Articles
MICROSATELLITE-BASED FINGERPRINTING OF WESTERN BLACKBERRIES FROM PLANTS, IQF BERRIES AND PUREE
Article number
859_6
Pages
73 – 80
Language
English
Abstract
The blackberry industry needs a reliable method to ensure trueness-to-type of blackberry products.
Microsatellite markers or simple sequence repeats (SSRs) are ideal for cultivar fingerprinting, paternity testing and identity certification.
Fingerprinting is valuable for variety identification, quality control and as a legal method to protect against infringement by competitors.
The objectives of this study were to develop a DNA extraction protocol and SSR-based identification for individually quick-frozen (IQF) Marion and Kotata whole berries and concentrate and to generate genetic fingerprints for 16 important western blackberry cultivars.
IQF berries and frozen concentrate of Marion and Kotata were generously provided by reliable commercial sources.
The FAST ID Kit worked better than two other DNA extraction methods for isolating DNA from IQF berries and from frozen and thawed concentrate.
Out of twenty-nine SSRs tested, ten polymorphic SSRs differentiated between Marion and Kotata leaves and were chosen for subsequent analyses.
SSR-based fingerprinting of individual IQF berries (using the receptacle for DNA extraction) revealed a mixture of Kotata and Marion berries in the commercial Marion bag while fingerprinting of frozen Marion concentrate identified Kotata in the small frozen puree sample evaluated and possible contamination from seed DNA. The ten SSRs differentiated between each of the 16 western cultivars included in this study.
In fact, one SSR marker, Rubus 275a was sufficient to distinguish these 16 cultivars.
In summary, blackberry can be reliably identified with SSR markers, using leaves and frozen berries as sources of DNA. Fingerprinting from concentrate does not appear reliable for identity certification due to possible contamination from seed DNA.
Microsatellite markers or simple sequence repeats (SSRs) are ideal for cultivar fingerprinting, paternity testing and identity certification.
Fingerprinting is valuable for variety identification, quality control and as a legal method to protect against infringement by competitors.
The objectives of this study were to develop a DNA extraction protocol and SSR-based identification for individually quick-frozen (IQF) Marion and Kotata whole berries and concentrate and to generate genetic fingerprints for 16 important western blackberry cultivars.
IQF berries and frozen concentrate of Marion and Kotata were generously provided by reliable commercial sources.
The FAST ID Kit worked better than two other DNA extraction methods for isolating DNA from IQF berries and from frozen and thawed concentrate.
Out of twenty-nine SSRs tested, ten polymorphic SSRs differentiated between Marion and Kotata leaves and were chosen for subsequent analyses.
SSR-based fingerprinting of individual IQF berries (using the receptacle for DNA extraction) revealed a mixture of Kotata and Marion berries in the commercial Marion bag while fingerprinting of frozen Marion concentrate identified Kotata in the small frozen puree sample evaluated and possible contamination from seed DNA. The ten SSRs differentiated between each of the 16 western cultivars included in this study.
In fact, one SSR marker, Rubus 275a was sufficient to distinguish these 16 cultivars.
In summary, blackberry can be reliably identified with SSR markers, using leaves and frozen berries as sources of DNA. Fingerprinting from concentrate does not appear reliable for identity certification due to possible contamination from seed DNA.
Authors
N.V. Bassil, M. Muminova, W. Njuguna
Keywords
individually quick-frozen (IQF), simple sequence repeat (SSR) markers, Rubus species
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