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Efficacy of fungicides to control Early
Blight genotypes
Field experiment AGV7205
A. Evenhuis & H.T.A.M. Schepers
This study was carried out by Wageningen University & Research and was commissioned and financed by UPL
Europe Ltd
Wageningen Plant Research is the collaboration of Wageningen University and the foundation Stichting
Wageningen Research.
Wageningen, November 2016
CONFIDENTIAL
Report 3750331500
A. Evenhuis & H.T.A.M. Schepers, 2016. Efficacy of fungicides to control Early Blight genotypes.
Wageningen Plant Research, Confidential Report 3750331500.
© 2016 Wageningen, Stichting Wageningen Research, Wageningen Plant Research, P.O. Box 16,
6700 AA Wageningen, The Netherlands; T +31 (0)317 48 07 00; www.wur.nl/plant-research
Chamber of Commerce no. 09098104 te Arnhem
VAT NL no. 8065.11.618.B01
Stichting Wageningen Research. All rights reserved. No part of this publication may be reproduced,
stored in an automated database, or transmitted, in any form or by any means, whether
electronically, mechanically, through photocopying, recording or otherwise, without the prior written
consent of the DLO Foundation.
Wageningen Research is not liable for any adverse consequences resulting from the use of data from
this publication.
Sjef van der Heijden
UPL Europe Ltd
The Centre
Birchwood Park
Warrington, WA3 6YN UK
Confidential Wageningen Plant Research Report 3750331500
Contents
Contents 3
1 Introduction 5
2 Methods and materials 6
2.1 Lay out 6
2.2 Treatments 6
2.3 Inoculation 6
2.4 Disease observations 7
2.5 Sampling and genotyping 7
2.6 Statistics 7
3 Results 8
3.1 Field assessments 8
3.3 Genotyping 10
4 Discussion and conclusions 11
4.1 Early Blight 11
4.2 Soil coverage 11
4.3 Genotyping 11
4.4 Conclusions 12
Experimental lay-out 13Annex 1
nVWA recognition 15Annex 2
Raw data disease severity (%) 16Annex 3
Raw data soil coverage (%) 17Annex 4
Raw data genotyping 18Annex 5
Weather data 19Annex 6
Confidential Wageningen Plant Research Report 3750331500 | 5
1 Introduction
Early Blight is caused by Alternaria solani. Although late blight, caused by Phytophthora infestans, is
the most important disease in potato production Early Blight is regularly found in potato fields in the
second half of the season. Farmers spray potatoes 2 to 4 times during a season to control Early Blight.
Specific fungicides most commonly used in the Netherlands are Signum, Amistar and recently also
Narita and Carial Star. Furthermore a number of fungicides used to control potato late blight show also
some efficacy to control Early Blight. Especially products based on mancozeb can be mentioned here.
Resistance of A. solani to azoxystrobin has been reported in the United States (Pasche and
Gudmestad, 2008). This resistance is associated with the F129L mutation. This mutation has also been
reported in Germany (Leiminger et al., 2014), and one isolate with the F129L mutation has been
found in the Netherlands (Evenhuis, et al., 2013). Until now no obvious lack of control of azoxystrobin
has been found in agricultural practise (Leiminger and Hausladen, 2013). However in a field
experiment conducted in Valthermond in the Netherlands in 2014 the efficacy of Amistar and Signum
to control Early Blight was less than expected. It is not known what caused this effect. Fungicide
resistance management of A. solani was not investigated so far in the Netherlands.
In this experiment the efficacy of fungicides to control a mixture of A. solani isolates in a field
experiment was tested by request of UPL.
6 | Confidential 3750331500
2 Methods and materials
2.1 Lay out
The experiment was set up in a randomised block design. Details are described in Appendix 1.
2.2 Treatments
Fungicide applications were carried out using a trial site sprayer with Airmix 110.04 nozzles. Nozzles
were hanging approximately 50 cm above the foliage. Sprayings were carried out based on 300 l/ha.
Potato plants were sprayed for the first time when they reached a height of 20-30 cm when the rapid
growth started. Specific sprays to control Early Blight started Mid-July. Haulm killing was carried out
on 30 September 2016, despite natural senescence of the crop. In Table 1 the fungicides used and
dose rates are presented.
Table 1 The fungicides used in the 2016 experiment and the applied dose rates.
Fungicide Active ingredient Dose rate
L or kg /ha
Narita difenoconazole 250 g/l 0.5
Signum pyraclostrobin 6.7% + boscalid 26.7% 0.2
Penncozeb 80 WP mancozeb 800 g /kg 2.0
Treatment A is the untreated control and H the reference treatment chosen. Spray strategies are
given in Table 2.
Table 2 Spray strategies and date of application.
Code Fungicide 12-7 19-7 26-7 2-8 9-8 16-8 23-8
A UTC - - - - - - -
B Signum 0.2 S - S - S - S
C Penncozeb 80 WP 2.0 P P P P P P P
D Penncozeb 80 WP 2.0 + Signum 0.2 P+S P P+S P P+S P P+S
E Narita 0.5 N - N - N - N
H Signum 0.2 or Narita 0.5 S - S - N - N
2.3 Inoculation
A selection of 2 A. solani isolates was made. AltNL03003 was isolated in 2003 and belongs to
genotype 1 and is wild type. AltNL15002 belongs to genotype 2 and possesses the F129L mutation.
Both isolates were grown on wheat kernels separately. A mixture of 95% wildtype and 5% F129L type
was inoculated. Infested kernels were broadcasted in the field on 15 July 2016.
Confidential 3750331500 | 7
2.4 Disease observations
Disease observations were carried out from mid-July. Depending on the disease development disease
observations were carried out with an interval of 1 to 2 weeks. The percentage necrotic foliage due to
A. solani per plot was estimated.
2.5 Sampling and genotyping
To assess Alternaria genotype and the presence of the F129L mutation leaves with Alternaria lesions
were collected. Per plot 8 leaflets with Alternaria lesions were sampled. Samples were taken four times
during the season in the last week of August and the first three weeks of September. Only the last
sample was genotyped. The samples were air dried and stored in Petri dishes until further processing.
At the laboratory monospore cultures were made on agar. Mycelium was scraped from the agar plate
after incubation. DNA was extracted from the mycelium. The genotype involved was assessed by
carrying out two PCR described by Pasche et al., 2004 and Lieminger et al., 2015. The PCR product
was extracted from the gel and sequenced. The nucleotide order was assessed and the presence of
the F129L mutation was established according to the publication of Lieminger et al., 2015.
2.6 Statistics
Analysis of variance on the parameters was made using GENSTAT 18th
Edition. The experiment was
carried out with four replications in a randomised block design. Each replication consisted of a plot.
Transformation of data was carried out when necessary.
8 | Confidential 3750331500
3 Results
3.1 Field assessments
The results are given in Table 3 and Table 4. On 22 July 2016 no Early Blight was found. A week later
on 29 July Early Blight was present in all plots at a low disease severity rate of 0.001% (data not
shown). From 5 August onwards the Early Blight epidemic started (Figure 1). After 17 September it
was not possible to assess Early Blight because it was too much entangled with natural senescence to
distinguish.
Phytotoxicity of the fungicides used was not observed. In this experiment the fungicides used were
crop safe.
Disease severity of all spray strategies was significantly lower than the untreated control. Based on
AUDPC, the efficacy of treatments D, E and H were significantly better than treatments B and C.
Table 3. Effect of fungicide spray treatments on Early Blight disease severity (%). Log 10 back
transformed Early Blight disease severity ratings.
Code 5-8 12-8 19-8 26-8 2-9 9-9 16-9 AUDPC
A 0.001 a 0.29 c 0.7 c 4.2 c 8.7 d 19.7 c 18.2 c 330 c
B 0.001 a 0.00 a 0.0 a 0.2 b 1.2 c 3.0 b 14.1 c 86 b
C 0.001 a 0.07 bc 0.0 ab 0.1 b 0.9 c 1.9 b 10.3 bc 68 b
D 0.001 a 0.03 abc 0.0 ab 0.0 a 0.0 a 0.1 a 5.1 b 21 a
E 0.001 a 0.02 abc 0.0 bc 0.1 b 0.2 b 0.2 a 1.2 a 12 a
H 0.001 a 0.01 ab 0.0 ab 0.1 b 0.1 b 0.2 a 1.5 a 10 a
1
) Values in columns followed by the same letter are not significantly different (P=0.05).
Figure 1 Early Blight disease severity (%)
Confidential 3750331500 | 9
On average soil coverage (%) of treatments B to H was significantly higher than the untreated control
(A). No further differences between average soil cover were found.
Table 4 Effect of spray treatments on soil coverage (%).
Code 2-9 9-9 16-9 2
23-9 2
Average
A 43 a1
21 a 3 a 1.0 a 17 a
B 73 b 53 b 17 b 2.1 ab 37 b
C 71 b 46 b 18 b 4.1 bc 36 b
D 74 b 54 b 21 b 5.7 c 39 b
E 61 b 43 ab 13 b 4.8 bc 31 b
H 70 b 53 b 15 b 6.2 c 37 b
1
) Values in columns followed by the same letter are not significantly different (P=0.05).
2
) Values on 16 & 23-9 are logit back transformed data.
10 | Confidential 3750331500
3.3 Genotyping
The results are presented in Table 5 and Figure 2. No significant shift of the Alternaria solani genotype
was found when Penncozeb was used compared to the untreated control. When Signum (B) or Signum
followed by Narita (H) was sprayed, significantly more GII F129L types were found compared to the
untreated control.
Table 5 Effect of spray treatments on frequency of Alternaria genotypes.
Code Strategy GI WT GI F129L GII WT GII F129L
A UTC 56 b 0 0 a 44 a
B Signum 7 a 0 3 b 90 b
C Penncozeb 52 b 0 0 a 48 a
H Signum / Narita 13 a 0 0 a 86 b
1
) Values in columns followed by the same letter are not significantly different (P=0.05).
Figure 2 Alternaria solani genotype distribution (%)
Confidential 3750331500 | 11
4 Discussion and conclusions
A field experiment was carried out in cultivar Agria. The Early Blight epidemic developed late despite
artificial inoculation. The weather in June was conducive for potato late blight and not so much for
Early Blight. Potato late blight was controlled with cover sprays using Revus 0.6 l/ha, Infinito 1.6 l/ha
and Ranman Top 0.5 l/ha. These fungicides have no known efficacy to control Early Blight.
Besides Alternaria spp. also Sclerotinia was found in the experiment. Sclerotinia severity was not
assessed. Nevertheless some treatments probably controlled Sclerotinia which together with Alternaria
and senescence account for soil coverage.
4.1 Early Blight
Early Blight disease severity in the untreated control was significantly higher than all other treatments
indicating that the inoculation was successful. Furthermore all spray strategies effectively controlled
Early Blight, although the efficacy varied between strategies.
Disease severity of treatment B and C increased more towards the end of the season compared to
treatments D, E and H. The last spray application was 23 August and the increase was observed from
9 September onwards. Interestingly when Penncozeb was added to Signum (or v.v.) the efficacy to
control Early Blight increased compared to the products used solo. However the efficacy to control
Early Blight was still less than using Narita or the combination Signum followed by Narita.
4.2 Soil coverage
Average soil coverage of all spray strategies was comparable and significantly higher than the
untreated control. On individual assessment data some minor differences between treatments
occurred.
4.3 Genotyping
The field was inoculated with wheat kernels with 2 Alternaria genotypes. Genotype I (GI) is wild type
and Genotype II (GII) possesses the F129L mutation. Predominantly these two genotypes were found
in the field. Only 1 isolate found was Genotype II, wild type. No Genotype I isolates were found
possessing the F129L mutation. In the untreated control 57% of the isolates were Genotype I and wild
type. The experiment was inoculated with 95% Genotype I wildtype. This could suggest that the
F129L mutation was already present in the natural A. solani population at the location of the field
experiment. Alternatively GII might be more aggressive than GI and therefore was found back more
frequently.
Leaves were picked at four times during the season. Alternaria solani was isolated only from the
leaves picked at the last sampling date. Leaves of the other sampling dates were stored under dry
conditions at room temperature and could be used for additional assessments.
When Signum or Signum followed by Narita was sprayed, significantly more GII F129L A. solani types
were found than in the untreated control and when Penncozeb was sprayed. This suggests that
selection towards GII F129L occurred under influence of spray strategies B and H. It is known that the
F129L mutation causes a reduced sensitivity to QoI fungicides. One of the active ingredients of
Signum is pyraclostrobin, a QoI fungicide.
12 | Confidential 3750331500
4.4 Conclusions
Phytotoxicity of the fungicides applied was not observed in this experiment. The used products are
crop safe.
Alternaria disease severity based on AUDPC was significantly higher in the untreated than all other
treatments.
Based on AUDPC, the efficacy of treatments D, E and H were significantly better than treatments
B and C.
On average soil coverage (%) of treatments B to H was significantly higher than the untreated
control.
No significant shift of the Alternaria solani genotype was found when Penncozeb was used
compared to the untreated control.
When Signum or Signum followed by Narita was sprayed, significantly more GII F129L types were
found compared to the untreated control.
Confidential 3750331500 | 13
Experimental lay-outAnnex 1
Site Lelystad, The Netherlands
Soil texture Sandy loam
Previous crop onion
Pre previous crop
Fertilization 500 kg KAS on 17 May and 300 kg KAS on 22 June
Variety: Agria
Planting date: 3 May 2016
Seed Rate: 2.500 kg/ha
Herbicide treatment: 24 May
Fungicide application: see paragraph 2 and normal practice
Alternaria treatments: see paragraph 2 and normal practice
Crop desiccation: Reglone (diquat dibromide 200g/l) 2.0 l/ha 30 September 2016.
Harvest: -
Tuber assessments: -
Gross plot dimensions: Four rows (0.75 m) of 11 m length
Net plot dimensions: Two rows (0.75) of 11 m length
Demo design: Four replications in a randomized block design
Lelystad
14 | Confidential 3750331500
Project:
Experiment: AGV7205 Alternaria genotypes
Location: Lelystad
Field: G86-4
8 16 24 32
C F G D
7 15 23 31
D A B G
6 14 22 30
H B E F
5 13 21 29
A H C B
4 12 20 28
E G D C
3 11 19 27
B C H E
2 10 18 26
G D F A
1 9 17 25
F E A H
1.5 < 3m > < 3m > < 3m > < 3m > < 3m > < 3m > 1.5
3750331500
N
I II III IV
Confidential 3750331500 | 15
nVWA recognitionAnnex 2
16 | Confidential 3750331500
Raw data disease severity (%)Annex 3
Confidential 3750331500 | 17
Raw data soil coverage (%)Annex 4
18 | Confidential 3750331500
Raw data genotypingAnnex 5
Confidential 3750331500 | 19
Weather dataAnnex 6
date T-avg T-max T-min rain radiation RV-min Wind
direction
Wind
speed
01-04-16 6.8 12.4 1.1 0 5.835 54 SSE 1.6
02-04-16 8.8 12.7 4.3 0 2.811 59 SSE 1.7
03-04-16 13.1 18.1 10.3 0.6 3.783 59 NE 2.7
04-04-16 12.3 14.6 8.9 6.8 3.358 61 E 2.8
05-04-16 9.9 12.1 8.5 0.6 2.26 76 SW 2.2
06-04-16 7.9 10.2 6.1 7.6 1.968 73 SW 5
07-04-16 7.2 9.2 5.3 0.4 3.779 71 SW 4.6
08-04-16 7.9 10.4 5.4 0.6 3.085 71 SE 2.1
09-04-16 8.7 14.5 3.2 0.2 4.43 46 ESE 2.7
10-04-16 9.5 13.6 6.7 1.2 5.779 64 NE 2.2
11-04-16 11.4 16.7 6.8 0 6.592 49 E 4
12-04-16 11.4 14.6 8 5.2 4.977 61 SW 2.7
13-04-16 9.7 14 5.1 2.2 4.047 66 SSW 1.6
14-04-16 9.7 14 5.6 0 3.791 62 E 1.3
15-04-16 9.6 11.9 8.4 4.4 2.401 80 SSE 2.3
16-04-16 8.6 10.6 6.1 5.8 4.979 70 NW 5.4
17-04-16 6.9 9.7 4.4 0.8 4.439 66 W 3.8
18-04-16 8 12 3.8 0 3.889 55 WSW 4.5
19-04-16 9.4 11 7.5 4 7.078 62 SW 15.3
20-04-16 7.8 10.8 4.9 0 6.904 60 NE 13.2
21-04-16 8.3 14.4 2.5 0 7.574 44 NNE 2.4
22-04-16 7.8 10.9 5.2 0 5.639 57 NNE 3.4
23-04-16 6.2 8.4 4.4 0.2 6.002 54 WNW 4.7
24-04-16 5 7.7 2.8 4.6 5.73 59 SE 5.8
25-04-16 4.9 7.6 2.5 4 2.788 68 WNW 4.2
26-04-16 4.3 7 2 8.4 3.36 77 NW 6.1
27-04-16 5.8 8.2 3.7 0.4 5.737 73 NNW 4.9
28-04-16 5.1 8.2 2.7 5.6 4.277 61 SSE 2
29-04-16 6.8 10.2 2.9 5 4.237 52 SSW 4.7
30-04-16 7.2 9.4 4.7 0.4 5.424 69 NNE 3.2
20 | Confidential 3750331500
date T-avg T-max T-min rain radiation RV-min Wind
direction
Wind
speed
01-05-16 7.5 11.2 3.3 0 8.126 63 NNW 2.5
02-05-16 10.5 17.1 3.2 0 6.75 36 SSW 2.5
03-05-16 10.3 12.3 7.6 1.2 7.365 57 W 4.1
04-05-16 10.1 15.1 3.5 0 7.354 42 ESE 1.4
05-05-16 12.8 18.4 5.8 0 8.425 30 ENE 2.3
06-05-16 16.6 23.9 9.3 0 8.115 33 ESE 2.3
07-05-16 19.5 26 12.1 0 8.186 28 ENE 2.1
08-05-16 19.6 24.8 12.6 0 8.428 25 ENE 3.6
09-05-16 19.5 24.6 13.7 0 8.075 36 ENE 3.6
10-05-16 18.8 22.5 14.5 0 3.86 44 NNE 2.1
11-05-16 19.8 25 15.2 0 7.243 35 ENE 1.9
12-05-16 19.4 24.8 13.7 0 8.174 34 NE 3.3
13-05-16 14.6 19.9 9.3 0 8.138 44 N 4.5
14-05-16 8.6 10.9 7.3 3 5.462 58 NW 6.7
15-05-16 8.4 10.7 6.6 7 5.947 61 WNW 6
16-05-16 9.3 11.6 7.1 0.4 5.334 64 WNW 3.9
17-05-16 12 16.5 8.7 0 6.039 54 S 2
18-05-16 13.1 17.5 7.3 0 4.184 52 E 2.3
19-05-16 14 18.4 10.3 0 5.038 62 S 1.6
20-05-16 13.1 16.5 10.6 1 2.534 69 WSW 2.8
21-05-16 16.8 22.6 9.7 0 5.506 50 SSE 2.7
22-05-16 15.9 18.7 13.2 8.4 2.679 70 NNE 1.8
23-05-16 12.1 13.4 11.4 17.6 2.329 86 NNW 6.2
24-05-16 10.7 12.1 10 0 4.308 76 NW 5.3
25-05-16 11.9 14.3 10.1 0.6 2.154 81 NW 0.7
26-05-16 13.7 17.2 10.5 0.4 6.127 58 NE 1.7
27-05-16 14.8 19.4 10.3 0 7.252 56 NNE 2.2
28-05-16 16.6 21 12.1 1.6 7.684 66 NNE 2.3
29-05-16 17 21.1 13.7 0.6 3.996 52 NNE 3.1
30-05-16 16.7 20 14.5 3.8 3.089 80 N 5.4
31-05-16 18.9 22.7 15.3 0 6.431 50 ESE 2.1
Confidential 3750331500 | 21
date T-avg T-max T-min rain radiation RV-min Wind
direction
Wind
speed
01-06-16 19.3 23.6 15.8 0 5.727 66 ENE 4.4
02-06-16 15.1 17.4 13.1 0 1.964 82 NNE 5.8
03-06-16 16.6 20.2 13.1 1.4 4.359 74 NW 2.3
04-06-16 20.9 25.6 15.9 0 7.601 55 NNE 2.3
05-06-16 18.1 20.9 13.6 0 9.082 70 NNE 3.7
06-06-16 16.4 21.7 11.2 0 9.128 65 NNE 2.8
07-06-16 18 22.6 12.7 0 7.692 58 N 3
08-06-16 14.3 16.3 12.1 0 6.105 72 N 3.2
09-06-16 14 16.1 12.1 0 9.187 63 N 4
10-06-16 14.3 17.7 10.8 0 7.744 37 NE 1.9
11-06-16 15.5 19.6 11.9 0 4.014 59 NNE 1.4
12-06-16 16.4 19.9 13.7 3.6 3.835 63 SSE 0.7
13-06-16 15.6 18.1 14.1 11.8 3.804 81 W 2
14-06-16 15 17.6 13.5 13 3.958 76 S 2.3
15-06-16 14.5 16.7 11.8 0.4 4.361 76 SW 1.7
16-06-16 15.5 18.9 11.7 0.4 7.239 58 WSW 1.5
17-06-16 15.3 17.3 13.4 10 5.734 75 NW 2.4
18-06-16 14.4 16.1 12.6 2.6 4.71 74 NW 3.3
19-06-16 14.4 17.2 12.1 0 7.379 62 SSE 3.3
20-06-16 14.8 16.1 14 20.8 1.817 79 W 4
21-06-16 16.8 19.4 13.7 6.2 4.194 78 WSW 2
22-06-16 18.6 23.4 13.6 0.2 7.33 67 ESE 2
23-06-16 20.6 25.9 18.1 12.4 5.444 75 SSE 1.7
24-06-16 19.1 21 15.6 0.2 5.317 75 N 2.4
25-06-16 15.2 17.7 12.5 5.6 3.704 80 WSW 0.9
26-06-16 14.7 17.6 11.6 0.2 5.265 68 SW 2.8
27-06-16 14.5 17.5 11.8 3.2 3.979 68 WNW 3
28-06-16 15.5 20.1 10.6 1.6 5.466 58 W 1.4
29-06-16 15.3 18.4 13.2 4.2 5.649 61 S 3.4
30-06-16 15.3 17.8 13.5 4.8 3.361 84 SSW 3.1
22 | Confidential 3750331500
date T-avg T-max T-min rain radiation RV-min Wind
direction
Wind
speed
01-07-16 16 17.5 14.3 3.2 1.821 84 SSW 3.7
02-07-16 15 16.6 12.9 1 7.48 59 SW 4.1
03-07-16 14.4 18.1 10.8 7.4 6.643 64 WNW 2.9
04-07-16 16.4 20.9 10.8 0 7.924 57 N 1.4
05-07-16 15 16.8 13.5 1 3.498 77 WNW 4
06-07-16 14.7 17.5 12.1 0.2 7.523 63 W 3.6
07-07-16 16 21.1 10.2 0 6.983 54 SSE 2.1
08-07-16 16 19 13.6 3.4 3.06 77 W 2.4
09-07-16 17.8 20.8 13.1 0.6 4.746 65 SSW 2.2
10-07-16 20.1 25 16 0 7.294 58 SSW 3.4
11-07-16 17.9 20.4 15.9 0 5.099 61 SW 4.9
12-07-16 16.3 18.8 14.5 17.6 4.893 75 SE 2.2
13-07-16 14.2 16.3 11.6 3.2 3.356 66 WNW 1.8
14-07-16 14.4 17 12.2 0 5.183 63 NW 4
15-07-16 15.8 19.5 12.7 0 7.468 60 W 3.2
16-07-16 17.5 19.1 13.8 0 3.029 83 S 2.3
17-07-16 18.6 21.1 16.4 4.2 8.149 65 NW 2.7
18-07-16 18.8 23.2 13.8 0 8.715 49 N 1.4
19-07-16 20.9 27.9 12.2 0 8.624 34 ENE 0.8
20-07-16 24.1 32.2 17.7 0 7.266 36 SW 2
21-07-16 21.1 25.1 16.9 0 7.465 65 NW 1.7
22-07-16 19.8 24 16.7 4.6 5.819 66 NNE 1.1
23-07-16 19 22.1 16.4 0 5.506 78 NNE 2
24-07-16 19.9 23.7 16.1 0 5.905 70 W 0.9
25-07-16 18.1 20.7 15.1 1.2 3.396 68 WNW 1.7
26-07-16 17.8 22.7 12.6 0 5.911 52 ENE 1.4
27-07-16 17.8 21 16.2 8.4 3.933 64 SSW 2
28-07-16 17.4 20.4 14.8 25.8 4.129 79 SSE 1.6
29-07-16 17.7 20.2 14.8 1.4 3.991 76 SW 2.8
30-07-16 17.5 20.4 15.3 9.4 4.493 62 SSE 1.8
31-07-16 16.2 19.1 13.7 0.2 5.557 66 WSW 2.3
Confidential 3750331500 | 23
date T-avg T-max T-min rain radiation RV-min Wind
direction
Wind
speed
01-08-16 16 19.5 11.6 0.2 6.365 56 WSW 1.9
02-08-16 15.5 17.5 13.2 1.2 2.62 81 SSW 1.7
03-08-16 18.5 20.5 16.8 5.2 3.068 74 SSW 3.8
04-08-16 17.2 20.1 14.7 0.2 4.029 64 SSW 3.9
05-08-16 16.9 20.1 13.5 0 5.576 65 W 2.6
06-08-16 16.3 20.3 13 0.4 5.93 60 WSW 2
07-08-16 16.7 21.6 11.6 0 3.396 75 SW 3
08-08-16 17.3 18.8 14.8 0 5.395 60 WNW 4.2
09-08-16 13.9 16.6 12.3 9.6 5.116 58 ESE 2.6
10-08-16 12.7 15.4 9.5 3.2 7.126 61 W 2.6
11-08-16 12.6 15.3 9.2 17 1.97 81 W 2.7
12-08-16 17.7 20.1 15.7 1.6 3.447 76 SSW 3.2
13-08-16 16.9 19.3 14.8 0 3.38 75 WSW 2.6
14-08-16 16 18.7 12.4 0.2 5.549 68 N 1.6
15-08-16 15.5 19.4 11.2 0 7.448 68 SE 1.9
16-08-16 15.4 18.7 11.5 0 6.712 64 NE 1.7
17-08-16 16.8 22.2 11.4 0 7.399 40 ENE 1.8
18-08-16 16.7 22.3 11.4 0 7.545 44 NE 1.8
19-08-16 17.8 24.4 12.1 3.8 4.898 49 SSW 1.6
20-08-16 17.8 22 14.6 0 5.605 49 S 3.2
21-08-16 15.7 18.1 14.3 21.6 2.871 80 SSW 3.5
22-08-16 17 20.4 14.7 2 2.299 75 SSW 3.1
23-08-16 20.6 26.2 15.8 0 6.315 47 ESE 1.4
24-08-16 22.9 29.9 16.5 0 6.415 41 E 1.7
25-08-16 23.8 30.9 17.9 0 6.314 46 N 1.2
26-08-16 20.8 22.3 18 0 5.234 72 N 1.9
27-08-16 19.9 24.1 15.5 5 5.336 54 SSE 2.4
28-08-16 19.9 22.9 18.2 2.2 1.433 68 SW 2.7
29-08-16 17.3 19 12.9 0.2 2.279 68 N 3.3
30-08-16 16.3 21.5 11.4 0 2.944 57 NNE 1.1
31-08-16 17.8 24.4 11.5 0 3.111 50 WSW 1.1
24 | Confidential 3750331500
date T-avg T-max T-min rain radiation RV-min Wind
direction
Wind
speed
01-09-16 17.3 20.4 13.6 0 2.277 66 WSW 1.8
02-09-16 16 20.5 12 3 2.434 66 WSW 2.1
03-09-16 17.6 21.1 14.2 2.6 3.106 68 SW 2.1
04-09-16 17.2 19.2 16.1 17.2 3.492 71 W 4.2
05-09-16 17.7 20.9 15.1 2.8 4.652 69 NW 2.6
06-09-16 18 23.2 13.4 0.2 4.373 68 ENE 0.8
07-09-16 19 24.9 13.2 0 5.695 57 E 0.6
08-09-16 19.4 26.1 13.2 0 5.49 53 WNW 2
09-09-16 17.9 22.4 13.7 0 4.63 56 SSE 1.5
10-09-16 18.2 24.6 12.4 0 5.22 48 SSW 1.6
11-09-16 17.9 20.3 16 1.2 2.141 77 ESE 1.1
12-09-16 20.7 27.7 15.3 0 5.422 46 NNE 1.2
13-09-16 23 30.8 16.5 0 5.153 41 ENE 1.2
14-09-16 23.1 30.1 17.2 0 5.122 46 ENE 1.3
15-09-16 21.4 28.1 16.1 0 4.722 46 W 1.6
16-09-16 17.6 20.3 15.5 3.8 2.567 70 SSE 2.5
17-09-16 16.4 19.7 12 0 1.977 73 NNE 1.2
18-09-16 16.3 18.8 14.1 0 4.406 63 NNE 2.2
19-09-16 14.8 18.2 12.6 0 2.75 56 NNW 0.8
20-09-16 13.8 17.7 9.4 0 3.774 62 ENE 0.7
21-09-16 15.4 19.8 11.5 0 3.913 45 E 1.1
22-09-16 14.5 19.4 9.1 0 3.124 59 S 0.9
23-09-16 14.9 18.7 10.3 0 4.374 60 SSW 1.9
24-09-16 14.9 22.1 8.9 0 4.324 51 SE 1.7
25-09-16 16.3 23.8 12.2 2.6 4.037 49 SW 2.7
26-09-16 14.2 18 10.3 0 3.11 65 NNW 1.1
27-09-16 13.9 19.8 8.3 0.2 4.595 45 SW 1.6
28-09-16 17.7 20.2 15.3 2 4.039 63 SW 4.7
29-09-16 15.4 16.8 13.4 9.4 0.962 77 WSW 4.8
30-09-16 13.7 16.7 11.9 0.8 3.034 60 SSW 2.4
Corresponding address for this report:
P.O. Box 16
6700 AA Wageningen
The Netherlands
T +31 (0)317 48 07 00
www.wur.nl/plant-research
Confidential Wageningen Plant Research
Report 3750331500
The mission of Wageningen University and Research is “To explore the
potential of nature to improve the quality of life”. Under the banner
Wageningen University & Research, Wageningen University and the
specialised research institutes of the Wageningen Research Foundation have
joined forces in contributing to finding solutions to important questions in the
domain of healthy food and living environment. With its roughly 30 branches,
5,000 employees and 10,000 students, Wageningen University & Research is
one of the leading organisations in its domain. The unique Wageningen
approach lies in its integrated approach to issues and the collaboration
between different disciplines.

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Efficacy of fungicides to control Early Blight genotypes

  • 1. Efficacy of fungicides to control Early Blight genotypes Field experiment AGV7205 A. Evenhuis & H.T.A.M. Schepers This study was carried out by Wageningen University & Research and was commissioned and financed by UPL Europe Ltd Wageningen Plant Research is the collaboration of Wageningen University and the foundation Stichting Wageningen Research. Wageningen, November 2016 CONFIDENTIAL Report 3750331500
  • 2. A. Evenhuis & H.T.A.M. Schepers, 2016. Efficacy of fungicides to control Early Blight genotypes. Wageningen Plant Research, Confidential Report 3750331500. © 2016 Wageningen, Stichting Wageningen Research, Wageningen Plant Research, P.O. Box 16, 6700 AA Wageningen, The Netherlands; T +31 (0)317 48 07 00; www.wur.nl/plant-research Chamber of Commerce no. 09098104 te Arnhem VAT NL no. 8065.11.618.B01 Stichting Wageningen Research. All rights reserved. No part of this publication may be reproduced, stored in an automated database, or transmitted, in any form or by any means, whether electronically, mechanically, through photocopying, recording or otherwise, without the prior written consent of the DLO Foundation. Wageningen Research is not liable for any adverse consequences resulting from the use of data from this publication. Sjef van der Heijden UPL Europe Ltd The Centre Birchwood Park Warrington, WA3 6YN UK Confidential Wageningen Plant Research Report 3750331500
  • 3. Contents Contents 3 1 Introduction 5 2 Methods and materials 6 2.1 Lay out 6 2.2 Treatments 6 2.3 Inoculation 6 2.4 Disease observations 7 2.5 Sampling and genotyping 7 2.6 Statistics 7 3 Results 8 3.1 Field assessments 8 3.3 Genotyping 10 4 Discussion and conclusions 11 4.1 Early Blight 11 4.2 Soil coverage 11 4.3 Genotyping 11 4.4 Conclusions 12 Experimental lay-out 13Annex 1 nVWA recognition 15Annex 2 Raw data disease severity (%) 16Annex 3 Raw data soil coverage (%) 17Annex 4 Raw data genotyping 18Annex 5 Weather data 19Annex 6
  • 4.
  • 5. Confidential Wageningen Plant Research Report 3750331500 | 5 1 Introduction Early Blight is caused by Alternaria solani. Although late blight, caused by Phytophthora infestans, is the most important disease in potato production Early Blight is regularly found in potato fields in the second half of the season. Farmers spray potatoes 2 to 4 times during a season to control Early Blight. Specific fungicides most commonly used in the Netherlands are Signum, Amistar and recently also Narita and Carial Star. Furthermore a number of fungicides used to control potato late blight show also some efficacy to control Early Blight. Especially products based on mancozeb can be mentioned here. Resistance of A. solani to azoxystrobin has been reported in the United States (Pasche and Gudmestad, 2008). This resistance is associated with the F129L mutation. This mutation has also been reported in Germany (Leiminger et al., 2014), and one isolate with the F129L mutation has been found in the Netherlands (Evenhuis, et al., 2013). Until now no obvious lack of control of azoxystrobin has been found in agricultural practise (Leiminger and Hausladen, 2013). However in a field experiment conducted in Valthermond in the Netherlands in 2014 the efficacy of Amistar and Signum to control Early Blight was less than expected. It is not known what caused this effect. Fungicide resistance management of A. solani was not investigated so far in the Netherlands. In this experiment the efficacy of fungicides to control a mixture of A. solani isolates in a field experiment was tested by request of UPL.
  • 6. 6 | Confidential 3750331500 2 Methods and materials 2.1 Lay out The experiment was set up in a randomised block design. Details are described in Appendix 1. 2.2 Treatments Fungicide applications were carried out using a trial site sprayer with Airmix 110.04 nozzles. Nozzles were hanging approximately 50 cm above the foliage. Sprayings were carried out based on 300 l/ha. Potato plants were sprayed for the first time when they reached a height of 20-30 cm when the rapid growth started. Specific sprays to control Early Blight started Mid-July. Haulm killing was carried out on 30 September 2016, despite natural senescence of the crop. In Table 1 the fungicides used and dose rates are presented. Table 1 The fungicides used in the 2016 experiment and the applied dose rates. Fungicide Active ingredient Dose rate L or kg /ha Narita difenoconazole 250 g/l 0.5 Signum pyraclostrobin 6.7% + boscalid 26.7% 0.2 Penncozeb 80 WP mancozeb 800 g /kg 2.0 Treatment A is the untreated control and H the reference treatment chosen. Spray strategies are given in Table 2. Table 2 Spray strategies and date of application. Code Fungicide 12-7 19-7 26-7 2-8 9-8 16-8 23-8 A UTC - - - - - - - B Signum 0.2 S - S - S - S C Penncozeb 80 WP 2.0 P P P P P P P D Penncozeb 80 WP 2.0 + Signum 0.2 P+S P P+S P P+S P P+S E Narita 0.5 N - N - N - N H Signum 0.2 or Narita 0.5 S - S - N - N 2.3 Inoculation A selection of 2 A. solani isolates was made. AltNL03003 was isolated in 2003 and belongs to genotype 1 and is wild type. AltNL15002 belongs to genotype 2 and possesses the F129L mutation. Both isolates were grown on wheat kernels separately. A mixture of 95% wildtype and 5% F129L type was inoculated. Infested kernels were broadcasted in the field on 15 July 2016.
  • 7. Confidential 3750331500 | 7 2.4 Disease observations Disease observations were carried out from mid-July. Depending on the disease development disease observations were carried out with an interval of 1 to 2 weeks. The percentage necrotic foliage due to A. solani per plot was estimated. 2.5 Sampling and genotyping To assess Alternaria genotype and the presence of the F129L mutation leaves with Alternaria lesions were collected. Per plot 8 leaflets with Alternaria lesions were sampled. Samples were taken four times during the season in the last week of August and the first three weeks of September. Only the last sample was genotyped. The samples were air dried and stored in Petri dishes until further processing. At the laboratory monospore cultures were made on agar. Mycelium was scraped from the agar plate after incubation. DNA was extracted from the mycelium. The genotype involved was assessed by carrying out two PCR described by Pasche et al., 2004 and Lieminger et al., 2015. The PCR product was extracted from the gel and sequenced. The nucleotide order was assessed and the presence of the F129L mutation was established according to the publication of Lieminger et al., 2015. 2.6 Statistics Analysis of variance on the parameters was made using GENSTAT 18th Edition. The experiment was carried out with four replications in a randomised block design. Each replication consisted of a plot. Transformation of data was carried out when necessary.
  • 8. 8 | Confidential 3750331500 3 Results 3.1 Field assessments The results are given in Table 3 and Table 4. On 22 July 2016 no Early Blight was found. A week later on 29 July Early Blight was present in all plots at a low disease severity rate of 0.001% (data not shown). From 5 August onwards the Early Blight epidemic started (Figure 1). After 17 September it was not possible to assess Early Blight because it was too much entangled with natural senescence to distinguish. Phytotoxicity of the fungicides used was not observed. In this experiment the fungicides used were crop safe. Disease severity of all spray strategies was significantly lower than the untreated control. Based on AUDPC, the efficacy of treatments D, E and H were significantly better than treatments B and C. Table 3. Effect of fungicide spray treatments on Early Blight disease severity (%). Log 10 back transformed Early Blight disease severity ratings. Code 5-8 12-8 19-8 26-8 2-9 9-9 16-9 AUDPC A 0.001 a 0.29 c 0.7 c 4.2 c 8.7 d 19.7 c 18.2 c 330 c B 0.001 a 0.00 a 0.0 a 0.2 b 1.2 c 3.0 b 14.1 c 86 b C 0.001 a 0.07 bc 0.0 ab 0.1 b 0.9 c 1.9 b 10.3 bc 68 b D 0.001 a 0.03 abc 0.0 ab 0.0 a 0.0 a 0.1 a 5.1 b 21 a E 0.001 a 0.02 abc 0.0 bc 0.1 b 0.2 b 0.2 a 1.2 a 12 a H 0.001 a 0.01 ab 0.0 ab 0.1 b 0.1 b 0.2 a 1.5 a 10 a 1 ) Values in columns followed by the same letter are not significantly different (P=0.05). Figure 1 Early Blight disease severity (%)
  • 9. Confidential 3750331500 | 9 On average soil coverage (%) of treatments B to H was significantly higher than the untreated control (A). No further differences between average soil cover were found. Table 4 Effect of spray treatments on soil coverage (%). Code 2-9 9-9 16-9 2 23-9 2 Average A 43 a1 21 a 3 a 1.0 a 17 a B 73 b 53 b 17 b 2.1 ab 37 b C 71 b 46 b 18 b 4.1 bc 36 b D 74 b 54 b 21 b 5.7 c 39 b E 61 b 43 ab 13 b 4.8 bc 31 b H 70 b 53 b 15 b 6.2 c 37 b 1 ) Values in columns followed by the same letter are not significantly different (P=0.05). 2 ) Values on 16 & 23-9 are logit back transformed data.
  • 10. 10 | Confidential 3750331500 3.3 Genotyping The results are presented in Table 5 and Figure 2. No significant shift of the Alternaria solani genotype was found when Penncozeb was used compared to the untreated control. When Signum (B) or Signum followed by Narita (H) was sprayed, significantly more GII F129L types were found compared to the untreated control. Table 5 Effect of spray treatments on frequency of Alternaria genotypes. Code Strategy GI WT GI F129L GII WT GII F129L A UTC 56 b 0 0 a 44 a B Signum 7 a 0 3 b 90 b C Penncozeb 52 b 0 0 a 48 a H Signum / Narita 13 a 0 0 a 86 b 1 ) Values in columns followed by the same letter are not significantly different (P=0.05). Figure 2 Alternaria solani genotype distribution (%)
  • 11. Confidential 3750331500 | 11 4 Discussion and conclusions A field experiment was carried out in cultivar Agria. The Early Blight epidemic developed late despite artificial inoculation. The weather in June was conducive for potato late blight and not so much for Early Blight. Potato late blight was controlled with cover sprays using Revus 0.6 l/ha, Infinito 1.6 l/ha and Ranman Top 0.5 l/ha. These fungicides have no known efficacy to control Early Blight. Besides Alternaria spp. also Sclerotinia was found in the experiment. Sclerotinia severity was not assessed. Nevertheless some treatments probably controlled Sclerotinia which together with Alternaria and senescence account for soil coverage. 4.1 Early Blight Early Blight disease severity in the untreated control was significantly higher than all other treatments indicating that the inoculation was successful. Furthermore all spray strategies effectively controlled Early Blight, although the efficacy varied between strategies. Disease severity of treatment B and C increased more towards the end of the season compared to treatments D, E and H. The last spray application was 23 August and the increase was observed from 9 September onwards. Interestingly when Penncozeb was added to Signum (or v.v.) the efficacy to control Early Blight increased compared to the products used solo. However the efficacy to control Early Blight was still less than using Narita or the combination Signum followed by Narita. 4.2 Soil coverage Average soil coverage of all spray strategies was comparable and significantly higher than the untreated control. On individual assessment data some minor differences between treatments occurred. 4.3 Genotyping The field was inoculated with wheat kernels with 2 Alternaria genotypes. Genotype I (GI) is wild type and Genotype II (GII) possesses the F129L mutation. Predominantly these two genotypes were found in the field. Only 1 isolate found was Genotype II, wild type. No Genotype I isolates were found possessing the F129L mutation. In the untreated control 57% of the isolates were Genotype I and wild type. The experiment was inoculated with 95% Genotype I wildtype. This could suggest that the F129L mutation was already present in the natural A. solani population at the location of the field experiment. Alternatively GII might be more aggressive than GI and therefore was found back more frequently. Leaves were picked at four times during the season. Alternaria solani was isolated only from the leaves picked at the last sampling date. Leaves of the other sampling dates were stored under dry conditions at room temperature and could be used for additional assessments. When Signum or Signum followed by Narita was sprayed, significantly more GII F129L A. solani types were found than in the untreated control and when Penncozeb was sprayed. This suggests that selection towards GII F129L occurred under influence of spray strategies B and H. It is known that the F129L mutation causes a reduced sensitivity to QoI fungicides. One of the active ingredients of Signum is pyraclostrobin, a QoI fungicide.
  • 12. 12 | Confidential 3750331500 4.4 Conclusions Phytotoxicity of the fungicides applied was not observed in this experiment. The used products are crop safe. Alternaria disease severity based on AUDPC was significantly higher in the untreated than all other treatments. Based on AUDPC, the efficacy of treatments D, E and H were significantly better than treatments B and C. On average soil coverage (%) of treatments B to H was significantly higher than the untreated control. No significant shift of the Alternaria solani genotype was found when Penncozeb was used compared to the untreated control. When Signum or Signum followed by Narita was sprayed, significantly more GII F129L types were found compared to the untreated control.
  • 13. Confidential 3750331500 | 13 Experimental lay-outAnnex 1 Site Lelystad, The Netherlands Soil texture Sandy loam Previous crop onion Pre previous crop Fertilization 500 kg KAS on 17 May and 300 kg KAS on 22 June Variety: Agria Planting date: 3 May 2016 Seed Rate: 2.500 kg/ha Herbicide treatment: 24 May Fungicide application: see paragraph 2 and normal practice Alternaria treatments: see paragraph 2 and normal practice Crop desiccation: Reglone (diquat dibromide 200g/l) 2.0 l/ha 30 September 2016. Harvest: - Tuber assessments: - Gross plot dimensions: Four rows (0.75 m) of 11 m length Net plot dimensions: Two rows (0.75) of 11 m length Demo design: Four replications in a randomized block design Lelystad
  • 14. 14 | Confidential 3750331500 Project: Experiment: AGV7205 Alternaria genotypes Location: Lelystad Field: G86-4 8 16 24 32 C F G D 7 15 23 31 D A B G 6 14 22 30 H B E F 5 13 21 29 A H C B 4 12 20 28 E G D C 3 11 19 27 B C H E 2 10 18 26 G D F A 1 9 17 25 F E A H 1.5 < 3m > < 3m > < 3m > < 3m > < 3m > < 3m > 1.5 3750331500 N I II III IV
  • 15. Confidential 3750331500 | 15 nVWA recognitionAnnex 2
  • 16. 16 | Confidential 3750331500 Raw data disease severity (%)Annex 3
  • 17. Confidential 3750331500 | 17 Raw data soil coverage (%)Annex 4
  • 18. 18 | Confidential 3750331500 Raw data genotypingAnnex 5
  • 19. Confidential 3750331500 | 19 Weather dataAnnex 6 date T-avg T-max T-min rain radiation RV-min Wind direction Wind speed 01-04-16 6.8 12.4 1.1 0 5.835 54 SSE 1.6 02-04-16 8.8 12.7 4.3 0 2.811 59 SSE 1.7 03-04-16 13.1 18.1 10.3 0.6 3.783 59 NE 2.7 04-04-16 12.3 14.6 8.9 6.8 3.358 61 E 2.8 05-04-16 9.9 12.1 8.5 0.6 2.26 76 SW 2.2 06-04-16 7.9 10.2 6.1 7.6 1.968 73 SW 5 07-04-16 7.2 9.2 5.3 0.4 3.779 71 SW 4.6 08-04-16 7.9 10.4 5.4 0.6 3.085 71 SE 2.1 09-04-16 8.7 14.5 3.2 0.2 4.43 46 ESE 2.7 10-04-16 9.5 13.6 6.7 1.2 5.779 64 NE 2.2 11-04-16 11.4 16.7 6.8 0 6.592 49 E 4 12-04-16 11.4 14.6 8 5.2 4.977 61 SW 2.7 13-04-16 9.7 14 5.1 2.2 4.047 66 SSW 1.6 14-04-16 9.7 14 5.6 0 3.791 62 E 1.3 15-04-16 9.6 11.9 8.4 4.4 2.401 80 SSE 2.3 16-04-16 8.6 10.6 6.1 5.8 4.979 70 NW 5.4 17-04-16 6.9 9.7 4.4 0.8 4.439 66 W 3.8 18-04-16 8 12 3.8 0 3.889 55 WSW 4.5 19-04-16 9.4 11 7.5 4 7.078 62 SW 15.3 20-04-16 7.8 10.8 4.9 0 6.904 60 NE 13.2 21-04-16 8.3 14.4 2.5 0 7.574 44 NNE 2.4 22-04-16 7.8 10.9 5.2 0 5.639 57 NNE 3.4 23-04-16 6.2 8.4 4.4 0.2 6.002 54 WNW 4.7 24-04-16 5 7.7 2.8 4.6 5.73 59 SE 5.8 25-04-16 4.9 7.6 2.5 4 2.788 68 WNW 4.2 26-04-16 4.3 7 2 8.4 3.36 77 NW 6.1 27-04-16 5.8 8.2 3.7 0.4 5.737 73 NNW 4.9 28-04-16 5.1 8.2 2.7 5.6 4.277 61 SSE 2 29-04-16 6.8 10.2 2.9 5 4.237 52 SSW 4.7 30-04-16 7.2 9.4 4.7 0.4 5.424 69 NNE 3.2
  • 20. 20 | Confidential 3750331500 date T-avg T-max T-min rain radiation RV-min Wind direction Wind speed 01-05-16 7.5 11.2 3.3 0 8.126 63 NNW 2.5 02-05-16 10.5 17.1 3.2 0 6.75 36 SSW 2.5 03-05-16 10.3 12.3 7.6 1.2 7.365 57 W 4.1 04-05-16 10.1 15.1 3.5 0 7.354 42 ESE 1.4 05-05-16 12.8 18.4 5.8 0 8.425 30 ENE 2.3 06-05-16 16.6 23.9 9.3 0 8.115 33 ESE 2.3 07-05-16 19.5 26 12.1 0 8.186 28 ENE 2.1 08-05-16 19.6 24.8 12.6 0 8.428 25 ENE 3.6 09-05-16 19.5 24.6 13.7 0 8.075 36 ENE 3.6 10-05-16 18.8 22.5 14.5 0 3.86 44 NNE 2.1 11-05-16 19.8 25 15.2 0 7.243 35 ENE 1.9 12-05-16 19.4 24.8 13.7 0 8.174 34 NE 3.3 13-05-16 14.6 19.9 9.3 0 8.138 44 N 4.5 14-05-16 8.6 10.9 7.3 3 5.462 58 NW 6.7 15-05-16 8.4 10.7 6.6 7 5.947 61 WNW 6 16-05-16 9.3 11.6 7.1 0.4 5.334 64 WNW 3.9 17-05-16 12 16.5 8.7 0 6.039 54 S 2 18-05-16 13.1 17.5 7.3 0 4.184 52 E 2.3 19-05-16 14 18.4 10.3 0 5.038 62 S 1.6 20-05-16 13.1 16.5 10.6 1 2.534 69 WSW 2.8 21-05-16 16.8 22.6 9.7 0 5.506 50 SSE 2.7 22-05-16 15.9 18.7 13.2 8.4 2.679 70 NNE 1.8 23-05-16 12.1 13.4 11.4 17.6 2.329 86 NNW 6.2 24-05-16 10.7 12.1 10 0 4.308 76 NW 5.3 25-05-16 11.9 14.3 10.1 0.6 2.154 81 NW 0.7 26-05-16 13.7 17.2 10.5 0.4 6.127 58 NE 1.7 27-05-16 14.8 19.4 10.3 0 7.252 56 NNE 2.2 28-05-16 16.6 21 12.1 1.6 7.684 66 NNE 2.3 29-05-16 17 21.1 13.7 0.6 3.996 52 NNE 3.1 30-05-16 16.7 20 14.5 3.8 3.089 80 N 5.4 31-05-16 18.9 22.7 15.3 0 6.431 50 ESE 2.1
  • 21. Confidential 3750331500 | 21 date T-avg T-max T-min rain radiation RV-min Wind direction Wind speed 01-06-16 19.3 23.6 15.8 0 5.727 66 ENE 4.4 02-06-16 15.1 17.4 13.1 0 1.964 82 NNE 5.8 03-06-16 16.6 20.2 13.1 1.4 4.359 74 NW 2.3 04-06-16 20.9 25.6 15.9 0 7.601 55 NNE 2.3 05-06-16 18.1 20.9 13.6 0 9.082 70 NNE 3.7 06-06-16 16.4 21.7 11.2 0 9.128 65 NNE 2.8 07-06-16 18 22.6 12.7 0 7.692 58 N 3 08-06-16 14.3 16.3 12.1 0 6.105 72 N 3.2 09-06-16 14 16.1 12.1 0 9.187 63 N 4 10-06-16 14.3 17.7 10.8 0 7.744 37 NE 1.9 11-06-16 15.5 19.6 11.9 0 4.014 59 NNE 1.4 12-06-16 16.4 19.9 13.7 3.6 3.835 63 SSE 0.7 13-06-16 15.6 18.1 14.1 11.8 3.804 81 W 2 14-06-16 15 17.6 13.5 13 3.958 76 S 2.3 15-06-16 14.5 16.7 11.8 0.4 4.361 76 SW 1.7 16-06-16 15.5 18.9 11.7 0.4 7.239 58 WSW 1.5 17-06-16 15.3 17.3 13.4 10 5.734 75 NW 2.4 18-06-16 14.4 16.1 12.6 2.6 4.71 74 NW 3.3 19-06-16 14.4 17.2 12.1 0 7.379 62 SSE 3.3 20-06-16 14.8 16.1 14 20.8 1.817 79 W 4 21-06-16 16.8 19.4 13.7 6.2 4.194 78 WSW 2 22-06-16 18.6 23.4 13.6 0.2 7.33 67 ESE 2 23-06-16 20.6 25.9 18.1 12.4 5.444 75 SSE 1.7 24-06-16 19.1 21 15.6 0.2 5.317 75 N 2.4 25-06-16 15.2 17.7 12.5 5.6 3.704 80 WSW 0.9 26-06-16 14.7 17.6 11.6 0.2 5.265 68 SW 2.8 27-06-16 14.5 17.5 11.8 3.2 3.979 68 WNW 3 28-06-16 15.5 20.1 10.6 1.6 5.466 58 W 1.4 29-06-16 15.3 18.4 13.2 4.2 5.649 61 S 3.4 30-06-16 15.3 17.8 13.5 4.8 3.361 84 SSW 3.1
  • 22. 22 | Confidential 3750331500 date T-avg T-max T-min rain radiation RV-min Wind direction Wind speed 01-07-16 16 17.5 14.3 3.2 1.821 84 SSW 3.7 02-07-16 15 16.6 12.9 1 7.48 59 SW 4.1 03-07-16 14.4 18.1 10.8 7.4 6.643 64 WNW 2.9 04-07-16 16.4 20.9 10.8 0 7.924 57 N 1.4 05-07-16 15 16.8 13.5 1 3.498 77 WNW 4 06-07-16 14.7 17.5 12.1 0.2 7.523 63 W 3.6 07-07-16 16 21.1 10.2 0 6.983 54 SSE 2.1 08-07-16 16 19 13.6 3.4 3.06 77 W 2.4 09-07-16 17.8 20.8 13.1 0.6 4.746 65 SSW 2.2 10-07-16 20.1 25 16 0 7.294 58 SSW 3.4 11-07-16 17.9 20.4 15.9 0 5.099 61 SW 4.9 12-07-16 16.3 18.8 14.5 17.6 4.893 75 SE 2.2 13-07-16 14.2 16.3 11.6 3.2 3.356 66 WNW 1.8 14-07-16 14.4 17 12.2 0 5.183 63 NW 4 15-07-16 15.8 19.5 12.7 0 7.468 60 W 3.2 16-07-16 17.5 19.1 13.8 0 3.029 83 S 2.3 17-07-16 18.6 21.1 16.4 4.2 8.149 65 NW 2.7 18-07-16 18.8 23.2 13.8 0 8.715 49 N 1.4 19-07-16 20.9 27.9 12.2 0 8.624 34 ENE 0.8 20-07-16 24.1 32.2 17.7 0 7.266 36 SW 2 21-07-16 21.1 25.1 16.9 0 7.465 65 NW 1.7 22-07-16 19.8 24 16.7 4.6 5.819 66 NNE 1.1 23-07-16 19 22.1 16.4 0 5.506 78 NNE 2 24-07-16 19.9 23.7 16.1 0 5.905 70 W 0.9 25-07-16 18.1 20.7 15.1 1.2 3.396 68 WNW 1.7 26-07-16 17.8 22.7 12.6 0 5.911 52 ENE 1.4 27-07-16 17.8 21 16.2 8.4 3.933 64 SSW 2 28-07-16 17.4 20.4 14.8 25.8 4.129 79 SSE 1.6 29-07-16 17.7 20.2 14.8 1.4 3.991 76 SW 2.8 30-07-16 17.5 20.4 15.3 9.4 4.493 62 SSE 1.8 31-07-16 16.2 19.1 13.7 0.2 5.557 66 WSW 2.3
  • 23. Confidential 3750331500 | 23 date T-avg T-max T-min rain radiation RV-min Wind direction Wind speed 01-08-16 16 19.5 11.6 0.2 6.365 56 WSW 1.9 02-08-16 15.5 17.5 13.2 1.2 2.62 81 SSW 1.7 03-08-16 18.5 20.5 16.8 5.2 3.068 74 SSW 3.8 04-08-16 17.2 20.1 14.7 0.2 4.029 64 SSW 3.9 05-08-16 16.9 20.1 13.5 0 5.576 65 W 2.6 06-08-16 16.3 20.3 13 0.4 5.93 60 WSW 2 07-08-16 16.7 21.6 11.6 0 3.396 75 SW 3 08-08-16 17.3 18.8 14.8 0 5.395 60 WNW 4.2 09-08-16 13.9 16.6 12.3 9.6 5.116 58 ESE 2.6 10-08-16 12.7 15.4 9.5 3.2 7.126 61 W 2.6 11-08-16 12.6 15.3 9.2 17 1.97 81 W 2.7 12-08-16 17.7 20.1 15.7 1.6 3.447 76 SSW 3.2 13-08-16 16.9 19.3 14.8 0 3.38 75 WSW 2.6 14-08-16 16 18.7 12.4 0.2 5.549 68 N 1.6 15-08-16 15.5 19.4 11.2 0 7.448 68 SE 1.9 16-08-16 15.4 18.7 11.5 0 6.712 64 NE 1.7 17-08-16 16.8 22.2 11.4 0 7.399 40 ENE 1.8 18-08-16 16.7 22.3 11.4 0 7.545 44 NE 1.8 19-08-16 17.8 24.4 12.1 3.8 4.898 49 SSW 1.6 20-08-16 17.8 22 14.6 0 5.605 49 S 3.2 21-08-16 15.7 18.1 14.3 21.6 2.871 80 SSW 3.5 22-08-16 17 20.4 14.7 2 2.299 75 SSW 3.1 23-08-16 20.6 26.2 15.8 0 6.315 47 ESE 1.4 24-08-16 22.9 29.9 16.5 0 6.415 41 E 1.7 25-08-16 23.8 30.9 17.9 0 6.314 46 N 1.2 26-08-16 20.8 22.3 18 0 5.234 72 N 1.9 27-08-16 19.9 24.1 15.5 5 5.336 54 SSE 2.4 28-08-16 19.9 22.9 18.2 2.2 1.433 68 SW 2.7 29-08-16 17.3 19 12.9 0.2 2.279 68 N 3.3 30-08-16 16.3 21.5 11.4 0 2.944 57 NNE 1.1 31-08-16 17.8 24.4 11.5 0 3.111 50 WSW 1.1
  • 24. 24 | Confidential 3750331500 date T-avg T-max T-min rain radiation RV-min Wind direction Wind speed 01-09-16 17.3 20.4 13.6 0 2.277 66 WSW 1.8 02-09-16 16 20.5 12 3 2.434 66 WSW 2.1 03-09-16 17.6 21.1 14.2 2.6 3.106 68 SW 2.1 04-09-16 17.2 19.2 16.1 17.2 3.492 71 W 4.2 05-09-16 17.7 20.9 15.1 2.8 4.652 69 NW 2.6 06-09-16 18 23.2 13.4 0.2 4.373 68 ENE 0.8 07-09-16 19 24.9 13.2 0 5.695 57 E 0.6 08-09-16 19.4 26.1 13.2 0 5.49 53 WNW 2 09-09-16 17.9 22.4 13.7 0 4.63 56 SSE 1.5 10-09-16 18.2 24.6 12.4 0 5.22 48 SSW 1.6 11-09-16 17.9 20.3 16 1.2 2.141 77 ESE 1.1 12-09-16 20.7 27.7 15.3 0 5.422 46 NNE 1.2 13-09-16 23 30.8 16.5 0 5.153 41 ENE 1.2 14-09-16 23.1 30.1 17.2 0 5.122 46 ENE 1.3 15-09-16 21.4 28.1 16.1 0 4.722 46 W 1.6 16-09-16 17.6 20.3 15.5 3.8 2.567 70 SSE 2.5 17-09-16 16.4 19.7 12 0 1.977 73 NNE 1.2 18-09-16 16.3 18.8 14.1 0 4.406 63 NNE 2.2 19-09-16 14.8 18.2 12.6 0 2.75 56 NNW 0.8 20-09-16 13.8 17.7 9.4 0 3.774 62 ENE 0.7 21-09-16 15.4 19.8 11.5 0 3.913 45 E 1.1 22-09-16 14.5 19.4 9.1 0 3.124 59 S 0.9 23-09-16 14.9 18.7 10.3 0 4.374 60 SSW 1.9 24-09-16 14.9 22.1 8.9 0 4.324 51 SE 1.7 25-09-16 16.3 23.8 12.2 2.6 4.037 49 SW 2.7 26-09-16 14.2 18 10.3 0 3.11 65 NNW 1.1 27-09-16 13.9 19.8 8.3 0.2 4.595 45 SW 1.6 28-09-16 17.7 20.2 15.3 2 4.039 63 SW 4.7 29-09-16 15.4 16.8 13.4 9.4 0.962 77 WSW 4.8 30-09-16 13.7 16.7 11.9 0.8 3.034 60 SSW 2.4
  • 25.
  • 26. Corresponding address for this report: P.O. Box 16 6700 AA Wageningen The Netherlands T +31 (0)317 48 07 00 www.wur.nl/plant-research Confidential Wageningen Plant Research Report 3750331500 The mission of Wageningen University and Research is “To explore the potential of nature to improve the quality of life”. Under the banner Wageningen University & Research, Wageningen University and the specialised research institutes of the Wageningen Research Foundation have joined forces in contributing to finding solutions to important questions in the domain of healthy food and living environment. With its roughly 30 branches, 5,000 employees and 10,000 students, Wageningen University & Research is one of the leading organisations in its domain. The unique Wageningen approach lies in its integrated approach to issues and the collaboration between different disciplines.