European and Mediterranean Plant Protection Organization Organisation Europenne et Mditerranenne pour la Protection des Plantes PP 1/3(4)

Efficacy evaluation of fungicides

Pseudoperonospora humuli

Specific scope This standard describes the conduct of trials for the efficacy evaluation of fungicides against Pseudoperonospora humuli, causing downy mildew of hop.

Specific approval and amendment First approved in 1977-09. evision approved in 1990-09. !ligned with revised standard te"t in 199#. evision approved in $000-09.

%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% 1. Experimental conditions 1.1 Test organisms, selection of crop and cultivar Test organism& Pseudoperonospora humuli '()(*+,-. ! susceptible cultivar of hop Humulus lupulus '+,./,- should preferably be used. The disease manifests itself in two forms, as primary infections or as secondary infections. 0t is normally necessary to carry out separate trials to evaluate products against these two forms of the disease. (rimary infections arise in spring from the systemically infected rootstoc1 and give rise to infected basal shoots with short internodes, which are referred to as primary spi1es. )econdary infections result from spores produced on the primary infections which spread to the new growth and give rise to secondary basal spi1es, stunting of the tips of the climbing shoots, angular leaf spots, infected inflorescences 'burr- and brown lesions on the cones. 1.2 Trial conditions The trial should be set up in the field, preferably in an area where the inoculum of the disease is high and uniformly distributed. 0f a product is to be tested for controlling secondary infections, the diseased basal spi1es should be removed prior to the first application if they are not evenly distributed in the trial area. +owever, in trial sites where there is a relatively uniform distribution of primary infections, it may be very useful to leave them undisturbed to provide reliable sources of inoculum. 2ultural conditions 'e.g. soil type, fertili3ation, tillage- should be uniform for all plots of the trial and should conform with local agricultural practice. The trial should form part of a trial series carried out in different regions with distinct environmental conditions and preferably in different years or growing seasons 'see *((4 )tandard (( 15161 2onduct and reporting of efficacy evaluation trials-. 9 1.3 Design and lay-out of the trial Treatments& test product's-, reference product and untreated control, arranged in a suitable statistical design. (lot si3e 'net-& for trials against primary infections, at least 10 plants. For smaller plants with fewer bines 1 'e.g. three bines per plant- or where the distribution of infection is not uniform, up to 70 plants may be needed. For trials against secondary infections, the gross plot si3e should consist of si" rows, so as to avoid border effects. The two central rows should be used for evaluation purposes and constitute the net plot. eplicates& normally at least 8, but e"ceptionally 9 if the net plot si3e is sufficiently large 'e.g. 70 plants-. For further information on trial design, see *((4 )tandard (( 1517$ :esign and analysis of efficacy evaluation trials. 2. Application of treatments 2.1 Test product(s) The product's- under investigation should be the named formulated product's- 'see *((4 )tandard (( 15161 2onduct and reporting of efficacy evaluation trials-. 2.2 Reference product The reference product should be a product 1nown to be satisfactory in practice under the agricultural, plant health and environmental 'including climaticconditions in the area of intended use. 0n general, type of action, time of application and method of application should be as close as possible to those of the test product.
1

The word ;bine; is used to mean the group of stems that have been trained to grow up one of the strings used to support the hop plant.

2.3 Mode of application !pplications should comply with good standard practice. $.9.1 Type of application The type of application 'e.g. a spray for secondary infections, or a drench or granules for primary infections- should be as specified for the intended use. $.9.$ Type of equipment !pplication's- should be made with e<uipment which provides an even distribution of product on the whole plot or accurate directional application where appropriate, e<uivalent to a good commercial practice. Factors which may affect efficacy 'such as operating pressure, no33le type, depth of incorporation- should be chosen in relation to the intended use. $.9.9 Time and frequency of application The number of applications and the date of each application should be as specified for the intended use. $.9.8 Doses and volumes The product should normally be applied at the dosage specified for the intended use. :oses higher or lower than the intended dose may be tested to determine the margin of effectiveness and crop safety. The dosage applied should normally be e"pressed in 1g or / of formulated product per ha. 0t may also be useful to record the dose in g of active substance per ha 'or per plant-. For sprays, data on concentration 'as = or g product per 100 /- and volume '/ ha1- should also be given. :eviations from the intended dosage should be noted. $.9.7 Data on other plant protection products 0f other plant protection products 'or any biocontrol agents- have to be used, they should be applied uniformly to all plots, separately from the test product and reference product. (ossible interference with these should be 1ept to a minimum. 3. Mode of assessment, measurements recording and

4n the date of application, meteorological data should be recorded which is li1ely to affect the <uality and persistence of the treatment. This normally includes at least precipitation 'type and amount in mm- and temperature 'average, ma"imum, minimum in >2-. !ny significant change in weather should be noted, and in particular its time relative to the time of application. Throughout the trial period, e"treme weather conditions, such as severe or prolonged drought, heavy rain, late frosts, hail, etc., which are li1ely to influence the results, should also be reported. !ll data concerning irrigation should be recorded as appropriate. 9.1.$ Edaphic data For soil-applied products especially, the following characteristics of the soil should be recorded& p+, organic matter content, soil type 'according to a specified national or international standard-, moisture 'e.g. dry, wet, waterlogged- and fertili3er regime. 3.2 Type, time and frequency of assessment The ??2+ growth stage of the crop at each date of application and assessment should be recorded. 9.$.1 Type (rimary infections& the percentage of infected shoots should be determined on at least 10 randomly selected and mar1ed plants from the central part of the plot. ,p to 70 plants per plot may be necessary if the distribution of infection is not sufficiently uniform. :iseased shoots should be removed after each assessment. )econdary infections& the percentage of infected leaves, flowers and cones should be determined on at least 10 randomly chosen and mar1ed bines from the central part of the plot at regular intervals during the growing period. 4nly the percentage of infested leaves, flowers and cones should be evaluated, not the severity of attac1 on the individual plant parts. 0n addition, the percentage of infected cones, or the degree of damage on the cones, should be determined at harvest. The percentage of plant parts attac1ed can be calculated directly by counting the number of plant parts evaluated and the number with symptoms. !lternatively, an estimate can be made using a scale of 0-100 or the scale in !ppendi" 0. 0t may be convenient to ma1e separate estimates for the level of attac1 on the lower, middle and the upper thirds of the plant. 9.$.$ Time and frequency (rimary infections& preliminary assessment @ust before applicationA 1st assessment 7 days after applicationA $nd assessment 18 days after applicationA 9rd assessment $1 days after application. Further assessments may be useful. )econdary infections& preliminary assessment @ust before applicationA further assessments before each subse<uent applicationA last assessment at harvest. 10

3.1 Meteorological and edaphic data 9.1.1 Meteorological data 4n the days before and after application, meteorological data should be recorded which is li1ely to affect the development of the crop and5or pest and the action of the plant protection product. This normally includes data on precipitation and temperature. !ll data should preferably be recorded on the trial site, but may be obtained from a nearby meteorological station.

!ssessment on cones at harvest& in each plot, at least 10 randomly selected and mar1ed bines 'see 1.1should be harvested. The percentage of infected cones or the degree of damage on 700 cones from each plot should be assessed using the scale in !ppendi" 00. 3.3 Direct effects on the crop The crop should be e"amined for the presence of phytoto"ic effects. 0n addition, any positive effects should be noted. The type and e"tent of such effects on the crop should be recorded and, if there are no effects, this fact should also be recorded. (hytoto"icity should be scored as follows& '1- if the effect can be counted or measured, it should be e"pressed in absolute figures A '$- in other cases, the fre<uency and intensity of damage should be estimated. This may be done in either of two ways& each plot is scored for phytoto"icity by reference to a scale, or each treated plot is compared with an untreated plot and = phytoto"icity estimated. 0n all cases, symptoms of damage to the crop should be accurately described 'stunting, chlorosis, deformation, etc.-. For further details, see *((4 )tandard (( 15197 (hytoto"icity !ssessment which contains sections on individual crops. 0t may be useful to record alpha acid content. 3.4 Effects on non-target organisms 9.8.1 Effects on other pests !ny observed effects, positive or negative, on the incidence of other pests should be recorded. 9.8.$ Effects on other non-target organisms !ny observed effects, positive or negative, on naturally occurring or introduced pollinators or natural enemies should be recorded. !ny observed effect, positive or negative, on ad@acent or succeeding crops should be recorded. !ny environmental effects should also be recorded, especially effects on wildlife. 3 .5 uantitative and qualitative recording of yield

Appendix ! scale such as the following may be used and should be described& 1 C no visible symptomsA $ C few, scattered spots on the leavesA 9 C D7 = of the leaves have spotsA 8 C 7-17 = of the leaves have spotsA 7 C 17-$7 = of the leaves have spotsA # C $7-70 = of the leaves, flowers and cones have large spotsA 7 C leaves, flowers and cones are partly deadA 6 C leaves, flowers and cones are mostly dead. Appendix ! scale such as the following may be used for the assessment on cones& 1 C without damageA $ C light damageA 9 C medium damageA 8 C severe damage.

Buantitative recording of yield is not re<uired, but any obvious deviations from normal <uality should be recorded. 4. Results The results should be reported in a systematic form and the report should include an analysis and evaluation. 4riginal 'raw- data should be available. )tatistical analysis should normally be used, by appropriate methods which should be indicated. 0f statistical analysis is not used, this should be @ustified. )ee *((4 )tandard (( 1517$ :esign and analysis of efficacy evaluation trials. 11