Yield loss and management of downy mildew on field pea in Alberta,Canada

Downy mildew of field pea (Pisum sativum) caused by Peronospora viciae f. sp. pisi has become widespread in the main field pea production areas of central Alberta. Field experiments were conducted at naturally-infested field sites over several years to assess the effect of seeding depth, seeding date, seed treatment and foliar fungicides on downy mildew incidence and severity, and to estimate the relationship between severity and yield loss. Downy mildew was shown to cause substantial yield loss on field pea. Even a moderately severe infestation reduced pod numbers by 65% and seed yield by 75%. The loss (pod number or seed yield) was best explained by a linear model (y = −2.3114x + 10.086; R² = 0.9441 and y = −2.5165x + 10.378; R² = 0.9533, respectively). Depth of seeding (range 3–7 cm) did not affect downy mildew. Similarly, seeding date (early, mid and late May) did not have a consistent effect on disease levels. Late seeding occasionally reduced downy mildew, but always resulted in low seed yield. Several seed treatment fungicides reduced downy mildew, and metalaxyl-based products produced the highest yield. Several foliar-applied fungicides, including pyraclostrobin, azoxystrobin and metalaxyl, reduced downy mildew severity, but the results were not consistent across years. We conclude that cultural practices may not be sufficient for effective management of downy mildew, and that metalaxyl-based fungicides applied as seed treatments or foliar sprays could represent the best control option until downy mildew resistant pea cultivars become available.     

Fungicidal activity of fluopicolide for suppression of Phytophthora capsici on squash

The efficacy of a new fungicide fluopicolide in suppression of Phytophthora blight caused by Phytophthora capsici was evaluated under laboratory and field conditions. Studies with 51 P. capsici isolates from vegetable crops in Georgia, USA, indicated that 5.9% of the isolates were resistant, 19.6% were intermediately sensitive, and 74.5% were sensitive to 100 μg ml⁻¹ of mefenoxam based on in vitro mycelial growth. EC₅₀ values of fluopicolide in inhibiting mycelial growth of 25 isolates, representing resistant, intermediately sensitive, and sensitive to mefenoxam, ranged from 0.05 to 0.35 μg ml⁻¹ with an average of 0.2 μg ml⁻¹ EC₅₀ values of fluopicolide in suppressing zoospore germination and sporangium production of the 25 isolates ranged from 1.1 to 4.5 μg ml⁻¹ and 0.3–9.0 μg ml⁻¹, respectively. Evaluation of a collection of 150 P. capsici isolates from vegetables and irrigation ponds found none of the isolates were resistant to 10 μg ml⁻¹ of fluopicolide. Field experiments were conducted to determine the efficacy and application methods of fluopicolide for control of P. capsici on squash in spring 2007 and 2009. Fluopicolide applied through drip irrigation or as a foliar spray at 86.6 or 115.4 g ha⁻¹ consistently provided significant disease reduction and increased squash yield. Results with fluopicolide were similar or slightly superior to mefenoxam applied at recommended rate. Fluopicolide applied at 57.7 g ha⁻¹ did not provide consistent satisfactory disease suppression. The results indicated that fluopicolide was effective in suppression of different stages of the life cycle of P. capsici and could be a viable alternative to mefenoxam for managing Phytophthora blight in squash production.     

Evaluation of systemic acquired resistance inducers for control of downy mildew on basil

Basil downy mildew, caused by Peronospora belbahrii Thines sp. nov., is a devastating foliar disease of fresh-cut basil first discovered in the U.S. in South Florida in 2007. Since then the pathogen has been found in over 20 U.S. states and has become a major threat to sweet basil production. In this study, acibenzolar-S-methyl (ASM, Actigard 50WG), DL-3-aminobutyric acid (BABA), isonicotinic acid (INA), salicylic acid (SA) and sodium salicylate (SS) were evaluated for their potential to control basil downy mildew in the greenhouse. Efficacy of these systemic acquired resistance (SAR) inducers varied in control of basil downy mildew depending on the rate, method and timing of application. Foliar sprays of ASM applied pre-, post- or pre- + post-inoculation at rates ranging from 25 to 400 mg l⁻¹ significantly (P = 0.05) reduced disease severity compared to the non-treated control in all experiments. ASM sprayed at 50 mg l⁻¹ three times on a weekly basis starting 3 and 7 days post- inoculation resulted in a 93.8 and 47.1% reduction in disease severity, respectively. Six weekly foliar sprays of BABA as pre- + post-inoculation at rates equal or higher than 125 mg l⁻¹ significantly suppressed downy mildew compared to the non-treated control. Foliar treatments of ASM or BABA followed by one or two post-inoculation sprays of a mixture of potassium phosphite (Prophyt) and azoxystrobin (Quadris) significantly improved efficacy for disease control. Sporangia counted on ASM treated leaves were significantly lower than leaves sampled from the non-treated control. ASM and BABA at concentrations lower than 1.0 mM did not inhibit sporangial germination in vitro. The effect of INA, SA and SS on disease reduction was generally inconsistent and not significant compared to the non-treated control.     

Fungitoxicity and synergism of mixtures of fluopicolide and pyraclostrobin against Phytophthora infestans

Fluopicolide and pyraclostrobin were new systemic fungicides with highly inhibitory activity on a broad spectrum of oomycetes, but so far the fungitoxicity of their mixtures on the different developmental stages of Phytophthora infestans and whether synergism exists have not been investigated. The joint-toxicity of the mixtures of fluopicolide with pyraclostrobin was determined against the different developmental stages (mycelial growth, zoospore release, cystospore germination and sporangial germination) of P. infestans and late blight on the leaf discs and the potted plants of potato, and whether the synergistic interaction exists in the mixtures of the two fungicides were evaluated in vitro and in vivo based on the synergistic ratios calculated with the Wadley formulas. The protective activity and curative activity against P. infestans and the efficacy duration in controlling potato late blight of the synergistic mixtures of the two fungicides were examined in this study. The results showed that mixtures of fluopicolide (F) and pyraclostrobin (P) at the ratios of 10:1 and 1:4 (F:P) exhibited synergistic interactions and had excellent inhibitory activity against almost all developmental stages of P. infestans. Synergistic interactions of the mixtures at the ratios of 1:4 and 10:1 (F:P) against the mycelial growth of metalaxyl-resistant isolates were more obvious than against the mycelial growth of metalaxyl-sensitive isolates. Synergistic interactions of the mixtures at the ratio of 1:4 (F:P) against the mycelial growth of the most of the tested isolates were more obvious than those of the mixtures at the ratio of 10:1 (F:P). The highest activity of the mixtures was observed against zoospore release at the ratio of 1:4 (F:P) with EC₅₀ value of 0.0044 μg ml⁻¹. The strongest synergistic interaction of the mixtures was observed against zoospore release at the ratio of 10:1 (F:P) with the synergistic ratio of 5.27. The mixtures of fluopicolide and pyraclostrobin at the ratios of 1:4 and 10:1 (F:P) showed synergistic interactions against P. infestans on the leaf discs and intact plants of potato. The EC₅₀ values for protective activity and curative activity of the mixture at the ratio of 10:1 (F:P) were 0.63 μg ml⁻¹ and 0.87 μg ml⁻¹, and EC₅₀ values of protective activity and curative activity of the mixture at the ratio of 1:4 (F:P) were 0.27 μg ml⁻¹ and 0.44 μg ml⁻¹, respectively. At 14 days after spraying of fungicide solutions, the disease control efficacy of the mixtures at the ratios of 10:1 and 1:4 (F:P) was still over 80% at the dosages of 50 μg ml⁻¹ and 100 μg ml⁻¹, and the disease control efficacy of the two mixtures remained at about 97% at the dosages of 200 μg ml⁻¹, indicating long-lasting effective control efficacy of the two fungicides against potato late blight.     

Sensitivity of Phytophthora parasitica to mandipropamid: In vitro determination of baseline sensitivity and in vivo fungitoxicity

Tobacco black shank caused by Phytophthora parasitica is a devastating soil-borne disease of tobacco in China. Control of tobacco black shank relies on numerous fungicide applications. A new carboxylic acid amide (CAA) fungicide, mandipropamid, was examined for its effects on various asexual developmental stages of P. parasitica in vitro and in tobacco seedlings. Although mandipropamid did not affect discharge of zoospores from sporangia, it strongly inhibited mycelial growth (mean EC₅₀ value of 0.0112 μg ml⁻¹), sporangia production (mean EC₅₀ value of 0.009 μg ml⁻¹), germination of encysted zoospores (mean EC₅₀ value of 0.014 μg ml⁻¹), and germination of sporangia (mean EC₅₀ value of 0.017 μg ml⁻¹). For protective activity in tobacco seedlings, various applications of mandipropamid were superior in reducing black shank compared to that of metalaxyl and of azoxystrobin; while for curative activity assay, 100 μg ml⁻¹ of mandipropamid exhibited the same efficacy as that of metalaxyl, and presented superior activity than that of azoxystrobin. In 2010 and 2011, 119 isolates of P. parasitica from Guizhou Province of China were characterized for the baseline sensitivity to mandipropamid by measuring mycelial growth. Values of effective concentrations for 50% mycelia growth inhibition varied from 0.0068 to 0.0285 μg ml⁻¹ and averaged 0.013 (±0.0045) μg ml⁻¹, with a unimodal distribution. This information will serve as a baseline for tracking future changes in sensitivities of P. parasitica populations to mandipropamid in China.     

Efficacy of azoxystrobin for the control of cucumber downy mildew (Pseudoperonospora cubensis) and fungicide residue analysis

During a five-year trial (2007–2011), the efficacy of azoxystrobin (Quadris, 250 g a.i. L⁻¹, Syngenta) in two doses (187.5 g a.i. ha⁻¹ and 250 g a.i. ha⁻¹) and chlorothalonil (Bravo 720-SC, 720 g a.i. L⁻¹, Syngenta) at a rate of 1.44 kg a.i. ha⁻¹ was tested for the control of cucumber downy mildew (CDM). Cultivars that were susceptible or resistant to CDM (Regal and Haros, respectively) were tested for their response to fungicide applications. Differences in both disease severity and yield of the cultivars among resistance levels and fungicide treatments were observed. A highly significant and negative correlation was obtained between AUDPC and yield. Higher yields can be achieved by planting more resistant cultivars in combination with lower doses of fungicides. This is an indication that CDM contributes significantly to yield losses in cucumber production in Serbia. While monitoring the degradation of azoxystrobin residues, a decrease in residue levels to 1.0 mg kg⁻¹ below the maximum residue level (MRL) was observed at the end of the pre-harvest interval (PHI).     

Assessment of phenotypic diversity of Plasmopara viticola on Vitis genotypes with different resistance

The lack of characterized isolates of Plasmopara viticola is a very limiting issue in the management of downy mildew of grapevine. Although molecular studies have previously confirmed a high diversity of this pathogen, there are still no phenotypically characterized pathotype strains or races available which could be used to study the mechanisms of interaction with host genotypes of different resistance. A leaf disk inoculation technique was used to assess the variability of reactions of six different Vitis genotypes infected with single sporangial clones of five field isolates of P. viticola from different geographical origins. The virulence of thirty P. viticola clones was characterized on grapevine cultivars (Müller-Thurgau, Regent and Cabernet Cortis) and wild species (Vitis vinifera ssp. sylvestris, Vitis rupestris and Vitis riparia) with different susceptibility to downy mildew. Five categories ranging from full susceptibility with profuse and not clearly restricted sporulation (type A) to complete resistance with no sporulation or necrotic reaction (type E) were defined to evaluate the pathogen phenotype. A high variation of pathotypes was found amongst the samples from different field accessions and an unexpected variability was observed even between the cloned strains from the same field. This also accounted for the fungicide tolerance against the two most common fungicides, metalaxyl-M (Phenyl Amide group) and dimethomorph (Carboxylic Acid Amide group), where clones showing high sensitivity and high tolerance to the one or both fungicides were found within the same field. The virulence patterns found within clones of single field isolates supports previous reports that numerous genotypes may be involved in the infection of a single plant or even a single leaf. Moreover, the study provides a methodology to identify and select specific host–pathogen combination suitable for future studies in mechanisms of grapevine downy mildew interaction.     

Baseline and differential sensitivity to mandipropamid among isolates of Peronophythora litchii, the causal agent of downy blight on litchi

Litchi downy blight caused by Peronophythora litchii is a devastating disease of litchi plants in China. Control of litchi downy blight requires numerous fungicide applications. A new carboxylic acid amide (CAA) fungicide, mandipropamid, was examined for its in vitro effects on multiple asexual stages of four single-sporangium P. litchii isolates and protective activity against downy blight in detached fruit assays. Though mandipropamid did not affect discharge of zoospores from sporangia, it strongly inhibited mycelial growth (mean EC₅₀ = 0.0048 μg ml⁻¹), sporangia production (mean EC₅₀ = 0.0032 μg ml⁻¹), germination of encysted zoospores (mean EC₅₀ = 0.0023 μg ml⁻¹), and germination of sporangia (mean EC₅₀ = 0.0061 μg ml⁻¹). On detached fruit, 0.39, 1.56 and 6.25 μg ml⁻¹ of mandipropamid were superior in reducing downy blight compared to metalaxyl and flumorph, however, the 25 μg ml⁻¹ application rate was necessary for all three CAA fungicides to completely inhibit the disease. In 2007, 100 isolates from Fujian, Guangdong, and Guangxi Provinces of China were characterized for the baseline sensitivity to mandipropamid. The isolates obtained from different provinces showed similar baseline sensitivities to mandipropamid. Baseline sensitivities formed a unimodal curve with mean EC₅₀ values of 0.0055 ± 0.0012 μg ml⁻¹ for inhibition of mycelial growth. The described baseline sensitivities of P. litchii populations will be useful for monitoring possible shifts in sensitivity to mandipropamid.     

Potassium phosphite for control of downy mildew of soybean

Downy mildew of soybean, caused by Peronospora manshurica, is widely spread throughout Brazil. The objective of this study was to evaluate the use of potassium phosphite to control this disease. Field experiments were conducted during the growing season of 2006/2007 and 2007/2008 in the state of Parana in southern Brazil. The experimental design consisted of completely randomized blocks in a factorial arrangement (4 × 2) with four replications. Four rates of potassium phosphite (0, 375, 750 and 1500 g P₂O₅ + K₂O ha⁻¹) were applied at two growth stages, V6 (fifth trifoliolate leaf) and R2 (full flowering), followed by one or two applications of pyraclostrobin and epoxiconazole (66.5 + 25 g a.i. ha⁻¹) at R3 (pod development) or R2 and R5.1 (10% of pod filling), mainly for the control of Asian soybean rust (Phakopsora pachyrhizi) and powdery mildew (Microsphaera diffusa). Field experiments were conducted to quantify the severity of downy mildew on leaves, nutrient content in leaf tissue (N, P and K), leaf area index (LAI), yield and seed weight. The maximum severity of downy mildew was observed at a growth stage of R5.3 (50% of pods were ripe), with 14% and 46% of the leaf area affected in 2006/2007 and 2007/2008, respectively. Also it was detected some effect of phosphite on Asian rust control but it was mostly in the trial of 2007/08 when the epidemic was very low (9.7-21.8% of severity). There was a linear reduction in the severity of downy mildew and a significant improvement in the LAI with an increase in the rate of phosphite applied. During the 2006/2007 growing season, a significant yield improvement was observed due to the application of the highest rate of phosphite. Two fungicide applications following phosphite application significantly improved the control of Asian soybean rust and powdery mildew, yield and seed weight when compared to a single fungicide application.     

Efficacy of fosetyl-Al,propamocarb, dimethomorph,cymoxanil, metalaxyl and metalaxyl-M in Czech Pseudoperonospora cubensis populations during the years 2005 through 2010

A total of 134 Czech Pseudoperonospora cubensis isolates originating from Cucumis sativus were used for fungicide resistance screening. Efficacy of six commonly used and registered fungicides was screened in Czech P. cubensis populations. The investigations covered the frequency and occurrence of sensitive, moderately resistant and resistant isolates to the individual fungicides during the period 2005–2010. Fosetyl-Al (Aliette 80 W) and propamocarb (Previcur 607 SL) were the most effective fungicides. All tested isolates were sensitive on all tested fosetyl-Al concentrations. However, some isolates expressed resistance (profuse sporulation) or moderate resistance (limited sporulation) to lower and/or even to recommended concentrations of propamocarb in the years 2006 and 2008–2010. Metalaxyl (Ridomil PLUS 48 WP) and metalaxyl-M (Ridomil Gold MZ 68 WP) were ineffective. Isolates collected in 2008 and 2009 showed large variation in moderate resistance or resistance even at high fungicide dosages. However, this result was not confirmed in 2010, when efficacy of these fungicides increased slightly, and 69% and 43% of isolates were controlled at the recommended concentration of metalaxyl-M and metalaxyl, respectively. Limited or profuse sporulation was observed only sporadically at high concentrations. Sensitivity of isolates to cymoxanil (Curzate K) differed among the studied years. While cymoxanil was ineffective in the years 2005–2008 and in 2010, 68% of isolates were controlled at the recommended concentration in 2009. During the years 2005 through 2010, a shift towards higher sensitivity to dimethomorph was observed at all screened concentrations.     

Characterization of fungicide resistant isolates of Phaeoacremonium aleophilum infecting grapevines in Spain

Fungicide resistance in Phaeoacremonium aleophilum, one of the most frequent fungal pathogens associated with grapevine trunk diseases, was investigated and found to exist in some isolates of the pathogen against a commercial formulation, Escudo^®. The effect of this compound and its two active substances, carbendazim and flusilazole, was first evaluated on the mycelial growth of P. aleophilum. Escudo^®-resistant isolates were estimated at a frequency of 24% in Spanish vineyards. Then, the two active substances were used individually to test their effect on mycelia growth of twelve single-spore cultures originating from six Escudo^®-resistant isolates. Flusilazole (DMI-triazole) did not inhibit mycelia growth of any single-spore cultures of P. aleophilum. Carbendazim (benzimidazole) used alone allowed the growth of the same single-spore cultures that were also resistant to Escudo^®. AFLP characterization of sensitive and resistant single-spore cultures showed genetic diversity within P. aleophilum isolates but no AFLP markers were associated with resistance. New primers set (L2/R1) were designed to partially amplify the exon 6 of the beta-tubulin gene of P. aleophilum. Two different point mutations resulted in glycine (GGC) or lysine (AAA) replacing the glutamic acid (GAG) at codon 198 of the beta-tubulin gene in some of the resistant single-spore cultures studied. Resistant single-spore cultures of P. aleophilum were shown to have different aggressiveness levels as sensitive single-spore cultures by inoculation of wood segments of Vitis vinifera in the presence and absence of fungicide.     

Downy mildew resistant/susceptible cucumber germplasm (Cucumis sativus L.) genetic diversity assessment using ISSR markers

Downy mildew is one of the major foliar diseases of cucumber in which the new resistant cucumber cultivars are necessary to control this disease. Before improving the new cultivar, knowing the information about genetic diversity of cucumber germplasm is necessary. Thus, the objective of this study is to evaluate the genetic diversity of downy mildew resistant/susceptible cucumber germplasm. Forty cucumber accessions were screened for downy mildew disease reaction and were classified into five classes which the average of percent of symptomatic leaf area ranged from 20.1% (highly resistant) to 95.0% (highly susceptible). Then, these cucumber germplasms were analyzed with 20 inter simple sequence repeat (ISSR) markers. A total of 17 ISSR primers (85%) could amplify in all 40 cucumber accessions with the percent of polymorphic bands of each primer ranged from 55.56% to 100%. The polymorphism information content (PIC) ranged from 0.12 to 0.45 with the mean of 0.25, while the resolving power (RP) averaged at 1.82 and ranged from 0.50 to 3.20. The phylogenetic tree and principal component analysis revealed the association between downy mildew disease reaction and country of the origin of cucumber germplasm which could help the cucumber breeders to select the parental lines for improving the new cultivars that resist to downy mildew in the future.     

Susceptibility to downy mildew (Plasmopara viticola) of different Vitis varieties

The susceptibility of different varieties of Vitis vinifera and other Vitis species to downy mildew (caused by Plasmopara viticola) in field, greenhouse and laboratory tests was compared over a period of three years. Different degrees of susceptibility were detected. The most susceptible V. vinifera varieties were ‘Treixadura’ and ‘Albariño; ‘Cabernet Sauvignon’, ‘Mencía’ and ‘Chasselas Doré’ were the least susceptible. The non-vinifera varieties showed no symptoms of downy mildew in the field. Surprisingly, in laboratory and greenhouse tests, the rootstock 110-R was much more susceptible to infection than S04. No relationship was seen between susceptibility and berry colour, or the time of sprouting or fruit ripening. Neither was any correlation seen between susceptibility and the condition of being a traditional or introduced variety. Better knowledge of the susceptibility to downy mildew of different varieties would allow for growers and breeders to select those that are more resistant.     

Cyperus difformis evolves resistance to propanil

Cyperus difformis L. is one of the worst weeds of rice world-wide and has evolved resistance to acetolactate synthase (ALS)-inhibiting herbicides in rice fields of California. Propanil use was intensified to control the widespread resistant biotypes. Rice growers have recently experienced poor control, suggesting resistance to this photosystem II-inhibiting herbicide may have evolved in C. difformis populations. The objectives of this study were to detect the presence of propanil resistance, to establish resistance levels, and to investigate involvement of enhanced herbicide detoxification as mechanism of resistance through the use of metabolic inhibitors. Four C. difformis populations collected in rice fields from the Sacramento Valley of California were confirmed resistant to propanil. This is the first case of such resistance outside the Poaceae and the first time C. difformis exhibits resistance to an herbicide mechanism of action other than ALS inhibition. Carbaryl and malathion applied individually in mixture with propanil had minor effects on herbicide toxicity suggesting metabolic detoxification was not a resistance mechanism. A resistant biotype produced more than 80% biomass after a propanil (6.7 kg a.i. ha⁻¹) and carbaryl (1.9 kg a.i. ha⁻¹) or propanil and malathion (1.0 kg a.i. ha⁻¹) treatment compared to <20% by a susceptible biotype, suggesting substantial resistance still persisted in spite of insecticide addition. Propanil-resistant plants were cross-resistant to bensulfuron-methyl, imazosulfuron, halosulfuron-methyl and penoxsulam, but susceptible to carfentrazone. The loss of propanil to control this important weed of rice underscores the fragility of herbicide-based weed control in monoculture rice. Integrated weed management approaches to decrease herbicide selection pressure are needed to mitigate the evolution of multiple-herbicide resistance in C. difformis of California rice.     

Sensitivity,resistance stability,and cross-resistance of Plasmopara viticola to four different fungicides

Baseline sensitivity to fungicides was determined in 105 Plasmopara viticola isolates using the floating leaf disk test. The mean EC₅₀ values were 0.372 ± 0.104, 0.604 ± 0.215, 0.306 ± 0.101, and 0.922 ± 0.209 μg mL⁻¹ for fluxapyroxad, benthiavalicarb-isopropyl, ametoctradin, and famoxadone, respectively, which we regarded as the baseline sensitivity to the four fungicides. Resistant mutants can be generated using a median effective concentration or a minimum inhibitory concentration to fluxapyroxad, benthiavalicarb-isopropyl, ametoctradin, and famoxadone respectively, but the resistance stability of resistant mutants from the minimum inhibitory concentration (MIC) was significantly lower than that from the median effective concentration (EC₅₀). In brief, high resistant mutants obtained was at the higher concentration, the resistance stability of the mutants recovered was more quickly. But lower resistant mutants obtained was at the lower concentration, the resistance stability of the mutants recovered was slower. Even if the resistance level of the mutants is low, it is more capable of stably heritable. These data indicate that the highest dosage is not used, resistance will still develop. In a correlation analysis, no cross-resistance to each other of these four fungicides was observed.     

Sensitivity to iprodione, difenoconazole and fludioxonil of Rhizoctonia cerealis isolates collected from wheat in China

Since the 1980s wheat sharp eyespot (WSE) caused mainly by Rhizoctonia cerealis has become one of serious diseases of wheat in China. In this study, the sensitivity of 89 R. cerealis isolates to different fungicides was evaluated using mycelial growth inhibition assays. The results showed that all R. cerealis isolates tested were sensitive to iprodione, difenoconazole and fludioxonil with mean EC₅₀ (effective concentration that results in 50% of mycelial growth inhibition) values of 0.419, 0.062 and 0.033 μg/ml, respectively. To evaluate the risk of fungicide resistance in R. cerealis, an attempt was made to induce resistant mutants in the laboratory. Although difenoconazole- and fludioxonil-resistant mutants were not obtained, we obtained seven independently iprodione-resistant (IR) mutants from 89 parental isolates. The EC₅₀ values for these IR mutants were greater than 100 μg/ml, whereas those for the original wild-type counterparts were less than 1 μg/ml. After having been subcultured on PDA for 10 generations, the IR mutants did not show any decrease in resistance to iprodione. Additionally, these IR mutants also showed resistance to fludioxonil but remained sensitive to difenoconazole. Osmotic sensitivity tests showed that the IR mutants were hypersensitive to osmotic stress generated by NaCl. Inoculation tests showed that all the IR mutants lost their ability to infect the host plant. Taken together, these results indicate that the current population of R. cerealis is sensitive to these fungicides and a fitness cost is associated with iprodione-resistant mutants of R. cerealis in both osmotic stress and pathogenicity. The information obtained in this study is useful in monitoring and managing fungicide resistance in R. cerealis populations in China.     

Detection and characterization of bismerthiazol-resistance of Xanthomonas oryzae pv. oryzae

The resistance to bismerthiazol in the Xanthomonas oryzae pv. oryzae population in China has not been clearly determined. Three bismerthiazol-resistant mutants (1-1-1, 2-1-1, 4-1-1) of X. oryzae pv. oryzae were induced on rice plants and the characteristics were compared with their parental strain ZJ173. After 10 subcultures on fresh bismerthiazol-free nutrient agar plates and after being inoculation onto and re-isolation from untreated rice plants two times, the mutants remained pathogenicity on rice plants treated with 300 μg/ml bismerthiazol, indicating that the in vivo-induced bismerthiazol resistance was stable in vivo. The in vitro characteristics of the bismerthiazol-resistant mutants and a wild-type bismerthiazol-sensitive strain (ZJ173) were also compared. The three mutants grew faster than ZJ173 during the logarithmic growth phase but at the same rate during the decline phase. Bismerthiazol had protective and curative activity against strain ZJ173 but not against the three mutants. To evaluate the current degree of bismerthiazol resistance in field populations of X. oryzae pv. oryzae in China, 781 isolates of X. oryzae pv. oryzae were obtained from 1419 leaf samples collected from Yunnan, Sichuan, Jiangsu, Anhui, Hubei, Guangdong, Hainan, and Hunan Provinces in China from 2007 to 2009. The in vivo sensitivity of 505 of the 781 isolates to bismerthiazol was determined by applying them to rice plants treated with 300 μg/ml bismerthiazol and evaluating blast severity. The tested strains were considered resistant, if they showed <70% inhibition relative to strain ZJ173. Of the 505 isolates, 62 were resistant to bismerthiazol. The resistance frequencies among field population ranged from 0 to 21.1%, differed among the provinces, and was 11.2, 20.0 and 10.4% in 2007, 2008 and 2009, respectively. Resistance determined in vivo was not related to resistance determined in vitro. The in vivo-induced bismerthiazol resistance identified in this study will serve as a base line for further studying bismerthiazol resistance in fields, and the current degree of bismerthiazol resistance in field populations of X. oryzae pv. oryzae in China determined would be useful for the rice growers for control of rice bacterial blight.     

Baseline sensitivity of Monilinia laxa from Greece to fenhexamid and analysis of fenhexamid-resistant mutants

Fenhexamid is a hydroxyanilide fungicide with excellent performance against Botrytis cinerea but also effective against Monilinia spp. which cause brown rot disease in apple and stone fruit. A total of 75 Monilinia laxa field isolates were utilized to determine baseline sensitivity while a number of fenhexamid-resistant laboratory mutants were used to evaluate the resistance risk associated with the longevity of the effectiveness of fenhexamid. Fenhexamid was found to be highly effective against all field isolates. EC₅₀ values ranged from 0.02 to 1 μg mL⁻¹ and were distributed unimodaly around an average of 0.1 μg mL⁻¹. M. laxa laboratory strains with moderate and high resistance levels to fenhexamid were isolated after UV mutagenesis. All fenhexamid-resistant strains showed parental sensitivity to carbendazim, iprodione, fludioxonil, pyraclostrobin, flusilazole and prochloraz. Interestingly, some of the mutant strains were also resistant to tridemorph, fenpropimorph and spiroxamine. Studies on fitness parameters of fenhexamid-resistant strains revealed a fitness cost on sporulation and pathogenicity but not on mycelial growth. These results suggest that fenhexamid should be a good alternative site-specific fungicide for the control of brown rot disease caused by M. laxa. However, appropriate anti-resistance strategies should be considered to ensure the successful commercial use of fenhexamid in the long run.     

Multiple-fungicide resistance to carbendazim,diethofencarb, procymidone,and pyrimethanil in field isolates of Botrytis cinerea from tomato in Henan Province,China

Botrytis cinerea, the fungal pathogen responsible for causing gray mold in tomatoes, frequently exhibits multiple fungicide resistance. In this study, the sensitivity to carbendazim (Car), diethofencarb (Die), procymidone (Prc), and pyrimethanil (Pyr) was determined for 263 isolates of B. cinerea from diseased fruit and leaves of tomato plants from 87 commercial greenhouses in Henan Province of China during 2013 and 2014. Of the isolates tested, 89%, 93%, 85%, and 86% were resistant to Car, Die, Prc, and Pyr, respectively. Twelve phenotypes were found having different sensitivity patterns, and multi-fungicide resistance was found to one, two, three or four fungicides. The percentage of multiple fungicide resistant isolates (Car^RDie^RPrc^RPyr^R) was 68%. Mycelial growth, spore production, and mycelial dry weight were not significantly different between resistant and sensitive phenotypes of the B. cinerea isolates. However, the virulence of Car^SDie^RPrc^SPyr^S isolates was higher than that of the other fungicide-resistance phenotypes. All of the carbendazim and diethofencarb resistant phenotypes carried a point mutation at codon 198 (E198A or E198K) or 200 (F200Y) in the β-tubulin gene, and all of the procymidone resistant phenotypes carried point mutations at codons 369 (Q369P) and 373 (N373S) in the Bos1 gene. These results provided important reference data for assessment of resistance risk of B. cinerea isolates from commercial tomato transplants in Henan Province, indicating the existence of a high frequency of resistance to carbendazim, diethofencarb, procymidone, and pyrimethanil revealing the necessity of a more integrated control of gray mold.     

Multilocation evaluation and selection of stable genotypes for reaction to downy mildew in pearl millet

Twenty-five pearl millet (Pennisetum typhoides (Burm. f.) Stapf and Hub. = P. americanum (L.) K. Schum.) genotypes were evaluated for resistance to downy mildew (Sclerospora graminicola (Sacc.) Schroet.) at 10 different locations in India and Africa during 1979 and 1980. The objective of this work was to identify genotypes with stable and generalized resistance. Genotypic differences in linear regression coefficients were found and regression lines for downy mildew incidence in the various genotypes tended to converge. It was concluded from this that selection on the basis of disease severity would enable genotypes that are superior at all disease levels to be selected. Seven genotypes, namely 700516, 700651, J-1486 × 700787-2-10, P-7, EB-83-2, NC-7174 and SSC-7218, showed low disease scores, average regressions and non-significant deviations, and therefore expressed stable, generalized resistance. Genotype 700251 was particularly suitable for high-disease environments. BJ-104, an important Indian hybrid cultivar, was very unstable and susceptible to downy mildew.