PAG-IX: APPLICATION OF MULTIPLEX SEQUENCE-TAGGED SITE MICROSATELLITES (STSMS) FOR DISTINCTNESS, UNIFORMITY AND STABILITY TESTING IN <I>BRASSICA NAPUS</I>

PAG-IX   Plant & Animal Genome IX Conference

Town & Country Hotel, San Diego, CA, January 13-17, 2001.

Workshop: Brassicas
W40

APPLICATION OF MULTIPLEX SEQUENCE-TAGGED SITE MICROSATELLITES (STSMS) FOR DISTINCTNESS, UNIFORMITY AND STABILITY TESTING IN BRASSICA NAPUS

LIVIA TOMMASINI1, Jacqueline Batley1, Robert J Cooke2, David Lee2, Chris Lowe2, Paolo Donini2, Carolyn Moule3, Martin Trick3, K. J. Edwards1

1 Crop Genetics Department, IACR-Long Ashton Research Station, University of Bristol, Long Ashton, Bristol BS41 9AF, UK.
2 National Institute of Agricultural Botany, Huntingdon Road, Cambridge CB3 OLE, UK.
3 Brassica and Oilseed Research Department, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK.

Newly bred varieties of crops must undergo distinctness, uniformity and stability testing to determine their eligibility for inclusion in the National List of Varieties before they can be marketed within the European Union. This testing constitutes the basis for intellectual property rights. Currently, phenotypic characters are used in DUS testing. However, the potential of STSMs is proving useful for variety testing. For the wider application and acceptability of molecular markers in DUS testing, it is essential to develop high throughput marker kits that can be used successfully irrespectively of the technology used. In this project we have developed multiplex STSMs sets for maximising the efficiency of variety testing. 15 STSMs loci were analysed in 10 varieties of Brassica napus. The potential of multiplex and single primer sets to test distinctness between varieties was assessed by cluster analysis and expressed using separation coefficient rates. All the varieties examined could be uniquely identified by combining the information of the 2 most polymorphic markers and by using STSMs data from single multiplex sets. Cluster and PCO analyses produced from 3 multiplex STSMs data brought together spring oilseed rape types in one group, winter oilseed rape types in another group and the forage rape variety in a third group. There is a suggestion that varieties linked by pedigree were grouped by PCO and cluster analysis, indicating that results from STSMs data reflect genetic relationships between varieties. Uniformity of variety profiles was evaluated. Preliminary data showed that some varieties were non-uniform at specific loci.

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