Use of Biochemical and Molecular Techniques in the DUS Examination and Cooperation between Relevant International Organizations

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1 ISTA Congress 2013 / Discussion Forum Discussion on the use and International standardization of DNA technologies in the area of seeds Use of Biochemical and Molecular Techniques in the DUS Examination and Cooperation between Relevant International Organizations Fumi Aihara Counsellor, UPOV Antalya, Turkey June 17,

2 1. Introduction to UPOV and the role of UPOV Technical Working Parties (TWPs) Overview 2. Overview of the DUS examination 3. Use of biochemical and molecular techniques in the DUS examination 4. Cooperation between relevant international organizations

3 1. Introduction to UPOV and the role of UPOV Technical Working Parties (TWPs) Overview 2. Overview of the DUS examination 3. Use of biochemical and molecular techniques in the DUS examination 4. Cooperation between relevant international organizations

4 UPOV Structure COUNCIL CONSULTATIVE COMMITTEE ENLARGED EDITORIAL COMMITTEE TECHNICAL COMMITTEE (TC) ADMINISTRATIVE AND LEGAL COMMITTEE (CAJ) Technical Working Party on Automation and Computer Programs (TWC) Technical Working Party for Agricultural Crops (TWA) Technical Working Party for Fruit Crops (TWF) Technical Working Party for Ornamental Plants and Forest Trees (TWO) Technical Working Party for Vegetables (TWV) Working Group on Biochemical and Molecular Techniques (BMT)

5 UPOV Structure Technical Working Party on Automation and Computer Programs Technical Working Party for Agricultural Crops Technical Working Party for Fruit Crops Technical Working Party for Ornamental Plants and Forest Trees Technical Working Party for Vegetables Working Group on Biochemical and Molecular Techniques Test Guidelines

6 UPOV Structure Technical Working Party on Automation and Computer Programs Technical Working Party for Agricultural Crops Technical Working Party for Fruit Crops Technical Working Party for Ornamental Plants and Forest Trees Technical Working Party for Vegetables Working Group on Biochemical and Molecular Techniques

7 Overview 1. Introduction to UPOV and the role of UPOV Technical Working Parties (TWPs) 2. Overview of the DUS examination 3. Use of biochemical and molecular techniques in the DUS examination 4. Cooperation between relevant international organizations

8 CONDITIONS FOR GRANTING A BREEDER S RIGHT Criteria to be satisfied NOVELTY DISTINCTNESS UNIFORMITY STABILITY DUS 8

9 DISTINCTNESS e.x. Apple: Fruit color UNIFORMITY A uniform variety A variety lacking uniformity e.x. Apple: Flower bud color STABILITY Stable variety The relevant characteristics of the variety do not change through the generations. Original material Generation 1 Generation 2 Generation N 9

10 Nature of the DUS Examination The DUS Test (field trial) 10

11 Molecular Techniques?

12 Overview 1. Introduction to UPOV and the role of UPOV Technical Working Parties (TWPs) 2. Overview of the DUS examination 3. Use of biochemical and molecular techniques in the DUS examination 4. Cooperation between relevant international organizations

13 Relevant UPOV documents and their status (1) UPOV/INF/17/1 (INFormation document) Guidelines for DNA Profiling: Molecular Marker Selection and Database Construction ( BMT Guidelines ) The purpose of this document (BMT Guidelines) is to provide guidance for developing harmonized methodologies with the aim of generating high quality molecular data for a range of applications. The BMT Guidelines are also intended to address the construction of databases containing molecular profiles of plant varieties [ ] UPOV/INF/18/1 (INFormation document) Possible Use of Molecular Markers in the Examination of Distinctness, Uniformity and Stability The purpose of this document is to provide guidance on the possible use of biochemical and molecular markers in the examination of Distinctness, Uniformity and Stability (DUS). [ ] Both documents have been adopted and publicized.

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16 UPOV/INF/18 POSSIBLE APPLICATION MODELS MODELS WITH A POSITIVE ASSESSMENT Characteristic-specific molecular markers Combining phenotypic and molecular distances in the management of variety collections Calibrated molecular distances in the management of variety collections MODELS WITHOUT A POSITIVE ASSESSMENT Use of molecular marker characteristics

17 Relevant UPOV documents and their status (2) TGP/15/1 Draft 4 (Technical Guidelines Protocol) Guidance on the Use of Biochemical and Molecular Markers in the Examination of Distinctness, Uniformity and Stability (DUS) [The purpose of this document is to provide guidance on the use of biochemical and molecular markers in the examination of Distinctness, Uniformity and Stability (DUS) on the basis of the models in document UPOV/INF/18 that have received a positive assessment and for which accepted examples have been provided.] The TC and CAJ have agreed with the latest draft to be discussed by the Council in October, 2013.

18 TGP/15 Guidance for the Examination of DUS Characteristic-specific molecular markers Combining phenotypic and molecular distances in the management of variety collections

19 APPLICATION MODELS (TGP/15/1 Draft 4) 1. Characteristic-specific molecular markers Molecular markers can be used as a method of examining DUS characteristics that satisfy the criteria for characteristics set out in the General Instruction, [ ] on the following basis: (a) the test for the marker is conducted on the same number of individual plants, with the same criteria for distinctness, uniformity and stability as for the examination of the characteristic by a bioassay; (b) there is verification of the reliability of the link between the marker and the characteristic; (c) different markers for the same characteristic are different methods for examining the same characteristic; (d) markers linked to different genes conferring expression of the same characteristic are different methods for examining the same characteristic; and (e) markers linked to different regulatory elements for the same gene conferring expression of the same characteristic are different methods for examining the same characteristic. 19

20 General Instruction Selection of characteristics The basic requirements that a characteristic should fulfill before it is used for DUS testing or producing a variety description are that its expression (TG/1/3: Section 4.2.1) : (a) results from a given genotype or combination of genotypes; (b) is sufficiently consistent and repeatable in a particular environment; (c) exhibits sufficient variation between varieties to be able to establish distinctness; (d) is capable of precise definition and recognition; (e) allows uniformity requirements to be fulfilled; (f) allows stability requirements to be fulfilled, meaning that it produces consistent and repeatable results after repeated propagation or, where appropriate, at the end of each cycle of 20 propagation.

21 1. Characteristic-specific molecular markers Molecular markers can be used as a method of examining DUS characteristics [ ] q Application in DUS testing (example of France) Characteristic-specific markers Mr. Joel Guiard Technical Committee March 20, 2013 Detection of gene Hs1 pro-1 controlling resistance to nematode Heterodera Schachtii in Sugar beet varieties (routinely used) Detection of genes Tm1, Tm2 and Tm2 2 controlling resistance to mosaic virus in Tomato varieties (supporting tool in case of doubt) Detection of genes controlling resistance to Meloidogyne incognita, Verticillium and Fusarium oxysporum in Tomato and resistance to mosaic virus in Lettuce (still under development) 21

22 APPLICATION MODELS (TGP/15/1 Draft 4) 2. Combining phenotypic and molecular distances in the management of variety collections A combination of phenotypic differences and molecular distances can be used to identify within the variety collection, those varieties which need to be compared with candidate varieties in order to improve the selection of Distinct plus varieties, on the following basis: (a) (b) (c) there is reliable information that the molecular distances are sufficiently related to phenotypic differences, such that the method selects varieties in the variety collection which are similar to the candidate varieties; and the method does not create an increased risk of not selecting a variety in the variety collection which needs to be compared to the candidate varieties in the field. 22

23 2. Combining phenotypic and molecular distances in the management of variety collections A combination of phenotypic differences and molecular distances can be used to identify within the variety collection [ ] q Application in DUS testing (example of France) This model is routinely used by GEVES on reference collections of: Mr. Joel Guiard Technical Committee March 20, 2013 Maize inbred lines Barley Lettuce Under development for other species Benefit: 20 to 40 % of reduction of reference varieties in the DUS trial except for Lettuce New development: use of SNP markers in case of Maize 23

24 Importance of Harmonized Approach within UPOV To facilitate cooperation in DUS testing e.g. purchase of DUS reports To establish internationally recognized variety descriptions (effective protection) 24

25 1. Introduction to UPOV and the role of UPOV Technical Working Parties (TWPs) Overview 2. Overview of the DUS examination 3. Use of biochemical and molecular techniques in the DUS examination 4. Cooperation between relevant international organizations

26 The following international organizations including UPOV and ISTA are working on the closely-related topics (molecular techniques applied for plants). UPOV: Techniques used in the DUS examination for Plant Breeder Rights (PBRs) ISTA: Laboratory protocols for seed testing ISO: International standardization of testing protocols OECD: The International Union for the Protection of New Varieties of Plants The International Seed Testing Association The International Organization for Standardization The Organization for Economic Co-operation and Development To provide international framework for the certification of high quality seeds

27 Coordination between International Organizations The UPOV Technical Committee (49 th session, March 20, 2013) received the following presentations from the relevant international organizations. Situation with regard to use of molecular techniques in relation to seeds in the International Organization for Standardization Mr. Michael Sussman (ISO) * Presented by Mr. Paul Zankowski (United States of America) Situation with regard to use of molecular techniques in the International Seed Association Ms. Rita Zecchinelli (ISTA) Situation with regard to use of molecular techniques in the Organisation for Economic Co-operation and Development Mr. Michael Ryan (OECD) (presentation slides available at: 27

28 Role of the Working Group on Biochemical and Molecular Techniques (BMT) of UPOV The BMT is a group open to DUS experts, biochemical and molecular specialists and plant breeders, whose role is to: (i) Review general developments in biochemical and molecular techniques; (ii) Maintain an awareness of relevant applications of biochemical and molecular techniques in plant breeding; (iii) Consider the possible application of biochemical and molecular techniques in DUS testing and report its considerations to the TC; (iv) If appropriate, establish guidelines for biochemical and molecular methodologies and their harmonization [ ]; (v) Consider initiatives from TWPs, for the establishment of crop specific subgroups [ ]; (vi) Develop guidelines regarding the management and harmonization of databases of biochemical and molecular information, in conjunction with the TWC; (vii) Receive reports from Crop Subgroups and the BMT Review Group; (viii) Provide a forum for discussion on the use of biochemical and molecular techniques in the consideration of essential derivation and variety identification.

29 The forty-ninth session of the Technical Committee of UPOV (TC/49) v The TC endorsed the initiative for a joint meeting with ISO, ISTA and OECD and including breeders, as a means of supporting the role of the BMT in relation to (i), (ii), (iv), (vi) and particularly (viii) above. v The TC agreed to propose to hold a coordinated meeting of the fourteenth session of the BMT (BMT/14) with meetings of other relevant international organizations in 2014, [ ] v The TC had previous approved the program for the fourteenth session of the BMT, including the dedication of a particular date ( Breeders Day ), for the items on the use of molecular techniques. 29

30 Thank you for your attention. Please visit UPOV website at: for more information.