STUDIES ON DEVELOPMENT OF GENIC-SSRs IN RASPBERRY (Rubus ellipticusSmith.) AND THEIR TRANSFERABILITY ACROSS RELATED SPECIES
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Date
2013
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UHF,NAUNI
Abstract
ABSTRACT
Rubus ellipticus Smith. commonly known as ‘Yellow Himalayan raspberry’ is an important
member of Rosaceae family with high medicinal importance having high . In the present study EST-SSR markers were datamined for R. ellipticusand were used for crosstransferability studies. EST sequences of R. ellipticus /Rubus were downloaded from NCBI website
(www.ncbi.nlm.nih.gov/nucest). Seven EST sequences for R.ellipticusand 3184 for other Rubusspecies, R.
ulmifoliusand R.idaeuswere obtained. ESTs containing SSR motifs were extracted out using an online tool,
SSRIT (www.gramene.org/db/searches/SSRtool). None of the R.ellipticusESTs contained any SSR motif,
so EST sequences obtained for R.ulmifoliusand R.idaeuswere used for SSR extraction. SSR primers were
designed from the EST-SSR containing sequences using PRIMER 3 software (www.frodo.wimit.edu/primer3/) and 20 primers were custom synthesized. SSR studies was carried out using ten Rubus
species (four R. ellipticus collections of different geographical origin, R. ulmifolius, R. hypargyrus, R.
panniculata, R.nutans, R.macilentusand R.strigosus). To study polymorphism and transferability among
the ten Rubus species, DNA was isolated from young leaves of all the ten species using CTAB method
(Doyle and Doyle, 1987). The polymorphism study among ten Rubus accessions was carried out with the 20
custom synthesized Rubus primers. All 20 primers showed amplification, with polymorphism of 98.36%.
The transferability studies were also carried out using already used 20 polymorphic peach primers, which
had shown transferability of 95% to four genotypes of apple and rose each, all belonging to Rosaceae
family. Jaccard’s similarity matrix was developed and dendrogram was generated using NTSYSpc ver.2.02h
to establish the percent similarity among the ten Rubus accessions. Two clusters ‘A’ and ‘B’ were obtained.
R. strigosusin cluster ‘A’ was found to diverge from rest of the nine accessions, all grouped under cluster
‘B,’ revealing high percentage of variability of R. strigosus from rest of the nine species. Maximum
similarity was found between R.ellipticusIII and R.macilentus. Thus EST-SSRs used in the present study
revealed a high level of polymorphism in the ten Rubusaccessions. Also interspecific and intergeneric cross
transferability was established among these accessions
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