www.eurofins.com Setting Standards

GMO –

Update on essential Topics for the Food and Feed Sector

Ulf Rathjens 08.10.2019

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Setting Standards

GMOs – Overview

Gene Promoter Terminator

Virus Plant /Bacterium Bacterium

Definition of a Genetically Modified Organism?

WHO „GMO: Organism whose genetic material (DNA) has been modified in a way that does not occur naturally”

Genetic engineering

? – Chemical mutagenesis

? – Cytoplasmatic male sterility (CMS)

? – Gene-editing techniques (e.g. CRISPR/Cas)

……

Discussions are ongoing

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Setting Standards

GMOs – Overview

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Genetically Modified Organisms – GMOs

Food / feed production

Plants Animals Bacteria Fungi Viruses

Commercialised genetically modified organisms/viruses

Approved for Commercialisation

Virus Picture: by Magnus Manske, http://en.wikipedia.org/wiki/User:Magnus_Manske; Fungi Picture: by Peter G Werner, http://en.wikipedia.org/wiki/User:Peter_G_Werner

Ornamental organisms

Setting Standards

Commercial Planting of GMOs

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Increase of GM Crops in Countries with Highest Production 1996-2018

• ˃ 90% of global planting area in USA, Brazil, Argentina, India, Canada, China (177,4 M ha out of a total of 191,7 M ha)

• EU:

• Only MON810 corn approved for commercial planting

• Most EU member states prohibited GMO planting

• 2018 Planting in Spain (0,12 M ha), Portugal

0

20

40

60

80

100

120

140

160

180

200

96 98 00 02 04 06 08 10 12 14 16 18

M ha

year

USA Brazil Argentina India Canada China

Setting Standards

Commercial Planting of GMOs

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Global Adoption Rates (%) for Principal Biotech Crops, 2018: by Crop (Million Hectares)

• >99% of the global GM planting area is shared by four crops: Maize, Soybean, Cotton, Canola

• In addition, regional commercial planting of GMOs from other plants • Sugarbeet, Alfalfa, Papaya,

Zucchini (Squash), Tomato, Carnation, Eggplant, Potato, Apple, Pineapple etc.

• Accidental release of GMO, contamination of seeds in the past years: • Maize, Rice, Linseed, Papaya,

Petunia

Figures based on ISAAA Brief 54: Global Status of Commercialized of Biotech/Crops 2018

0

50

100

150

200

Maize Soy Canola Cotton

M ha

Crop Type

non-GMGM

Setting Standards

Commercial Planting of GMOs

Please do not hesitate to contact us for our actual customer information on GM-Soy, GM-Corn, GM-Canola, which are updated in regular intervals and free of charge

UlfRathjens@eurofins.de

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Setting Standards

Detection Methods: PCR

GMO Testing Procedure at Eurofins GeneScan

1 2

Extraction control

1 2

Samples

Homogenisation control

Weigh-In control

Duplicate Extraction

Quality control of extracted DNA Type (high molecular / low molecular)

and amount PCR

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Setting Standards

Detection Methods: PCR

The Polymerase Chain Reaction (PCR)

1st cycle: 1 molecule 2nd cycle: 2 molecules

3rd cycle: 4 molecules 10th cycle: 512 molecules

20th cycle: 524,288 molecules 40th cycle: 5.5 *1011 molecules

50th cycle: 5.6 *1014 molecules

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Setting Standards

Detection Methods: PCR

ISO 24276 Eurofins GeneScan Duplicate Extraction Required Standard Procedure

Negative extraction control Per extraction series Standard Control

Positive extraction control In regular time intervals Standard Control

Positive DNA target control Required Standard Control

PCR reagent control Required Standard Control

PCR inhibition control Recommended / required Standard Control

Evaluation of raw data

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Setting Standards

Detection Methods: PCR Elements of a GMO Testing Strategy

• Screening methods: Detection of wide-spread elements used in plant transformation / PCR targets present in a large number of GMO (e.g. 35S promoter / NOS terminator / AgroBorder)

• Identification:

Modification-specific detection / PCR targets presents in a little number of GMO Event-specific detection (targeting junctions of DNA elements) / PCR targets presents in one specific GMO, only

• Quantification: Determination of relative GMO content by comparing GMO DNA content with total species DNA content

• Plant species detection / quantification: Detection of a species-specific gene, important i.e. in the case of botanical impurities in the sample

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Setting Standards

Detection Methods: PCR GMO Testing: Basic Approach

GMO content not expected in the sample

Qualitative Screening for the presence of GMOs

Identification of GMOs

Quantification of the GMO content

e.g. exclusion of non-approved GMO, depending on risk assessment of possible presence, or identification of approved GMO

check for labelling requirements

scope of screening depends on ingredients

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Setting Standards

Detection Methods: PCR GMO Testing: Basic Approach

GMO content expected in the sample

Qualitative screening for the presence of unexpected /

unapproved GMOs

Identification of GMOs

Quantification of the GMO content

Direct quantification of expected GMOs

plus

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Setting Standards

Broad and Cost Efficient Testing

customer demand

special requirements e.g. labelling

potential botanical impurities

formulation / raw materials

accidental release of

GMOs

Das Bild kann zurzeit nicht angezeigt werden.

Das Bild kann zurzeit nicht angezeigt werden.

legal framework in

country of destination

commercial planting

in the country of origin

Selection of testing strategy

Combination

of PCR targets

should cover relevant

GMOs

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Setting Standards

• Planted in more than 8 countries

• Global planting area 95.9 M ha

• 15 approved events in the EU (food / feed)

• 1 with 0.1% tolerance EU (feed)

• 2 pending EU approvals

• > 30 different events described

• 10 events commercially planted as single or stacked event in any country worldwide

(Status: 20.09.2019)

Facts & Figures – GM Soybean

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Setting Standards

Facts & Figures – GM Soybean

Planting of GM soybean – GM and non-GM planting area (Countries with more than 1 M ha GM soybean planting, 2018)

0

5

10

15

20

25

30

35

40

USA BrazilArgentina Paraguay Canada

Uruguay Bolivia

M ha

Country

non-GM

GM

(Status: 20.09.2019)

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Setting Standards

Ukraine • “Rumours” on illegal planting: GM soy 40 – 70% of soy

planting area • USDA GAIN Report UP1725; 11/16/2017: “60 – 70

percent of soybeans […] produced for export is genetically engineered”

Romania • Commercial planting of GM soy before EU membership

(2006: ~ 100.000 ha) • Still positive findings

….. Is GM soy grown in my sourcing region? Commercially / illegal? No growing but import of GM-soy? Which GM-soy event is of risk in detail (grown, imported)? ….

commercial planting

in the country of origin

Facts & Figures – GM Soybean

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PCR Screening for GM Soybean – 1/2 (extract)

„….detected by PCR on …“: GM-Soy Plant.(1) 35S NOS ABII CsVMV CV127 305423

GTS 40-3-2 + + +

MON89788 + +

MON87701 + +

MON87708 + +

MON87705 + +

A2704-12 + +

A5547-127 + +

CV127 + +

FG72 + +

MON87769 + +

(1): commercial planting as single or stacked event within the last 10 years in any country worldwide

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„SoyScreen“: •35S-promoter •nos-terminator •AgroBorder II-element •CsVMV modification •event CV127 •event 305423

GM Soybean – Detection

Setting Standards

Setting Standards

PCR Screening for GM Soybean – 2/2 (extract)

„SoyScreen“: •35S-promoter •nos-terminator •AgroBorder II-element •CsVMV modification •event CV127 •event 305423

„….detected by PCR on …“: GMO Plant.(1) 35S NOS ABII CSVMV CV127 305423

305423 + +

356043 +

DAS-68416-4 +

DAS-44406-6 + +

DAS-81419-2 +

MON87751 +

SYHT02 + +

MON87712 + +

A2704-21 + G94-19, G-168, G94-6, G94-1 + +

A5547-35 + +

HB4 +

DBN-09004-6 +

GMB151 +

GM Soybean – Detection

(1): commercial planting as single or stacked event within the last 10 years in any country worldwide

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CONFIDENTIAL AND PROPRIETARY - © Eurofins Scientific (Ireland) Ltd, 2015. All rights reserved. Any use of this material without specific permission of an authorised representative of Eurofins Scientific SE is strictly prohibited.

GM Soybean - Detection

Screening Coverage of Commercialised GM Soybean Events

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45,4

5%

63,6

4%

81,8

2%

100,

00%

0%

20%

40%

60%

80%

100%

positive negative Recommended Soy Screening: • 35S-promoter • nos-terminator • AgroBorder II-element • CsVMV modification • event CV127 • event 305423

Eurofins Firewall Seminar: GMO - Update on essential Topics

Setting Standards

GM Corn - Detection

Screening Coverage of Commercialised GM Corn Events 95

,65%

95,6

5%

95,6

5%

100,

00%

0%

20%

40%

60%

80%

100%

positive negative

„CornScreen“: •35S-promoter •nos-terminator •AgroBorder II-element

•event DAS-40278-9 •event VCO01981-5 •LY038 modification

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Setting Standards

AquAdvantage®-Salmon – the First GM Animal Approved for Food

• Developed by AquaBounty Technologies

• Approved for food in Canada and the US

• Sold for human consumption

in Canada in 2017 / 18

Assay for the detection of genetically modified salmon available

Excursus - Genetically Modified Salmon

GMOs – Update

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Setting Standards

Thank you for your attention

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Setting Standards

Contact

Eurofins GeneScan Engesserstrasse 4 79108 Freiburg Germany T: +49 171 8154583 E: UlfRathjens@eurofins.de

Ulf Rathjens

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