Molecular identification using DNA barcoding and pharmacognostical evaluation of Justicia beddomei (C. B. Clarke) Bennet

Nagalekshmi Rajeswariammal Senthilkumar Palaniappan   

Open Access   

Published:  May 15, 2024

DOI: 10.7324/JAPS.2024.172730

Justicia beddomei (C.B. Clarke) Bennet has been utilized for many years in the traditional medical system. This study aimed to facilitate the identification of J. beddomei (C.B. Clarke) Bennet using trnHpsbA DNA barcode region, NCBI database, and pharmacognostical features of the plant parts. Genomic DNA was extracted from plant leaf samples and polymerase chain reaction amplification and DNA sequence determination were performed. The contigs DNA sequence 319 bp was analyzed using the similarity basic search method BLASTn. The trnHpsbA barcode region of 319 bp contigs sequence showed 100% similarity with the standard sequence of J. beddomei with the Accession No. MK347214.1 from the NCBI database. Microscopical studies of the different parts of the plant facilitate the identification and differentiation of J. beddomei from its morphologically similar and confusing plant Justicia adhatoda L. Justicia beddomei can be easily identified by the flower and its inflorescence arrangement. The other identification features are the presence of cystoliths in the mesophyll region of the leaf, and the presence of colored content and crystals in the powder microscopic analysis. The outcome of the present detailed microscopic study of the stem, leaf, and flower of J. beddomei is of great value in the identification of the powdered samples and their adulterants.

Keyword:     Justicia beddomei (C.B. Clarke) Bennet trnH–psbA barcode NCBI database pharmacognostical studies molecular identification DNA sequence


Rajeswariammal N, Palaniappan S. Molecular identification using DNA barcoding and pharmacognostical evaluation of Justicia beddomei (C. B. Clarke) Bennet. J Appl Pharm Sci. 2024. Online First.

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

HTML Full Text


1. Khan S, Mirza KJ, Zainulabdin M. Authentication of the medicinal plant Sennaangustifolia by RAPD profiling. Saudi J Biol Sci. 2011;18(3):287–92. doi:

2. Rastogi S, Pandey MK, Prakash J, Sharma A, Singh GN. Veterinary herbal medicines in India. Pharmacogn Rev. 2015;9(18):155–63. doi:

3. Grover JK, Yadav S, Vats V. Medicinal plants of India with anti-diabetic poential. J Ethnopharmacol. 2002;81(1):081–100. doi:

4. Bahuguna Y, Zaidi S, Kumar N, Rawat K. Standardization of polyherbal marketed formulation Triphala Churna. Research and reviews. J Pharmacogn Phytochem. 2014;2(3):028–35.

5. Santhosh Kumar JU, Krishna V, Seethapathy GS, Ganesan R, Ravikanth G, Shaanker RU. Assessment of adulteration in raw herbal trade of important medicinal plants of India using DNA barcoding. 3 Biotech. 2018 Mar;8(3):135. doi:

6. Tilney PM, Wyk VBE. The value of antomy in pharmacognosy and forensic studies. South Afr J Bot. 2010;76(2):404. doi:

7. Hebert PD, Cywinska A, Ball SL, deWaard JR. Biological identifications through DNA barcodes. Proc Biol Sci. 2003;270(1512):313–21. doi:

8. De Mattia F, Bruni I, Galimberti A, Cattaneo F, Casiraghi M, Labra M. A comparative study of different DNA barcoding markers for the identification of some members of Lamiacaea. Food Res Int. 2011;44(3):693–702. doi:

9. Amin S, Ghosh S, Biswas B, Arifuzzaman M, Azad M, Siddiki A. Molecular identification of four medicinal plants using DNA barcoding approach from Chittagong, Bangladesh. J Adv Biotechnol Exp Ther. 2020;3(3):268. doi:

10. Hou D, Song J, Shi L, Ma X, Xin T, Han J, et al. Stability and accuracy assessment of identification of traditional Chinese materia medica using DNA barcoding: a case study on Flos Lonicerae Japonicae. Biomed Res Int. 2013;2013:549037.

11. Handa SS. Indian efforts for quality control and standardization of herbal drugs/products. Proceedings of the 1st joint workshop on quality control and standardization of traditional medicine—Indo-China experience. Lucknow, India: National Botanical Research Institute; 2004. vol. 8, pp 008–010.

12. Goverment of India Ministry of Ayush. Pharmacopoeia commission for Indian medicine and homeopathy. Ghaziabad, India: Goverment of India Ministry of Ayush; 2004.

13. Warrier PK, Nambiar VP, Ramankutty C. Indian medicinal plants. Chennai, India: Orient Longman Ltd.; 1993. vol. 268, 272–3 pp. doi:

14. Prabhukumar KM, Beegam F, George S, Balachandran I. Revisiting the taxonomy of Justicia beddomei (Acanthaceae). Phytotaxa. 2018;350(1):71. doi:

15. Karthikayan S, Murthy S, Sanjappa SM. Flowering plants of India, dicotyledons (Acanthaceae-Avicenniaceae) 1. Kolkata, India: Botanical Survey of India; 2009. 166–4 pp.

16. Prabhukumar KM, Robi AJ, Hareesh VS, Balachandran I. Justicia gambleana (Acanthaceae): a new species from Kerala, India. Kew Bull [Internet]. 2016;71(3):39. doi:

17. Nandhini S, Ilango K. Comparative study on pharmacognostical, phytochemical investigations and quantification of vasicine content in the extracts of Adhatoda vasica Nees and Adhatoda beddomei CB Clarke. Pharmacogn J. 2020;12(4):884–96. doi:

18. Karthikayan S, Murthy S, Sanjappa SM. Flowering plants of India, Dicotyledons (Acanthaceae-Avicenniaceae). Kolkata, India: Botanical Survey of India; 2009. 366 p.

19. Sasidharan N. Biodiversity documentation for Kerala 6. Flowering plants. Thrissur, India: Kerala Forest Research Institute; 2004.

20. SunilKumar KN, Ramya A. A review on critically endangered species of Acanthacea: Justicia beddomei (Clarke) Bennet. An immune booster. J Med Plants. 2020;8:94–8. doi:

21. Vijayakumar AS, Jeyaraj M, Kolanchinathan P, Selvakumar M, Praveenkumar D. Molecular identification of the medicinal plant Justicia gendarussa (burm) f. Using ITS gene. Life Sci Inform Publ. 2019;5:663–74. doi:

22. Sang T, Crawford D, Stuessy T. Chloroplast DNA phylogeny, reticulate evolution, and biogeography of Paeonia (Paeoniaceae). Am J Bot. 1997;84(8):1120. doi:

23. Tate JA, Simpson BB. Paraphyly of Tarasa (Malvaceae) and diverse origins of the polyploid species. Syst Bot. 2003;28:723–37. doi:

24. Neerakkal I, Rajagopal R, Vijayakumar KR. Influence of inanimate shade on growth of rooted Adalodakam (Adhatoda beddomei C.B.Clarke) cuttings. J Trop Agric. 2005;43:91–3.

25. Haleshi C, Sringeswara AN, Danapur V. Pharmacognostic study of Justicia beddomei (C. B. Clarke) Bennet. Eur J Med Plants. 2020;31(9):83–8. doi:

26. Pathak MR, Mohamed AA, Farooq M. DNA barcoding and identification of medicinal plants in the Kingdom of Bahrain. Am J Plant Sci. 2018;9(13):2757–74. doi:

27. Hall TA. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser. 1999;41:95–8. doi:

28. Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994;22(22):4673–80. doi:

29. Ross HA, Murugan S, Li WL. Testing the reliability of genetic methods of species identification via simulation. Syst Biol. 2008;57(2):216–30. doi:

30. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990;215(3):403–10. doi:

31. Government of India 2, Ministry of Health and Family Welfare. New Delhi, India: Controller of Publications; 1996.

32. Oshi A. Pharmacognostical study of Justicia adhatoda Linn. Int J Herb Mede. 2014;1:14. Available from:

33. Reddy MP, Shantha TR, Rao VR, Venkateshwarlu G. Pharmacognostical and physicochemical evaluation on the flowers of Justicia adhatoda L. Res J Pharmacogn Phytochem. 2015;7(2):73. doi:

34. Datta Kalme RK. Pharmacognostic characterization of stem and root of Adhatoda zeylanica Medicus. J Pharm Sci Res. 2012;3(11):4264–9. doi:

Article Metrics
17 Views 3 Downloads 20 Total



Related Search

By author names