Antifungal vapour-phase activity of allyl-isothiocyanate against Penicillium expansum on pears

The fungitoxicity of allyl-isothiocyanate (AITC) vapour against Penicillium expansum , the causal agent of blue mould on pears (cvs Conference and Kaiser), was evaluated. The best control of blue mould was obtained by exposing fruits for 24 h in a 5 mg L⁻¹ AITC-enriched atmosphere, the extent of control depending on the inoculum density. Lesion diameter was inversely related to AITC concentration. In treated fruits the percentage of infected wounds increased with conidial concentration, with fewer than 20% affected at 1 × 10³ conidia mL⁻¹ to almost 80% at 1 × 10⁶ conidia mL⁻¹. In comparison, >98% of wounds were infected in untreated fruits irrespective of conidial concentration. AITC treatments were effective up to 24 h after inoculation for Conference and 48 h for Kaiser. AITC treatments also controlled a thiabendazole-resistant strain of P. expansum , reducing the incidence of blue mould by 90% in both cultivars. The use of AITC produced from pure sinigrin or from Brassica juncea defatted meal may be an economically viable alternative to synthetic fungicides against P. expansum .     

TRANSLOCATION OF 14C-ASSIMILATES AND 3-AMINO-1,2,4-TRIAZOLE AND ITS METABOLITES IN AGROPYRON REPENS

Summary. Translocation of 3-amino-1,2,4-triazole-5-¹⁴C (¹⁴C-aminotriazole) was compared to that of ¹⁴C-assimilates in couch grass ( Agropyron repens (L.) Beauv.) at three different growth stages.
Assimilates of ¹⁴CO₂ were translocated from the treated shoot to other shoots and rhizomes at the 2–3-leaf and 3–4-leaf stages of development. Much less labelled material was translocated into untreated shoots at the 5-leaf stage. More ¹⁴C-assimilates were translocated to the roots than to untreated shoots at all developmental stages. The translocation patterns of ¹⁴C-aminotriazole and ¹⁴C-assimilates were similar.
Two metabolites, A and B, were formed from ¹⁴C-aminotriazole, which chromatographed identically to previously described metabolites in Cirsium arvense (L.) Scop. A was further metabolized into B. Labelled aminotriazole and its two metabolites were translocated throughout the plants. Metabolite A was phytotoxic when concentrated and re-applied to couch grass, but its properties were not those of Unknown II from Cirsium arvense. They were the same as those of Unknown III. Whether or not metabolite A and Unknown III are identical was not established.
Migration des substances assimilables marquées au ¹⁴ C, du 3 amino-1,2,4-triazole et de ses métabolites chez Agropyron repens     

Berry infection and the development of bunch rot in grapes caused by Aspergillus carbonarius

The effects of different levels of inoculum of Aspergillus carbonarius and time of inoculation on berry infection and the development of aspergillus bunch rot on grapevines (cv. Sultana) were studied under field conditions. Inflorescences at full bloom were inoculated with aqueous spore suspensions of A. carbonarius containing 0 or 1 × 10⁶ spores mL⁻¹ in 2004/05 and 0, 1 × 10² or 1 × 10⁵ spores mL⁻¹ in 2005/06. In both years, the incidence of infection in inoculated berries was significantly higher than in uninoculated berries. Incidence of infection in berries from veraison until harvest was higher than at earlier stages of bunch development (berry set to berries that were still hard and green). Inoculation of bunches at veraison did not significantly increase A. carbonarius infection prior to harvest, at harvest, 6 days after harvest or when berries were over-ripe. Bunches inoculated at harvest did not significantly increase infection 6 days after harvest or when berries were over-ripe. Aspergillus carbonarius was isolated more frequently from the pedicel end (53·1%) than from the middle section (37·5%) and distal end (35·0%) of berries that were inoculated with 10⁵ spores mL⁻¹.     

PCR-based specific and sensitive detection of Pectobacterium carotovorum ssp. carotovorum by primers generated from a URP-PCR fingerprinting-derived polymorphic band

A 24-mer primer pair was generated by sequencing a URP-PCR fingerprinting-derived polymorphic band that is uniquely shared in Pectobacterium carotovorum ssp . carotovorum strains (Pcc). The primer set (EXPCCF/EXPCCR) amplified a single band of expected size (0·55 kb) from genomic DNA obtained from 29 Pcc strains and three Pectobacterium carotovorum ssp. wasabiae (Pcw) strains, but not from other P. carotovorum subspecies atrosepticum , betavasculorum or odoriferum , or from other Erwinia spp. or bacterial genera. The Rsa I digestion profile of the amplified bands divided Pcc strains into five groups with a unique profile from Pcw strains. First-round PCR detected between 5 × 10² and 1 × 10³ colony forming units (CFU) mL⁻¹ and detection sensitivity was increased to as few as 2–4 CFU mL⁻¹ after second-round (nested) PCR. This PCR protocol was used directly to detect Pcc strains in infected plant tissues.     

Sensitivity of Sclerotinia homoeocarpa to demethylation-inhibiting fungicides in Ontario, Canada, after a decade of use

In late 2003, nine populations of Sclerotinia homoeocarpa in Ontario Canada (seven of which had been previously sampled in early 1994, prior to the registration of sterol demethylation-inhibiting (DMI) fungicides for turf disease control in Canada) were sampled and tested for sensitivity to propiconazole. Four of the nine populations had not been treated with DMI fungicides during the intervening years, and isolates from these locations were sensitive to propiconazole (geometric mean EC₅₀ values of 0·005–0·012 µ g mL⁻¹, compared with 0·005–0·008 µ g mL⁻¹ for the original 1994 populations). Among the five populations from 2003 that had been exposed to DMI fungicides, mean EC₅₀ values were significantly greater, ranging from 0·020 to 0·048 µ g mL⁻¹. A significant correlation of determination was found between estimated number of fungicide applications and log EC₅₀ ( R ² = 0·832, P  = 0·0001), and the equation predicted that 42·3 applications of propiconazole would be needed to bring a sensitive population (EC₅₀ < 0·01  µ g mL⁻¹) to a resistant level (EC₅₀ > 0·10  µ g mL⁻¹). Fungicide sensitivity vs. duration of fungicide efficacy was also tested, and it was found that isolates with decreased sensitivity were able to more quickly overcome the inhibitory effects of fungicide application, reducing the duration of control from 3 weeks to 2 weeks.     

Interaction of a bioherbicide and glyphosate for controlling hemp sesbania in glyphosate-resistant soybean

The bioherbicidal fungus, Colletotrichum truncatum (Schwein.) Andrus & Moore, was tested at different inoculum concentrations alone and in combination with, prior to or following treatment with different rates of glyphosate ( N -[phosphonomethyl]glycine) (Roundup Ultra) for the control of hemp sesbania ( Sesbania exaltata [Raf.] Rydb. ex A.W. Hill) in Roundup Ready soybean field plots. Colletotrichum truncatum and glyphosate were applied in all pair-wise combinations of 0, 1.25, 2.5, 5.0, and 10.0 × 10⁶ spores mL⁻¹ (i.e. 3.125, 6.25, 12.5, and 25 × 10¹¹ spores ha⁻¹), and 0.15, 0.30, 0.60, and 1.2 kg ha⁻¹, respectively. Weed control and disease incidence were enhanced at the two lowest fungal and herbicidal rates when the fungal spores were applied after glyphosate treatment. The application of the fungus in combination with or prior to glyphosate application at 0.30 kg ha⁻¹ resulted in reduced disease incidence and weed control regardless of the inoculum's concentration. At the highest glyphosate rates, the weeds were controlled by the herbicide alone. These results suggest that it might be possible to utilize additive or synergistic herbicide and pathogen interactions to enhance hemp sesbania control.     

Visual and PCR assessment of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) cultivars

Methods to assess light leaf spot ( Pyrenopeziza brassicae ) on winter oilseed rape cultivars were compared in laboratory, controlled-environment and field experiments. In controlled-environment experiments with seedling leaves inoculated at GS 1,4, the greatest differences in percentage area affected by P. brassicae sporulation were observed with inoculum concentrations of 4 × 10³ or 4 × 10⁴ spores mL⁻¹, rather than 4 × 10² or 4 × 10⁵ spores mL⁻¹, but older leaves had begun to senesce before assessment, particularly where they were severely affected by P. brassicae . In winter oilseed rape field experiments, a severe light leaf spot epidemic developed in 2002/03 (inoculated, September/October rainfall 127·2 mm) but not in 2003/04 (uninoculated, September/October rainfall 40·7 mm). In-plot assessments discriminated between cultivars best in February/March in 2003 and June in 2004, but sometimes failed to detect plots with many infected plants (e.g. March/April 2004). Ranking of cultivar resistance differed between seedling experiments done under controlled-environment conditions and field experiments. The sensitivity of detection of P. brassicae DNA extracted from culture was greater using the PCR primer pair PbITSF/PbITSR than using primers Pb1/Pb2. P. brassicae was detected by PCR (PbITS primers) in leaves from controlled-environment experiments immediately and up to 14 days after inoculation, and in leaves sampled from field experiments 2 months before detection by visual assessment.     

Estimating relative crop yield loss resulting from herbicide damage using crop ground cover or rated stunting, with maize and sethoxydim as a case study

The research goal was to determine whether crop damage from herbicides measured early in the growing season soon after treatment could be used to estimate relative crop yield loss. Percentage stunting was rated visually and percentage crop ground cover (i.e. percentage of the ground surface covered by vegetation) was determined from video photographs taken 2–4 weeks after sethoxydim-susceptible maize ( Zea mays L.) was sprayed with sethoxydim at various rates plus crop oil concentrate. Averaged over 3 years, relative percentage maize yield was a negative sigmoidal function of relative sethoxydim rates from 0.065× to 0.5×, where the 1× rate was 420 g a.i. ha⁻¹ ( r ² = 0.80). Relative maize yield was positively linearly related to percentage crop ground cover and negatively linearly related to rated percentage stunting averaged over 3 years. Linear regression models of relative maize yield vs. percentage maize ground cover explained only slightly more data variability ( r ² = 0.86) than did rated stunting ( r ² = 0.82) over 3 years. The advantages and disadvantages of rated stunting and crop ground cover as scientific measurements are discussed.     

Production of conidia by Peronosclerospora sorghi on sorghum crops in Zimbabwe

Factors affecting the production of conidia of Peronosclerospora sorghi , causing sorghum downy mildew (SDM), were investigated during 1993 and 1994 in Zimbabwe. In the field conidia were detected on nights when the minimum temperature was in the range 10–19°C. On 73% of nights when conidia were detected rain had fallen within the previous 72 h and on 64% of nights wind speed was < 2.0 m s⁻¹. The time period over which conidia were detected was 2–9 h. Using incubated leaf material, conidia were produced in the temperature range 10–26°C. Local lesions and systemically infected leaf material produced 2.4–5.7 × 10³ conidia per cm². Under controlled conditions conidia were released from conidiophores for 2.5 h after maturation and were shown to be well adapted to wind dispersal, having a settling velocity of 1.5 × 10⁻⁴ m s⁻¹. Conditions that are suitable for conidia production occur in Zimbabwe and other semi-arid regions of southern Africa during the cropping season.     

Effects of nitrogen, phosphorus, potassium and calcium nutrition on strawberry anthracnose

The effects of a range of concentrations of four nutrients – nitrogen, phosphorus, potassium and calcium – in fertilizer solutions on the severity of anthracnose on strawberry cv. Nyoho cultivated under a noncirculation hydroponics system were determined after inoculation with Colletotrichum gloeosporioides . Crop growth and tissue nitrogen, phosphorus, potassium and calcium contents of the entire above-ground parts of the plant were also investigated. Elevated nitrogen and potassium concentrations in the fertilizer solution increased disease severity in contrast to phosphorus and calcium. Treatment with either NH₄ or NO₃ nitrogen was not significantly different. The dry weight of the strawberry plants increased significantly with elevated concentrations of nitrogen ( R ² = 0·9078) and phosphorus ( R ² = 0·8842), but was not influenced by the elevated amounts of potassium ( R ² = 0·8587) and calcium ( R ² = 0·6526) concentrations.     

Factors influencing infection of mechanical wounds by Fusarium circinatum on Monterey pines (Pinus radiata)

Pitch canker, caused by the fungus Fusarium circinatum , is a disease affecting many pine tree species. In California, Pinus radiata (Monterey pine) is the principal pine host affected by pitch canker. This investigation into factors affecting infection frequency by F. circinatum of P. radiata examined the influence of: (i) wound size; (ii) relative humidity; (iii) time of inoculation; (iv) inoculum density; and (v) wound age. Wounded branches sustained significantly more infections when large-diameter (1·6 mm) rather than small-diameter (0·5 mm) wounds were made. Infection frequencies tended to be higher at 100% RH than at ambient humidity, although these differences were not statistically significant. Infection frequencies were significantly higher on branches inoculated after 17·00 h than on branches inoculated before noon. Infection frequencies were significantly higher for wounded branches spray-inoculated with 5 × 10⁷ rather than 1 × 10⁷ spores mL⁻¹. Infection frequencies of pruning wounds declined as wounds aged.     

Current status of biological control of paddy weeds in Vietnam

Rice is a staple food in Vietnam and accounts for > 7.7 × 10⁶ cultivated ha, which provide 35.5 × 10⁶ t of rice, of which 4.2 × 10⁶ t were exported in 2004. The enlargement of the cropping area and the enhancement of rice yield have rapidly increased the amount of agrochemicals, including herbicides, in crop production in Vietnam. From 1990–2003, the percentage of herbicides in total pesticides has increased ≈ 10-fold to 30.2%. In addition, the improper use of herbicides caused environmental hazards, unsafe agricultural products, and human health problems. Biological management integrated with traditional weed control techniques might help to reduce the dependence on synthetic herbicides and build eco-friendly, sustainable agricultural production in Vietnam. This paper reviews the efforts in establishing a strategy for biological management of weeds that was conducted in recent years by Vietnamese weed scientists. This has included cropping system management, water and soil management, integrated pest management, and utilization of plant allelopathy as major components of the strategy. Many plants with strong allelopathic potential can be a source for biological weed suppression and soil fertility improvement. The utilization of allelopathic properties in rice might also help to provide new rice cultivars with weed-suppressing characteristics.     

Biological control of damping-off of sugar beet by Pseudomonas putida applied to seed pellets

Pseudomonas putida 40RNF applied to seed pellets reduced the occurrence of Pythium damping-off of sugar beet. A density of 6 × 10⁷ 40RNF per pellet reduced Pythium damping-off from 70 to 26% when seeds were sown in artificially infested soil (250 propagules Pythium ultimum per g dry soil). The efficacy of 40RNF was dependent on its density in the seed pellet (in the range 2 × 10⁴–6 × 10⁸ per pellet) and on the number of propagules of Pythium in soil. 40RNF declined to or stabilized at approximately 1 × 10⁶ per pellet 3 days after planting, and this was independent of the inoculum density. This indicated that the crucial steps resulting in damping-off of sugar beet caused by Pythium ultimum must occur within 3–4 days of sowing. 40RNF reduced pericarp colonization by P. ultimum by 43% 48 h after planting and caused a 68% decrease in the number of sporangia of P. ultimum in the surrounding soil (0.0–5.0 mm). P. putida 40RNF also reduced pre and post-emergence damping-off (from 69.5 to 37.5%) caused by indigenous populations of Pythium species in an infested soil and this was as effective as the fungicide hymexazol (69.5 to 40%).     

Factors affecting the control of Pythium ultimum damping-off of sugar beet by Pythium oligandrum

Application of Pythium oligandrum to a soil-based compost as a mycelial suspension (5 × 10² CFU g⁻¹ of dry compost) and oospore alginate pellets (10⁵ oospores/g of dry compost) controlled pre- and postemergence damping-off of sugar beet caused by Pythium ultimum to a level similar to metalaxyl seed treatment. Oospore seed treatments and aqueous suspensions of oospores applied to compost failed to control disease. Problems in the use of P. oligandrum oospore inocula for the control of damping-off were highlighted. It was shown that treatment of oospores with cellulase (20 g L⁻¹) increased germination approximately three-fold in comparison to untreated spores. Untreated and cellulase pretreated oospores were subsequently evaluated as seed treatments for their ability to control damping-off of sugar beet. The highest rate of pretreated oospores (10⁴ oospores/seed) gave levels of emergence and establishment in infested compost that were not significantly different from the uninfested controls, whereas seed treatment with untreated oospores gave no significant reduction in disease. In a trial carried out in a controlled environment to assess the effect of pH (4.5–8.0), P. oligandrum (10⁴ cellulase pretreated oospores/seed) was shown to control pre- and postemergence damping-off of sugar beet at pH 7.0 and 7.5 only.     

Factors influencing infection of eucalypts by Cylindrocladium pteridis

The pattern of Cylindrocladium pteridis adhesion, germination and penetration in eucalypt leaves was assessed using scanning electron microscopy. The effects of inoculum concentration, leaf wetness period, plant age and branch position of cylindrocladium leaf blight and defoliation severity were assessed in greenhouse studies using two Eucalyptus grandis × E. urophylla hybrid clones. Penetration occurred through stomata, and there was no difference in the number of penetrations between young and old leaves. Percentage leaf area with lesions and defoliation increased with the increase in inoculum concentration (1 × 10² to 10⁵ conidia mL⁻¹), duration of leaf wetness period (6 to 48 h) and plant age (60 to 180 days). Branch position in plants also significantly affected the percentage leaf area with lesions and defoliation, the latter variable being significantly higher at the stem base. The highest values of lesion area were also observed on leaves at the stem base in both clones. The Pearson correlation between defoliation and leaf area with lesions was significant in all experiments ( r  > 0·9) indicating a high association between these two variables.     

The incorporation of pathogen spores into rain-splash droplets: a modelling approach

Simple, theoretical, physical principles and existing experimental data were used to derive an analytical model to describe the incorporation of plant pathogen spores into splash droplets. Data were obtained from experiments on splash dispersal of spores of Pseudocercosporella herpotrichoides (cereal eyespot), Pyrenopeziza brassicae (oilseed rape light leaf spot) and Septoria nodorum (wheat glume blotch). In these experiments, incident drops of diameter 4–5 mm were allowed to fall onto spore suspensions 0.5 mm deep with 1.2 × 10⁵ to 6.5 × 10⁵ spores/mL. The analytical model was constructed as the product of three functions of droplet diameter which described, respectively, the frequency distribution of droplet diameters, the proportion of droplets carrying spores and the mean number of spores in spore-carrying droplets in each diameter category. The frequency distribution of droplet sizes was described by a log-normal distribution, the proportion of droplets carrying spores was described by an exponential function and the adimensional spore concentration in spore-carrying droplets was described by a power law. The cumulative proportions of spores in droplets in diameter categories of increasing diameter were calculated to compare observed and fitted data.     

Effect of 2,6-dichloroisonicotinic acid, its formulation materials and benzothiadiazole on systemic resistance to alternaria leaf spot in cotton

A wettable powder (WP) formulation providing 5–25 μg mL⁻¹ of 2,6-dichloroisonicotinic acid (INA) and 15–75 μg mL⁻¹ of WP applied to cotton cotyledons significantly increased the resistance of the next two leaves to challenge inoculation by Alternaria macrospora . The wettable powder alone at 15–75 μg mL⁻¹ had a lesser effect. A wettable granule (WG) formulation supplying 35 μg mL⁻¹ of benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH) and 35 μg mL⁻¹ of WG, applied as a cotyledonary treatment, significantly reduced the formation of lesions on the subsequent two leaves when challenged with A. macrospora . The WG control had no effect. Each treatment except for the WG control also raised the activities of β-1,3-glucanase in unchallenged leaf and stem tissue. Each of the components of the wettable powder without INA applied to cotyledons raised enzyme activities in the next leaves. Individual components, as suspensions of silicic acid and kaolin and solutions of the detergent Attisol II, the wetting agent Ultravon W300 and pure INA, applied to cotyledons increased the resistance of the next leaves to A. macrospora . The responsiveness of cotton to BTH and to each of the components of formulated INA is discussed in relation to knowledge of the effects of BTH and INA on other plants and to possible ways in which the other components of the wettable powder may affect the process of signalling for systemic resistance to disease.     

Growth and photosynthesis of four invasive aquatic plant species in Europe

Crassula helmsii , Hydrocotyle ranunculoides , Ludwigia grandiflora and Myriophyllum aquaticum are four well known invasive aquatic plants in European waters. In this study, plant growth at different nutrient availabilities, regeneration capacity and photosynthesis were investigated. Results show high relative growth rates (RGR) of the species of up to 0.132 ± 0.008 g g⁻¹ dry weight (dw) day⁻¹ ( H. ranunculoides ) and a significant increase in RGR with increasing nutrient availability. All species show a high regeneration capacity and the ability to form new shoots from single nodes, even though it differs between the species. Ludwigia grandiflora and M. aquaticum also show regeneration from single leaves. Species differed in maximal amounts, and in temperature and light optima of net assimilation rates: H. ranunculoides leaves reach maximum photosynthetic rates of up to 3500 μmol CO₂ × h⁻¹ g⁻¹ dw, L. grandiflora (leaves) up to 2200 μmol CO₂ × h⁻¹ g⁻¹ dw, M. aquaticum (shoots) 400   μmol CO₂ × h⁻¹ g⁻¹ dw and C. helmsii (shoots) up to 200 μmol CO₂ × h⁻¹ g⁻¹ dw. Hydrocotyle ranunculoides , L. grandiflora and M. aquaticum preferred high light intensity and high temperatures, whilst C. helmsii was negatively affected by intense sunlight. Summarising, it can be assumed that at least H. ranunculoides , L. grandiflora and M. aquaticum can grow well under current and likely future central European climate conditions.     

Influence of water management, application timing and temperature on efficacy of MTB-951, a mycoherbicide using Drechslera monoceras to control Echinochloa crus-galli L.

MTB-951 is a potential mycoherbicide using a fungal plant pathogen ( Drechslera monoceras ) isolated from native Echinochloa species in Japan. Conidia of this pathogen were used as the active ingredient and its herbicidal performance was examined in a greenhouse. The efficacy of MTB-951 on Echinochloa crus-galli L. was higher in deep water (7–9 cm) than in relatively shallow water (3–5 cm). In a postemergence application, the efficacy decreased as the leaf stage of E. crus-galli proceeded between the 1 and 2.5 leaf stage. For example, the control ratio (%) of E. crus-galli was 95% when applied at the 1 leaf stage, and 72% at the 2.5 leaf stage in 5 cm water. Generally, mycoherbicidal efficacy was less when applied pre-emergence rather than postemergence. Efficacy was also influenced by the duration of submergence in deep water. For example, when water depth was kept at 5 cm for more than 7 days after application and then decreased down to 3 cm, the efficacy was high. However, when the water depth was kept at 5 cm for less than 7 days, the efficacy was low. Efficacy was lower under high temperatures (35°C/25°C, day/night) than under low temperatures (25°C/15°C, day/night). Water management, application timing and temperature are important factors on herbicidal efficacy of MTB-951 to control E. crus-galli .     

Genetic control of resistance to fenpropimorph in Ustilago maydis

Fenpropimorph-resistant mutants of Ustilago maydis were obtained at high frequency (30 × 10⁻⁶) after UV-irradiation followed by selection on media containing fenpropimorph (50 μ g mL⁻¹). Genetic analysis of 30 such mutants resulted in the identification of two unlinked chromosomal loci, the U/fpm -1 locus with two allelic genes ( U/fpm- 1A and U/fpm -1B) and the U/fpm -2 locus. The mutant genes U/fpm- 1A and U/fpm -2 are responsible for high resistance levels (Rf: 75–100 or 257–286 based on MICs or ED_50s, respectively), while the U/fpm -1B mutation gives only a small reduction (approximately 7–10-fold) in fenpropimorph sensitivity. Cross-resistance studies with other SBIs showed that the major gene ( U/fpm- 1A and U/fpm -2) mutants were cross-resistant to the related compound fenpropidin (Rf: 15–20 or 53–66 based on MICs or ED_50s values, respectively) and to tridemorph (Rf: 5 or 7.1–9.5 based on MICs or ED_50s values, respectively), but not to the inhibitors of steps of ergosterol biosynthesis preceding the Δ¹⁴-reductase. The minor gene ( U/fpm -1B) mutants also had low-level resistance (approximately 5-fold) to tridemorph and to fenpropidin, but in contrast with the major gene mutants they were 2–10 times more sensitive to the triazoles studied (triadimefon, triadimenol, propiconazole and flusilazole) and to the pyridine, pyrifenox.
Studies of the fitness of U. maydis mutants showed that in major gene mutants, resistance was not associated with changes in growth rate in liquid culture or pathogenicity on young maize plants. The minor gene mutation reduced significantly the growth rate in liquid culture and the pathogenicity, either in homozygous or heterozygous condition in dikaryotic mycelium.