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ISSR marker based DNA profiling studies in Rauwolfia species

Gulab Khan Rohela, Prasad Bylla, Sreenu Pendli, Korra Rajender, Christopher Reuben Thammidala


In this present study, a protocol was developed for the DNA profiling of medicinally important Rauwolfia species (05 species) by ISSR-PCR method. Initially genomic DNA was isolated from the fresh leaves (1 gram) of 24 months old Rauwolfia tetraphylla, R.serpentina, R.pentaphylla, R.vomitaria and R.micrantha. The content of genomic DNA isolated from 1 gram of fresh leaves of these five species ranges from 1.6 to 3.4 µg. Out of six inter simple sequence repeats (ISSR) primers used, HY3 primer has given the good amplification with distinct bands which were reproducible. The range of DNA bands produced by the HY3 primer were between 300 bp to 1500 bp among the five species. A dendrogram was constructed on the basis of similarity matrix data by unweighted pair group method with arithmetic average (UPGMA) cluster analysis using indigenous software. Average genetic similarity generated by HY3 has revealed 38% similarity between species of Sub cluster-A and species of Sub cluster-B which is consistent with their geographical distribution i.e. Sub cluster-A species R.tetraphylla and R.serpentina inhabit peninsular India, Sub cluster- B species R.micrantha and R. vomitoria inhabit the Himalayan foot hills. The developed protocol of DNA profiling in Rauwolfia species by ISSR-PCR method could be used efficiently for the differentiation and identification of the Rauwolfia species at molecular level.


Rauwolfia, DNA Profiling, ISSR-PCR, Dendrogram Analysis

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