3 Biotech

, 8:161 | Cite as

Molecular identification of Saraca asoca from its substituents and adulterants

  • Satisha Hegde
  • Archana Saini
  • Harsha Vasudev Hegde
  • Sanjiva D. Kholkute
  • Subarna Roy
Original Article


Saraca asoca (Roxb.) De Wilde is an important medicinal plant from the Western Ghats of India, traditionally used in treatment of various gynecological disorders. Increasing commercial demand and decreasing numbers has resulted in this plant becoming endangered with crude drug materials being extensively substituted/adulterated with other plant species. The present study was undertaken with the objective of development and evaluation of multivariate cluster analysis of ISSR fingerprints against rbcL-based DNA barcodes as tool to understand the relationships and to differentiate common adulterants and substituents from S. asoca. ISSR-based Hierarchical Cluster Analysis was carried out on 41 samples of S. asoca and 5 each of the 5 common substituent/adulterant plants and the clustering patterns were evaluated against DNA-sequence-based barcoding of rbcL region of their plastids. Factorial analysis and Principal Coordinate Analysis revealed distinct groups of genetic pools of respective taxa thereby confirming the utility of ISSR fingerprinting as a useful tool for differentiation between the genuine and the adulterants/substituents. NCBI-BLAST search on DNA barcode rbcL region confirmed the results of ISSR assays. Therefore, our study demonstrated the utility of simple, cost-effective method of ISSR fingerprinting coupled with rbcL barcoding in differentiating this important medicinal plant from its common adulterants/substituents.

Graphical Abstract


Detection DNA barcoding Identification ISSR Phylogenetics rbcL 



Inter Simple Sequence Repeat


Ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit


Principal Coordinate Analysis


Hierarchical Cluster Analysis


Cophenetic Correlation Coefficient


Unweighted Pair Group Method with Arithmetic Mean



Authors are thankful to the Director-in-Charge, ICMR-NITM, formerly RMRC, Belagavi, India, and Indian Council of Medical Research (Government of India), for supporting this study. Authors are also thankful to Dr. Vidya S. Gupta, Biochemical Science Division, National Chemical Laboratory, Pune, for technical suggestions and Bio-Medical Informatics Centre of ICMR at NITM, Belagavi, for informatics support. Authors extend thanks to Pramod Kumar and Jotiba B. Palekar, ICMR-NITM, Belagavi, Anil Bisht (Dehradun), Jayendrasinh Chavda (Gujarat) and Aparna J (Kerala), India, for help during sample collection. SH is grateful to ICMR for funding.

Compliance with ethical standards


This study was funded by The Indian Council of Medical Research, New Delhi (Grant No. 45/53/2013/BMS/TRM; IRIS ID: 2013-18920).

Conflict of interest

All authors declare that there is no conflict of interest.

Ethical approval

Not applicable.

Supplementary material

13205_2018_1175_MOESM1_ESM.pptx (1.6 mb)
Fig. 1a: ISSR amplification profile from genomic DNA of 41 individuals of Saraca asoca; M: 100+500bp Molecular weight markers; NC: Negative  control. Fig. 1b: ISSR amplification profile from genomic DNA samples of BV (1-5), SR (1-5), PL (1-5), MF (1-5) and TO (1-5); M: 100+500bp Molecular weight markers; NC: Negative  control (PPTX 1666 kb)
13205_2018_1175_MOESM2_ESM.doc (75 kb)
Supplementary material 2 (DOC 75 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.ICMR-National Institute of Traditional Medicine, Indian Council of Medical Research, Department of Health Research, Government of IndiaBelagaviIndia
  2. 2.KLE Academy of Higher Education and Research (Deemed-to-be-University), Dr. Prabhakar Kore Basic Science Research CentreBelagaviIndia

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