Assessment of glucosinolate-derived isothiocyanates as potential natural antifungal compounds against citrus sour rot disease agent Geotrichum citri-aurantii

In this study, the antifungal effects of six different isothiocyanate (ITCs) compounds (methyl, allyl, butyl, ethyl, benzyl and 2-phenylethyl ITCs) were investigated to be use against the citrus sour rot disease caused by Geotrichum citri-aurantii in vitro and semi-commercial (in vivo) conditions. Antifungal activities of the vapour phases of different ITC compounds were examined on the arthroconidia germination and mycelial growth of G. citri-aurantii. Mycelial growth of G. citri-aurantii was inhibited in a concentration-dependent way. The minimum inhibitory concentrations of benzyl, methyl, allyl and ethyl ITCs on mycelial growth were 0.06, 0.08, 0.10 and 0.10 µl/L, respectively. Arthroconidia germination of G. citri-aurantii was completely inhibited by benzyl, methyl, allyl and ethyl ITCs at concentrations of 0.05, 0.07, 0.07 and 0.07 µl/L, respectively. Light microscopy observations revealed that the ITC compounds, at completely inhibiting concentrations, caused considerable morphological changes in the fungal hyphae. Under in vivo conditions, the average rotting area caused by G. citri-arantii was inhibited 100% by ethyl, methyl and allyl ITC compounds at concentrations of 8.0, 12.0 and 12.0 µl/L, respectively. Results suggest that ITC’s may be useful and effective natural antifungal compounds to control the citrus sour rot disease agent.     

Effect of temperature and water activity on in vitro germination of Monilinia spp.

Aims:  This study evaluated the effect of temperature (0–38°C) and water activity ( a _w: 0·87–0·99) on the lag phase prior to germination and the percentage of germination over time for Monilinia laxa , Monilinia fructicola and Monilinia fructigena .
Methods and Results:  More than 80% of viable conidia germinated at 25°C and 0·99 a _w within 2 h for M. fructicola and M. fructigena and 4 h for M. laxa . There was no germination at 38°C, and all three Monilinia spp. germinated at 0°C. At the lowest a _w (0·87), none of the Monilinia spp. was able to germinate at any of the incubation temperatures studied. Whereas at 0·90 a _w, conidia were only able to germinate at 15, 25 and 30°C for the three species studied, except for M. fructicola at 15°C. In contrast, at 0·95, 0·97 and 0·99 a _w, germination occurred at all studied temperatures less 38°C. Generally, the lag phase was longer at low levels of a _w (0·90–095), and differences were more evident as temperatures were far from the optimum (0–5°C).
Conclusions:  Germination and lag phase period were markedly influenced by temperature and a _w, and in general when conditions of temperature and a _w were suboptimal, the lag phase was longer and the percentage of germination was lower.
Significance and Impact of the Study:  Knowledge of the germination requirements of this fungus is important in order to understand their behaviour in natural situations and to provide baseline data required for the construction of new prediction models. Our study might be used to develop a predictive model to understand and control the disease caused by Monilinia spp.     

Bovicin HC5 reduces thermal resistance of Alicyclobacillus acidoterrestris in acidic mango pulp

Aims:  To test the effect of bovicin HC5 against vegetative cells and endospores of Alicyclobacillus acidoterrestris DSMZ 2498 in synthetic media and in acidic mango pulp.
Methods and Results:  Alicyclobacillus acidoterrestris was grown in synthetic medium at 40°C and pH 4·0. The effect on vegetative cells was assayed by adding bovicin HC5 to synthetic medium (40–160 AU ml⁻¹) or to mango pulp (100 AU ml⁻¹) at various pH values and determining the effect on growth (OD_600nm) and viable cell number, respectively. The effect of bovicin HC5 on spore germination and thermal sensitivity of A. acidoterrestris was tested in mango pulp (pH 4·0) containing 80 AU ml⁻¹ of bovicin HC5. Bovicin HC5 was bactericidal against vegetative cells of A. acidoterrestris at different pH values and showed sporicidal activity against endospores of this bacterium. When spores of A. acidoterrestris were heat treated in the presence of bovicin HC5, D -values decreased 77% to 95% compared to untreated controls at temperatures ranging from 80 to 95°C.
Conclusion:  Bovicin HC5 was bactericidal and sporicidal against A. acidoterrestrsi DSMZ 2498.
Significance and Impact of the Study:  These results indicated that bovicin HC5 has potential to prevent spoilage of acidic fruit juices by thermocidophilic spore-forming bacteria.     

The combined effects of high pressure and nisin on germination and inactivation of Bacillus spores in milk

Aims:  The aim of this work was to investigate the germination and inactivation of spores of Bacillus species in buffer and milk subjected to high pressure (HP) and nisin.
Methods and Results:  Spores of Bacillus subtilis and Bacillus cereus suspended in milk or buffer were treated at 100 or 500 MPa at 40°C with or without 500 IU ml⁻¹ of nisin. Treatment at 500 MPa resulted in high levels of germination (4 log units) of B. subtilis spores in both milk and buffer; this increased to >6 logs by applying a second cycle of pressure. Viability of B. subtilis spores in milk and buffer was reduced by 2·5 logs by cycled HP, while the addition of nisin (500 IU ml⁻¹) prior to HP treatment resulted in log reductions of 5·7 and 5·9 in phosphate buffered saline and milk, respectively. Physical damage of spores of B. subtilis following HP was apparent using scanning electron microscopy. Treating four strains of B. cereus at 500 MPa for 5 min twice at 40°C in the presence of 500 IU ml⁻¹ nisin proved less effective at inactivating the spores of these isolates compared with B. subtilis and some strain-to-strain variability was observed.
Conclusions:  Although high levels of germination of Bacillus spores could be achieved by combining HP and nisin, complete inactivation was not achieved using the aforementioned treatments.
Significance and Impact of the Study:  Combinations of HP treatment and nisin may be an appealing alternative to heat pasteurization of milk.     

Survival and growth of Cronobacter species (Enterobacter sakazakii) in wheat-based infant follow-on formulas

Aims:  To determine the survival and growth characteristics of Cronobacter species ( Enterobacter sakazakii ) in infant wheat-based formulas reconstituted with water, milk, grape juice or apple juice during storage.
Methods and Results:  Infant wheat-based formulas were reconstituted with water, ultra high temperature milk, pasteurized grape or apple juices. The reconstituted formulas were inoculated with Cronobacter sakazakii and Cronobacter muytjensii and stored at 4, 25 or 37°C for up to 24 h. At 25 and 37°C, Cronobacter grew more (>5 log₁₀) in formulas reconstituted with water or milk than those prepared with grape or apple juices ( c. 2–3 log₁₀). The organism persisted, but did not grow in any formulas stored at 4°C. Formulas reconstituted with water and milk decreased from pH 6·0 to 4·8–5·0 after 24 h, whereas the pH of the formulas reconstituted with fruit juices remained at their initial pH values, c. pH 4·8–5·0.
Conclusions:  Cronobacter sakazakii and C. muytjensii can grow in reconstituted wheat-based formulas. If not immediately consumed, these formulas should be stored at refrigeration temperatures to reduce the risk of infant infection.
Significance and Impact of the Study:  The results of this study will be of use to regulatory agencies and infant formula producers to recommend storage conditions that reduce the growth of Cronobacter in infant wheat-based formulas.     

Production and partial characterization of lipases from a newly isolated Penicillium sp. using experimental design

Aims:  The objective of this work was to investigate the lipase production by a newly isolated Penicillium sp . , using experimental design technique, in submerged fermentation using a medium based on peptone, yeast extract, NaCl and olive oil, as well as to characterize the crude enzymatic extracts obtained.
Methods and Results:  Lipase activity values of 9·5 U ml⁻¹ in 96 h of fermentation was obtained at the maximized operational conditions of peptone, yeast extract, NaCl and olive oil concentrations (g l⁻¹) of 20·0, 5·0, 5·0 and of 10·0 respectively. The partial characterization of crude enzymatic extract obtained by submerged fermentation showed optimum activity at pH range from 4·9 to 5·5 and temperature from 37°C to 42°C. The crude extract maintained its initial activity at freezing temperatures up to 100 days.
Conclusions:  A newly isolated strain of Penicillium sp . used in this work yielded good lipase activities compared to the literature.
Significance and Impact of the Study:  The growing interest in lipase production is related to the potential biotechnological applications that these enzymes present. New lipase producers are relevant to finding enzymes with different catalytic properties of commercial interest could be obtained, without using genetically modified organisms (GMO).     

Sensitivity of wild and genetic sexing strain Ceratitis capitata to high-temperature disinfestation of Valencia oranges

Abstract:  The purpose of this research was to investigate post-harvest heat treatment of Valencia oranges as an effective disinfestation protocol (fast, no fruit damage) for the Mediterranean fruit fly Ceratitis capitata (Wiedemann). Forced high-temperature air was applied under the following conditions: (a) exposure to air temperature of 56°C, for fast increase of temperature in the interior of the fruit, (b) reduction in air temperature to 47°C when the fruit centre reached 47°C and (c) maintenance of fruits in the chamber for another 30 min. Relative humidity in the treatment chamber was kept between 50% and 65% during treatment. Forced air at 47°C applied for 30 min on eggs before hatch or late third instar larvae (the most heat-tolerant stages) resulted in complete kill. Egg and larval sensitivity to high temperature differed between a wild strain and a laboratory genetic-sexing strain based on white pupa mutation. In this strain males emerge from brown pupae and females from white pupae. In particular, mature eggs from the wild strain were significantly more temperature resistant than eggs from the laboratory strain. Exposure of Valencia oranges of a diameter of 7–7.5 cm to 56°C forced air for about 86 and 99 min was required to increase temperature to 47°C at 1.5 cm depth and the fruit centre respectively. Treated oranges showed no substantial peel or interior deterioration, or change in colour and taste when kept at 25°C and 50–60% RH for a period of up to 1 month following treatment. Treatment in 1% O₂ atmosphere, produced by flushing of CO₂ into the treatment chamber, resulted in about 1°C reduction in killing temperature and faster increase in temperature inside the fruit to a lethal level.     

The influence of the rate of temperature change on the activation of dormant seeds of Rumex obtusifolius L.

1. One temperature shift from 20 to 30°C in darkness induces 30–40% germination in Rumex obtusifolius seeds. The same germination percentages are found with heat treatment varying between 1 and 6h duration, indicating that the total heat sum of the temperature shift is not important.
2. Germination is greatly enhanced by three consecutive heat shifts of 1h at 30°C separated by 1h periods at 20°C.
3. The seeds are activated to a small extent after a slow warming (+2°Ch^–1) from 20 to 30°C, followed by incubation for 1h at 30°C. Germination is much higher after rapid heating (+10°Ch^–1) to 30°C, followed by 1h incubation at this temperature. Repeated fast heating treatments on four consecutive days enhances germination. Moderately rapid heatings (+3·3°Ch^–1) give intermediate results.
4. The rate of cooling does not influence the germination percentage. Cooling alone cannot induce germination.
5. Heating alone from 15 to 25°C without cooling also activates germination. In this temperature range the seeds are more activated by rapid warming than by slow warming.
6. The ecological relevance of the response to different warming rate is discussed. The insensitivity of seeds to a slow warming might keep deeply buried seeds in a dormant stage.     

In vitro and in vivo antagonism of actinomycetes isolated from Moroccan rhizospherical soils against Sclerotium rolfsii: a causal agent of root rot on sugar beet (Beta vulgaris L.)

Aims:  To evaluate the ability of the isolated actinomycetes to inhibit in vitro plant pathogenic fungi and the efficacy of promising antagonistic isolates to reduce in vivo the incidence of root rot induced by Sclerotium rolfsii on sugar beet.
Methods and Results:  Actinomycetes isolated from rhizosphere soil of sugar beet were screened for antagonistic activity against a number of plant pathogens, including S.   rolfsii . Ten actinomycetes out of 195 screened in vitro were strongly inhibitory to S. rolfsii . These isolates were subsequently tested for their ability to inhibit sclerotial germination and hyphal growth of S. roflsii . The most important inhibitions were obtained by the culture filtrate from the isolates J-2 and B-11, including 100% inhibition of sclerotial germination and 80% inhibition of hyphal growth. These two isolates (J-2 and B-11) were then screened for their ability to protect sugar beet against infection of S. rolfsii induced root rot in a pot trial. The treatment of S. rolfsii infested soil with a biomass and culture filtrate mixture of the selected antagonists reduced significantly ( P  ≤ 0·05) the incidence of root rot on sugar beet. Isolate J-2 was most effective and allowed a high fresh weight of sugar beet roots to be obtained. Both antagonists J-2 and B-11 were classified as belonging to the genus Streptomyces species through morphological and chemical characteristics as well as 16S rDNA analysis.
Conclusion:  Streptomyces isolates J-2 and B-11 showed a potential for controlling root rot on sugar beet and could be useful in integrated control against diverse soil borne plant pathogens.
Significance and Impact of the Study:  This investigation showed the role, which actinomycete bacteria can play to control root rot caused by S.   rolfsii , in the objective to reduce treatments with chemical fungicides.     

Heat resistance of Cronobacter species (Enterobacter sakazakii) in milk and special feeding formula

Aim:  To determine D - and z -values of Cronobacter species ( Enterobacter sakazakii ) in different reconstituted milk and special feeding formula and the effect of reconstitution of powdered milk and special feeding formula with hot water on the survival of the micro-organism.
Methods and Results:  Five Cronobacter species (four C. sakazakii isolates and C. muytjensii ) were heated in reconstituted milk or feeding formula pre-equilibrated at 52–58°C for various times or mixed with powdered milk or feeding formula prior to reconstitution with water at 60–100°C. The D -values of Cronobacter at 52–58°C were significantly higher in whole milk (22·10–0·68 min) than in low fat (15·87–0·62 min) or skim milk (15·30–0·51 min) and significantly higher in lactose-free formula (19·57–0·66 min) than in soy protein formula (17·22–0·63 min). The z -values of Cronobacter in reconstituted milk or feeding formula ranged from 4·01°C to 4·39°C. Water heated to ≥70°C and added to powdered milk and formula resulted in a > 4 log₁₀ reduction of Cronobacter .
Conclusions:  The heat resistance of Cronobacter should not allow the survival of the pathogen during normal pasteurization treatment. The use of hot water (≥70°C) during reconstitution appears to be an effective means to reduce the risk of Cronobacter in these products.
Significance and Impact of the Study:  This study supports existing data available to regulatory agencies and milk producers that recommended heat treatments are sufficient to substantially reduce risk from Cronobacter which may be present in these products.     

Antimicrobial properties of plant essential oils and essences against five important food-borne pathogens

The antimicrobial properties of 21 plant essential oils and two essences were investigated against five important food-borne pathogens, Campylobacter jejuni, Salmonella enteritidis, Escherichia coli, Staphylococcus aureus and Listeria monocytogenes. The oils of bay, cinnamon, clove and thyme were the most inhibitory, each having a bacteriostatic concentration of 0·075% or less against all five pathogens. In general, Gram-positive bacteria were more sensitive to inhibition by plant essential oils than the Gram-negative bacteria. Campylobacter jejuni was the most resistant of the bacteria investigated to plant essential oils, with only the oils of bay and thyme having a bacteriocidal concentration of less than 1%. At 35 °C, L. monocytogenes was extremely sensitive to the oil of nutmeg. A concentration of less than 0·01% was bacteriostatic and 0·05% was bacteriocidal, but when the temperature was reduced to 4 °C, the bacteriostatic concentration was increased to 0·5% and the bacteriocidal concentration to greater than 1%.     

Antifungal activity of Moroccan medicinal plants against citrus sour rot agent Geotrichum candidum

Aims: The aim of this work was to find an alternative to the chemical fungicides currently used in the control of Geotrichum candidum, the causal agent of citrus sour rot. Methods and Results: Minimal inhibition concentrations (MIC) and minimal fungicidal concentrations (MFC) were determined using agar dilution method. The methanol extracts of Cistus villosus, Ceratonia siliqua and Halimium umbellatum exhibited strong antifungal activity with MIC values ranged between 0·156 and 1·25 mg ml^?1, and MFC values ranged between 2·5 and 5 mg ml^?1. Incidence of sour rot was lowered to 0·00, 3·33 and 11·66% when mandarin fruit was treated with C. villosus, C. siliqua and H. umbellatum methanol extracts at 50 mg ml^?1, respectively, compared with 95% in the control. Conclusions: Cistus villosus, C. siliqua and H. umbellatum methanol extracts successfully reduced the disease incidence caused by G. candidum, and no phytotoxic effects were recorded on citrus fruit. Significance and Impact of the Study: These findings suggest that C. villosus, C. siliqua and H. umbellatum plants may be useful and effective agents for control of citrus sour rot. Such natural products therefore represent a sustainable alternative to the use of synthetic fungicides.     

Effects of temperature on metal tolerance and the accumulation of Zn and Pb by metal-tolerant fungi isolated from urban runoff treatment wetlands

Aims:  To investigate the ability of two fungi to accumulate Zn and Pb, the effect of temperature on their metal tolerance and possible mechanisms involved in metal accumulation.
Methods and results:  Beauveria bassiana and Rhodotorula mucilaginosa isolated from constructed wetlands receiving urban runoff were grown in modified glycerol asparagine medium containing elevated levels of Zn and Pb at 30°C. Beauveria bassiana accumulated up to 0·64% of available Zn and 8·44% of Pb. The corresponding values for R. mucilaginosa were up to 2·05% for Zn and 16·55% for Pb. Radial growth of colonies grown at 4° and 30°C on agar containing Zn or Pb indicated that metal tolerance was not seriously affected by a decrease in temperature. Transmission electron microscopy and emission dispersion x-ray spectrophotometry suggested that the mechanism of resistance in B. bassiana may be associated with the precipitation of Pb (possibly in the form of oxalates).
Conclusion:  The processes of biosorption could potentially occur throughout the year with both living and dead cells able to accumulate metals.
Significance and Impact of the Study:  Identified precipitation processes could be an important mechanism in metal removal in wetland substrates serving as long-term storage sinks.     

Evaluation of commercial essential oil samples on the growth of postharvest pathogen Monilinia fructicola (G. Winter) Honey

Aim: To assess the effect of several commercial essential oils samples Australian lemon myrtle (Backhousia citriodora), cinnamon bark (Cinnamomum zeylanicum), oregano (Origanum vulgare), thyme oil (Thymus vulgaris), clove bud (Eugenia caryophyllata), valerian (Valeriana officinalis) and Australian tea tree oil (Melaleuca alternifolia) on mycelium growth and spore germination of Monilinia fructicola. The effectiveness of lemon myrtle essential oil as a fumigant for the control of brown rot in nectarines was evaluated. Methods and Results: Monilinia fructicola exhibited a different level of sensitivity to each tested essential oil with results suggesting that the essential oils provide excellent control of the pathogen with respect to mycelium growth and spore germination at very low concentrations, whereas for others higher concentrations are needed to reduce significant fungal growth. In vivo application of lemon myrtle essential oil effectively reduced the incidence of M. fructicola on noninoculated fruit. Fumigation of nectarines following inoculation did not reduce the incidence of brown rot in comparison with the inoculated control treatment. No evidence of phytotoxicity on the fruit was recorded. Conclusions: Lemon myrtle essential oil exhibited the strongest antifungal activity against M. fructicola, in vitro and to a lesser extent, under in vivo conditions. Significance and Impact of the Study: The results demonstrate that lemon myrtle essential oil, in particular, has potential as an antifungal agent to control M. fructicola.     

Change in susceptibility of satsuma mandarin fruit to sour rot pathogen (Geotrichum candidum citrus race) with relation to biochemical changes during maturation and storage

Change in susceptibility of satsuma mandarin fruit (Citrus unshiu) cultivar “Miyagawawase” to sour rot pathogen was studied with relation to biochemical changes during maturation and storage. The susceptibility of the fruit decreased with the advancement of maturity and was relatively constant during storage at 25°C for 4 wk. The young, green fruit that contained less total soluble solid, sugars and polyphenol, but more citric acid and water contents than mature, yellow fruit was more susceptible to sour rot pathogen. The susceptibility was correlated with total soluble solid, citric acid, sugars and water contents, but not with polyphenol. The results suggested that the difference in susceptibility during maturation was influenced to some extent by several constituents of fruit, although they may not be the only factors involved in susceptibility.     

Modelling the growth of Weissella cibaria as a function of fermentation conditions

Aims:  To investigate the effect of pH, water activity ( a _w) and temperature on the growth of Weissella cibaria DBPZ1006, a lactic acid bacterium isolated from sourdoughs.
Methods and Results:  The kinetics of growth of W. cibaria DBPZ1006 was investigated during batch fermentations as a function of pH (4·0–8·0), a _w (0·935–0·994) and temperature (10–45°C) in a rich medium. The growth curve parameters (lag time, growth rate and asymptote) were estimated using the dynamic model of Baranyi and Roberts (1994. A dynamic approach to predicting bacterial growth in food. Int J Food Microbiol 23, 277–294). The effect of pH, a _w and temperature on maximum specific growth rate (μ_max) were estimated by fitting a cardinal model. μ_max under optimal conditions (pH = 6·6, a _w = 0·994, T  = 36·3°C) was estimated to be 0·93 h⁻¹. Minimum and maximum estimated pH and temperature for growth were 3·6 and 8·15, and 9·0°C and 47·8°C, respectively, while minimum a _w was 0·918 (equivalent to 12·2% w/v NaCl).
Conclusions:  Weissella cibaria DBPZ1006 is a fast-growing heterofermentative strain, which could be used in a mixed starter culture for making bread.
Significance and Impact of the Study:  This is the first study reporting the modelling of the growth of W. cibaria , a species that is increasingly being used as a starter in sourdough and vegetable fermentations.     

Characterization of Metarhizium species and varieties based on molecular analysis, heat tolerance and cold activity

Aims:  The genetic relationships and conidial tolerances to high and low temperatures were determined for isolates of several Metarhizium species and varieties.
Methods and Results:  Molecular-based techniques [AFLP and rDNA (ITS1, ITS2 and 5·8S) gene sequencing] were used to characterize morphologically identified Metarhizium spp. isolates from a wide range of sources. Conidial suspensions of isolates were exposed to wet heat (45 ± 0·2°C) and plated on potato dextrose agar plus yeast extract (PDAY) medium. After 8-h exposure, the isolates divided clearly into two groups: (i) all isolates of Metarhizium anisopliae var. anisopliae ( Ma-an ) and Metarhizium from the flavoviride complex ( Mf ) had virtually zero conidial relative germination (RG), (ii) Metarhizium anisopliae var. acridum ( Ma-ac ) isolates demonstrated high heat tolerance ( c . 70–100% RG). Conidial suspensions also were plated on PDAY and incubated at 5°C for 15 days, during which time RGs for Ma-an and Ma-ac isolates were virtually zero, whereas the two Mf were highly cold active (100% RG).
Conclusions:  Heat and cold exposures can be used as rapid tools to tentatively identify some important Metarhizium species and varieties.
Significance and Impact of the Study:  Identification of Metarhizium spp. currently relies primarily on DNA-based methods; we suggest a simple temperature-based screen to quickly obtain tentative identification of isolates as to species or species complexes.     

Nonisothermal heat resistance determinations with the thermoresistometer Mastia

Aims:  To design and build a thermoresistometer, named Mastia, which could perform isothermal and nonisothermal experiments.
Methods and Results:  In order to evaluate the thermoresistometer, the heat resistance of Escherichia coli vegetative cells and Alicyclobacillus acidoterrestris spores was explored. Isothermal heat resistance of E. coli was characterized by D _60°C = 0·38 min and z =  4·7°C in pH 7 buffer. When the vegetative cells were exposed to nonisothermal conditions, their heat resistance was largely increased at slow heating and fast cooling rates. Isothermal heat resistance of A. acidoterrestris was characterized by D _95°C = 7·4 min and z =  9·5°C in orange juice. Under nonisothermal conditions, inactivation was reasonably well predicted from isothermal data.
Conclusions:  The thermoresistometer Mastia is a very suitable instrument to get heat resistance data of micro-organisms under isothermal and nonisothermal treatments.
Significance and Impact of the Study:  The thermoresistometer Mastia can be a helpful tool for food processors in order to estimate the level of safety of the treatments they apply.     

Effects of Temperature on Germination of Peronospora parasitica Conidia and Infection of Brassica oleracea

The effect of temperature on germination of a South African isolate of Peronospora parasitica , and infection of Brassica oleracea was studied. The optimum condition for germination was 20° C at 100% relative humidity. The percentage germination obtained was 80–98% and 70–80% between 15 and 25° C at 100% relative humidity, after a 12 and 6h incubation period, respectively. Optimum temperature for germ tube growth was also 20° C. The temperature range for maximum infection of seedlings of a highly susceptible cabbage cultivar and subsequent disease development in vitro was 15–25° C and 90–100% infection was achieved after 48 h of incubation. At<15°C and 26–30° C infection percentage was decreased to 40–50% and 35–40%, respectively. No disease incidence was recorded at temperatures above 35° C. A scanning electron microscope study of the infection process showed that penetration of cotyledons by germ tubes was mostly via stomata and occasionally directly through the cuticle. Results are discussed in relation to the need for future studies of P. parasitica in South Africa.     

Effect of certain inorganic chemicals on growth and spore germination of Alternaria tenuis auct., the fungus causing core rot of mandarin oranges in India

Summary Laboratory studies to determine the tolerance of an isolate ofAlternaria tenuis causing a black core rot of mandarin oranges (Citrus reticulata) have shown that presence of low amounts of manganese, zinc, iron, copper, boron, and molybdenum in a synthetic medium stimulated growth and sporulation of the fungus. Concentrations of 20 ppm or more of copper sulphate, boric acid, and ammonium molybdate, however, caused significant reduction of growth. Conidia of the fungus were very sensitive to even low amounts of copper, zinc, and iron. Copper sulphate was most toxic to spore germination followed, in order of inhibitory effect, by zinc sulphate, ferrous sulphate, and boric acid. Ammonium molybdate and manganese sulphate were stimulatory to spore germination at 5 ppm. There was a positive correlation between percentage of spore germination and average length of germ tubes in different concentrations of various chemicals. The possibility of using copper sulphate, zinc sulphate, and ferrous sulphate as spray materials for trees and as dip for ripe fruits to prevent black core rot are discussed.Assoc. Professor and Research Assistant, respectively. Publication under journal series No. 27 from Experiment Station, U.P. Agricultural University.