Systemicity and persistence of cymoxanil in mixture with oxadixyl and mancozeb against Phytophthora infestans and Plasmopara viticola

Uptake and translocation of the fungicide cymoxanil was studied using tomato and potato plants infected with Phytophthora infestans and grape plants infected with Plasmopara viticola. Translocation of cymoxanil was mainly translaminar and acropetal from roots and stems to leaves to an extent similar to that of the phenylamide fungicide oxadixyl. Basipetal translocation from upper to lower leaves, as well as lateral movement, was also observed but to a lesser extent. Plant parts that grew after application were protected against P. infestans and P. viticola because of systemic activity up to 7 days by cymoxanil alone and up to 14 days when cymoxanil was used as part of a three-way mixture with oxadixyl and mancozeb. Drench application of cymoxanil and a simultaneous spray application of mancozeb against P. infestans and P. viticola resulted in a significant synergistic interaction. The spray application of the three-way mixture oxadixyl/mancozeb/cymoxanil was more effective against both sensitive and phenylamide-resistant strains of P. infestans and P. viticola than the individual components alone, representing significant levels of synergistic interactions. Synergy ratios were higher for resistant than for sensitive strains of both pathogens.     

A comparison of the biochemical and physiological effects of the systemic fungicide cyprofuram with those of the related compounds metalaxyl and metolachlor

Strains of Phytophthora megasperma insensitive to the fungicide metalaxyl showed limited cross-resistance to the structurally related fungicide cyprofuram and to metolachlor, a herbicide with antifungal properties. The three compounds each inhibited nucleic acid synthesis, but the wide variation in resistance factors and in the degree of nucleic acid inhibition at the concentrations inhibiting mycelial growth by 50% indicated that cyprofuram and metolachlor possess alternative mechanisms of toxicity. Studies with protoplasts suggested that cyprofuram has a direct effect on the plasma membrane which may be important against those resistant strains where nucleic acid synthesis is least affected. Metolachlor also was extremely effective in causing protoplast lysis. Sporangia production by P. palmivora was stimulated by low concentrations of cyprofuram, and germination was delayed. As with other ‘acylalanine-type’ fungicides, inhibition of nucleic acid synthesis is considered to be the main mode of action. Some degree of cross-resistance between cyprofuram and other acylalanines is likely to occur in the field but the dual mode of action of cyprofuram might delay its onset. The effect of increased sporogenesis on the development of resistance is more difficult to assess.     

Effects of azoxystrobin, difenoconazole, polyoxin B (polar) and trifloxystrobin on germination and growth of Alternaria alternata and decay in red delicious apple fruit

Alternaria alternata is the predominant fungal pathogen responsible for moldy-core in red delicious strains of apple. In this study, we report on the effects of the polyoxin B compound Polar, the strobilurin fungicides, azoxystrobin and trifloxystrobin, and the sterol inhibitor difenoconazole on spore germination, mycelial growth and fruit decay on detached fruits caused by A. alternata. Germination was most sensitive to polyoxin B and trifloxystrobin among the tested compounds. The 50% and 95% effective concentration (EC₅₀, and EC₉₅), values for in vitro inhibition of conidial germination of the fungus were lowest for polyoxin B and trifloxystrobin, ranging from <0.01 to 0.15 μg/ml and 180 μg/ml, respectively. Germination was least sensitive to difenoconazole and azoxystrobin (EC₅₀ and EC₉₅ values ranging from 25 to 72 μg/ml and 720 μg/ml, respectively). Mycelial growth in vitro of A. alternata was most sensitive to difenoconazole (EC₅₀ and EC₉₅ values of 0.8 and 12 μg/ml, respectively) and least sensitive to both strobilurin fungicides (EC₉₅>1000 μg/ml). Decay formation by A. alternata on mature detached fruits was most affected by trifloxystrobin and azoxystrobin (EC₅₀ and EC₉₅ values of 0.015–0.087 μg/ml and 8 μg/ml, respectively), intermidiate in sensitivity to Polyoxin B (EC₅₀ and EC₉₅ from 1 to 33 μg/ml, respectively), and difenoconazole was the least effective (EC₅₀ and EC₉₅ from 20 to 490 μg/ml, respectively). Polyoxin B applied at 24 or 48 h post-inoculation inhibited fruit decay development. Difenoconazole and trifloxystrobin were less effective and azoxystrobin was ineffective when applied at 48 h post inoculation. The activity of azoxystrobin, difenoconazole, Polyoxin B and trifloxystrobin on one or more stages of the life cycle of A. alternata and on decay development in fruits suggests that these compounds potentially could provide control of moldy-core disease in apple.     

New sources of resistance to Phytophthora megakarya identified in wild cocoa tree populations of French Guiana

Cocoa black pod rot, a disease caused by oomycetes of the genus Phytophthora, causes substantial yield losses throughout the world, particularly in Africa with the very aggressive species Phytophthora megakarya. In order to reduce the impact of that pathogen, priority is given to genetic control through more resistant cultivars, and breeders are seeking sources of resistance in wild cocoa trees. Wild cocoa trees were surveyed in French Guiana between 1985 and 1995, leading to the collection of abundant plant material from more than 200 mother trees originating from five river basins. We present here the results of tests to assess resistance to the species P. megakarya (a species only existing in Africa), conducted at CIRAD in Montpellier, France, on circa 40 genotypes collected in the Camopi river basin, along with approximately 20 genotypes from other populations (Kérindioutou, Borne 7, Euleupousing, Pina and Oyapok). The strain used for artificial inoculation was NS269, isolated in Cameroon. Seven cacao clones were classified as “highly resistant” and 29 as “resistant”, some of which displayed greater resistance to P. megakarya than the reference resistant clone IMC 47. This study suggests that the wild material from French Guiana could play a significant role in controlling P. megakarya in Africa and also Phytophthora palmivora in all cocoa-producing zones.     

Effects of chrysophanol, parietin, and nepodin of Rumex crispus on barley and cucumber powdery mildews

Three substances that show antifungal activity against Blumeria graminis f. sp. hordei were isolated from roots of Rumex crispus and identified as chrysophanol, parietin, and nepodin. The substances were tested for plant disease control activity in vivo against six plant pathogenic fungi. All specifically reduced the development of barley powdery mildew. The concentrations required for 50% disease control were 4.7 μg/ml for chrysophanol, 0.48 μg/ml for parietin, and 20 μg/ml for nepodin. These agents showed both curative and protective activity against barley powdery mildew. Chrysophanol (100 μg/ml) and nepodin (400 μg/ml) were more effective than the fungicides fenarimol (30 μg/ml) and polyoxin B (100 μg/ml), under glasshouse conditions, against cucumber powdery mildew, which is caused by Podosphaera xanthii. Parietin (30 and 10 μg/ml) reduced the development of cucumber powdery mildew as efficiently as fenarimol (30 μg/ml) and more effectively than polyoxin B (100 μg/ml).     

Two decades of monitoring irish populations of<Emphasis Type="Italic">Phytophthora infestans</Emphasis> for phenylamide resistance

Summary Between 1981 and 1998,Phytophthora infestans populations in 878 potato crops in Ireland were monitored for phenylamide resistance. The incidence of resistance was very high in 1981 following intensive use of metalaxyl for blight control from 1977 onwards. It decreased rapidly in the absence of phenylamide use between 1981 and 1984. Following the reintroduction of phenylamide based fungicides in 1985, there was a rapid increase in phenylamide resistance following the widespread curative use of these fungicides. From 1990 onwards there was greater adherence among growers to the use of phenylamides in accordance with an antiresistance strategy and resistance distribution again decreased. During the 18-year period of the study, resistance was always lower at the beginning of the season and increased as the season progressed. Following the reintroduction of the phenylamide based fungicides in 1985 there was a steady increase in phenylamide use up to 1986 followed by a continuous decrease in use up to 1990. Since then there has been a steady increase in phenylamide use. They are currently used on some 60% of crops in Ireland and are still providing acceptable levels of disease control.     

Quantification of synergistic interactions of fungicides against Plasmopara and Phytophthora

Three fungicides with different modes of action—oxadixyl (phenylamide; systemic), mancozeb (dithiocarbamate; protectant), and cymoxanil (penetrant)—were tested alone and in different combinations against Plasmopara viticola on grapes and Phytophthora infestans on tomatoes. The fungicidal activities against sensitive and phenylamide-resistant strains were compared. The fungicidal activity of the mixture oxadixyl/mancozeb decreased significantly with increasing levels of resistance, whereas the mixture oxadixyl/mancozeb/cymoxanil was almost equally active against all strains. The level of synergistic interaction of fungicide mixtures was found to be dependent on the proportion of the single components in the mixture as well as on the activity of the individual compounds against the fungal strains. Cymoxanil-containing mixtures showed equally high or even higher levels of synergism against resistant compared with sensitive strains, whereas in oxadixyl/mancozeb mixtures synergism was lower in resistant situations. The methods used are suitable for finding those combinations of fungicides which give rise to maximum synergistic interactions: this was achieved with the uses of isobolograms, which were adapted to three-component mixtures leading to three-dimensional figures.     

Effect of antagonistic Fusarium spp. and of different commercial biofungicide formulations on Fusarium wilt of basil (Ocimum basilicum L.)

The ability of antagonistic Fusarium spp. to control Fusarium wilt of basil caused by F. oxysporum f. sp. basilici was tested under glasshouse conditions from 1994 to 1996. Fusarium oxysporum antagonistic strain 251/2, a protoplast fusion-derived hybrid coded FI-11 and F. moniliforme strains TF4 and TF4 RB were able to decrease (P = 0.05) the incidence of Fusarium wilt significantly. Biocontrol was consistent, especially when the antagonists were applied by mixing a talc preparation of propagules in the substrate (10⁵ CFU/ml of substrate) 1–2 weeks before sowing or at sowing. Seed coating with the biocontrol strains (10⁸ CFU/g of seed) did not generally provide sufficient disease containment, whereas the combination of substrate treatment and seed coating did not improve wilt control in comparison with substrate treatment alone. Two commercial formulations, based on antagonistic F. oxysporum strains and one containing the antagonistic strain K61 of Streptomyces griseoviridis, were generally not sufficiently effective at the tested dosages.     

马铃薯晚疫病菌对瑞毒霉与霜脲氰的交互抗性

2008~2011年在黑龙江省共分离了41个马铃薯晚疫病菌,并测定了这些菌株对瑞毒霉和霜脲氰的敏感性和交互抗性。瑞毒霉测定结果显示6个(14.63%)菌株显示为敏感性,11个(26.83%)菌株显示为中抗,24个(58.54%)菌株显示为抗性。这说明黑龙江省分离的晚疫病菌已对瑞毒霉产生抗性。对于霜脲氰,EC50值为0.0630~2.1289μg/mL,最不敏感菌株是最敏感菌株的33.79倍,敏感性基线值为0.1879μg/mL。交互抗性分析结果显示,未产生交互抗性的菌株为73.16%,产生交互抗性菌株为26.84%。以上结果说明霜脲氰和瑞毒霉不能交叉使用。     

Algicidal effect of 3-(3-indolyl)butanoic acid, a control agent of the bacterial wilt pathogen, Ralstonia solanacearum

The chemical control agent 3-(3-indolyl)butanoic acid, previously reported as a control agent for the bacterial wilt pathogen Ralstonia solanacearum, was shown to suppress the growth of green algae during hydroponic culture of tomato. The algicidal activity of the compound was effective at 10 μg/ml, completely preventing generation of green algae under non-shaded greenhouse conditions. The algicidal effect was mainly due to suppression of the growth of motile unicellular algal cells tentatively identified as Chlamydomonas spp., which are commonly occurring in the hydroponic solution and vigorously multiply to form an algal mat on the sponge supports. The compound has potential as a non-phytotoxic algicide for hydroponically cultured crop plants.     

Comparison of clofentezine and hyxythiazox with cyhexatin for integrated control of European red mite, Panonychus ulmi (Koch)

The ovicides clofentezine and hexythiazox were shown to be compatible with integrated control of European red mite, Panonychus ulmi (Koch), in a replicated field trial conducted over two growing seasons in an apple orchard at Batlow, New South Wales, Australia. Reduced concentrations of clofentezine (5 and 10 g a.i. per 100 litres water—normally 20 g) applied as foliar sprays to run-off, stabilized the predator/prey interaction between P. ulmi and Typhlodromus pyri Scheuten more effectively than low rates of cyhexatin (5 and 10 g a.i. per 100 litres water—normally 20 g). There was no evidence that the effectiveness of clofentezine or hexythiazox would destablize integrated control by eliminating P. ulmi and starving T. pyri. Populations of T. pyri persisted in low numbers on the trees in the absence of P. ulmi or other arthropod food sources on leaves.     

Fungicidal control of metalaxyl-insensitive strains of Bremia lactucae on lettuce

Strains of B. lactucae insensitive to 0.01 g a.i. l⁻¹ metalaxyl have been detected in all the lettuce-growing districts of South Australia. Despite this, applications of a formulated mixture of 0.25 g a.i. l⁻¹ metalaxyl and 2 g a.i. l⁻¹ mancozeb every 7–10 days controlled downy mildew in field experiments, where similar applications of either metalaxyl or mancozeb alone were ineffective. Protectant schedules of either 0.3 g a.i. l⁻¹ dimethomorph or 2–2.4 g a.i. l⁻¹ phosphonic acid also controlled downy mildew, but were less effective than the metalaxyl/mancozeb formulation. Mixtures of either dimethomorph or phosphonic acid with mancozeb enabled the time between the sprays to be increased to 14 days without significantly reducing control. Fungicide applications increased marketable yield by 60% compared with that of the unsprayed plants. Drenching seedlings with 4 g a.i. l⁻¹ phosphonic acid controlled downy mildew for at least 14 days and should prevent the spread of the disease from plant production areas.     

Studies of mechanisms of metalaxyl fungitoxicity and resistance to metalaxyl

The mode of action of the systemic fungicide metalaxyl and possible changes in Phytophthora spp. leading to resistance to metalaxyl were investigated. Uptake of labelled uridine and thymidine into RNA and DNA were reduced by sub-lethal amounts of metalaxyl. There was no evidence that metalaxyl caused a build-up of nucleotides or blocked the DNA template. Metalaxyl metabolites were absent from the fungus and the medium; the unchanged fungicide is probably the primary toxic agent. Washing metalaxyl-treated mycelium with water removed about 80% of the fungicide but the rest was firmly bound. Metalaxyl-resistant and sensitive strains of Phytophthora had similar growth rates in vitro, took up similar amounts of fungicide and incorporated uridine into RNA at similar rates. The mechanism of resistance remains obscure: subtle alterations to nucleic acid synthesis may be involved but other possibilities remain.     

Effect of pythium root rot on yield of white bean, Phaseolus vulgaris L.

The effects of Pythium paroecandrum and P. ‘spherical’ on seed yield of white bean (Phaseolus vulgaris L.) were determined under field conditions. Fenaminosulf (Lesan) applied as a soil drench selectively suppressed Pythium populations in soil and increased seed yields by 47% in 1976 and by 33% in 1977. Fumigation of soil with DD-MIT (Vorlex), a commercial mixture of methyl isothiocyanate, dichloropropenes and dichloropropanes, reduced populations of Pythium and Fusarium in soil and increased seed yields by 79% in 1976 and by 29% in 1977. The combination of fenaminosulf and DD-MIT produced the greatest reductions in soil populations of Pythium and Fusarium and increased seed yields by 65%. Pythium paroecandrum and P. ‘spherical’ were associated with feeder-root necrosis and were the only bean-root pathogens detected at the experimental site. In this study, losses in seed yield of white bean due to Pythium root rot were estimated to be 44% in 1976 and 39% in 1977.     

Patterns of cross-resistance to fungicides in Botryotinia fuckeliana (Botrytis cinerea) isolates from French vineyards

Field strains of Botryotinia fuckeliana were isolated from diseased grape berries which were collected at the harvest, between 1993 and 1997, from French vineyards located in Alsace, Armagnac, Bordeaux, Champagne and Loire Valley. According to the inhibitory effects of fungicides towards spore germination, germ-tube elongation and mycelial growth, several phenotypes could be characterized. Thus, two types of benzimidazole (e.g. carbendazim, thiabendazole)-resistant strains were detected; negative cross-resistance towards diethofencarb, diphenylamine and dicloran was found only in one type. Most dicarboximide (e.g. iprodione, procymidone, vinclozolin)-resistant strains were also weakly resistant to aromatic hydrocarbon fungicides (e.g. chloroneb, dicloran, tolclofos-methyl) but remained sensitive to phenylpyrroles (e.g. fenpiclonil, fludioxonil). However, in some other dicarboximide-resistant strains, resistance was observed either as being restricted to dicarboximides or as extending weakly to phenylpyrroles. Towards anilinopyrimidines (e.g. cyprodinil, mepanipyrim, pyrimethanil), three resistant phenotypes have been identified. In the most resistant one, resistance was restricted to anilinopyrimidines, whereas in the others, resistance extended to various other groups of fungicides including dicarboximides, phenylpyrroles, sterol biosynthesis inhibitors (e.g. tolfanate, prochloraz, tebuconazole) and the hydroxyanilide derivative, fenhexamid. According to the pattern of cross-resistance, two phenotypes were characterized within these multifungicide-resistant strains. In vitro tests conducted with fenpropimorph, fenpropidin and spiroxamine, revealed the existence of strains highly susceptible to these sterol Δ¹⁴- reduction inhibitors. This property was correlated with a decreased sensitivity towards fenhexamid.     

Resistance of Botrytis cinerea Pers. and Plasmopara viticola (Berk. & Curt.) Berl. and de Toni to fungicides in French vineyards

The control of Botrytis cinerea and Plasmopara viticola, two of the most important parasites on grapevines, is achieved by using fungicides. Recently several highly active compounds have been introduced but with some of them, failures of disease control have been observed in French vineyards. These phenomena are due to the build-up of resistance to benzimidazoles (benomyl, carbendazim, thiophanate-methyl) or to di-carboximides (iprodione, procymidone, vinclozolin) in Botrytis cinerea and to phenylamides (metalaxyl, ofurace) in Plasmopara viticola. The characteristics of the various resistant strains (levels of resistance, patterns of cross-resistance, fitness) are described. In 1982 and 1983 there has been monitoring for resistance in most French vineyards: the tests used and the results with regard to the frequencies of resistant strains are discussed.     

Resistance to the legume pod borer, Maruca vitrata Fabricius, and the probable modalities involved in wild Vigna

A large number of accessions belonging to selected wild Vigna species namely V. unguiculata subspecies dekindtiana, V. oblongifolia, and V. vexillata were evaluated using choice (DCAT) and no-choice (NCFT) laboratory feeding bioassays to determine their resistance to the pod borer, Maruca vitrata Fabricius. The most resistant accessions belonged to V. vexillata, followed by those from V. oblongifolia, with a few outstanding exceptions from V. unguiculata. Even though the latter were the least resistant, they provided the greatest variability among the tested accessions. In the DCAT, the cultivated cowpea line, IT84S-2246, which was used as the susceptible control was always preferred for feeding by the larvae (feeding index [FI] of 1.10–2.60) than the V. unguiculata accessions (FI ranged from 0–0.52). Against the resistant control (itself belonging to V. vexillata), only a few accessions were less preferred, most of these being either V. vexillata or V. oblongifolia. Larvae lost weight on the most resistant from all three species, because they either did not feed, or fed very little. Weight gain was positively correlated with seed damage (r = 0.82, P < 0.01) and the amount of frass produced (r = 0.91, P < 0.01). Seed damage was also positively correlated with the amount of frass produced (r = 0.64, P < 0.05). On the basis of these results, we believe that both antibiosis (post-ingestive effects) and antixenosis (deterrence to boring into the pods to feed) modalities of resistance are involved. The results are discussed further in relation to the origin, domestication and use of these accessions in cowpea improvement.     

A study on soil solarization and combined with fumigant application to control Phytophthora crown blight (Phytophthora capsici Leonian) on peppers in the East Mediterranean region of Turkey

Field trials conducted in the pepper growing area ( çel/Turkey) showed that solarization alone and combined with a reduced dose of methyl bromide (40 g/m²), were effective in controlling crown blight disease caused by Phytophthora capsici. Solarization was achieved by covering plots with a clear polyethylene sheet, 0.03 mm thick for 8 weeks. The temperature reached 47 °C and 35 °C at 5 cm and 30 cm soil depth, respectively, an average of 3–8 °C higher than those at equivalent depths in the untreated plots. The average incidence of the disease in solarization plus a reduced dose of methyl bromide, the recommended dose of methyl bromide, solarized and untreated plots in 1991 and 1992 were 17.6 and 13.3%; 20.8 and 16.8%; 24.1 and 19.7%; 39.8 and 42.9%, respectively. All treatments significantly reduced disease incidence.     

Recovery of Pantoea ananatis, causal agent of center rot of onion, from weeds and crops in Georgia, USA

Center rot of onion, caused by Pantoea ananatis, has been a problem of sweet onions in Georgia since 1997. A polymerase chain reaction (PCR) protocol was developed to screen for populations of P. ananatis on plant surfaces. Plant samples producing a positive PCR reaction were targeted for further processing to culture the bacterium. In a survey of the Vidalia onion-growing region of Georgia, we detected and cultured P. ananatis from 25 asymptomatic weed species, which included commonly occurring weeds such as crabgrass (Digitaria sanguinalis), Florida beggarweed (Desmodium tortuosum), Florida pusley (Richardia scabra), sicklepod (Cassia obtusifolia), Texas Millet (Panicum texanum), tall verbena (Verbena bonariensis) and yellow nutsedge (Cyperus esculentus). In addition, the bacterium was recovered from crop plants such as Bermuda grass (Cynodon dactylon), cowpea (Vigna unguiculata) and soybean (Glycine max). Based on the field survey and tests with strains held in storage in the Coastal Plain Experiment Station culture collection, we concluded that P. ananatis was in Georgia prior to 1997, and was distributed widely on weeds and crops throughout southern Georgia.     

霜疫霉侵染后荔枝果皮结构变化的扫描电镜观察

霜疫霉病严重危害荔枝果实的产量和质量,其病原菌是霜疫霉(Phytophthora litchii Chen ex Ko et al.)。以4×10~4cfu/mL霜疫霉孢子悬浮液分别接种抗病品种‘黑叶’和感病品种‘桂味’的果实,通过扫描电子显微镜观察接种0、1、6、12、24、48、72、96 h后荔枝外果皮显微结构的变化。扫描电镜观察发现,‘黑叶’果皮的龟裂片平整,主要组成单元为微小乳突状结构,其中间微微凸起且结构致密紧实;‘桂味’果皮的裂片谷较深,主要组成单元为微小蜂窝状结构,其中间凹陷且壁薄挺立。随着贮藏时间的延长,2个品种外果皮均出现不同程度的损伤,但‘黑叶’比‘桂味’损伤较轻。用相同浓度霜疫霉孢子接种后,‘桂味’果皮附着孢子的数量是‘黑叶’的近5倍,说明‘桂味’果皮显微结构更易于霜疫霉孢子的附着和侵染。接种霜疫霉后,病原菌侵染加剧荔枝外果皮结构的崩解,但‘黑叶’外果皮崩解速度较慢。在‘黑叶’上霜疫霉的孢子萌发、菌丝生长和孢子囊形成都晚于‘桂味’,说明其对霜疫霉的生长有一定的抑制作用。‘黑叶’和‘桂味’果皮显微结构的差异是其对霜疫霉表现不同抗性的重要原因。