14 I EUROPEAN SEED I EUROPEAN-SEED.COM a variety is treated differently than a vegetatively propagated crop for which all plants belonging to one variety have identical genotypes. This information will have an effect on the number of samples to be analysed. After a final research plan is completed, sampling of all plant material to be included is done. It is of crucial importance that the sampling should be independent. To function as legal proof, a bailiff might be involved to take the suspected samples at the potential infringers growing site. For the genetic analysis two main questions must be considered: 1. How many markers are needed (which DNA technology is best to use)? and, 2. Which reference framework is relevant (which con- trol samples should be included into the test to compare with)? To answer this last question, Naktuinbouw developed some guidelines for the selection of samples in a Variety Tracer anal- ysis. Samples should be representative for the diversity within the species (all relevant morphological characteristics should be present in unbiased way). Some closely related samples should be included. This can be parental lines with seedlings and, when present, known and accepted mutant varieties with the original variety. Choices on which technology to use highly depend on the crop involved and on what information is publically available. For many species sequence information and crop-specific SNP or SSR markers are available. When no DNA information is available a random marker technology like AFLP can be applied. The interest in variety identity tests increases; the number of Variety Tracer projects grows every year. (85 projects in 2017). The projects vary from simple identity questions to detect mix- ups to complicated infringement cases involving many parties. In nearly all infringement cases that were subject of the specialized PBR court in The Hague, Naktuinbouw was involved as independ- ent expert and the Variety Tracer report served as evidence. FUTURE PERSPECTIVE – TECHNOLOGICAL DEVELOPMENTS With the release of Next Generation Sequencing (NGS) tech- nology (in 2005), a real sequencing revolution has taken place. NGS machines enable massive parallel sequencing, increas- ing sequencing data output for decreasing costs. NGS is often referred to as High Through-put Sequencing (HTS). To increase the resolution and distinguishable power of DNA markers used in Variety Tracer projects, Naktuinbouw now invests to replace the old technologies like AFLP and SSR by SNP genotyping based on High Through-put Sequencing. So, Variety Tracer 2.0 is currently developed. There are three phases in the process of developing a SNP genotyping method for a particular species: 1. the identifica- tion of SNPs and selection of a SNP panel that is representative for the genetic diversity within the species; 2. development of a (sequence based) method to determine the SNP genotype of multiple varieties in a high throughput and cost effective way; 3. storage of all SNP genotypes in databases that can be used for variety identification purposes including infringements. Using these new technologies will enable Naktuinbouw to increase the accuracy and reliability of the genetic research for a lower price. Variety Tracer 2.0 will be better and cheaper to be applied for a plant world without piracy. Editors’ Note: Hedwich Teunissen is Molecular Biologist at Naktuinbouw In a Variety Tracer project often a combination of morphological side-by-side growing trials and genetic research is applied. “Putting in place good regulations and a strong IP rights system for plants cannot rule out piracy in the world of plants.” – Hedwich Teunissen