Comparison of fungicides for the control of Rhizoctonia solani causing datnping-off of mung bean (Phaseohis aureus)

Of 41 fungicides tested in the laboratory, copper carbonate, copper sulphate, mercuric chloride, Agrosan GN, quintozene, kasugamycin, carboxin, pyracar-bolid, carbendazim, chloroneb, benomyl, Ohric, RH 893 (2-n-octyl-4-isothiazole-3-one) and Terrazole were most inhibitory to the mycelial growth of Rhizoctonia solani on Czapek's agar plates and had EC₅₀ values of less than 1 μg a.i./ml, while copper oxychloride, Udonkor, zineb, ziram, F 319 (3-hydroxy-5-methyl isoxazole) and anilazine were much less toxic, ziram being least inhibitory with an EC₅₀ of 214 μg a.i./ml. Of 17 fungicides tested in the greenhouse as seed treatments, thiabendazole, carbendazim, benomyl, thiophanate-methyl, dichlozoline and Ohric gave 80–90% control of damping-off of mung bean seedlings. A single soil drench with thiophanate-methyl and two drenches with benomyl gave about 90% disease control, More seedlings with R. solani infection survived when thiophanate-methyl was used as a post-inoculation soil drench than when benomyl or chloroneb were used.     

Variation of Fungicide Resistance in Czech Populations of Pseudoperonospora cubensis

During the growing seasons between the years 2001 and 2004, 98 isolates of Pseudoperonospora cubensis from nine regions of Czech Republic were collected and screened for tolerance/resistance to the three frequently used fungicides (propamocarb, fosetyl‐Al, metalaxyl). Fungicides were tested in five different concentrations, using a floating disc bioassay. Fungicide effectiveness varied considerably. Propamocarb appeared most effective and all the isolates collected in the years 2001–2003 were found sensitive to all tested concentrations [607–9712 μg active ingredient (a.i.)/ml]. In 2004, some strains with increased resistance to propamocarb were detected. These strains were characterized by tolerance at the lowest concentrations (607 μg a.i./ml, eventually on 1214 μg a.i./ml); however, they were controlled by 2428 μg a.i./ml. Fosetyl‐Al was effective at the recommended concentration of 1600 μg a.i./ml against all isolates. However, the occurrence of isolates (collected in 2001) which sporulated at low concentrations (400 and 800 μg a.i./ml) indicated that the selection for tolerance occurs in the pathogen population. Nevertheless, this phenomenon was not confirmed with the P. cubensis isolates collected between the years 2002 and 2004. Metalaxyl was found ineffective, because 97% of the isolates showed the resistance to the recommended concentration (200 μg a.i./ml), and the other 3% of isolates expressed tolerant response. The majority of the isolates showed profuse and/or limited sporulation at higher concentrations (400 and 800 μg a.i./ml). A substantial shift to highly metalaxyl resistant strains was evident in the Czech P. cubensis populations during 2001–2004.     

Laboratory tests of fungicides against Peronospora parasitica (Pers. ex Fr.) Fr

Methods were investigated of culturing Peronospora parasitica on detached cotyledons in plastic box moist chambers in constant-temperature cabinets. Light was essential and a temperature of 15 °C was most suitable for infection and survival of cotyledons. Using such cotyledons the protectant and eradicant actions of various fungicides against P. parasitica were compared with those of zineb. Captafol, Daconil 2787, dichlofluanid and propineb were markedly superior to zineb as protectants, being highly effective at 0·025 % a.i. None of these compounds (at 0·2 % a.i.) checked fungal development when applied 5 h after inoculation, during which time the fungus had penetrated the cotyledons, and it was concluded that the fungicides had thus no internal effect. Internal fungistasis was shown, however, by a maneb-nickel sulphate fungicide and also by nickel sulphate. The latter (at 0·4% NiSO₄.6H₂O) gave some disease control when applied up to 72 h after inoculation and appeared to be translocated within the tissue of the cotyledon.     

Effect of fungicides on growth,germination and cuticle-degrading enzyme production by Lecanicillium muscarium

Lecanicillium muscarium Zare and Gams (previously known as Verticillium lecanii) is a well-known pathogen of arthropods. The influence of six fungicides on growth, sporulation, conidial germination and cuticle-degrading enzyme production by L. muscarium was investigated under laboratory conditions. The maximum reduction in vegetative growth, sporulation and conidial germination in relation to the control treatment was observed for propiconazole, whereas mancozeb and chlorothalonil caused the lowest reduction in these parameters. Propiconazole and pyraclostrobin caused higher reduction in enzyme activities (chitinase, Pr1, Pr2 and lipase) at all three concentrations (10, 100 and 1000 µg/ml), whereas low reduction in enzyme activities was caused by chlorothalonil and mancozeb when used at 10 µg/ml. The data presented can be used for future recommendations of these fungicides in Integrated Pest Management (IPM) programmes where L. muscarium is an important control agent.     

Evaluation of fungicides for control of South American leaf blight of Hevea brasiliensis

Of 43 fungicides tested in vitro, 19 showed strong, seven moderate and 17 weak inhibition of germination of conidia and ascospores of Microcyclus ulei. The formation of lesions on Hevea brasiliensis leaf discs was also suppressed by the first category of fungicides as well as by the five adjuvants tested. Ascospores were not released when perithecia were treated with urea, thiabendazole or alcoholic mercury chloride at 10.00, 0.10, 1.00 g/1 respectively; other fungicides had no such inhibitory effect. In field trials, thiophanate methyl (0.07% a.i.) and benomyl (0.025% a.i.) were most effective in controlling leaf infection, followed by chlorothalonil (0.15% a.i.) and mancozeb (0.32% a.i.). Benomyl suppressed conidial sporulation, whereas one application of thiophanate methyl (0.14% a.i.) to perithecia inhibited ascospore release; half of this concentration applied to conidial lesions or pycnidia caused the perithecia formed subsequently to abort. Thiophanate methyl thus shows promise for SALB control and elimination and benomyl may be valuable as a supplement in later rounds of spraying to control conidial sporulation. After 6 days of showery rain (2 mm for 17 min per day), water collected from sprayed leaves still gave complete inhibition of spore germination. However, inhibition was markedly reduced after 6 days of heavy rain (over 8 mm for 24 min per day).     

Dynamics of Free Pisatin Elicitor Accumulation in Infection-droplets of Monilinia fructicola on Pea Endocarp*

Rapid elicitor accumulation in the infection-droplet of the pea-M. fructicola interaction began between 2 and 3 h after inoculation. Rapid accumulation of pisatin began between 2 and 3 h, however, low levels (0.06–0.1 μg/ml) were detected in the infection-droplet as early as 1–2 h following inoculation with the fungus. The pisatin concentration reached levels inhibitory to the fungus between 6 and 12 h (ca. 1–5 μg/ml) and the ED₅₀ value of 10 μg pisatin/ml for mycelial growth of M. fructicola was attained after 14 h. Elicitor activity in infection-droplets after 4 and 18 h was a function of inoculum concentration and pisatin accumulation in diffusate after 40 h was a function of elicitor concentration (linear doseresponse curve). However, when elicitor was applied at zero time, the rate of pisatin accumulation over the first 12 h was indistinguishable from that which occurred when M. fructicola conidia were applied to the endocarp and elicitor accumulated with time. The initial rate of pisatin accumulation therefore appears to be dependent on the pea pod and independent of any time delays associated with conidial germination and elicitor accumulation. However, the final pisatin concentration which accumulated in the infection-droplet was dependent on the, dose' of elicitor irrespective of the nature and timing of the elicitor treatment. The presence of elicitor activity was demonstrated in the interaction in space and time where and when these molecules could function as elicitors of pisatin in vivo.     

Compatability of Metarhizium anisopliae var. anisopliae with chemical pesticides

The effects of various insecticides on the mycelial growth, sporulation and conidial germination of Metarhizium anisopliae var. anisopliae isolate E₉ were studied in the laboratory. Chlorpyrifos was the most toxic organophosphate to mycelial growth and sporulation at all concentrations. Temephos, malathion and leptophos were highly toxic to sporulation while malathion was the most inhibitory to germination. The carbamates, carbofuran, methomyl and oxamyl were moderately toxic to mycelial growth and sporulation while oxamyl had an adverse effect on germination. The pyrethroids (pyrethrin, permethrin and resmethrin) and the insect growth regulators (diflubenzuron and methoprene) were not inhibitory to the various developmental stages of isolate E₉. The chlorinated hydrocarbons (chlordane, lindane and toxaphene) were more deleterious than all other insecticide groups tested. Among the fungicides, benomyl and maneb produced the greatest inhibition.     

Dracaena Leaf Proliferosis, a Newly Recorded Disease Affecting Dracaena sanderiana in Egypt

Dracaena leaf proliferosis is a newly reported disease affecting Dracaena sanderiana in Egypt. A cause and effect relationship between this disease and the fungus,Fusarium proliferatum var. minus has been established. In addition to D. sanderiana the fungus was found to be pathogenic, when tested in the laboratory, to several other members of the family Liliaceae. While the in vitro growth of the fungus is optimum at 25 °C, symptom expression is best at 30°C. Twelve fungicides were tested for their in vitro effect on fungal growth. Benlate, Rubigan, Saprol, Cercobin and Vitavax-200 came first on the list and inhibited growth at 2.5, 12.0, 55.0, 75.0, and 94.0 μg/ml, respectively. Although, Benlate was the most effective fungicide in this respect it failed to demonstrate similar effect on disease development when applied to plants artificially inoculated with the fungus. Fungal growth was completely inhibited on PDA medium by a bacterium belonging to Bacillus sp. but when the bacterium at a concentration of 1 × 10¹¹ cell/ml was applied 24 h before, at the same time with, or 24 h after inoculation no control of the disease was achieved. Naturally-infected plants could, however, be freed from infection when subjected to a hot air treatment at 35 ± 5 °C during day time and 25 ± 5° C at night for 3 months.     

Greenhouse and field evaluations of potassium phosphonate: the control of Phytophthora foot rot of black pepper in Vietnam

Phytophthora foot rot of black pepper caused by Phytophthora capsici is a major disease of black pepper throughout production areas in Vietnam. The disease causes collar, foot and tap root rots and eventual death of the infected vine. Potassium phosphonate was evaluated for the control of this disease in greenhouse and field trials. In greenhouse trials three-month-old vines treated with phosphonate by soil drenching (10–20 g a.i./l) and then inoculated with P. capsici mycelium (2% v/v soil) had significantly less foot rot compared to vines grown in non-treated soil. In field trials mature vines were treated with phosphonate at 50–100 g a.i/pole soil drenching or 10 g a.i./l by root infusion. After 10 days root, stem and leaf specimens were removed for bioassay by inoculation with 5 ml of P. capsici zoospores suspension (10⁶–10⁸ spores/ml). Soil drenching with phosphonate inhibited the colonisation of pathogen on excised leaf, stem and root tissues, significantly more than phosphonate root infusion. Our study provides further evidence supporting the efficacy of potassium phosphonate in the management of black pepper foot rot caused by P. capsici. The excised leaf and stem bioassay used in this study is a rapid and useful technique for testing the efficacy of systemic fungicides in controlling this disease.     

灰飞虱体内类酵母共生菌异常毕赤酵母的分离培养及其对杀菌剂的敏感性

【目的】随着杀虫剂等化学农药的大规模使用,灰飞虱Laodelphax striatellus的抗药性不断增强,防治效果下降,因此寻找新的防治方法尤为重要。本研究探索使用杀菌剂来减少灰飞虱体内共生菌的数量,以降低灰飞虱的活力,为"抑菌防虫"提供理论依据。【方法】对灰飞虱体内的类酵母共生菌(yeast-like symbiotes,YLS)进行离体培养并鉴定,测定离体培养YLS对不同浓度不同种类杀菌剂的敏感性,将对其抑制作用明显的杀菌剂喷施于麦苗,待灰飞虱取食后统计灰飞虱死亡率、体重,并通过荧光定量PCR检测体内共生菌的数量变化。【结果】离体培养得到1株YLS,经碳源同化实验和18S r DNA分子鉴定,将该菌株归为异常毕赤酵母Pichia anomala,以变性梯度凝胶电泳(denatured gradient gel electrophoresis,DGGE)技术验证了该菌株在灰飞虱体内的存在。对杀菌剂敏感性的测定结果显示,70%丙森锌和50%戊唑醇+25%肟菌酯对P.anomala抑制效果明显,而氟吡菌胺+霜霉威盐酸盐(687.5 g a.i./L)与40%嘧霉胺对P.anomala的抑制效果较差。用对P.anomala抑制效果明显的两种杀菌剂(70%丙森锌和50%戊唑醇+25%肟菌酯)以800 mg/L浓度喷施麦苗,灰飞虱取食喷施麦苗后的死亡率分别达到46.7%与63.3%,显著高于对照,而体重比对照组显著低。荧光定量检测结果表明,灰飞虱体内3种共生菌(Noda菌、季也蒙毕赤酵母和异常毕赤酵母)的数量在使用了各种杀菌剂后均显著降低。【结论】结果说明,体外喷施杀菌剂对灰飞虱体内共生菌有抑制作用,进而对灰飞虱的生长存活产生影响,这为杀菌剂与杀虫剂的协同使用防治灰飞虱奠定基础。     

In vitro evaluation of six fungicides on radial mycelial growth and regrowth of Fusarium pallidoroseum isolated from castor (Ricinus communis) in Samaru,Nigeria

Six fungicides at three rates (1.5x, 1.0x and 0.5x mg a.i/ml) were evaluated on radial growth and and regrowth of mycelia of Fusarium pallidoroseum isolated from castor (Ricinus communis) in vitro. It was observed that the fungicides (Benomyl, Benomyl + Thiram, Mancozeb, Metalaxyl-m + Thiomethoxan + Difenconazol, Tricyclazole and Carbendazim + Mancozeb) at all the concentrations tested inhibited mycelial growth and regrowth of the fungus. Benomyl, Benomyl + Thiram and Tricyclazole completely inhibited mycelia growth of fungus at 1.5x, 1.0x and 0.5x mg a.i/ml. Metalaxyl-m + Thiomethoxan + Difenconazole, Carbendazim + Mancozeb partially inhibited radial growth and re-growth of mycelia only at 1.5x mg a.i/ml, not at 1.0x and 0.5x mg a.i/ml. The inhibitory effect of all the fungicides on mycelia growth and re-growth was greatest at 1.x5 mg a.i/ml.     

Some effects of differently-acting nematicides on the cereal cyst-nematode (Heterodera avenae) and on the appearance of ''scorch'' in spring wheat on light loamy sand

In fungitoxicity tests against Phytophthora cinnamomi on Chamaecyparis lawsoniana cv. Ellwoodii, a drench of furalaxyl (1000 mg a.i./l) applied to the compost in which 1-yr-old plants were growing, 1 wk before they were inoculated with 650 000 zoospores, controlled disease for at least 12 months. With an inoculum dose of 650 zoospores/plant, furalaxyl at 500 mg a.i./l controlled disease even when inoculation was 12 wk after fungicide treatment. Aluminium tris (ethyl phosphonate) (2000 mg a.i./l) applied as a drench 1 wk before inoculation with 650 000 zoospores/plant did not prevent root infection but delayed foliar symptoms for 9 months: the same treatment, using etridiazole (500 mg a.i./l) only slightly reduced disease incidence. When applied as a single drench 2 days before inoculation, prothiocarb (2000 mg a.i./l) and cuprammonium compounds (200 mg a.i./l) were much less effective than furalaxyl (1200 mg a.i./l), sodium ethyl phosphonate (1500 mg a.i./l), aluminium tris (ethyl phosphonate) (1500 mg a.i./l) or etridiazole (500 mg a.i./l). However, a drench of furalaxyl at 1000 mg a.i./l, aluminium tris (ethyl phosphonate) at 2000 mg a.i./l or etridiazole at 500 mg'a.i./l did not eradicate P. cinnamomi from compost containing infected root debris. Pre-planting drenching of the compost was ineffective. All fungicide treatments were non-phototoxic to 1-yr-old C. lawsoniana cv. Ellwoodii. These results are of special relevance to the control of P. cinnamomi on container-grown woody ornamentals.     

Antibacterial effect of honey on the in vitro and in vivo growth of Escherichia coli

The study investigated the antibacterial effect of honey against pathogenic Escherichia coli. Honey showed inhibitory activity against the growth of E. coli (ATCC 25922) in agar plate assay. In liquid culture (48 h, 37 °C) the growth rate of bacterial cells decreased in the presence of honey (9.6 × 10⁵ c.f.u./ml) compared with sucrose (2.87 × 10⁸ c.f.u./ml). Rats fed with honey and orally inoculated with E. coli excreted significantly (P < 0.05) less bacterial cells in faeces compared to controls. Animals acclimatized to feeding of honey prior to E. coli inoculation showed a significant decrease in excreted bacterial load compared with the group provided with honey after bacterial inoculation. Consumption of honey also enhanced the concentration of short chain fatty acids in the intestine of rats (83 mM) compared with the control group (44.5 mM). The results show that honey possessed significant antibacterial activity against E. coli under in vitro and in vivo conditions, and indicate the potential benefit of consumption of honey regularly on the microbiological constitution of animals feeding on it.     

Responses of diamondback moth to diverse entomopathogenic fungi collected from non-agricultural habitats – Effects of dose,temperature and starvation

We evaluated the virulence of Beauveria bassiana and Metarhizium isolates from soil collected across different vegetation types in Queensland, against chlorantraniliprole-resistant and insecticide-susceptible diamondback moth (DBM) larvae. Host insecticide resistance status had no effect on susceptibility to the pathogens when conidia were topically applied to larvae in the laboratory, and one B. bassiana isolate was significantly more virulent to larvae than the others (seven days after inoculation). The influence of temperature (15, 20, 25 or 30 °C): (i) at the point of host inoculation with conidia and (ii) when the pathogens had already initiated infection and were proliferating in the host haemocoel, was determined experimentally for its influence on virulence, disease progression, and sporulation. Temperature at inoculation had a greater effect on host insect mortality than it did when the fungus was already proliferating in the host haemocoel. The rearing temperature of hosts prior to inoculation had a greater effect on host susceptibility to disease than starvation of the larvae at the time of inoculation. Our results also show that each fungal isolate has its own temperature relations and that these can vary considerably across isolates, and at different points in the pathogen life cycle (germination and cuticular penetration versus growth in the host haemocoel). Temperature also had an idiosyncratic effect, across isolates and across the variables typically used to assess the potential of fungal entomopathogens as biological control agents (time to death, mortality and sporulation rates). This study demonstrates that in addition to pathogenicity and virulence, the temperature relationships of each fungal isolate when infecting insects needs to be taken into account if we are to understand their ecology and use them effectively in pest management.     

Enhancement of L-glutamate oxidase production in liquid fermentation of Streptomyces sp. by calcium addition

Summary The effect of calcium ions on production of extracellular L-glutamate oxidase (GluOx) in liquid fermentation of Streptomyces sp. N1 was investigated. By supplementing a relatively large amount of Ca²⁺ (15–40 mM) to the medium, the GluOx production was significantly enhanced, although the microbial growth was inhibited to some degree. For the first time, the highest production (i.e. 6.5 U/ml) and productivity (i.e. 0.25 U/ml/h) of GluOx as ever reported (i.e., 1.2 U/g medium after 144 h solid state fermentation and 0.59 U/ml after 96 h liquid fermentation) were achieved with addition of 30 mM Ca²⁺.     

Control of Fusarium solani rot of potato tubers with fungicides

Tecnazene (up to 33 ppm) and dichloran (up to 500 ppm) had little effect on germination of spores or growth of Fusarium solani isolated from and causing a rot of potato tubers; they also did not decrease rotting when applied to wounds later inoculated with the pathogen. Benomyl and thiabendazole (up to 500 ppm) also had little effect on spore germination but did greatly decrease growth at 5 ppm. A pronounced pink coloration developed in cultures growing slowly in the presence of benomyl; a similar though less striking effect appeared in agar cultures containing thiabendazole. Benomyl suspended in water or diluted with Fuller's earth gave good control of rotting when applied to wounds inoculated later with F. solani. Still better control was obtained with thiabendazole; dusts containing 1% a.i. substantially decreased rots and those containing 10 % a.i. gave almost complete control when applied to wounds shortly before inoculation. Thiabendazole was also very effective when used 24 h after inoculation and a fair measure of control was obtained when it was applied 24 h later. Benomyl and thiabendazole placed on apparently intact surfaces of tubers caused tissue 5 mm deep to become toxic to F. solani 10 days later, and, unexpectedly, this tissue prevented spore germination. Fuller's earth alone substantially decreased rotting. The results obtained suggest that dusts containing thiabendazole have some promise for the control of Fusarium rots of potato tubers, especially of early crops.     

The effect of sublethal doses of rifampin on the sporulation of Clostridium botulinum.

Sublethal doses of rifampin (0-005 mug/ml), added to vegetatively growing cultures of a sporogenic mutant of Clostridium botulinum at inoculation time or after 4 h, resulted in a decrease of growth and in blockage of spore formation. But when rifampin was added 6 to 24 h after inoculation, normal growth and sporulation occurred, indicating that the time of addition was critical and that rifampin was most effective on rapidly dividing, exponential-phase cells. Ultrastructural studies showed that when rifampin was added at the time of inoculation, endospore development was blocked at stage III. During subsequent incubation (greater than 10 h) the cells lost their rigidity, and lysis of the mother cell was followed by that of the forespore. When the cultures were treated with rifampin at 4 h, about 40% of the cells were blocked at stage III and about 60% reached stages IV and V. Some showed excessive elongation and contained developing spores at each pole. They appeared to be derived from two daughter cells unable to form a division septum because of a specific inhibitory effect of rifampin on division. It would seem, therefore, that two daughter cells which are genetically coded to form endospores will do so irrespective of the development of a division septum, and the spores are formed at the 'old' polar regions.     

Effect of gramicidin C on the formation and germination of Bacillus brevis var. GB (P+-variant) spores]

The effect of gramicidin C added to the medium at various periods of cultivation in concentrations of 20, 40 and 100 gamma/ml on sporulation of P+-variant of Bac. brevis var. GB was studied. The most effective increase in the sporulation rate and percentage of the cells germinating into the spores was observed on addition of the antibiotic to the medium in amounts of 20 and 40 gamma/ml in 13 hours of the culture development. The amount of gramicidin C during sporulation decreased and partially passed into the spores which did not differ after germination from those of P+-variant grown on the synthetic medium with glucose and without preliminary addition of the antibiotic. Addition of gramicidin C in an amount of 100 gamma/ml at the end of the lag phase, i.e. 4 hours after the culture inoculation suppressed sporulation and had no effect on growth of the cells of its own producing organism.     

Some effects of benomyl and carbendazim on Aphis fabae and Acyrthosiphon pisum on field bean (Vicia faba)

Field bean plants were treated with benomyl in a glasshouse, then nymphs of Aphis fabae and Acyrthosiphon pisum were caged on the second pinnate leaf. Soil drenches at concentrations of 150 and 75 μg benomyl/ml or above increased mortality of A. fabae and A. pisum respectively; 250 μg a.i./ml increased mortality and decreased progeny production of alate A. fabae. The effect on mortality persisted for at least 16 days after treatment. Foliar sprays increased mortality at concentrations of 75 μg a.i./ml and above (A. fabae). Field populations of A. pisum were reduced when bean plants were drenched or sprayed at a concentration of 250 μg a.i./ml and A. fabae populations were reduced by drenches but not by foliar sprays. A commercial formulation of carbendazim (Bavistin) increased aphid mortality whereas the formulation medium did not. Under the experimental conditions, benomyl affected the distribution of both species on young bean plants but did not induce a repellent effect; aphids preferred untreated leaves. Mortality and preference tests, and a field experiment, indicated that A. pisum was affected more than A. fabae.