A blackberry (Rubus L.) expressed sequence tag library for the development of simple sequence repeat markers
Kim S Lewers1, Chris A Saski, Brandon J Cuthbertson
1USDA-ARS, Beltsville Agricultural Research Center, Genetic Improvement of Fruits and Vegetables Lab, Bldg, 010A, BARC-West, 10300 Baltimore Ave., Beltsville, MD 20705-2350, USA. kim.lewers@ars.usda.gov
BMC Plant Biology
|June 24, 2008
Summary
Researchers developed the first blackberry expressed sequence tag (EST) library, creating molecular markers for improved breeding. This work enables the development of genetic maps for marker-assisted breeding in blackberries.
Area of Science:
- Plant genetics
- Molecular biology
- Bioinformatics
Background:
- Novel repeat-fruiting blackberry cultivars necessitate advanced breeding tools.
- Existing blackberry breeding relies on morphological markers, lacking molecular genetic resources.
- There is a need for genetic maps, molecular markers, and sequences for cultivated blackberry.
Purpose of the Study:
- To develop the first blackberry expressed sequence tag (EST) library.
- To design and test simple sequence repeat (SSR) markers derived from ESTs.
- To lay the groundwork for molecular marker-assisted breeding in blackberry.
Main Methods:
- Generated a cDNA library of 18,432 clones from 'Merton Thornless' blackberry leaves.
- Annotated 3,000 genes, identifying those involved in energy, cell structure, and defense.
- Designed 673 primer pairs from ESTs containing SSRs and tested 33 pairs for polymorphism.
Main Results:
- The EST library contained over 18,000 clones, with 3,000 genes annotated.
- 673 SSR primer pairs were designed from the EST sequences.
- 10 of 33 tested primer pairs revealed an average of 1.9 polymorphic PCR products between two cultivars.
Conclusions:
- The developed EST-SSR markers show potential for generating a comprehensive genetic map.
- This resource could enable association of molecular markers with desirable phenotypic traits.
- The findings support the advancement of molecular marker-assisted breeding in blackberry, complementing existing methods.
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