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Biotechnology: A tool towards a more

sustainable potato production?

September 1st 2015 Kick-Off Potato Europe, Doornik

Dr. ir. Bart Van Droogenbroeck

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Content

1. Introduction – Biotechnology? GMO?

– GM Crops

2. Challenges in potato production – Potato diseases

– Processing – Quality - Health

3. Recent developments − Disease resistance : Late blight

− Combination of mulitple traits

4. Conclusions

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1. INTRO - BIOTECHNOLOGY?

Definitions - basics

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Biotechnology? A definition Biotechnology studies and makes use of bacteria, plants, animals or other living organisms, or parts of it, to alter or produce food, materials, pharmaceuticals or other products of interest.

Initially classical biotechnology used traditional methods

to use plants and animals to produce products and use bacteria, yeast and fungi to produce e.g. bread, wine, cheese, beer

Modern biotechnology goes beyond these methods: it

applies the knowledge about characteristics of bacteria, plants and animals for the production of useful products by changing the DNA, the code of all genetic information.

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GMO?

• GMO= Genetically Modified Organism

• European definition of a ‘GMO’ , according to Dir 2001/18/EC:

“… means an organism, with the exception of human beings, in which the genetic material has been altered in a way that does not occur naturally by mating and/or natural recombination;.…”

Genetic material = DNA

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Modern biotech = DNA

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DNA Protein

The genes in our DNA encode for proteins

- Proteins execute all processes in cells/tissues off living creatures - Proteins or built with AA, 20 different ones - Genetic code determines which AA is encoded

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Examples of some fundamental techniques

1. Cutting and pasting DNA (1972)

Using enzymes as scissors (= restriction-enzymes) and paste (= ligases)

Biotechnology?

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Examples of some fundamental techniques

1. Cutting and pasting DNA (1972)

development of the first GMO, a GM bacteria

Biotechnology?

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Examples of some fundamental techniques

1. Cutting & pasting DNA (1972)

2. Reading DNA = sequencing (1977)

3. Amplification of DNA (1980)

Using enzyme (polymerase) to copy DNA

Biotechnology?

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Biotech Evolution 1982: First biotech medicin, insuline, produced by GM bacteria 1983: First GM plant, Gent, Prof. Van Montagu 1994: First GM tomato on the market, VS, FlavrSavr

2012: First commercial gene therapy (cancer)

2015: Synthetic biology – R&D ‘assembly’ of organisms

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1. INTRO – GM CROPS Methods & Global developments

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How are GM crops made?

both methods result in the random insertion of the DNA of interest in the host its genome

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How are GM crops made?

New Breeding Technologies Fast evolution – recently developed methods:

• Cisgenesis • Oligonucleotide Directed Mutagenesis (ODM) • Zinc Finger Nuclease Technology • Grafting • Agro-infiltration • RNA-dependent DNA methylation (RdDM) • Reverse breeding • Synthetic genomics

More sophisticated: • Specific integration in genome possible • Specific, targeted changes possible, down to 1 bp changes

Hardly or impossible to discriminate new genetically engineered organisms from wild type organisms

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Example of a recently developed method

Oligonucleotide Directed Mutagenesis

www.cibus.com

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Three generations of GM crops

First generation:

• Food crops– ‘input traits’ – advantages for growers

• GM crops in field today

(>180 mio ha, 18 mio landbouwers, 28 landen)

• Typical examples:

– ‘Bt’ crops - insect-resistance - Corn (borer), cotton bollworm, eggplant …

– ‘Roundup Ready’ crops - herbicide tolerance, soy, corn, sugarbeet

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First generation GM crops

19 consecutive years of growth Recently faster growth in developing world Important crops: soybean, maize, cotton, canola

http://www.isaaa.org

Mio Hectare

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http://www.isaaa.org

Mio Hectare

Most important trait = herbicide tolerance Increase of GM crops with combined traits

First generation GM crops

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http://www.isaaa.org

Production mainly in N&S- America and Asia Negligible in EU: 1 crop = 100.000 ha insect-resistant corn (Spanje,

Portugal, Tsjechië, Roemenië & Slovakije)

First generation GM crops

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Three generations of GM crops

First generation GM Potato products:

• Bt Potato – resistant to potato beetle (1995)

• Potato resistant to beetle + Potato Leaf Roll Virus (PLRV) - (1998)

• Potato resistant to beetle + PLRV + Potato Virus Y (PVY) – (1999)

All GE potato varieties were discontinued by the developer in March of 2001 and since have not been sold to farmers for planting

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Second generation:

• Food & non-food crops– output traits, advantage for grower/processor/consumer

• Typical example :

– ‘golden rice’ with increased vitA content – R&D phase

Three generations of GM crops

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Three generations of GM crops

Second generation GM Potato products:

• Amflora Potato – with altered starch composition, resulting in improved processing characteristics

BASF withdrew the product in 2012, citing opposition to the technology

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Three generations of GM crops

Third generation :

• Molecular Farming = the production of valuable proteins in crops specifically designed for this goal = plant as bioreactor

• Food/non-food crops – focus on advantages for consumer/patiënts

• First commercial products on market – lot of R&D – Therapeutic protein produced in cultures of carrot cells = cheaper

www.protalix.com

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2. CHALLENGES POTATO PRODUCTION

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Challenges Potato production

According to 2015 Beijing World Potato congress:

• Potato Mechanization • Water use • Integrated crop managment • Disease resistance • Varietal development • Storage and processing quality • Innovation in process & product development

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• Late blight is biggest problem

Challenges - Disease resistance

• Virus resistance: PVX, PVY, PLRV, … • Fungal resistance: Alternaria, … • Nematode resistance: potato cyst nematode, …

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Impact of late blight on Belgian potato production

• Caused by fungus Phytophthora infestans

• More virulent strains appearing since ’80

• 12-15 fungicide applications/season

(up to 20 in wet season)

• Economic loss in Belgium ≈ 55 mio euro – Fungicide applications

– Yield - and storage loss

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Resistant varieties are best solution

>20 R-genes known from wild relatives in Mexico, Andes

R&D projects in WUR (The Netherlands), TSL and JHI (UK), CRA-W (Belgium), CIP (Peru)…

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Late blight resistance in potato

Qualitative resistance based on gene-for-gene interaction

Phytophthora has avr genes Potato has/has not R genes

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Resistant varieties are best solution

• R-genen can be introduced in two ways :

Via conventional breeding:

• Crossing between two plants

• Selection on trait(s) of interest in offspring

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Conventional breeding takes time…

Faster and more targeted using biotechnology

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Why GM approach?

Biotech allows faster and more targeted improvement of crops

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GM approach is fast & targeted

• Introduction resistance in one step

• Preserves all other variety characteristics

• Introduction of multiple (R-)genes at the same time is possible

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Late blight resistance in potato Important ongoing Late blight Research projects • DurPh project (WUR- Netherlands): Durable Resistance against

Phytophthora through cisgenic marker-free modification – Cisgenic LB resistant Desirée potato development

– Classical and molecular Breeding – interaction with conventional and organic sector

• Gerephyti project (CRA-W, Belgium): Improving and managing potato resistance to Phytophthora infestans in order to develop ecologically intensive agriculture – Classical and molecular breeding approach

– R-gene discovery

– Cisgenic potato development

• Bintje+ project (UGent-VIB-ILVO) (visit ILVO booth for more info)

– Cisgenic approach to develop multiple R-gene LB resistant Bintje potatoes

• Company Simplot (US): Development of Innate potatoes – Cisgenic LB resistance and other traits in Russet Burbank

• TSL Potato Partnership Project (UK) – GM approach – combining multiple traits in Maris Piper potatoes

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Why cisgenic? • Cisgenic (vs. transgenic) = only DNA from crossable relatives –

conventional breeding

EFSA Opinion 2012: “…similar hazards can be associated with cisgenic and conventionally bred plants, while novel hazards can be associated with intragenic and transgenic plants.”

‘Higher’ public acceptance

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FIELD TRIAL WETTEREN

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Potatoes in the Wetteren Trial (2011-2012)

Wageningen UR – DuRPh: in Desirée background:

– sto1 R-gene lines (> S. stoloniferum)

– vnt1.1 R-gene lines (> S. venturii)

– sto1 + vnt1.1. + blb3 R-genen lines (> S. bulbocastanum)

‘Fortuna’ (BASF):

− blb1 +blb2 R- genes (> S. bulbocastanum)

WT Desirée and other reference varieties

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Trial set up (2011-2012)

= Maize

= Nicola

= Bintje

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Field trial Wetteren - 2011

Mix of Pi isolates from

Belgian fields

27 juli 2011

No Pi fungicide treatment

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3 aug 2011

GM lines with 1, 2 or 3 R-genen show resistance

Results field trial Wetteren - 2011

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Conventional varieties Nicola, Bintje, Fontane, Desiré & Agria are sensitive

3 Aug 2011

Results field trial Wetteren - 2011

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Organic varieties:

Bionica = sensitive

Sarpo Mira = resistant

3 aug 2011

Results field trial Wetteren - 2011

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Gevoelig Resistent

Results field trial Wetteren - 2012

Results 2012 confirm those observed in 2011: GM lines survive late blight without fungicide treatments

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Conclusions Field trial Wetteren

• Transformation of the susceptible potato variety Desiree with single or multiple late blight (R) resistance genes is possible.

• All selected events were significantly more resistant than Desiree wild type, both in field trials in The Netherlands and Belgium.

• Plants with single R genes showed a lower level of resistance than plants with R gene stacks.

• The events harbouring three late blight R genes were immune until the end of the growing season.

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Results field trial Wetteren - published

Available via http://www.sciencedirect.com/science/article/pii/S0261219415300739# or upon simple request to the authors

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Follow up project - Bintje+ (2014-2019)

• With support of Flemish Government, Boerenbond en Belgapom

• Development of cisgenic multiple late blight resistant Bintje = ~ 50 % of acreage in Belgium

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Current status Bintje+ project (2014-2019)

• First GG Bintje+ transformants selected in greenhouse

• Restistant lines selected by PCR and leaf assyas

• Field trials planned 2017-2018

BintjePLUS

UGent, ILVO en VIB binden de strijd aan tegen Phytophthora

Phytophthora is nog altijd de grootste plaag in onze aardappelteelt. UGent, ILVO en VIB leggen hun nieuwste kennis en expertise samen om dit probleem op een duurzame en robuuste manier te verhelpen. Ze zullen het populaire aardappelras Bintje tegen de ziekte wapenen door er meerdere natuurlijke resistentiegenen in te brengen.

Wat weten we? We weten welke de meest voorkomende en meest agressieve isolaten van Phytophthora zijn in onze streek. We weten uit de veldproef die we in Wetteren uitvoerden dat we resistentiegenen moeten stapelen voor een goede en duurzame resistentie. En we hebben vandaag een hele reeks resistentiegenen beschikbaar om dit te realiseren.

Wat gebeurt er nu? We introduceren met gentechnologie verschillende combinaties van natuurlijke resistentiegenen tegen Phytophthora in Bintje. Die genen worden vervolgens volgens de geldende standaarden op hun werkzaamheid getest. We kiezen de allerbest werkende combinatie van minimaal drie resistentiegenen om finaal zonder zogenoemde selectiemerker in Bintje binnen te brengen. Daardoor bezit BintjePLUS alleen extra genetisch materiaal afkomstig uit de aardappelfamilie. Hij bevat geen soortvreemd genetisch materiaal. Hij is met andere woorden cisgeen.

Het transformatieproces in beeld Van links naar rechts: (1) ongetransformeerde Bintje, (2) explantaten met callus, (3) overzetten op vers medium, (4) scheutjes, (5) geregenereerde transformanten

BintjePLUS; een project voor en samen met de Belgische aardappelsector

Foto rechts: de veldproef met gemodificeerde Désirée aardappelen in Wetteren toonde aan dat het stapelen van resistentiegenen goed werkt.

Beter voor mens en milieu? Het telen van resistente aardappelen is een veel betere manier om Phytophthora te lijf te gaan dan ze vroeg te rooien of ze met fungiciden te bespuiten. Aardappelen met een genetisch ingebouwde drie- of viervoudige resistentie behoeven niet of nauwelijks bescherming door fungiciden tegen Phytophthora. Dat is milieuwinst. Dit betekent ook een fikse besparing op de kosten voor fytoproducten. De teelt wordt gemakkelijker. De risico’s op opbrengstverliezen dalen sterk.

Waarom Bintje? Bintje is het belangrijkste ras in onze aardappelteelt. Het is een aardappel ‘van bij ons’, perfect voor het maken van echte frieten. Maar Bintje is ook heel gevoelig voor Phytophthora. Gentechnologie maakt het mogelijk om in één stap een duurzaam resistente Bintje te maken zonder verlies van zijn kwaliteiten.

De cisgene BintjePLUS aardappelen bevatten natuurlijke resistentiegenen tegen Phytophthora, afkomstig van wilde familieleden van onze cultuuraardappel.

De teelt van duurzaam resistente aardappelen zal het fungicidegebruik in deze aardappelen zeer fors doen dalen met milieuwinst en kostenbesparing in de gewasbescherming tot gevolg.

Behoud van de kwaliteiten van het Bintje ras. Daar gaat het om. Bintje is lekker, kan lang bewaard worden en heeft goede verwerkingseigenschappen.

Voor wanneer? Het ontwikkelen van Bintje aardappelen met een duurzame resistentie tegen Phytophthora kost tijd. De onderzoekers van UGent, ILVO en VIB hopen in 2017 een eerste veldproef met cisgene Bintjes te kunnen uitvoeren. Daarna duurt het nog een aantal jaren om er het BintjePLUS ras uit te ontwikkelen.

1 2 3 4 5

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Challenges (2) - Varietal development & storage and processing quality

• Cold storage = accumulation of reducing sugars

Upon high-temperature processing: reducing sugars react with free AA = brown colour, bitter taste + elevated levels of acrylamide (carcinogen)

• Reduced browning and black spots from bruising

During storage potatoes get bruised due to mechanical pressure + oxidation leads to brown colouring

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Recent developments in GM potato

• Commercial: - Simplot “Innate “potatoes - US :

• Innate 1.0 : Bruise resistant and lowered acrylamide levels - entry in to market

• Innate 2.0 : 1.0 + late blight resistance (1 gene) + cold storage traits

• Innate 3.0 : 2.0 + extra late blight resistance genes + PVY resistance

Eerste generatie Innate aardappelen VS - http://www.simplotplantsciences.com/

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Recent developments in GM potato

• R&D project- UK - US:

Research funding (2015 – 2019) to develop a new potato which could be better for the environment, healthier for consumers and cheaper to produce has been approved.

http://www.tsl.ac.uk/news/new-potato-at-the-sainsbury-laboratory/

Combination of 5 traits using 8 genes in Maris Piper: - Late blight resistance (3 genes ) +potato cyst nematode resistance (2 genes) + lower levels of reducing sugars and asparagine and lower bruise damage sensitivity (3 genes)

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BIOTECHNOLOGY: A TOOL TOWARDS A MORE SUSTAINABLE POTATO PRODUCTION?

Conclusies

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Conclusions

Conventional and organic potato breeding can benefit from biotechnology (molecular breeding, Pi monitoring) and GM approach to speed up classical breeding projects in the R&D phase (cfr DuRPh and Gerephyti project)

Soil management, crop rotation, responsible use of pesticides and disease resistances and other GAP are as important for biotech crops as for non-GM crops

Biotech/GM products that eventually appear on the (EU) market can absolutely be regarded as safe, both for humans, animals and the environment

Biotech development goes fast – regulatory approval procedures in EU are

lagging behind

GM potatoes are not the only and ultimate solution for a sustainable potato production, but could be part of it, together with best practices from conventional and organic farming

GM potatoes have the potential to contribute to a more sustainable production by using less surface, water, energy and pesticides

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Thank you for your attention!

Questions? Remarks?

Instituut voor Landbouw- en Visserijonderzoek

Burg. Van Gansberghelaan 115 9820 Merelbeke – België

T + 32 (0)9 272 28 00 F +32 (0)9 272 28 01

T&vat@ilvo.vlaanderen.be www.ilvo.vlaanderen.be

Contact: Dr. ir. Bart Van Droogenbroeck T + 32 (0)9 272 28 39 bart.vandroogenbroeck@ilvo.vlaanderen.be