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Rauwolfia tetraphylla L., is an important medicinal plant in Apocynaceae family and is recognized as an alternative source to Rauwolfia serpentina L., in terms of anti-hypertensive alkaloid production i.e. reserpine. In view of this, the present study is conducted to estimate the reserpine content in different parts (leaf, stem and root) of field grown plants (2 years old), tissue cultured plantlets (R1) (two months old) and cell suspensions cultures (two months old with and without precursor feeding) of R. tetraphylla by using high performance liquid chromatography (HPLC) technique. Overall maximum content of reserpine (in %) was estimated from the root samples. Roots of field grown plants has recorded high percent of reserpine (0.39%) followed by roots of tissue cultured plantlets (0.35%) and root callus based cell suspension cultures (0.38 %) which was fed with precursor amino acid (100 mg/L of tryptophan). In control type of root callus based cell suspension cultures, reserpine content was quantified as 0.14%; by precursor feeding (100 mg/L of tryptophan) it was enhanced to 0.38%. In conclusion, the reserpine content (0.35 and 0.38%) produced by the roots of tissue cultured plantlets (R1) and 100 mg/L tryptophan fed root callus based cell suspensions was comparable to that of the reserpine content (0.39%) of root parts of field grown plants. The present study demonstrates the reserpine production by in vitro cell suspension cultures throughout the year without sacrificing the medicinal plants.

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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.