New Plant Breeding Techniques Group 4 Others

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Barbara Welch
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WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES 12-13

Maria Lusser Joint Research Centre, European Commission Workshop

1 WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN New Plant Breeding Techniques Group 4 Others Maria Lusser Joint Research Centre, European Commission Workshop Comparative situation of new plant breeding techniques September 2011, Seville, Spain 1

2 Techniques WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 2 Grafting on GM rootstock Agro-infiltration Agro-infection

3 Grafting WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 3 Graft = chimeric organism consisting of above ground vegetative component of one plant (scion) attached to rootstock of another plant Grafting of non-gm scion onto GM rootstock Improved characteristics of rootstock (e.g. rooting capacity, resistance to soil-borne diseases) Change of the gene expression in the scion due to movement of specific proteins and/or RNA from the rootstock leading to new trait Tool for studying the movement of macromolecules in the plant and the silencing and expression of genes

Grafting WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES 12-13 SEPTEMBER

4 Grafting WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 4 Non-transgenic scion Transgenic rootstock Classification of fruits?

5 Agro-infiltration sensu stricto WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 5 Locally applied in non-germline tissues, mostly leaves Infiltration with suspension of Agrobacterium containing the desired genes Local, transient gene expression at high levels Screening (selection) for plants with valuable traits, e.g. pathogen resistance Use in research context, e.g. to study plant-pathogene interaction Production platform for high value recombinant proteins

SEPTEMBER 2011 SEVILLE, SPAIN 6 Classification of progeny?

6 Agro-infiltration and Agro-infection WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 6 Classification of progeny? Detached plant part infiltrated Intact plant locally infiltrated

7 Agro-infection or Agro-inoculation WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 7 Applied in non-germline tissues, mostly leaves Infiltration with construct containing the foreign gene in a full-length virus vector Facilitates the spreading and expression o the target gene in the whole plant

8 Commercial pipeline WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 8 Grafting Survey - crops and traits not disclosed research phase or phase I Workshop - crops are close (5 years) to release on the market PHASE I: Gene optimisation, crop transformation PHASE II: Trait development, pre-regulatory data, large-scale transformation PHASE III: Trait integration, field testing, regulatory data generation (if applicable) PHASE IV: Regulatory submission (if applicable), seed bulk-up, premarketing

9 Commercial pipeline WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 9 Grafting Notifications of field trials (corr. to phase III) - Grafting on GM rootstock: grape (resistance to grape fanleaf virus (GFLV) citrange (modified plant architecture) sweet orange (tolerance to Phytophtora citophtora) apple/pear (improved rooting ability) - Grafting of GM scion: apple (resistance to fungi)

10 Commercial pipeline WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 10 Agro-infiltration Survey - Research on crops such as potatoes (trait not disclosed), rape seed (trait not disclosed), and lettuce (test for resistance to downy mildew) Source: Lusser M. et al., New Plant breeding Techniques: State-of-the art and prospects for commercial development, JRC Technical Report EUR EN (2011)

11 Changes in the genome WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 11 Grafting on GM rootstock Changes in the genome are targeted to root tissue Intended manipulation of gene expression (new trait) in the scion can be achieved through movement of specific proteins and/or RNA from roots to scion, e.g. RdDM Possible unintended changes: - genetic exchange restricted to graft site - unintended manipulation of gene expression through movement of specific proteins and/or RNA from roots to scion

12 Changes in the genome WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 12 Agro-infiltration and Agro-infection Temporary expression of specific coding sequences No DNA integration in the plant genome Possible unintended changes: Agro-infiltration: - Integration of T-DNA cannot be excluded Agro-infection: - RNA viruses cause spreading of the gene construct - Since spread via RNA molecules, no integration in plant genome

13 Changes in the genome WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 13 Source: Glandorf B. et al., Evaluation of changes in the genome in plants through application of new plant breeding techniques in JRC Technical Report New plant Breeding Techniques, EUR EN

14 Detection/Identification WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 14 Grafting In the whole plant including the GM rootstock, the transformation can be detected and identified with PCR-methods RNA molecules, proteins and metabolites moving to the non-gm scion lead to modified transcriptom (however, analysis currently technically not feasible) DNA sequence of fruits from non-gm scion is not modified and detection and identification through PCR consequently not possible

15 Detection/Identification WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 15 Agro-infiltration and Agro-infection During the experimental phase, transiently present DNA fragments can be detected by PCR T-DNA is not inserted in germline tissue Progeny of the infiltrated plant does not carry the transformation. Detection and identification not possible in the progeny. Source: New plant breeding techniques: Challenges for detection and identification report of the New Techniques Task force (NTTF) in JRC Technical Report New plant Breeding Techniques, EUR EN (2011)

16 WORKSHOP COMPERATIVE SITUATION OF NEW PLANT BREEDING TECHNIQUES SEPTEMBER 2011 SEVILLE, SPAIN 16 Copyright: European Union, 2011

Plant Breeding and Propagation

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