Articles
DEVELOPMENT OF SEQUENCED TAGGED MICROSATELLITE SITE (STMS) MARKERS IN AZALEA
Article number
546_21
Pages
193 – 197
Language
Abstract
A genomic library was constructed from DNA of two azalea genotypes: a Belgian pot azalea R. simsii hybrid Mevr.
Van Belle and a Chinese R. simsii from Daoxian.
An enrichment of microsatellite containing sequences was performed as in Van de Wiel et al. (1999). Fragments were sequenced and primers were designed that allow the amplification of the microsatellite repeat.
About 220 microsatellite containing clones were selected from the enrichment procedure.
Mainly dinucleotide repeats and some trinucleotide repeats were found.
The selected primers were tested in a small set of reference varieties to check their value (specificity and polymorphic rate) and to set up the PCR-conditions.
Five primer pairs have been tested, two of them gave a specific and polymorphic pattern.
They were further screened by radioactive PCR on a selection of 5 plants from the azalea breeders gene pool which included the two genotypes used library construction.
These 2 STMS markers uniquely identified the 5 plants.
Van Belle and a Chinese R. simsii from Daoxian.
An enrichment of microsatellite containing sequences was performed as in Van de Wiel et al. (1999). Fragments were sequenced and primers were designed that allow the amplification of the microsatellite repeat.
About 220 microsatellite containing clones were selected from the enrichment procedure.
Mainly dinucleotide repeats and some trinucleotide repeats were found.
The selected primers were tested in a small set of reference varieties to check their value (specificity and polymorphic rate) and to set up the PCR-conditions.
Five primer pairs have been tested, two of them gave a specific and polymorphic pattern.
They were further screened by radioactive PCR on a selection of 5 plants from the azalea breeders gene pool which included the two genotypes used library construction.
These 2 STMS markers uniquely identified the 5 plants.
Authors
J. Dendauw, J. De Riek, P. Arens, E. Van Bockstaele, B. Vosman, M. De Loose
Keywords
Rhododendron species, Belgian pot azalea, microsatellite enrichment, dinucleotide and trinucleotide repeats
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