Genetic improvement of grapevine: WHY ? Melané A Vivier Institute for Wine Biotechnology Stellenbosch University

Why am I here trying to answer this question? Because we, together with a large international community, supported by numerous industries and other funding agencies, are trying to ask and answer scientific questions regarding the most important fruit crop of the world, namely Vitis vinifera (grapevine) We focus on understanding the genetic potential of the species, using molecular and cellular biology tools We have developed (bio)technologies that can lead to genetically improved grapevines, aligned with industry needs We have applied for an experimental field trial with transgenic grapes, which sparked much debate

Media reports GM Grapes earn wrath of growers (Sunday Times) SA Scientists seek go-ahead for field trials of GM grapes (Cape Times) Wingerdinstituut se GM proef sal ‘n eerste wees (Die Burger) GM grape trials could affect SA wine sales (Farmer’s Weekly, letter) Frankenfoods’ from a madman’s lab. Gene grapes no barrel of fun (Son) GM wingerdplante ontlok reaksie (Landbou Burger) No to GMO in the wine industry (WOSA) Protests at Stellenbosch transgenic grapevine experiment (Grape news) Genetically modified wine on the cards (Cape Times)

So, are we interested in “contaminating” the environment, creating “weedy” grapevines or causing harm to wine-drinking customers, or the wine-industry in general? Aims of the programme To know more about grapevine and how it “works”…… To find solutions to support environmentally friendly production…..less pesticides  To find solutions to the impact of the changing climate…..stress-tolerance  To find novel ways to improve quality and wholesomeness

Towards environmentally friendly production and stress tolerance….. A grape gene (Vvpgip), present in multiple copies protects against fungal pathogens A grape gene (VvNCED), protects against water stress

The process involved to generate a transgenic grapevine and the evaluations needed Photographs obtained from the work of Dr. Khrishnan Vasanth, IWBT Somatic embrogenesis Cryopreservation Genetic transformation Regeneration

GRAPEVINE TISSUE CULTURE GRAPEVINE TRANSFORMATION EVALUATION OF TRANSGENICS The process and evaluations

The specifics about the contained experimental vineyard Why did we apply? –To obtain a mature, fruit-bearing GMO vineyard for scientific purposes What would it contain and where will it be? –Grapevine plants with inserted reporter and antibiotic genes (the latter as selectable marker) alongside non-transgenic grapevines –The transgenic material will be grafted on non-transgenic rootstocks on the Welgevallen experimental farm of Stellenbosch University Objectives? –To assess the reporter gene activity over several seasons and to assess the agronomical performance of the transgenic material alongside conventional grapevines

On Welgevallen experimental farm 930 m 2, fenced, restricted admission 238 US Vit 8-7 (NON-TRANSGENIC) Rootstocks planted – 100 used for first trial Virus-free (molecular analyses) Trellis and irrigation systems for Chardonnay and Sultana established Structure for super fine netting to cover trial Normal viticultural practices The experimental vineyard

Transgenic grapevine plants 100 plants in first round trial –6 Lines Chardonnay, 6 lines Sultana, 4 controls of each –5 plants of each line Random block design Monitoring: at least five seasons Management plan included

Public notification Public notification with details of proposed field trial: –IWBT and DVO staff and students –SU Research Development SU management Ethical committee –IWBT Website –SU Bulletin –Winetech and DFPT – to industry role players –Local media: Eikestadnuus Die Burger Cape Argus The wording of the notification was not optimal…..

Reaction to public notification 2 Official objections within allowed period (Biowatch and African Centre for Biosafety); 1 Subsequent objection from Biowatch Several messages from the public Replies to all official objections, a further press release in collaboration with SA Wine Industry and significant correspondence Interviews with the media: –Newspapers –Electronic journals / newsletters –Magazines –Radio stations (SABC) Not factual reporting in all cases!

General concerns General aspects: 1.General anti-GMO 2.Environmental contamination / Biosafety 3.Antibiotic Resistance Genes: nptII Wine-industry specific aspects: 1.Activities done without support of SA wine industry 2.Impact of the possible commercialisation of GM grapevine and wine

Understanding the specifics about Vitis will clarify many misconceptions Grapevines are not indigenous to South Africa –The first Vitis species arrived with the settlers! Grapevines have no close relatives in South Africa, meaning that there is no plant species with which it could interbreed

Misconceptions……… Pollen flow and seed dispersal Domesticated grapevine flowers are functionally bisexual and largely self- pollinated Grapevine is not planted from seed; it is vegetatively propagated –In SA certified plant material is strictly controlled in the supply chain

Misconceptions………. Seed dispersal and seedling volunteers Grapevines are planted in “manicured” vineyards which are intensively worked Seedling volunteers are scarce, very visible and routinely removed through normal viticultural practices

Understanding the specifics about Vitis will clarify many misconceptions The concept of “true-to-typeness” (in viticultural terms)…… –In the wine industry, the varietal names and characteristics are important marketing attributes –The viticultural performance, as well as the varietal characteristics in the vineyard and in the resulting wine are important factors to consider –This has a direct impact on the prospect of GMO vines and wines and therefore needs to be assessed rigorously Scientific advances provides the possibility to also perform genetic and transcriptomic profiles of GMO vines in comparison with non-transgenic vines

Environmental contamination Possible release of transgenes into the environment –Pollination of grapevine: largely by self-pollination, no wild relatives of grapevine in SA, no related plant species to hybridise Flowers in the trial will be covered with bags –Seed dispersal: Seedlings do not normally survive in uncultivated habitats; Volunteer seedlings controlled by routine viticultural practices. The trial site will be covered with a superfine net and fruit will be bagged –Soil contamination: The transgenic plants will be grafted on non-transgenic rootstocks, frequency of horizontal gene transfer from plants to microorganisms very low under normal circumstances, transgenes already present in very large amounts in environment, transgenes do not give competitive advantage. Rigorous management of fallen leaves and other plant material; Environmental impact monitored General concerns

Perspectives The challenge is perhaps to strike a balance between strict control and monitoring of GMO materials and their release, while not hampering the progress of science Thank you