The interactive effects of temperature and osmotic potential on the growth of marine isolates of <Emphasis Type="Italic">Fusarium solani</Emphasis>

The mycelial growth of 18 Fusarium solani strains isolated from sea beds of the south-eastern coast of Spain was tested on potato-dextrose-agar adjusted to different osmotic potentials with either KCl or NaCl (−1.50 to −144.54 bars) in 10 °C intervals ranging from 15 to 35 °C. Fungal growth was determined by measuring colony diameter after 4 days of incubation. Mycelial growth was maximal at 25 °C. The quantity and frequency pattern of mycelial growth of F. solani differ significantly at 15 and 25 °C, with maximal growth occurring at the highest water potential tested (−1.50 bars); and at 35 °C, with a maximal mycelial growth at −13.79 bars. The effect of water potential was independent of salt composition. The general growth pattern of F. solani showed declining growth at potentials below −41.79 bars. Fungal growth at 35 °C was always higher than that grow at 15 °C, of all the water potentials tested. Significant differences observed in the response of mycelia to water potential and temperature as main and interactive effects. The viability of cultures was increasingly inhibited as the water potential dropped, but some growth was still observed at −99.56 bars. These findings could indicate that marine strains of F. solani have a physiological mechanism that permits survival in environments with low water potential. The observed differences in viability and the magnitude of growth could indicate that the biological factors governing potential and actual growth are affected by osmotic potential in different ways.     

The effect of water potential on soil-borne diseases of wheat seedlings

Under glasshouse conditions, Fusarium culmorum caused more injury to wheat seedlings in soil at 33% saturation (approximately – 1 bar) than at 66% (–0·1 bar). The same effect occurred with inoculated seeds raised on soil moisture tension tables at –0·1 and –0·5 bar, at 20° and at 15°C. In controls, spontaneous infection by soil-borne F. culmorum was unaffected by soil water potential. Inoculated F. nivale, at 127deg; and at 15°C, showed a similar, but smaller, effect to that with inoculated F. culmorum. Water potential in this range did not affect disease incidence with inoculated Gaeumannomyces graminis, but disease was more severe in the drier soil. On tension tables, seedlings did not consistently develop faster under wetter conditions, and factors other than host growth rate probably played a part in limiting fungal attack.     

Effects of water potential on spore germination and viability of <Emphasis Type="Italic">Fusarium</Emphasis> species

Germination of macroconidia and/or microconidia of 24 strains of Fusarium solani, F. chlamydosporum, F. culmorum, F. equiseti, F. verticillioides, F. sambucinum, F. oxysporum and F. proliferatum isolated from fluvial channels and sea beds of the south-eastern coast of Spain, and three control strains (F. oxysporum isolated from affected cultures) was studied in distilled water in response to a range of water potentials adjusted with NaCl. (0, −13.79, −41.79, −70.37, −99.56 and −144.54 bars). The viability (UFC/ml) of suspensions was also tested in three time periods (0, 24 and 48 h). Conidia always germinated in distilled water. The pattern of conidial germination observed of F. verticilloides, F. oxysporum, F. proliferatum, F. chlamydosporum and F. culmorum was similar. A great diminution of spore germination was found in −13.79 bars solutions. Spore germination percentage for F. solani isolates was maximal at 48 h and −13.79 bars with 21.33% spore germination, 16% higher than germination in distilled water. F. equiseti shows the maximum germination percentage in −144.54 bars solution in 24 h time with 12.36% germination. This results did not agree with those obtained in the viability test were maximum germination was found in distilled water. The viability analysis showed the great capacity of F. verticilloides strains to form viable colonies, even in such extreme conditions as −144.54 bars after 24 h F. proliferatum colony formation was prevented in the range of −70.37 bars. These results show the clear affectation of water potential to conidia germination of Fusaria. The ability of certain species of Fusarium to develop a saprophytic life in the salt water of the Mediterranean Sea could be certain. Successful germination, even under high salty media conditions, suggests that Fusarium spp. could have a competitive advantage over other soil fungi in crops irrigated with saline water. In the specific case of F. solani, water potential of −13.79 bars affected germination positively. It could indicate that F. solani has an special physiological mechanism of survival in low water potential environments.     

The laboratory germination of crisp lettuce seeds under moisture stress

Seeds of the crisp lettuce cultivar Pennlake were germinated using all combinations of six ‘initial’ solutions of polyethylene glycol 6000 (PEG) with osmotic potentials ranging from 0 to -8 bars and seven ‘secondary’ solutions of PEG with osmotic potentials ranging from 0 to -10 bars, to which seeds were moved after 24 or 48 h in the ‘initial’ solution. The number of seeds germinating decreased at more negative osmotic potentials of both ‘initial’ and ‘secondary’ solutions but there was an interaction between germination temperature and the osmotic potential of the ‘initial’ solution. At an ‘initial’ solution osmotic potential of 0 bars germination at 20°C exceeded that at 10°C. As the osmotic potential of the ‘initial’ solution decreased germination at 20°C decreased more than at 10°C so that at the more negative osmotic potentials germination at 10°C exceeded that at 20°C. However seeds ungerminated after 14 days germinated normally when transferred back to water, so that the average final germination was 99.5%. The results suggest that major fluctuations in soil water potential in a seedbed are unlikely to influence seed germination per se provided that a period of 24 to 48 h at 0 bars tension is available at some time. The timing of such a period relative to sowing will have a considerable effect on the time of germination and hence the time of emergence. It is concluded that factors other than the direct effect of soil moisture content on germination are involved in reducing seedling emergence under fluctuating soil moisture conditions in the field.     

Trichoderma harzianum antagonistic activity and competition for seed colonization against seedborne pathogenic fungi of sunflower

This study investigated the antagonistic effects of Trichoderma harzianum isolate (TRIC8) on mycelial growth, hyphal alteration, conidial germination, germ tube length and seed colonization by the seedborne fungal pathogens Alternaria alternata, Bipolaris cynodontis, Fusarium culmorum and F. oxysporum, the causes of seedling rot in over 30% of sunflowers. The antagonistic effect of TRIC8 on mycelial growth of pathogens was evaluated on dual culture that included two inoculation assays: inoculation of antagonist at 48 h before pathogen (deferred inoculation) and inoculation at the same time with pathogen (simultaneous inoculation). TRIC8 inhibited mycelial growth of the fungal pathogens between 70·67 and 76·87% with the strongest inhibition seen with deferred inoculation. Alterations in hyphae were observed in all pathogens. Conidial germination of F. culmorum was inhibited by most of the fungal pathogens (38·28%) by TRIC8. Inhibition of germ tube length by the antagonist varied from 31·83 to 37·67%. In seed colonization experiments, TRIC8 was applied in combination with each pathogen to seeds of a sunflower genotype that is highly tolerant to downy mildew. Seed death was inhibited by TRIC8 and the antagonist did not allow growth of A. alternata, B. cynodontis and F. culmorum on seeds and inhibited the growth of F. oxysporum at the rate of 58·32%.     

Effect of osmotic potential on the growth of Gaeumannomyces graminis and Phialophora spp.

The linear growth of 10 isolates each of Gaeumannomyces graminis var. graminis, G. graminis var. tritici and Phialophora graminicola and five isolates each of G. graminis var. avenae and a lobed-hyphopodiate Phialophora sp. was studied on osmotically adjusted agar at 20 °C. While most isolates of G. graminis var. avenae ceased growing at osmotic potentials of -60 bars (1 bar = 10⁵ Pa), six out of 10 isolates of G. graminis var. tritici grew at that potential. The growth of all isolates of G. graminis var. tritici and var. avenae ceased at -70 bars. In contrast, four out of 10 isolates of P. graminicola grew at -70 bars, but all stopped growing at -80 bars. Most of the isolates of G. graminis var. graminis and the lobed-hyphopodiate Phialophora sp. grew at -70 bars while three out of 10 isolates of G. graminis var. graminis and one out of five isolates of the lobed-hyphopodiate Phialophora sp. were capable of growth at -80 bars. None of the fungi grew at -90 bars. Detailed studies of the growth of two or three isolates each of the five fungi at 10, 20, 30 and 35 °C were carried out on osmotic agar controlled by the addition of either sodium chloride or potassium chloride. In general, similar reductions in growth occurred with decreasing osmotic potential regardless of the solute used. At 10 and 20 °C., all three isolates of P. graminicola showed optimal growth at about -5 bars while the other fungi grew fastest at -1²middot; bars. At 30 °C., one isolate of the lobed hyphopodiate Phialophora sp. and two isolates each of P. graminicola, G. graminis var. tritici and G. graminis var. avenae grew optimally at osmotic potentials of -10 to -15 bars. The other isolate of the Phialophora sp. and two isolates of G. graminis var. graminis studied grew optimally at the highest potential (-1·2 bars). However, at 35 °C the last three fungi exhibited optimal growth at osmotic potentials of-10 to -20 bars. The ecological significance of these results is discussed in relation to cross-protection against the take-all fungi by the avirulent fungi.     

The importance of water potential range tolerance as a limiting factor on Trichoderma spp. biocontrol of Sclerotinia sclerotiorum

Biological control of fungal phytopathogens is often more variable in efficacy compared with disease suppression achieved by conventional pesticide use. Matching the environmental range of a potential biocontrol agent with that of the target phytopathogen is necessary if consistent disease suppression is to be achieved under field conditions. Strains of Trichoderma that could parasitise sclerotia of Sclerotinia sclerotiorum had their spore germination and mycelial growth (five strains) and ability to parasitise sclerotia (two strains) tested under a range of water potentials under laboratory conditions. Relative mycelial growth and germination of all strains decreased with decreasing osmotic and matric potentials, with matric potential having a greater impact on growth and germination over the range examined. Trichoderma harzianum LU698 mycelial growth was the least affected by decreasing osmotic potential than the other isolates, and Trichoderma atroviride LU141 growth least affected by decreasing matric potential. The germination of LU698 and LU144 was also generally less affected by decreasing osmotic potential, although generally decreasing matric potential had the greatest affect on the germination of LU698 along with T. atroviride LU132. Soil treatments of LU698 and Trichoderma asperellum LU697 reduced sclerotial viability in all but the lowest soil water potential tested, with LU698 being most effective at ?0.1 and ?0.3 MPa after 28 days and LU697 most effective at ?0.01 and ?1.5 MPa after 28 days. We conclude that differences in the tolerance of potential biocontrol agents to changing water potential is an important experimental factor to consider when assaying biocontrol or making predictions of biocontrol efficacy in the field.     

Influence of Water Potential on Growth, Enzyme Secretion and In Vitro Enzyme Activities of Trichoderma harzianum at Different Temperatures

The influence of water potential on linear mycelial growth, secretion, and the in vitro activities of enzymes β-glucosidase, cellobiohydrolase, β-xylosidase, exochitinase, and chymotrypsin of Trichoderma harzianum strain T66 was studied at different temperatures. Nearly linear correlation was found between water potential and colony growth rate at both 25°C and 10°C, with higher growth rates at the higher temperature and higher water potentials. The amounts of enzyme secretion depended on the water potential and not on the quality of salt (NaCl or KCl) used as osmoticum. Enzyme activities were significantly affected by water potential. Significant enzyme activities were measured for most of the enzymes even at −14.800 megapascal (MPa), which is below the water potential where mycelial growth ceased. These results suggest the possibility of using mutants with improved xerotolerance for biocontrol purposes in soils with lower water potentials. Received: 5 October 1999 / Accepted: 22 November 1999     

Plumieride from Allamanda cathartica as an inhibitory compound to plant pathogenic fungi

Allamanda leaf extract (Allamanda cathertica) was made in water at room temperature (25?± 2?°C) as well as in a number of less polar to highly polar solvents like methylene chloride, benzene, chloroform and ethyl acetate at their boiling point, that means, at refluxing temperature (40?± 2?°C). Methylene chloride, benzene, chloroform, ethyl acetate and water extracts were applied to determine their growth inhibition against Phomopsis vexans, Phytophthora capsici, Fusarium oxysporum, Rhizoctonia solani and Sclerotium rolfsii. Results of these extracts showed that refluxing methanol, ethanol and ethyl acetate extracts of Allamanda were statistically similar for inhibition of mycelial growth of all fungi tested. But effect of 50% ethanol extract is different; it inhibited 100% mycelial growth of P. vexans, P. capsici and F. oxysporum; 83.33% of R. solani and 88.63% of S. rolfsii. Effort was also made to find out the compound in Allamanda to be responsible for such antifungal activity. Thin layer chromatography (TLC) of Allamanda extracts showed the presence of a number of compounds having polarity very high to low. The R_f values of compounds in 37–42 fractions were calculated and from these six fractions, crystals were separated. These crystals were more or less white. Melting point of these crystals was determined by ordinary and digital melting point apparatus that ranged from 145.5–162 C. Structural determination of the compound was done by Infra-red (IR) spectral study. The finger print region was 700–1400?cm^?1. The strong band at 1612.4, 1633.6, 1693.4, 1655 and 2850.6?cm^?1 indicated the presence of conjugated double bond (–C=C–C=C–), non-conjugated double-bond (–C=C–C–C–C=C–), carbonyl group attached to carbon–carbon double (–CO–C=C), ester (–COOR) and C–H stretching, respectively. Mass spectra of separated compounds gave molecular weight 470. All these characters are typical to pumieride as described previously. Again, In vitro screening of plumieride against P. vexans, P. capsici, F. oxysporum, R. solani and S. rolfsii were found effective in inhibiting radial mycelial growth of these fungi at 1:2 w/v concentration.     

Water and temperature relations of softrot bacteria: growth and disease development

The effect of water availability and the temperature of the growth substrate on growth and disease development of softrot bacteria were studied using artificial media and plant material. Water availability was measured as the osmotic potential of a solution (ψ_osm) and was assessed for solutions of PEG4000 and KNO₃ as artificial osmotica and for plant tissue of chicory heads. Growth of softrot bacteria was found at water potentials from ψ= -0.12 MPa to ψ= -8.0 MPa but the lag phase of the growth curve increased with decreasing water potential. The relative growth rates of the three softrot pathogens showed a sigmoidal relationship with water potential, the relative growth rates decreasing rapidly at water potentials lower than ψ= -1.5 MPa. The water potential of harvested chicory heads decreased with storage time of the harvested crop but was still within the growth limits for softrot bacteria. In relation to temperature, the relative growth rate of Erwinia carotovora subsp. carotovora (Ecc) was highest at 10°C, of Erwinia carotovora subsp. atroseptica (Eca) at 15°C and of Pseudomonas marginalis (Pm) at 5°C. Chicory heads of two cultivars, Rumba and Salsa, inoculated with Ecc, had a significantly higher disease severity at 30°C (0.72 for Rumba and 0.47 for Salsa) than at lower or higher temperatures. In conclusion, temperature and water availability during forcing of chicory were not factors limiting populations of softrot bacteria. Possibilities for crop protection thus only avail during chicory root storage. During storage a high death rate combined with a low growth rate of the softrot bacteria may result in a decrease of bacterial populations below the minimum densities needed for infection during the forcing of chicory heads.     

EFFECT OF WATER POTENTIAL ON A MICROCOLEUS (CYANOPHYCEAE) FROM A DESERT CRUST1

The effect of water potential, on the growth and photosynthesis of a species of Microcoleus forming a desert crust was determined, using both osmotic and matric variations in water potential. The alga was quite sensitive to moisture stress, partial inhibition of growth being observed at -7 bars, and complete inhibition at -18 bars. Photosynthesis was markedly inhibited at -18 bars, and virtually completely at, -28 bars (water potential of seawater) and lower. The alga was more sensitive to matric reduction in water potential than osmotic. By comparisons of these results with those obtained with other algae, it is concluded that this desert crust alga is not especially adapted to grow and photosynthesize at low water potentials, although it shows considerable ability to survive severe drought conditions.     

A Comparison of the Effects of Temperature on the Growth of Fusarium oxysporum f. sp. narcissi in Solid and Liquid Media

The effect of different temperatures on mycelial growth of Fusarium oxysporum f. sp. narcissi in liquid and on solid medium was compared. Differences between optimum and maximum temperatures using these substrates were considerable and, could lead to different interpretations of the parasitic growth of the pathogen in vascular tissues, for example, in liquid culture Fusarium grew at 45°C whereas it did not on solid media. Initially F. oxysporum f. sp. narcissi may, be encouraged to grow faster in warm air drying systems than those using lower temperatures. The rates of diffusion between 17°C and 35°C are not markedly different. The speed with which moisture is removed from the internal tissues of the remaining roots is governed more by air flow over the bulbs than by temperature.     

Damping-off of Some Cucurbitaceous Crops in Saudi Arabia with Reference to Control Methods

Isolation of seed borne fungi from sweet melon (najed and red Queen varieties) and vegetable marrow (squash) using PDA, Plain agar media and blotters revealed the presence of Alternaria alternata (Fr.) Keissler, Fusarium semitectum var,majus from Najed melon, A. alternata (Fr.) Keissler, F. sambucinum Fuckel from Red Queen melon and A. chlamydospora Mouchacca, Cephalosporium sp., and F. oxysporum schlecht from squash. Pathogenicity tests showed, that all these fungi were highly pathogenic on their respective hosts. The optimum temperature for its growth ranged from 25–30°C and the optimum pH was 6.0. In pot trials, seed dressing with Banrot, Bavistin and Topsin-M at the rate of 4 g/kg seed was superior in controlling the damping-off of melon and squash. These fungicides were effective in inhibiting mycelial growth, spore germination and development of the isolated fungi. Hot water treatment at 55 C/20 min or solar heating (av. 37°C) for 90 min were next to fungicides.     

Partial Characterization of the Antimicrobial Activity of Streptomyces antimycoticus FZB53

The paper describes experiments aimed at evaluating the sensitivity of different fungi, most of them plant pathogens and bacteria towards Streptomyces antimycoticus FZB53, a biocontrol agent that, when applied as a seed treatment, in previous studies has shown good activity against different seed‐borne fungal diseases. When incorporated into agar media, the filtrate from shake cultures of S. antimycoticus FZB53 inhibited the mycelial growth or spore germination, respectively, of a broad spectrum of fungi. The most sensitive of the fungi tested was Fusarium culmorum. The inhibitory activity could be removed from the culture filtrate by extraction with ethyl acetate. When ethyl acetate extracts of the pellet and supernatant obtained by centrifugation of the shake culture were added to the agar medium, inhibition of mycelial growth of F. culmorum was restored, especially with the extracts of the pelleted biomass. Autoclaving of the culture filtrate reduced the inhibition of F. culmorum but completely eliminated the inhibitory activity against Fusarium graminearum. Among the bunt fungi tested, spore germination of Tilletia tritici was more sensitive to the culture filtrate of S. antimycoticus FZB53 than spore germination of Ustilago avenae and U. tritici. Separation by thin layer chromatography (tlc) and spraying with different reagents showed that ethyl acetate extracts from shake cultures or biomass scraped from agar media contained several hydrophobic metabolites. When eluted from the tlc‐plates, the material from one of the spots had strong antifungal activity against spore germination of T. tritici and mycelial growth of F. culmorum, respectively. Ethyl acetate extracts from biomass of S. antimycoticus FZB53 prevented the growth of the tested Gram‐positive bacteria, namely Clavibacter michiganensis and different species of Bacillus. The results indicated that these bacteria were at least as sensitive towards the metabolites of S. antimycoticus FZB53 as F. culmorum. The tested Gram‐negative bacteria were not affected.     

Rapid Diagnosis of Soya Bean Root Rot Caused by Fusarium culmorum Using a Loop‐Mediated Isothermal Amplification Assay

We report a rapid diagnosis of soya bean (Glycine max L.) root rot caused by Fusarium culmorum, using a loop‐mediated isothermal amplification (LAMP) assay. We used the CYP51C gene sequence to design LAMP assay primers specific for F. culmorum. The LAMP assay amplified the target gene efficiently in 60 min at 63°C. The sensitivity of the assay was 100 pg/μl of genomic DNA. Among the tested soya bean pathogens, a positive colour (sky blue) was only observed in the presence of F. culmorum with the addition of hydroxynaphthol blue (HNB) dye prior to amplification, whereas other species isolates showed no colour change. Suspected diseased soya bean samples collected in the field from Jiangsu, Shandong and Anhui provinces and Beijing were diagnosed successfully using the LAMP assay reported here. This study provides a new and readily available method for rapid diagnosis of soya bean root rot caused by F. culmorum.     

Glasshouse studies on the effect of low temperature on infection of perennial ryegrass seedlings by Fusarium nivale

In glasshouse experiments, isolates of Fusarium nivale, applied as mycelial macerates, caused marked reductions in the number and length of roots, and in the height and fresh weight of S. 321 perennial ryegrass seedlings. Spore inocula failed to cause infection. Damage by F. nivale was increased when inoculation was immediately followed by four alternate 12 h periods in a growth cabinet at 0°C and in a warm glasshouse at c. 17°C, succeeded by a further 11 days in the same glasshouse. Although inoculated seedlings receiving no cold treatments initially showed damage, regrowth of roots and some recovery of the shoots occurred in a fortnight. Exposure of the plants after that time to four intermittent 12 h periods at 0°C, however, arrested their recovery and caused further root damage. The implication of these results in the importance of low temperature in the winterkill syndrome is discussed.     

Characterization of chitinase from Streptomyces luridiscabiei U05 and its antagonist potential against fungal plant pathogens

Streptomyces luridiscabiei U05 was isolated from wheat rhizosphere. It produced chitinase, which showed in vitro antifungal properties. The crude enzyme inhibited the growth of Alternaria alternata, Fusarium oxysporum, F. solani, Botrytis cinerea, F. culmorum and Penicillium verrucosum. The chitinase enzyme of the molecular weight of 45 kDa was purified using affinity chromatography of chitin. Streptomyces luridiscabiei U05 produced different chitinolytic enzymes. The highest enzyme activity was observed with the use of 4‐MU‐(GlcNAc)_, which points to the presence of an β‐N‐acetylhexosaminidase. The optimum activity was obtained at 35–40°C and pH 7–8. The enzyme showed thermostability at 35–40°C during 240 min of preincubation and lost its activity at 50°C and 60°C in 60 min. The chitinase activity from S. luridiscabei U05 was strongly inhibited by Hg²⁺ and Pb²⁺ ions, and sodium dodecyl sulphate (SDS). The Ca²⁺, Cu²⁺ and Mg²⁺ ions stimulated the activity of the enzyme.     

Pathogenic potential of root-knot nematode Meloidogyne incognita and root-rot fungus Fusarium solani on chilli (Capsicum annuum L.)

Damaging threshold levels of root-knot nematode Meloidogyne incognita and root-rot fungus Fusarium solani on plant growth parameters, viz., plant length, fresh and dry weight of chilli were determined by conducting their pathogenicity trials in pot experiments. The results revealed a significant reduction in the plant growth parameters at and above the inoculum level of about 1000?J₂ per plant of M. incognita and the highest reduction was recorded at 8000?J₂ per plant. Significant reduction in plant growth parameters was recorded at 1.00?g mycelial mat of F. solani per plant, while the highest reduction was observed at 8.00?g mycelial mat per plant. The damaging threshold level was 1000?J₂ per plant of M. incognita and 1.00?g mycelial mat of F. solani.     

The Effect of Soil Water Regimes on Leaf Water Potential, Growth and Development of Soybeans

The growth and development of soybeans (Glycine max L. cv. Amsoy) was studied at soil matric potentials of ?0.1 to ?1.0 bars. Chlorophyll, photosynthesis, and leaf nitrogen per plant was greatest at ?4 bars leaf water potential. Leaf area, number of internodes, plant height and dry weight of vegetative parts declined as leaf water potential decreased from ?2 to ?19 bars. Nitrogen content and nitrate reductase activity per g fresh weight determined the percentage protein of individual seeds but nitrogen content and nitrate reductase activity per plant determined the amount of total seed protein. The protein synthesized in the seed changed little in amino acid composition with changes in leaf water potential. Leaf water potentials above or below ?4 bars decreased yield, total protein and total lipid but plants produced the largest percentage of individual seed protein at ?19 bars leaf water potential.     

Production of Antifungal Chitinase by Aspergillus niger LOCK 62 and Its Potential Role in the Biological Control

Aspergillus niger LOCK 62 produces an antifungal chitinase. Different sources of chitin in the medium were used to test the production of the chitinase. Chitinase production was most effective when colloidal chitin and shrimp shell were used as substrates. The optimum incubation period for chitinase production by Aspergillus niger LOCK 62 was 6?days. The chitinase was purified from the culture medium by fractionation with ammonium sulfate and affinity chromatography. The molecular mass of the purified enzyme was 43?kDa. The highest activity was obtained at 40?°C for both crude and purified enzymes. The crude chitinase activity was stable during 180?min incubation at 40?°C, but purified chitinase lost about 25?% of its activity under these conditions. Optimal pH for chitinase activity was pH 6–6.5. The activity of crude and purified enzyme was stabilized by Mg²⁺ and Ca²⁺ ions, but inhibited by Hg²⁺ and Pb²⁺ ions. Chitinase isolated from Aspergillus niger LOCK 62 inhibited the growth of the fungal phytopathogens: Fusarium culmorum, Fusarium solani and Rhizoctonia solani. The growth of Botrytis cinerea, Alternaria alternata, and Fusarium oxysporum was not affected.