation several infection sites on Karee/Lr35 displayed necrosis, confirming a hypersensitive response, whereas similar necrotic lesions were observed only for a limited number of infection sites on Karee at that sampling stage. The possible role of chitinases and β‐1,3‐glucanases in the hypersensitive defence response of wheat to leaf rust is discussed.

Antifungal activity of chitinases produced by some fluorescent pseudomonads against Colletotrichum falcatum Went causing red rot disease in sugarcane

Chitinase production and growth of certain fluorescent pseudomonads isolated from sugarcane rhizosphere on different subtrates were studied. When chitin was substituted for glycerol in King's B medium, 3 of the 4 strains showed enhanced bacterial multiplication. Bacterial cells grown on chitin-containing medium showed enhanced antifungal activity against Colletotrichum falcatum Went causing red rot disease in sugarcane. Chitinase production was significantly higher when chitin was amended to King's B medium. Higher chitinase production was also recorded when fluorescent pseudomonad strains were grown in the medium containing crab-shell chitin. Cell-free bacterial culture filtrate from chitin-containing medium significantly inhibited mycelial growth of the pathogen. These cell-free conditioned media contained 3 to 7 polypeptides. Western blot analysis revealed five isoforms of chitinase with molecular masses of 47, 36, 32, 20 and 18.5 kDa. A possible role of chitinases in red rot disease management is discussed.