CAPS and DHPLC Analysis of a Single Nucleotide Polymorphism in the Cytochrome b Gene Conferring Resistance to Strobilurins in Field Isolates of Blumeria graminis f. sp. hordei

A single nucleotide polymorphism in the wheat powdery mildew (Blumeria graminis f. sp. tritici) cytochrome b gene is responsible for resistance to inhibitors of the quinol outer binding site of the cytochrome bc₁ complex (QoI) fungicides. Analysis of a partial sequence of the cytochrome b gene from field isolates resistant and sensitive to QoI fungicides revealed the same point mutation in barley powdery mildew (B. graminis f. sp. hordei). Analysis of 118 and 40 barley powdery mildew isolates using a cleaved amplified polymorphic sequence assay and denaturing high performance liquid chromatography, respectively, confirmed that this single nucleotide polymorphism also confers resistance to QoI fungicides in barley powdery mildew.     

Efficacy of Milsana®, a Formulated Plant Extract from Reynoutria sachalinensis, against Powdery Mildew of Tomato (Leveillula taurica)

The efficacy of Milsana^® VP 1999 and 2000 (a formulated plant extract of Reynoutria sachalinensis), known to induce resistance to powdery mildew on cucumbers, was tested against Leveillula taurica (Lév.) Arn. on greenhouse tomato. In four out of five trials, Milsana^® achieved a disease reduction ranging from 42.2 to 64.6%. In one trial only, its efficacy was exceptionally low (23%). Application rates and disease pressure proved to be the main factors affecting the level of control. Milsana^® was significantly less effective than fungicides (alternated DMIs and penconazole) in situ. In contrast, Milsana^® was equally effective to wettable sulphur indicating that its effect was rather preventive than curative. The level of efficacy achieved by either Milsana^® or fungicides did not result in a significant increase of yield. Laboratory tests showed that Milsana^® (VP 1999) had a direct effect on conidial germination. Whether this effect significantly contributes to its field efficacy, remains to be elucidated. Overall, results indicate that Milsana^® could play an important role in disease management of powdery mildew in organic and low input tomato production.     

Greenhouse evaluation of chemicals for control of powdery mildews

2,6-Dinitro-4-s-alkylphenols were found to protect apple foliage against powdery mildew more effectively than isomeric 2,4-dinitro-6-s-alkylphenols; regression lines for four para-alkyl compounds were of similar slope but were much steeper than those for two ortho-alkyl compounds (including dinocap phenol). The ED₅₀ and ED₉₅ values of the most active compound studied, 2,6-dinitro-4-(I-ethylhexyl)phenol, were in the ratios to those of dinocap phenol of 1:13 and 1:125, respectively, and the protective action of this compound was greater than the curative, especially at the higher ED values. Protection of barley seedlings against powdery mildew was also greater by I-ethyl- or I-propyl-alkyl compounds than by I-methylalkyl or n-alkyl isomers. For phytotoxic and acaricidal actions, the ortho-alkyl isomers are more effective than para-alkyl. Nevertheless, the acaricidal activity of dinocap phenol is exceeded by that of the isomeric 2,4-dinitro-6-(I-ethylhexyl) phenol. The control of these powdery mildew diseases given by commercial products, supposed to be based on dinocap, cannot be accounted for by the activity shown for dinocap phenol. It can, however, be accounted for by the activities of 2,6-dinitro-4-(I-ethylhexyl)- and -(I-propylpentyl)-phenols, now known to be present in commercial products in larger amounts than dinocap phenol itself. The phytotoxic and acaricidal actions of such products, however, are mainly due to the ortho-octyldinitrophenols present; in view of the small proportion that is dinocap phenol, the acaricidal activity is likely to be due almost entirely to the other ortho-octyl isomers. It is suggested that the common name dinocap be retained for the mixture of dinitrooctylphenols now known to be present in commercial products, and that the two main groups be differentiated as 2,4-dinocap and 2,6-dinocap, respectively. The advantages for powdery mildew control of a product based on 2,6-dinocap are discussed.     

Glasshouse experiments on apple scab

Experiments with fungicide mixtures used protectively and curatively against apple scab showed no advantage from adding phenylmercury chloride (PMC) to certain protective fungicides. Sulphur fungicides greatly impaired the curative activity of PMC, especially in mixtures applied 24 hr. after infection, but captan showed little and dodine acetate no such effect. PMC, however, contributed appreciably to the curative effectiveness of mixtures with captan or dodine acetate. Commercially prepared mixtures of dispersible sulphur with phenylmercury dimethyldithiocarbamate and of zineb with mancozeb gave promising results, and an experimental mixture of dodine/glyodin acetates at low rates was effective protectively and especially curatively. Dodine acetate, dichlofluanid, lime-sulphur, and isobutyl-o-coumarate showed little translocated activity against scab when applied after infection, and PMC, which earlier showed strong translocated activity when applied in summer before infection, was much less effective when applied in spring 24 hr. after infection. Uncontrolled powdery mildew early in the scab-incubation period greatly reduced the establishment of scab infection on the test plants (clonal root-stocks); methods of mildew control in such experiments are suggested.     

Evaluation of microcapsule trunk injections for the control of apple scab and powdery mildew

A 3‐year field trial was conducted using established apple cv. Crown Gold and English oak (Quercus robur L.) to assess the efficacy of eight fungicides applied via microcapsule trunk injection against the foliar pathogens apple scab (Venturia inaequalis) and powdery mildew (Phyllactinia sp). In both apple cv. Crown Gold and English oak, the fungicide myclobutanil was not taken up when microcapsules were inserted into the tree vascular system at the root flare. Disease severity in injected trees, excluding myclobutanil, was lower over the following two growing seasons compared to water‐injected controls indicating seven of the eight fungicides used in this study provided a significant degree of protection against scab and powdery mildew infection. A difference in the magnitude of pathogen control achieved was recorded between fungicides. Of the fungicides tested, penconazole, pyrifenox and carbendazim significantly reduced disease severity and significantly increased leaf chlorophyll (Fv/Fm) and SPAD values as a measure of tree vitality and chlorophyll content, respectively, in both apple cv. Crown Gold and English oak over two growing seasons after microcapsule injection. Based on the results of this investigation, it is suggested that these three fungicides be used in preference to thiabendazole, fosetyl‐aluminium, triadimefon and propiconazole for the control of apple scab and powdery mildew where outbreaks of these foliar pathogens are problematic.     

Control of powdery mildew in okra using cultural filtrates of certain bio-agents alone and mixed with penconazole

This study was carried out to evaluate the culture filtrates of certain bio-agents (Epicoccum nigrum, Epicoccum minitans, Epicoccum sp., Trichoderma harzianum, Trichoderma viride and Bacillus pumilus) alone and mixed with penconazole against powdery mildew in okra. The results showed that the culture filtrate of E. nigrum induced the highest efficacy against powdery mildew relative to other treatments in both tested seasons. Moreover, culture filtrate of E.nigrum mixed with the tested fungicide gave higher efficiency against powdery mildew than use of the fungicide alone. The efficacy of the tested culture filtrates against powdery mildew due to the presence of different antifungal compounds identified by (GC–MS) analysis. This study suggests the ability of using the culture filtrates of the tested bio-agents as alternative of fungicides to control powdery mildew in okra. Furthermore, the results implied the possibility to minimise the use of fungicides by mixing with the microbial culture filtrates.     

Antimicrobial efficacy,disease inhibition and phenolic acid-inducing potential of chloroform fraction of cow urine

High-performance liquid chromatographic (HPLC) analysis of chloroform fraction (CF) of cow urine showed rich pool of phenolic acids. Antifungal and antibacterial bioassays of CF have shown its tremendous efficacy against some fungal plant pathogens as well as human pathogenic bacteria at very low concentrations. The CF also inhibited powdery mildew (Erysiphe cichoracearum) of balsam (Impatiens balsamania) under field conditions during pre- and post-inoculation treatments. HPLC analysis of pre- and post-inoculation-treated plant leaves indicated that CF induced phenolic acid synthesis as compared to control. The results revealed that CF of cow urine has the potential for controlling some important human diseases. The result on balsam powdery mildew is a good signal that CF may also be effective against other plant diseases in the field.     

Efficacy of Foliar Application of Phosphates in Controlling Powdery Mildew Fungus on Field-Grown Winegrapes: Effects on Cluster Yield and Peroxidase Activity in Berries

Seven foliar applications of 0.025M K₂HPO₄ and KH₂PO₄ (both plus Tween 20) and the commercial systemic fungicides, Dorado (Pyrifenox) 480 EC, Penconazole EC and Benomyl, were applied at 14-day intervals starting at 10-cm shoot length on field-grown Chardonnay winegrapes. Both phosphates and systemic fungicides inhibited development of powdery mildew fungus (Uncinula necator, Schw., Burr.) on fruit clusters, as compared with untreated control vines. Diseases, everity on clusters of plants treated with K₂HPO₄ and fungicides was 0.3 and 0.2, respectively, as compared with 1.3 on control clusters (on a 0–4 scale), for the first rating, conducted 10 days after the fifth application of fungicides and phosphates, Five days after the last application, disease severity was 3.5 on non-treated control clusters and 0.3 and 0.8 on clusters treated with Dorado and K₂HPO₄, respectively. Powdery mildew infection remarkably reduced the weight of non-treated control clusters as compared with Dorado and phosphate treated clusters. Phosphate treatment caused an increase (3-fold) of peroxidase activity in the soluble fraction of non-infected control berries. A remarkable peroxidase enhancement was detected in the soluble (8-fold) and ionically bound (2-fold) fractions from the phosphate-treated and infected berries. Results indicate that phosphates can be used as foliar fertilizers for disease control in the field and that peroxidase might, be involved in the defense process.     

Effects of elevated atmospheric CO2 and temperature on the management of powdery mildew of zucchini

The impact of combined environmental factors, such as temperature and CO₂, on the control of the powdery mildew of zucchini, caused by Podosphaera xanthii, and of different control measures has been studied on plants grown in phytotrons. Five experimental trials were conducted, and the powdery mildew severity of both treated and untreated zucchini plants was found to be significantly affected by the interaction between temperature (three different regimes: 16–18; 18–22; 22–26°C), CO₂ (two concentrations: 400–450 and 800–850 ppm) and the treatments. However, at the end of the trials, the efficacy of all the products was not affected by the different, tested environmental conditions. Sulphur consistently provided the highest disease control (75%–85% efficacy). Among the resistant inducers that were tested, calcium oxide was the most effective, in terms of powdery mildew control under all the conditions tested in phytotrons, reducing disease severity from 46% to 61%. Foliar applications of phosphite (14%–28% efficacy), Ampelomyces quisqualis (12%–23% efficacy) and potassium silicate (13%–24% efficacy) only slightly reduced the disease severity for all the tested temperature regimes and CO₂ concentrations, compared to the untreated control. The results obtained under our experimental conditions show that a possible increase in CO₂ concentration and temperature, which is expected for the next few years, should not influence the efficacy of the tested resistance inducers or of sulphur against powdery mildew on zucchini. Moreover, the suppressive effect of calcium oxide is in light of its possible use in greenhouses for zucchini powdery mildew control under 400–450 ppm of CO₂ and under enriched condition of 800–850 ppm of CO₂.     

Effect of combination between inducer resistance chemicals and systemic fungicides on management of sweet melon downy mildew

The efficiency of some inducer resistance chemicals (IRCs) like bion, chitosan, humic acid and salicylic acid as well as the fungicides like Folu-Gold, Galben Copper, Previcure-N and Redomil Gold Mancozeb on management of sweet melon downy mildew, caused by Pseudoperonospora cubensis was evaluated in vitro and in vivo. Also, the efficiency of the alternation between the sprayed two fungicides and IRCs on management of the disease and the produced fruit yield and its total soluble solids (TSS) under field conditions were assessed. The inhibitory effect of the IRCs and the tested fungicides on sporangial germination of P. cubensis resulted in a significant reduction in the germinated sporangia. In addition, IRCs were less effective than the fungicides. Disease management revealed the same trend of the in vitro experiment when they sprayed fungicides on sweet melon plants artificially inoculated with the sporangia of the causal fungus under greenhouse conditions. Furthermore, under field conditions, spraying sweet melon plants with the two tested fungicides was the most efficient in decreasing the disease and increasing fruit yield and its TSS, to somewhat, followed by the alternation between them and the tested IRCs. In addition, IRCs treatments showed the lowest efficiency in this regard.     

The effect of fungicides, irrigation and plant density on the development of Peronospora sparsa, the cause of downy mildew in rose and blackberry

Downy mildew of rose (Rosa spp.) and blackberry (Rubus fructicosus), caused by Peronospora sparsa, can cause significant losses in production. In a series of experiments, fungicides with different modes of action to the commonly used phenylamide‐based products were examined for efficacy on both crops. Cymoxanil + mancozeb + oxadixyl and fluazinam gave good downy mildew control on both rose and blackberry. On outdoor, container‐grown rose, high volume sprays of fosetyl‐aluminium were also effective, but on young micropropagated blackberry plants, application as a drench treatment was better than as a spray. Good control was also achieved on blackberry with chlorothalonil and with metalaxyl in formulation with either thiram or mancozeb. There was no evidence of loss of control with phenylamide fungicides due to resistance. Irrigation regimes were also examined on blackberry and sub‐irrigation of plants in propagation on a sand bed led to significantly lower disease levels than those found where plants had been irrigated by overhead sprinklers. Reducing the density of container‐grown rose plants had a measurable effect on the progress of downy mildew, though this was small and temporary when compared with the effect of fungicide treatment. Fungicide programmes which gave season‐long control of rose downy mildew resulted in significantly improved extension growth.     

Control of pearl millet downy mildew caused by Sclerospora graminicola with systemic fungicides in an artificially-contaminated plot

Among four fungicides, viz. metalaxyl (two formulations), fosetyl-Al, pro-pamocarb and cyomaxanil tested in vitro against sporangial germination inhibition of Sclerospora graminicola, cyomaxanil was found to be most inhibitory. In an artificially contaminated plot, when used as seed treatment or foliar spray for the control of downy mildew of pearl millet, only metalaxyl was effective. Metalaxyl 25 (Ridomil) and metalaxyl 35 (Apron) seed treatments protected the pearl millet plants from downy mildew up to 30 days. As a foliar spray, metalaxyl 25 used once at 20 days or twice after 20 and 38 days of plant growth gave less disease at harvest time. Seed treatment (metalaxyl 25 or 35) followed by one metalaxyl 25 spray was found to be effective in controlling the downy mildew. These treatments improved the growth of plants and yield significantly.     

Evaluation of fungicides and biocontrol products for the control of Phytophthora root rot of hydrangeas

Abstract

Phytophthora root rot is one of the most important diseases in almost all hydrangeas of nursery production. In this study, the efficacy of fungicides and biocontrol products against Phytophthora root rot of hydrangea was assessed in greenhouse and field experiments. Treatments used in field or greenhouse experiments were RootShield PLUS⁺, MBI110, IT-5103, Grotab, OxiPhos, TerraClean 5.0?+?TerraGrow program, Segovis, Pageant Intrinsic, Empress Intrinsic and Subdue Maxx. Pots/plots were inoculated with Phytophthora nicotianae grown on rice grains, sterilised rice grains were used for negative controls. After the trials, plant growth data (total plant weight, root weight, plant height, plant width) were recorded, and roots were assessed for disease severity using a scale of 0–100%. The treatments most effective in reducing Phytophthora root rot severity were Segovis, Empress Intrinsic, Subdue Maxx, TerraClean 5.0?+?TerraGrow program in both greenhouse and field experiments. This study will help nursery producers make proper management decisions by using recommended fungicides and biocontrol products of this study in a rotation or alone to manage Phytophthora root rot of hydrangea.     

A novel technique for greenhouse evaluation of plant extracts for management of downy mildew of sorghum

Antipathogenic potential of 16 plants was evaluated in the form of aqueous extracts against Peronosclerospora sorghi, causing downy mildew of sorghum. A novel method was developed in which conidial suspension and plant extracts were mixed individually and allowed to stand for 5?min and then used to inoculate the host by sprout-dip method. The sprouts thus inoculated were grown in pots, and disease incidence was observed. Three extracts (Parthenium hysterophorus, Duranta repens and Oxalis latifolia) suppressed the disease at par with chemical fungicide (Mancozeb 75%). In form of spray, D. repens extract with tween-20 adjuvant (1.9% disease) performed closest behind mancozeb (0% disease), while negative control showed 21% disease incidence. Organic management of airborne inoculum of downy mildew of sorghum is feasible and preferable as compared to the use of chemical fungicides, considering human and environmental health concerns. Use of water extract keeps the technology simple so that it can be directly prepared and used by farmers. Short listing of three most effective water extracts would help in self-reliance of farmers, reducing their dependence on commercial products.     

Isolation and characterization of Bacillus subtilis KS1 for the biocontrol of grapevine fungal diseases

Bacillus subtilis KS1 was isolated from grape berry skin as a biological control agent against grapevine fungal diseases. KS1 was identified as a new strain of B. subtilis according to morphological, biochemical, and genetic analyses. In vitro bioassay demonstrated that KS1 suppressed the growth of Botrytis cinerea (the casual agent of grape grey mold) and Colletotrichum gloeosporioides (the casual agent of grape ripe rot). The biocontrol activity of KS1 against grapevine fungal diseases in vineyards was evaluated over a 3-year span (from 2007 to 2009). Downy mildew, caused by Plasmopara viticola, was reduced on berry skins and leaves by treatment with KS1. The KS1 genome possesses ituD and lpa-14 genes, both of which play a role in iturin A production followed by iturin A production in the culture. In contrast, mutants lacking both genes lost the antagonistic activity against B. cinerea and C. gloeosporioides and the activity in iturin A production, suggesting that the antagonistic activity of KS1 against grapevine fungal pathogens may depend on iturin A production. As KS1 showed tolerance to various chemical pesticides, chemical pesticides could be applied before and/or after KS1 treatment in vineyards. Due to its potential as a biological control agent against grape downy mildew, KS1 is expected to contribute to the further improvement of integrated pest management systems and to potentially reduce the amount of chemical fungicides applied in vineyards.     

Inhibition of Fungal Plant Pathogens by Synergistic Action of Chito-Oligosaccharides and Commercially Available Fungicides

Chitosan is a linear heteropolymer consisting of β 1,4-linked N-acetyl-D-glucosamine (GlcNAc) and D-glucosamine (GlcN). We have compared the antifungal activity of chitosan with DP_n (average degree of polymerization) 206 and F _A (fraction of acetylation) 0.15 and of enzymatically produced chito-oligosaccharides (CHOS) of different DP_n alone and in combination with commercially available synthetic fungicides, against Botrytis cinerea, the causative agent of gray mold in numerous fruit and vegetable crops. CHOS with DP_n in the range of 15–40 had the greatest anti-fungal activity. The combination of CHOS and low dosages of synthetic fungicides showed synergistic effects on antifungal activity in both in vitro and in vivo assays. Our study shows that CHOS enhance the activity of commercially available fungicides. Thus, addition of CHOS, available as a nontoxic byproduct of the shellfish industry, may reduce the amounts of fungicides that are needed to control plant diseases.     

Antifungal activity of ethanolic extract of Archu (Rheum emodi) on powdery mildew (Erysiphe cichoracearum) and its role in the induction of resistance in balsam (Impatiens balsamania)

Rheum emodi, vernacularly known as Archu, is one of the important high altitude medicinal plants widely distributed in Himalayan regions. Though widely used in Ayurveda for curing various human diseases, its use in plant diseases is limited. Ethanolic extract of Rheum rhizome was assayed against spore germination of Alternaria solani, Heliminthosporium penniseti and Curvularia palliscens. The inhibition of spore germination was concentration dependent. Maximum inhibition was obtained at 4000 and 5000 ppm followed by 3000, 2000 and 1000 ppm. However, the extract was highly effective in the pre-inoculation treatment against powdery mildew (Erysiphe cichoracearum) of balsam (Impatiens balsamania) under field conditions. High performance liquid chromatographic (HPLC) analysis of balsam leaves showed increased synthesis of phenolic acids, which has been correlated with induced resistance in inhibiting the disease intensity of balsam powdery mildew.     

Resistance inducers applied alone or in association with fungicide for the management of leaf rust and brown eye spot of coffee under field conditions

The use of resistance inducers is a promising development in the management of plant diseases, owing to their ability to control a broad spectrum of pathogens and improve the efficacy of fungicides. This study evaluates different sources of phosphonates (potassium, manganese, copper), a formulation prepared from the by‐products of the coffee industry (Greenforce CuCa), as well as the effects of their application, alone or in association with fungicide, in the management of two important coffee fungal diseases in Brazil: leaf rust and brown eye spot, caused by Hemileia vastatrix and Cercospora coffeicola, respectively. The effect of these products on defoliation, productivity and chemical composition of coffee beans (content of trigonelline, chlorogenic acid, caffeine and total soluble solids) was evaluated. Among all the alternative products tested individually, potassium phosphonate (P₂O₅—33.6% + K₂O—29%) stood out, particularly for rust control, which was similar to the results of fungicide treatments. Treatments with fungicide, Greenforce CuCa and cuprous oxide, individually, caused less plant defoliation. Regarding the chemical composition of the coffee beans, the manganese phosphonate treatment showed the highest values for trigonelline, chlorogenic acid, caffeine and total soluble solid content. The results of this study show that resistance inducers can be useful in disease management, may come to eventually replace traditional fungicides and can also contribute to the beverage quality.     

Fungicide resistance in cucurbit downy mildew – methodological, biological and population aspects

Cucurbit downy mildew, caused by Pseudoperonospora cubensis, is among the most devastating diseases of cucurbitaceous plants. In spite of improved cultural practices and breeding for resistant cultivars, chemical control is still a very important tool to manage the disease. During the last several decades, many fungicides from various chemical classes have been developed. The occurrence of strains of P. cubensis resistant/tolerant to some fungicides encouraged research of this phenomenon. The first part of this article summarises the many different methodological approaches such as field trials, in vitro testing on active plant tissues or molecular diagnoses developed for the detection of resistant/tolerant strains of P. cubensis, as well as methods to collect and maintain pathogen isolates. The second part outlines the commonly used fungicides to control P. cubensis and their features like systemicity, biological and biochemical mode of action and translocation behaviour within plants. The last part deals with geographical aspects such as first appearance of resistance problems, distribution of resistance, temporal development of resistance under selection pressure by a fungicide, fitness of resistant subpopulations in competition with sensitive ones in the absence of a fungicide, as well as genetic and molecular sources of resistance.