Articles
INTER-SIMPLE SEQUENCE REPEAT (ISSR) MARKERS IN HAZELNUT
Article number
845_19
Pages
159 – 162
Language
English
Abstract
Inter-simple sequence repeat (ISSR) markers are generated by PCR amplification of DNA using a single primer composed of a microsatellite sequence that is usually anchored at the 3- or 5- end by two to four arbitrary, often degenerate, nucleotides.
ISSR markers are rapid, simple, inexpensive, and highly reproducible due to their primer length and to the high stringency achieved by the annealing temperature.
Since microsatellite or simple sequence repeat (SSR) loci are abundant in plant genomes, ISSR primers often produce multiple bands which may be useful for genotyping and mapping, and also developing SSR markers.
We used 100 ISSR primers from the University of British Columbia to amplify DNA of eight hazelnut (Corylus avellana) accessions.
Of these primers, 66 produced distinct and polymorphic bands, 11 produced smears or faint bands, and the remaining 23 did not amplify at all.
All 15 of the AT-repeat primers failed to amplify.
Primers in the first category produce markers that are useful for genetic studies.
The sequences of the bands can also be used to develop SSR markers.
Sixty ISSR primers that gave distinct polymorphic bands in the initial screening were used to amplify DNA of the two parents and six seedlings from our mapping population: three resistant seedlings with S3 and three susceptible seedlings with S8. Eleven markers scored in the entire population fit the expected segregation ratio of 1:1 and were assigned to linkage groups.
Two markers, 815-540 and 823-600 were placed on linkage group 5S. Marker 815-540 is particularly interesting as it maps close to the S-locus.
We plan to clone and sequence this fragment.
Based on the primer sequence (CTCTCTCTCTCTCTCTG) we expect CT/GA repeats at the ends.
We plan to obtain the sequences of the flanking regions and then develop an SSR marker closely linked to the S-locus.
ISSR markers are rapid, simple, inexpensive, and highly reproducible due to their primer length and to the high stringency achieved by the annealing temperature.
Since microsatellite or simple sequence repeat (SSR) loci are abundant in plant genomes, ISSR primers often produce multiple bands which may be useful for genotyping and mapping, and also developing SSR markers.
We used 100 ISSR primers from the University of British Columbia to amplify DNA of eight hazelnut (Corylus avellana) accessions.
Of these primers, 66 produced distinct and polymorphic bands, 11 produced smears or faint bands, and the remaining 23 did not amplify at all.
All 15 of the AT-repeat primers failed to amplify.
Primers in the first category produce markers that are useful for genetic studies.
The sequences of the bands can also be used to develop SSR markers.
Sixty ISSR primers that gave distinct polymorphic bands in the initial screening were used to amplify DNA of the two parents and six seedlings from our mapping population: three resistant seedlings with S3 and three susceptible seedlings with S8. Eleven markers scored in the entire population fit the expected segregation ratio of 1:1 and were assigned to linkage groups.
Two markers, 815-540 and 823-600 were placed on linkage group 5S. Marker 815-540 is particularly interesting as it maps close to the S-locus.
We plan to clone and sequence this fragment.
Based on the primer sequence (CTCTCTCTCTCTCTCTG) we expect CT/GA repeats at the ends.
We plan to obtain the sequences of the flanking regions and then develop an SSR marker closely linked to the S-locus.
Publication
Authors
K. Gürcan, S.A. Mehlenbacher, V. Cristofori
Keywords
filbert, fingerprinting, mapping, inter-simple sequence repeat
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