Evaluation of the efficacy for reducing copper use against downy mildew control in organic Mediterranean viticulture

Protection against downy mildew in organic viticulture is exclusively based on the use of copper products but these cause environmental concerns. The aim of this work was to assess a control strategy under Mediterranean climatic conditions with the goal of reducing the quantities of copper allowed under organic viticulture. The control strategy was based on the use of copper oxychloride with varying doses and intervals between their applications. The efficiency of the control was assessed based on the incidence and severity of the disease in grape leaves and bunches. The products used ensured effective control under moderate disease pressure. It was found that during seasons with a low pressure of downy mildew attack, it is possible to contain the fungus respecting the ceiling prescribed by the organic regulations. Otherwise, when the pressure of the fungus is high, in the case of humid and rainy seasons, it is not possible to respect the limit. Organic growers can minimize the use of copper in organic viticulture by using copper-alternatives when downy mildew infection is intermediate or low.     

Contribution of molecular studies to botanical epidemiology and disease modelling: grapevine downy mildew as a case-study

After being accidentally introduced from the USA at the end of the 19^th century, downy mildew caused by Plasmopara viticola (Berk. et Curt.) Berlese et De Toni became one of the most damaging diseases affecting Vitis vinifera in Europe. Downy mildew causes both direct and indirect losses and can lead to severe reduction of yield. Our understanding of the life cycle and epidemiology of P. viticola has been recently altered by molecular studies that revealed that the overwintering inoculum (i.e., the oospores) does more than initiate disease, as was previously thought. A mechanistic model was developed for predicting the entire chain of processes leading to primary infections, and this primary infection model was linked to other models of secondary infection cycles. The model for primary infections defines the length of the primary inoculum season and a seasonal oospore dose consisting of several cohorts of oospores that progressively mature. The model was evaluated by means of Bayesian analysis in both Italy and eastern Canada, and showed high sensitivity, specificity, and accuracy both for potted plants and vineyards. Fungicide applications are necessary to control downy mildew because preventive agronomic practices are not very effective, including host resistance. The use of warning systems based on weather-driven models leads to a reduction in the use and cost of chemicals and a reduction in their environmental impact.     

不同葡萄品种对霜霉病的田间苗期抗性评价

对36个葡萄品种(品系)苗期对霜霉病的田间抗病性进行了评价, 结果表明, 不同葡萄品种(品系) 对葡萄霜霉病的抗性存在明显差异, 绝大多数欧美杂交种的抗性要强于欧亚种葡萄, 供试的10个欧美杂种中, ‘瑞峰无核’对葡萄霜霉病表现为高抗; ‘DEMIR’、‘龙宝’、‘红富士’、‘黑奥林’、‘峰后’5个品种表现为中抗。     

Evidence for sporangial dispersal leading to a single infection event and a sudden high incidence of grapevine downy mildew

In a German vineyard (PRO) between 10 and 13 June 2004, the incidence of grapevine downy mildew ( Plasmopara viticola ) increased abruptly from 0 to 99%. Infected vines bore on average between 45 and 60 lesions each, corresponding to about 220 000 lesions ha⁻¹ in a non-aggregated distribution. A second vineyard (FUT), approximately 50 m distant from PRO, had been inoculated 3 weeks before the abrupt increase in incidence of disease in PRO. Using microsatellites to ascertain the sources of inoculum and likelihood and extent of interplot spread from FUT to PRO, 555 samples were collected and 20 unique genotypes were identified, of which one caused 80% of the sampled lesions in both vineyards. Three genotypes responsible for 95% of the lesions in FUT and PRO were identified as the genotypes originally established through earlier inoculations in FUT. This is the first report of definitive and quantitative evidence of sporangial migration up to 130 m in a single infection event. The utility of molecular tools to address practical epidemiological issues in this pathosystem is illustrated. The results of this study provide an example of how P. viticola was able to rapidly colonize European vineyards after the pathogen was introduced from North America in 1878.     

Involvement of phenolic compounds in the resistance of grapevine callus to downy mildew (Plasmopara viticola)

Callus tissue of different grapevines (Vitis spp.) was inoculated withPlasmopara viticola. Short, highly-branched hyphae with necrosis, and long hyphae with heavy sporulation were observed on resistant and susceptible callus respectively. Thin-layer chromatography and spectrophotometric analysis showed that resistant callus contained greater quantities of gallocatechin derivatives than susceptible callus. Regression analysis between the field disease rating of each variety and its gallocatechin derivatives content indicated 92.2% correlation. Histochemical studies showed that, after infection withP. viticola, flavonoids appeared in the superficial cell walls of the callus, to a lesser degree on susceptible callus than on resistant callus. At a late stage of infection, the superficial cells of resistant callus were suberized, which did not occur in susceptible callus. This study showed that the preformed gallocatechin derivatives, the induced flavonoids and suberized superficial cells might play a role in the resistance of grapevine callus tissue to this fungus.Abbreviations CallusI Callus ofV. riparia var. Gloire de Montpellier - CallusV Callus ofV. vinifera var. Grenache - TLC Thin Layer Chromatography - var. variety - GAD Gallic acid derivatives - GD gallocatechin derivatives - RC resistant callus - SC susceptible callus     

An economic assessment of the impact on the Western Australian viticulture industry from the incursion of grapevine downy mildew

Journal of Plant Diseases and Protection - Grapevine downy mildew (Plasmopara viticola) was first detected in commercial vineyards in the Swan Valley region of Western Australia (WA) in 1998 and...     

暗黑链霉菌PY-1活性产物分析及其对葡萄霜霉病田间防效评价

为探究暗黑链霉菌Streptomyces atratus PY-1对葡萄霜霉病菌Plasomopara viticola的抑制作用,通过扫描电镜观察PY-1发酵滤液对葡萄霜霉病菌孢子囊的影响,检测PY-1的活性物质种类,并测定其对葡萄霜霉病的田间防效。结果表明,PY-1发酵滤液能够导致葡萄霜霉病菌孢子囊和孢子囊梗出现褶皱、破裂和畸形,进而使其丧失侵染功能。PY-1菌株代谢产物中包含几丁质酶、蛋白酶、嗜铁素、1-氨基环丙烷-1-羧酸脱氨酶、氰化氢、吲哚乙酸,不含纤维素酶。PY-1菌株对葡萄霜霉病具有较好的田间防效,发酵原液对葡萄霜霉病的田间中期防效可达到89.17%以上,末期防效达86.28%以上,比52.5%噁唑菌酮·霜脲氰2 000倍液的防效略低,但显著高于58%甲霜·锰锌1 000倍液的防效;PY-1菌株发酵液稀释700倍后对葡萄霜霉病的末期防效与甲霜锰锌1 000倍液防效相当。表明PY-1菌株具有研制防治葡萄霜霉病生防制剂的潜力。     

葡萄霜霉病抗病性鉴定方法及品种抗病性测定

本文通过对葡萄霜霉病抗病性鉴定方法进行优化,建立了一种更为快捷、方便、可靠的鉴定方法,并对32个葡萄品种对霜霉病抗病性进行了鉴定,为葡萄抗病品种的选育和应用提供依据。结果显示:以田间混合的霜霉病菌为接种体采用叶盘法鉴定葡萄品种的抗病性更加快捷、方便。供试的32个葡萄品种对葡萄霜霉病的抗性存在显著差异。其中免疫品种有‘康拜尔早生';高抗的有‘阳光玫瑰'、‘美乐'、‘Ms27-31'等5个品种;中抗的有‘贝达'、‘小芒森'、‘2E-16-2'等6个品种;低抗的有‘瑞都红玉'、‘早黑宝'、‘摩尔多瓦'等10个品种;感病的有‘里扎马特'、‘玫瑰香'、‘香妃'等10个品种。     

The reliability of leaf bioassays for predicting disease resistance on fruit: a case study on grapevine resistance to downy and powdery mildew

This study was designed to assess the reliability of grapevine leaf bioassays for predicting disease resistance on fruit in the field. The efficacy of various grapevine quantitative trait loci (QTLs) for conferring resistance to downy and powdery mildew was evaluated in bioassays and in a 2‐year field experiment for downy mildew. The resistance genes studied were inherited from Muscadinia rotundifolia (Rpv1 and Run1) and from American Vitis species through cv. Regent (QTLRgP and QTLRgD). In bioassays, genotypes carrying Run1 blocked powdery mildew development at early stages. Genotypes combining Run1 with QTLRgP displayed no greater level of resistance. For downy mildew, genotypes carrying Rpv1 and/or QTLRgD were more resistant than the susceptible cv. Merlot, and showed a high level of leaf resistance in the field (<10% severity). Disease levels on bunches were much higher than those on leaves, with a high variability between Rpv1 genotypes (1–48%). A Bayesian decision theory framework predicted that an OIV‐452 threshold of 5 in leaf bioassays allowed accurate selection of grapevine genotypes (P = 0·83) with satisfactory disease severity on bunches. Therefore, this study validates that the use of early bioassays on leaves, as currently performed by grapevine breeders, ensures a satisfactory level of resistance to downy mildew of bunches in the field.     

Response of sorghum accessions from Chad and Uganda to natural infection by the downy mildew pathogen,Peronosclerospora sorghi in Mexico and the USA

Journal of Plant Diseases and Protection - In this study, 78 accessions from Chad, Central Africa and 20 photoperiod insensitive accessions from Uganda, East Africa were evaluated for downy mildew...     

Development of a PCR test to detect the downy mildew causal agent Plasmopara halstedii in sunflower seeds

Plasmopara halstedii , the causal agent of downy mildew of sunflower, is an obligate parasite but viable sporangia and oospores of the pathogen may be found in a quiescent state in seeds of sunflower and therefore may be transported with sunflower seeds in international commercial exchanges. In order to prevent the spread of this pathogen, especially the introduction of potentially new races, an efficient method to analyse sunflower seed samples is required. In this study, a P. halstedii -specific polymerase chain reaction (PCR) test was developed based on the ribosomal large sub unit (LSU) DNA. The forward (PHAL-F) and reverse (PHAL-R) PCR primers were designed from two polymorphic regions of LSU. After screening 22 isolates of P. halstedii corresponding to different races and countries and 32 other oomycete, deuteromycete and ascomycete isolates, the PHAL-F/R primers amplified only a single PCR band of c. 310 bp from P. halstedii . The PHAL-F/R PCR test could detect as little as 3 pg of P. halstedii genomic DNA per 20  µ L reaction volume and enabled the direct detection of P. halstedii in 35 g sunflower seed samples without the need for any prior biological baiting step. An internal amplification control (IAC) was developed to help discriminate against false negative samples due to the potential presence of inhibitory compounds in DNA extracts. The test was successfully used on samples of naturally contaminated seeds. These new molecular tools should be of great interest for quarantine seed testing purposes.     

Risks to the aquatic ecosystem from the application of Metarhizium anisopliae for locust control in Australia

Laboratory tests of Metarhizium anisopliae var acridum Driver & Milner, at a dose of 1.3 x 10(6) conidia ml-1, had no adverse effects on nymphs of mayfly, Ulmerophlebia sp or 8-week-old fry of the rainbow fish, Melanotaenia duboulayi Castelnau. This dose was toxic to the cladoceran, Ceriodaphnia dubia Richard, causing 100% mortality in 48 h. When this test was repeated at doses of up to 6.7 x 10(3) conidia ml-1, there was only 5% mortality after 192 h. Spraying of artificial water sources with a very high dose of the fungus as an aqueous spray resulted in 80-130 conidia ml-1 at 15 cm depth in the first 24 h after spraying. The conidia rapidly settled out and were absent from the top 15 cm layer of water after about 50 h. A similar experiment using the oil formulation as used in field control resulted in a 2- to 20-fold lower level of conidia in the water. Finally, sampling actual water sources in spray areas revealed a very low level of contamination of the water, with a maximum mean level of 29 conidia ml-1 in the first 24 h after treatment. Thus the level of conidia likely to enter water during control campaigns is a small fraction of that required to kill cladocerans, the only sensitive non-target organism tested. It is concluded that the biopesticide is very unlikely to pose any hazard to aquatic organisms.     

Evaluation of the Francome MkII exhaust nozzle sprayer to apply oil-based formulations of Metarhizium flavoviride for locust control

Conidia of the fungus Metarhizium flavoviride were formulated in a paraffinic oil, ‘Shellsol’ T, and sprayed using the Francome MkII exhaust nozzle sprayer. Germination of the conidia collected from the spray was reduced by 30% as compared to unsprayed conidia. However, in bioassays, there was no detectable difference in virulence with conidia collected from the spray samples and unsprayed formulation. This indicated that, despite the recorded reduction in the concentration of active conidia, the efficacy of the formulation remained unchanged after passing through the exhaust nozzle sprayer. The droplet size spectra produced by the sprayer were investigated using the Malvern series 2600cc particle size analyser. The optimum droplets for locust control produced by this sprayer were generated by the number 1 nozzle (internal diameter 2·5 mm) with the number 1 restrictor ring (internal diameter 12.5 mm) sprayed at a pressure of 0·2 bar. The droplets thus produced had a volume median diameter of 58 μm when the nozzle protruded between 1 and 2 mm above the level of the restrictor ring. Of the droplets in the spray plume created by these conditions, 33% were between 50 and 100 μm, a range recommended as an achievable optimum for the ultra-low-volume application of Metarhizium flavoviride. The role of the exhaust nozzle sprayer as a tool for the application of M. flavoviride for locust control is discussed with reference to other vehicle-mounted ultra-low-volume sprayers. © 1997 SCI     

Characterization of a resistance locus (Pfs-1) to the spinach downy mildew pathogen (Peronospora farinosa f. sp. spinaciae) and development of a molecular marker linked to Pfs-1

Downy mildew is a destructive disease of spinach worldwide. There have been 10 races described since 1824, six of which have been identified in the past 10 years. Race identification is based on qualitative disease reactions on a set of diverse host differentials which include open-pollinated cultivars, contemporary hybrid cultivars, and older hybrid cultivars that are no longer produced. The development of a set of near-isogenic open-pollinated spinach lines (NILs), having different resistance loci in a susceptible and otherwise common genetic background, would facilitate identification of races of the downy mildew pathogen, provide a tool to better understand the genetics of resistance, and expedite the development of molecular markers linked to these disease resistance loci. To achieve this objective, the spinach cv. Viroflay, susceptible to race 6 of Peronospora farinosa f. sp. spinaciae, was used as the recurrent susceptible parent in crosses with the hybrid spinach cv. Lion, resistant to race 6. Resistant F(1) progeny were subsequently backcrossed to Viroflay four times with selection for race 6 resistance each time. Analysis of the segregation data showed that resistance was controlled by a single dominant gene, and the resistance locus was designated Pfs-1. By bulk segregant analysis, an amplified fragment length polymorphism (AFLP) marker (E-ACT/M-CTG) linked to Pfs-1 was identified and used to develop a co-dominant Sequence characterized amplified region (SCAR) marker. This SCAR marker, designated Dm-1, was closely linked ( approximately 1.7 cM) to the Pfs-1 locus and could discriminate among spinach genotypes that were homozygous resistant (Pfs-1Pfs-1), heterozygous resistant (Pfs-1pfs-1), or homozygous susceptible (pfs-1pfs-1) to race 6 within the original mapping population. Evaluation of a wide range of commercial spinach lines outside of the mapping population indicated that Dm-1 could effectively identify Pfs-1 resistant genotypes; the Dm-1 marker correctly predicted the disease resistance phenotype in 120 out of 123 lines tested. In addition, the NIL containing the Pfs-1 locus (Pfs-1Pfs-1) was resistant to multiple races of the downy mildew pathogen indicating Pfs-1 locus may contain a cluster of resistance genes.     

The response of oilseed rape (Brassica napus ssp. oleifera) accessions with different glucosinolate and erucic acid contents to four isolates of Peronospora parasitica (downy mildew) and the identification of new sources of resistance

A total of 101 Brassica napus ssp. oleifera accessions with seed differing in glucosinolate and erucic acid contents were screened for resistance to four isolates of Peronospora parasitica at the cotyledon stage. Two groups of accessions with different resistance factors were identified. Lines that were homogeneous for resistance were selected from seedling populations of accessions that exhibited a heterogeneous reaction to some isolates. The resistance of one group differs from that of cv, Cresor, the only oilseed rape cultivar reported to have an isolate-specific gene for resistance to P. parasitica. The isolate specificity of the second group was identical to that of cv, Cresor, A comparison of the response of host accessions which expressed moderate to full susceptibility at the cotyledon stage, with no clear differential response to any of the four P. parasitica isolates, indicated that those with high glucosinolate and high erucic acid contents (12 accessions) were slightly but significantly less susceptible than those with high glucosinolate and low erucic acid (19 accessions), or low glucosinolate and low erucic acid contents (28 accessions). The mean differences between accessions with low erucic acid but differing in glucosinolate content were inconsistent. The last result was further confirmed by investigating the expression of resistance to three isolates of P. parasitica at three different seedling growth stages among 11 accessions of oilseed rape with seeds low in erucic acid but differing in glucosinolate content.     

Use of natural pyrethrum to control the red swamp crayfish Procambarus clarkii in a rural district of Italy

BACKGROUND: The crayfish Procambarus clarkii inflicts severe ecological and economic damages in Europe. To develop an efficient method for its control, four experiments were carried out to assess the impact of natural pyrethrum (i.e. Pyblast) on crayfish: (1) the 24 h LC₁₀₀ and LC₅₀ were quantified on crayfish; (2) the breakdown time of the 24 h LC₁₀₀ was assessed using Daphnia magna as a bioindicator; the effects of 24 h LC₁₀₀ on crayfish were investigated by applying the biocide into burrows (3) and in a drainage channel (4). RESULTS: Pyblast concentrations of 0.05 and 0.02 mg L^?1 corresponded to 24 h LC₁₀₀ and LC₅₀ respectively. The concentration of 0.05 mg L^?1 broke down after 72 h, whereas 0.02 mg L^?1 did not cause any significant mortality in D. magna after 24 h. However, 0.05 mg L^?1 had no effect on crayfish when introduced into the burrows, but led to a mortality of 95% when applied in the water. CONCLUSION: Experimental evidence is provided for the efficacy of Pyblast to control invasive crayfish. Obviously, before its use on a large scale, further studies are needed to find a concentration that will achieve the target 100% mortality with the shortest recovery time of the environment. Copyright © 2012 Society of Chemical Industry