Research Center Isagro S.p.A. Via Fauser 28,

28100 Novara (NO) (Italy)

FINAL REPORT

E01-16 Evaluation of the efficacy of two different formulations (BB and BC4) of Glyphosate in post-emergence application in greenhouse in comparison to standard (GLIFO) 10th May 2016

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Data Requirements

UNI EN ISO 9001:2008

Date

10th May 2016

Research Center Isagro S.p.A. Via Fauser, 28

28100 Novara (NO) Italy

Test Facility Sponsor Research Center SOITEM S.r.L ISAGRO S.p.A. Via Roberto Cozzi, 34 Via Fauser, 28 20125 Milano (MI) 28100 Novara (NO) Italy Italy

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1. Compliance Statement The present study was carried out in compliance with:

• UNI EN ISO 9001:2008

Herbicide Evaluation Manager

.............................................................................

date...... /...... /......

Research Center - ISAGRO S.p.A

Via Fauser, 28 – 28100 Novara (NO) - ITALY tel.: +39-0321-693648/ fax.: +39-0321-693636 / e-mail: ibondoni@isagro.it

Biological Efficacy Assessment Head

.............................................................................

date...... /...... /......

Research Center - ISAGRO S.p.A

Via Fauser, 28 – 28100 Novara (NO) - ITALY tel.: +39-0321-693712 / e-mail: rliguori@isagro.com

Sponsor Representative .............................................................................

date...... /...... /......

dr. Alfredo Rota SOITEM S.r.l

Via Roberto Cozzi, 34 – 20125 – Milano (MI) - ITALY tel.: +39-02-64741824 /e-mail: a.rota@soitem.it

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2. Statement of Confidentiality This report contains proprietary information of the sponsor, which must be kept strictly secret and not be disclosed to anyone, except the employees of the testing facility for experimental purposes only. No information related to the study or to the test substance(s) may be revealed or released to any third part without prior notification and authorization of the sponsor.

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3. Content

1. COMPLIANCE STATEMENT ............................................................................ 2

2. STATEMENT OF CONFIDENTIALITY ........................................................... 3

3. CONTENT ............................................................................................................... 4

4. OBJECTIVES ......................................................................................................... 5

5. MATERIALS AND METHODS ........................................................................... 5

5.1 Trial treatments .............................................................................................. 5 5.2 Trial and Site information .............................................................................. 6

5.3 Application information ................................................................................. 7

5.4 Assessment method ........................................................................................ 8

5.5 Statistical analysis .......................................................................................... 8

6. RESULTS AND CONCLUSIONS ........................................................................ 9

6.1 Efficacy on AMARE ...................................................................................... 9

6.2 Efficacy on ALOMY ................................................................................... 12

6.3 Efficacy on ECHCG .................................................................................... 15

6.4 Conclusions .................................................................................................. 18

7. APPENDIX ............................................................................................................ 19

7.1 Meteorological data ..................................................................................... 19

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4. Objectives To evaluate the efficacy of two different formulations of Glyphosate (BB and BC4 signed) in comparison to reference standard (GLIFO signed) on 3 different weeds in post emergence application (BBCH 13-14).

5. Materials and methods

5.1 Trial treatments

Trt Treatment Form Form Form Rate No. Type Name Conc Unit Type Density Rate Unit 1 CHK Untreated Check 2 HERB GLIPHO REF 360 g/L SL 1 45 g ai/ha 3 HERB GLIPHO REF 360 g/L SL 1 90 g ai/ha 4 HERB GLIPHO REF 360 g/L SL 1 180 g ai/ha 5 HERB GLIPHO REF 360 g/L SL 1 360 g ai/ha 6 HERB GLIPHO REF 360 g/L SL 1 720 g ai/ha 7 HERB GLIPHO BB 360 g/L SL 1 45 g ai/ha 8 HERB GLIPHO BB 360 g/L SL 1 90 g ai/ha 9 HERB GLIPHO BB 360 g/L SL 1 180 g ai/ha

10 HERB GLIPHO BB 360 g/L SL 1 360 g ai/ha 11 HERB GLIPHO BB 360 g/L SL 1 720 g ai/ha 12 HERB GLIPHO BC4 360 g/L SL 1 45 g ai/ha 13 HERB GLIPHO BC4 360 g/L SL 1 90 g ai/ha 14 HERB GLIPHO BC4 360 g/L SL 1 180 g ai/ha 15 HERB GLIPHO BC4 360 g/L SL 1 360 g ai/ha 16 HERB GLIPHO BC4 360 g/L SL 1 720 g ai/ha

Additional Treatment Information Type CHK = Check or Untreated HERB = Herbicide Treatment Name Untreated Check, , , = Not treated| Form Unit g/L = grams active ingredient per litre formulated product (same as ga/l) Form Type SL = soluble concentrate|Liquid|||A clear to opalescent liquid to be applied as a solution of the active ingredient after dilution in water. Rate Unit g AI/ha = Grams Active Ingredient per Hectare (US=g AI/A)|D Replications: 4, Untreated treatments: 1, Conduct under GLP/GEP: Yes (GEP with no protection), Design: NON Randomised, Treatment units: Treated 'Plot' experimental unit size, Dry Form. Unit: %, Treated 'Plot' experimental unit size Width: 1 meters, Treated 'Plot' experimental unit size Length: 1 meters, Application volume: 200 L/ha, Mix size: 40 mL, Format definitions: G-All7.def, G-All7.frm

Applications: 1 applications were carried out starting from BBCH 13-14. For details see the Application Description section in the present report.

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5.2 Trial and Site information

General Trial Information

Study Director:IVAN BONDONI Title:DS

Investigator:GABRIELE PIAZZON Title:TECNICHAL

Discipline:H herbicide

Trial Status:F one-year/final Trial Reliability:HIGH Initiation Date:Mar-23-2016 Planned Completion Date:Apr-12-2016

Completion Date:Apr-12-2016

Trial Location

City:Novara Country:ITA Italy State/Prov.:Novara

Postal Code:28100 Climate Zone:EPOMED EPPO Mediterranean

Contacts

Study Director:IVAN BONDONI Title:DS Organization:ISAGRO S.p.A

Address:VIA FAUSER 28 Phone No.:0321693648 City+State/Prov:NOVARA Mobile No.:3669080811

Postal Code:28100 E-mail:ibondoni@isagro.com Country:ITA Italy

Investigator:GABRIELE PIAZZON Title:TECNICHAL Organization:ISAGRO S.p.A.

Address:VIA FAUSER 28 Phone No.:0321693644 City+State/Prov:NOVARA

Postal Code:28100 E-mail:gpiazzon@isagro.com Country:ITA Italy

Pest Description

Pest 1 Type:W Code:ECHCG Echinochloa crus-galli

Common Name:Cockspur(grass)

Pest 2 Type:W Code:ALOMY Alopecurus myosuroides

Common Name:Blackgrass

Pest 3 Type:W Code:AMARE Amaranthus retroflexus

Common Name: Common amaranth Site and Design

Treated Plot Width:10 cm Site Type:GREENH greenhouse Treated Plot Length:10 cm

Treatments: 16

Replications:4 Study Design:NONRAN Non-Randomized

% Slope:0.0

Untreated Arrangement:ADJACENT control side by side of each treated plot

Soil Description

Description Name:BEA Soil % Sand:62 % OM:1.4 Texture: SL sandy loam

% Silt:24 pH:6.9 Soil Name: Origin - Salerano sul Lambro % Clay:14 CEC:7.87 Fert. Level: P poor

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5.3 Application information

Application Description A

Application Date: Mar-8-2016 Appl. Start Time: 9:00 Appl. Stop Time: 10:00 Application Method: SPRAY Application Timing: POSPOS Application Placement: POTP10 Applied By: G.PIAZZON Air Temperature, Unit: 24 C % Relative Humidity: 45 Soil Temperature, Unit: 15 C Soil Moisture: 70 %

Pest Stage At Each Application

A Pest 1 Code, Type, Scale: ECHCG W Stage Majority, Percent: 14 100 Pest 2 Code, Type, Scale: ALOMY W Stage Majority, Percent: 14 100 Pest 3 Code, Type, Scale: AMARE W Stage Majority, Percent: 14 100

Application Equipment

A Appl. Equipment: HERBICIDE MA Operation Pressure, Unit: 3 BAR Nozzle Type: XR8001VS Spray Volume, Unit: 200 L/ha Propellant: AIRFAN

Schedule for Mar-1-2016 to Apr-12-2016 Number Planned Trial Task Done Description Timing Completed Assigned to 1 Mar-8-2016 E01-16 Treat - A Yes Mar-8-2016 Mar-8-2016 G.PIAZZON 2 Mar-11-2016 E01-16 Assess Yes DAT3 Mar-11-2016 Mar-11-2016 G.PIAZZON 3 Mar-15-2016 E01-16 Assess Yes DAT7 Mar-15-2016 Mar-15-2016 G.PIAZZON 4 Mar-22-2016 E01-16 Assess Yes DAT14 Mar-22-2016 Mar-22-2016 G.PIAZZON 5 Mar-29-2016 E01-16 Assess Yes DAT21 Mar-29-2016 Mar-29-2016 G.PIAZZON

Photo 1: GLIPHO REF dilution (3.6 g a.i/l) in comparison to BB and BC4 (same concentration)

GLIPHO BB

3,6 g a.i/l of water

GLIPHO BC4

3,6 g a.i/l of water

GLIPHO REF

3,6 g a.i/l of water

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5.4 Assessment method The efficacy of the herbicide action on each weed species was estimated, using a rating scale from 0 to 100%, as follows:

- The % damaged plant surface was assessed (death, necrosis, size reduction, thinning etc.);

- Each treated plot was compared with the relative plant/weed population assessed in the untreated pot.

5.5 Statistical analysis Data from the assessments were analysed by analysis of variance (ANOVA) with software ARM 9.0 from Gylling Data Management. If consistent with the statistical distribution of data, data were automatically transformed as reported in the result tables. When Analysis of Variance (AOV) treatment was significant [P(F)≤0.05], differences between means were determined by the Duncan’s New MRT test at the Observed Significant Level (OSL) (P≤ 0.05).

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6. Results and Conclusions 6.1 Efficacy on AMARE Table 1: Evaluation of efficacy on AMARE 3,7,14 and 21 DAT

Pest Type W Weed W Weed W Weed W WeedPest Code AMARE AMARE AMARE AMAREPest Scientific Name Amaranthus ret> Amaranthus ret> Amaranthus ret> Amaranthus ret>Pest Name Common amaranth Common amaranth Common amaranth Common amaranthPart Assessed PLANT C PLANT C PLANT C PLANT CAssessment Date Mar-11-2016 Mar-15-2016 Mar-22-2016 Mar-29-2016Assessment Type CONTRO CONTRO CONTRO CONTROAssessment Unit % % % %Number of Subsamples 1 1 1 1SE Group No. 30 33 36 36Days After First/Last Applic. 3 3 7 7 14 14 21 21Trt-Eval Interval 3 7 14 21ARM Action Codes Trt Treatment Rate No. Name Rate Unit 3 6 9 12

1Untreated Check 2GLIPHO REF 45g ai/ha 2.5f 67.5ef 88.8bc 88.8bc 3GLIPHO REF 90g ai/ha 2.5f 75.0cde 91.3ab 93.8ab 4GLIPHO REF 180g ai/ha 12.5cd 90.0ab 95.0ab 95.0ab 5GLIPHO REF 360g ai/ha 15.0bcd 91.3a 100.0a 100.0a 6GLIPHO REF 720g ai/ha 20.0ab 95.0a 100.0a 100.0a 7GLIPHO BB 45g ai/ha 0.0f 50.0g 63.8d 62.5e 8GLIPHO BB 90g ai/ha 5.0ef 60.0fg 81.3c 80.0cd 9GLIPHO BB 180g ai/ha 10.0de 86.3abc 93.8ab 96.3ab

10GLIPHO BB 360g ai/ha 17.5abc 92.5a 100.0a 100.0a 11GLIPHO BB 720g ai/ha 22.5a 93.8a 100.0a 100.0a 12GLIPHO BC4 45g ai/ha 2.5f 57.5fg 67.5d 71.3de 13GLIPHO BC4 90g ai/ha 2.5f 72.5de 86.3bc 87.5bc 14GLIPHO BC4 180g ai/ha 5.0ef 77.5b-e 93.8ab 93.8ab 15GLIPHO BC4 360g ai/ha 12.5cd 83.8a-d 98.8a 100.0a 16GLIPHO BC4 720g ai/ha 20.0ab 95.0a 100.0a 100.0a

LSD P=.05 6.21 11.86 8.72 9.15Standard Deviation 4.35 8.30 6.10 6.40CV 43.46 10.48 6.73 7.01 Treatment F 12.479 12.918 14.677 13.140Treatment Prob(F) 0.0001 0.0001 0.0001 0.0001

Pest Type W, Weed, G-BYRW7, G-WedStg = Weed or volunteer crop Pest Code AMARE, Amaranthus retroflexus, = IE Part Assessed PLANT = plant C = Crop is Part Rated Assessment Type CONTRO = control / burndown or knockdown Assessment Unit % = percent Means followed by same letter do not significantly differ (P=.05, Duncan's New MRT) Mean comparisons performed only when AOV Treatment P(F) is significant at mean comparison OSL.

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Graph 1: Evaluation of efficacy of GLIPHO REF in comparison to BB and BC4 on AMARE at 3 days after treatment

Graph 2: Evaluation of efficacy of GLIPHO REF in comparison to BB and BC4 on AMARE at 21 days after treatment (at the end of the test).

0

20

40

60

80

100

45 90 180 360 720

% E

ffic

acy

RATE g a.i./ga

Efficacy on AMARE - 3 DAT

GLIPHO REF

GLIPHO BB

GLIPHO BC4

0

20

40

60

80

100

45 90 180 360 720

% E

ffic

acy

Rate g a.i./ha

Efficacy on AMARE - 21 DAT

GLIPHO REF

GLIPHO BB

GLIPHO BC4

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Photo 2: Efficacy on AMARE (Amaranthus retroflexus) – 21DAA

UTC GLIPHO REF 360 g a.i/ha

GLIPHO REF 720 g a.i/ha

GLIPHO BB

360 g a.i/ha

GLIPHO BC4 360 g a.i/ha

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6.2 Efficacy on ALOMY Table 2: Evaluation of efficacy on ALOMY 3,7,14 and 21 DAT

Pest Type W Weed W Weed W Weed W WeedPest Code ALOMY ALOMY ALOMY ALOMYPest Scientific Name Alopecurus myo> Alopecurus myo> Alopecurus myo> Alopecurus myo>Pest Name Blackgrass Blackgrass Blackgrass BlackgrassPart Assessed PLANT C PLANT C PLANT C PLANT CAssessment Date Mar-11-2016 Mar-15-2016 Mar-22-2016 Mar-29-2016Assessment Type CONTRO CONTRO CONTRO CONTROAssessment Unit % % % %Number of Subsamples 1 1 1 1SE Group No. 25 31 34 34Days After First/Last Applic. 3 3 7 7 14 14 21 21Trt-Eval Interval 3 7 14 21ARM Action Codes AS Trt Treatment Rate No. Name Rate Unit 1 4 7 10

1Untreated Check 2GLIPHO REF 45g ai/ha 0.0 d 0.0g 0.0g 0.0f 3GLIPHO REF 90g ai/ha 0.0 d 0.0g 5.0fg 5.0ef 4GLIPHO REF 180g ai/ha 0.0 d 7.5fg 10.0ef 10.0de 5GLIPHO REF 360g ai/ha 1.3 cd 12.5f 15.0e 15.0d 6GLIPHO REF 720g ai/ha 6.3 bc 37.5d 47.5d 57.5b 7GLIPHO BB 45g ai/ha 3.4 cd 7.5fg 12.5ef 12.5de 8GLIPHO BB 90g ai/ha 6.3 bc 7.5fg 12.5ef 15.0d 9GLIPHO BB 180g ai/ha 12.2 ab 42.5cd 55.0c 57.5b

10GLIPHO BB 360g ai/ha 14.6 ab 72.5b 93.8a 97.5a 11GLIPHO BB 720g ai/ha 14.6 ab 80.0a 97.5a 100.0a 12GLIPHO BC4 45g ai/ha 0.0 d 10.0f 10.0ef 17.5d 13GLIPHO BC4 90g ai/ha 3.4 cd 12.5f 12.5ef 17.5d 14GLIPHO BC4 180g ai/ha 3.4 cd 25.0e 42.5d 47.5c 15GLIPHO BC4 360g ai/ha 14.6 ab 45.0c 80.0b 92.5a 16GLIPHO BC4 720g ai/ha 17.2 a 75.0ab 97.5a 98.8a

LSD P=.05 1.37t 7.07 7.20 8.57Standard Deviation 0.96t 4.94 5.04 6.00CV 41.31t 17.05 12.79 13.98 Treatment F 7.601 129.857 212.489 162.022Treatment Prob(F) 0.0001 0.0001 0.0001 0.0001

Pest Type W, Weed, G-BYRW7, G-WedStg = Weed or volunteer crop Pest Code ALOMY, Alopecurus myosuroides, = IE Part Assessed PLANT = plant C = Crop is Part Rated Assessment Type CONTRO = control / burndown or knockdown Assessment Unit % = percent ARM Action Codes AS = Automatic square root transformation of X+0.5 Means followed by same letter do not significantly differ (P=.05, Duncan's New MRT) t=Mean descriptions are reported in transformed data units, and are not de-transformed. Mean comparisons performed only when AOV Treatment P(F) is significant at mean comparison OSL.

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Graph 3: Evaluation of efficacy of GLIPHO REF in comparison to BB and BC4 on ALOMY at 3 days after treatment

Graph 4: Evaluation of efficacy of GLIPHO REF in comparison to BB and BC4 on ALOMY at 21 days after treatment (at the end of the test)

0

20

40

60

80

100

45 90 180 360 720

% E

ffic

acy

Rate g a.i./ha

Efficacy on ALOMY - 3 DAT

GLIPHO REF

GLIPHO BB

GLIPHO BC4

0

20

40

60

80

100

45 90 180 360 720

% E

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acy

Rate g a.i./ha

Efficacy on ALOMY - 21DAT

GLIPHO REF

GLIPHO BB

GLIPHO BC4

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Photo 3: Efficacy on ALOMY (Alopecurus myosuroides )– 21DAA

UTC GLIPHO REF 360 g a.i/ha

GLIPHO REF 720 g a.i/ha

GLIPHO BB

360 g a.i/ha

GLIPHO BC4 360 g a.i/ha

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6.3 Efficacy on ECHCG

Table 3: Evaluation of efficacy on ECHCG 3,7,14 and 21 DAT

Pest Type W Weed W Weed W Weed W WeedPest Code ECHCG ECHCG ECHCG ECHCGPest Scientific Name Echinochloa c> Echinochloa c> Echinochloa c> Echinochloa c>Pest Name Junglerice Junglerice Junglerice JunglericePart Assessed PLANT C PLANT C PLANT C PLANT CAssessment Date Mar-11-2016 Mar-15-2016 Mar-22-2016 Mar-29-2016Assessment Type CONTRO CONTRO CONTRO CONTROAssessment Unit % % % %Number of Subsamples 1 1 1 1SE Group No. 26 32 35 35Days After First/Last Applic. 3 3 7 7 14 14 21 21Trt-Eval Interval 3 7 14 21ARM Action Codes Trt Treatment Rate No. Name Rate Unit 2 5 8 11

1Untreated Check 2GLIPHO REF 45g ai/ha 0.0d 0.0f 0.0d 0.0g 3GLIPHO REF 90g ai/ha 0.0d 2.5f 2.5d 2.5g 4GLIPHO REF 180g ai/ha 0.0d 2.5f 7.5d 5.0g 5GLIPHO REF 360g ai/ha 2.5d 5.0f 32.5c 32.5f 6GLIPHO REF 720g ai/ha 12.5abc 45.0bc 52.5b 62.5d 7GLIPHO BB 45g ai/ha 2.5d 2.5f 5.0d 7.5g 8GLIPHO BB 90g ai/ha 2.5d 2.5f 5.0d 7.5g 9GLIPHO BB 180g ai/ha 7.5bcd 25.0d 40.0c 47.5e

10GLIPHO BB 360g ai/ha 12.5abc 42.5c 60.0b 90.0b 11GLIPHO BB 720g ai/ha 15.0ab 67.5a 98.8a 100.0a 12GLIPHO BC4 45g ai/ha 2.5d 2.5f 5.0d 5.0g 13GLIPHO BC4 90g ai/ha 5.0cd 5.0f 10.0d 10.0g 14GLIPHO BC4 180g ai/ha 7.5bcd 17.5e 40.0c 40.0ef 15GLIPHO BC4 360g ai/ha 15.0ab 50.0b 52.5b 80.0c 16GLIPHO BC4 720g ai/ha 20.0a 63.8a 98.8a 100.0a

LSD P=.05 7.07 6.80 8.82 8.78Standard Deviation 4.94 4.76 6.17 6.15CV 70.63 21.38 18.14 15.63 Treatment F 6.896 107.755 118.394 154.141Treatment Prob(F) 0.0001 0.0001 0.0001 0.0001

Pest Type W, Weed, G-BYRW7, G-WedStg = Weed or volunteer crop Pest Code ECHCG, Echinochloa crus-galli, = IE Part Assessed PLANT = plant C = Crop is Part Rated Assessment Type CONTRO = control / burndown or knockdown Assessment Unit % = percent Means followed by same letter do not significantly differ (P=.05, Duncan's New MRT) Mean comparisons performed only when AOV Treatment P(F) is significant at mean comparison OSL.

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Graph 5: Evaluation of efficacy of GLIPHO REF in comparison to BB and BC4 on ECHCG at 3 days after treatment

Graph 6: Evaluation of efficacy of GLIPHO REF in comparison to BB and BC4 on ECHCG at 21 days after treatment (at the end of the test)

0

20

40

60

80

100

45 90 180 360 720

% E

ffic

acy

Rate g a.i./ha

Efficacy on ECHCG - 3 DAT

GLIPHO REF

GLIPHO BB

GLIPHO BC4

0

20

40

60

80

100

45 90 180 360 720

% E

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Rate g a.i./ha

Efficacy on ECHCG - 21 DAT

GLIPHO REF

GLIPHO BB

GLIPHO BC4

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Photo 3: Efficacy on ECHCG (Echinochloa crus-galli)– 21DAA

UTC GLIPHO REF 360 g a.i/ha

GLIPHO REF 720 g a.i/ha

GLIPHO BB

360 g a.i/ha

GLIPHO BC4 360 g a.i/ha

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6.4 Conclusions The trial was carried out in greenhouse to evaluate the efficacy of GLIPHO REF in comparison to two new formulations of same active ingredient, signed GLIPHO BB and GLIPHO BC4. For each product were tested 5 rate (45,90,180,360 and 720 g a.i./ha) and 4 replicates per thesis. These products were applied on three different weeds:

• one dicotyledonous plant: AMARE (Amaranthus retroflexus) • two monocotyledonous plants: ECHCG (Echinochloa crus-galli) and ALOMY

(Alopecurus myosuroides) The application was performed when the pants were at growth stage (BBCH13-14: 3-4 leaves unfolded). At 3, 7, 14 and 21 days after application the plants were observed in order to monitor their conditions and assess the efficacy. After 3 days there no differences in terms of speed of action and efficacy between GLIPHO REF and the other product on AMARE but the action of REF on monocots (ALOMY and ECHCH) seems to be less noticeable than BB and BC4. After 21 days were clear the dose response of tested product on the different weeds:

• AMARE: GLIPHO REF seems to be better than BB and BC4 only at lowest rate (45 and 90 g a.i./ha), instead at highest rate there no difference in terms of efficacy.

• ALOMY: the efficacy profile of BB and BC4 was the same for each thesis. The performance of BB and BC4 was higher than REF at all rate tested.

• ECHCG: the efficacy profile of BB and BC4 was the same for each thesis. The performance of BB and BC4 was higher than REF at all rate tested.

In conclusion, the efficacy profile of BB and BC4 is slightly better than the GLIPHO REF.

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7. Appendix 7.1 Meteorological data

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

16.0

18.0

20.0

22.0

24.0

26.0

Typical Temperature Trend during the test

Greenhouse n° 1 - first test week