Skip to main content
SpringerLink
Log in

In vitro propagation and assessment of genetic fidelity of Blepharispermum subsessile DC.: an endangered medicinal plant of India

  • Research Articles
  • Published:
Vegetos Aims and scope Submit manuscript

Abstract

Blepharispermum subsessile is an important threatened medicinal plant of India. This plant has been enlisted in ‘endangered’ category in different states of India due to over-exploitation and unregulated trading. Thus, urgent attention is needed for its conservation. This study presents an efficient in vitro plant propagation protocol for B. subsessile using axenic shoot tip and confirmation of genetic fidelity of the regenerated plants. Axenic shoot tips were inoculated on MS medium supplemented with different concentrations and combinations of plant growth regulators for shoot multiplication. Highest number of shoots per explant (48.1) were found on MS medium augmented with 3.0 mgl−1 meta Topolin (mT) and 1.5 mgl−1 Zeatin (Z). Ninety-five percent in vitro shoots were rooted on ½ MS medium augmented with 0.5 mgl−1 Indole-3-butyric acid (IBA). All rooted plantlets were successfully acclimatized and about 70% plants survived. The genetic fidelity of in vitro propagated plants was assessed by ISSR markers and confirmed their genetic integrity with that of the mother plant. This study could be used for conservation of B. subsessile.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Data availability

All data generated or analysed during this study are included in this published article.

References

  • Agarwal SK, Verma S, Singh SS, Kumar S, Keshri G (1999) Antifertility chromene from Blepharispermum subsessile. Fitoterapia 70:435–437

    Article  CAS  Google Scholar 

  • Behera S, Barik DP, Naik SK (2017) Micropropagation of Symplocos racemosa Roxb., a threatened medicinal tree of India. Curr Sci 113(4):555–558

    Google Scholar 

  • Behera S, Kamila PK, Rout KK, Barik DP, Panda PC, Naik SK (2018) An efficient plant regeneration protocol of an industrially important plant, Hedychium coronarium J. Koenig and establishment of genetic and biochemical fidelity of the regenerants. Ind Crop Prod 126:58–68

    Article  CAS  Google Scholar 

  • Behera S, Kar SK, Rout KK, Barik DP, Panda PC, Naik SK (2019) Assessment of genetic and biochemical fidelity of field-established Hedychium coronarium J. Koenig regenerated from axenic cotyledonary node on meta-topolin supplemented medium. Ind Crop Prod 134:206–215

    Article  CAS  Google Scholar 

  • Behera S, Rout KK, Panda PC, Naik SK (2020) Production of non-embryogenic synthetic seeds for propagation and germplasm transfer of Hedychium coronarium J Koenig. J Appl Res Med Arom Plant 19:100271

    Google Scholar 

  • Behera S, Monalisa K, Meher RK, Mohapatra S, Das PK, Madkami SK, Naik PK, Naik SK (2022) Phytochemical fidelity and therapeutic activity of micropropagated plant Curcuma amada Roxb.: an important medicinal herb. Ind Crop Prod 176:11440

    Article  Google Scholar 

  • Behera S, Chouhan VBS, Monalisa K, Mehera RK, Kar SK, Pati K, Bansode VV, Nedunchezhiyan M, Verma AK, Naik SK, Naik PK (2024) In vitro plant regeneration, genetic and biochemical fidelity and anticancer activity of anthocyanin rich purple flesh sweet potato var. Bhu Krishna. S Afr J Bot 166:332–343

    Article  CAS  Google Scholar 

  • Chakrabarty D, Datta SK (2008) Micropropagation of gerbera: lipid peroxidation and antioxidant enzyme activities during acclimatization process. Acta Physiol Plant 30:325–331

    Article  CAS  Google Scholar 

  • Dolker D, Behera S, Justine AK, Kumari V, Pati PK (2024) Production of large-scale genetically identical and phytochemically stable in vitro plants of Rhodiola imbricata using meta-Topolin and liquid culture medium. Plant Cell Tiss Organ Cult 156:1–17

    Article  Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Evenor D, Reuveni (2004) Micropropagation of Achillea filipendulina cv.’Parker. Plant Cell Tiss Organ Cult 79(1):91–93

    Article  CAS  Google Scholar 

  • Gomez KA, Gomez KA (1984) Statistical procedures for agricultural research, 2nd edn. Wiley, New York

    Google Scholar 

  • Hartmann HT, Kester DE (1979) Plant propagation: principles and practices, 4th edn. Prentice Hall of India. Ltd., p 407

    Google Scholar 

  • He J, Qi T, Yang J, Xu Q, Zou L, Ma Y (2023) Development of an efficient micropropagation protocol for Nematanthus wettsteinii using leaf and shoot-tip explants. In Vitro Cell Dev Biol-Plant 59:1–9

    Article  Google Scholar 

  • Kalidass C, Nayak P, Nayak S (2016) Effect of plant growth regulators and explant types on micropropagation of an endangered medicinal plant Blepharispermum subsessile DC. Plant Sci Res 38(1 & 2):74–82

    Google Scholar 

  • Kilic MU, Uysal H (2023) Establishment of in vitro cultures and investigation of micropropagation possibilities in shoot tip explants of different mastic tree (Pistacia lentiscus L.) Genotypes. Biol Bull 50(5):931–945

    Article  Google Scholar 

  • Kulkarni MM, Nagasampagi BA, Deshpande SG, Sharma RN (1987) Five chromenes from Blepharispermum subsessile. Phytochemistry 26(11):2969–2971

    Article  CAS  Google Scholar 

  • Manokari M, Mehta SR, Priyadharshini S, Badhepuri MK, Dulam S, Jayaprakash K, Cokul MR, Dey A, Rajput BS, Shekhawat MS (2021) Meta-Topolin mediated improved micropropagation, foliar micro-morphological traits, biochemical profiling, and assessment of genetic fidelity in Santalum album L. Ind Crops Prod 171:113931

    Article  Google Scholar 

  • Mezaka I, Klavina D, Kalane L, Kronberga A (2023) Large-scale in vitro propagation and ex vitro adaptation of the endangered medicinal plant Eryngium maritimum L. Horticulturae 9:271

    Article  Google Scholar 

  • Modi AR, Patil G, Kumar N, Singh AS, Subhash N (2012) A simple and efficient in vitro mass multiplication procedure for Stevia rebaudiana Bertoni and analysis of genetic fidelity of in vitro raised plants through RAPD. Sugar Tech 14:391–397

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Naik SK, Behera S, Rath SK, Patra JK (2020) A comprehensive scientific overview of Blepharispermum subsessile DC. (Asteraceae), a conservation concern medicinal plant with promising pharmaceutical potential. Indian J Trad Knowl 19(1):208–217

    Google Scholar 

  • Nayak P, Kalidass C (2016) In vitro regeneration of Blepharispermum subsessile DC: an endangered medicinal plant of Odisha, India using cotyledon explants. Plant Tiss Cult Biotechnol 26(2):255–266

    Article  Google Scholar 

  • Nayak SA, Kumar S, Satapathy K, Moharana A, Behera B, Barik DP, Acharya L, Mohapatra PK, Jene PK, Naik SK (2013) In vitro plant regeneration from cotyledonary nodes of Withania somnifera (L.) Dunal and assessment of clonal fidelity using RAPD and ISSR markers. Acta Physiol Plant 35:195–203

    Article  CAS  Google Scholar 

  • Nowakowska K, Pacholczak A (2020) Comparison of the effect of meta-topolin and benzyl adenine during Daphne mezereum L. micropropagation. Agronomy 10(12):1994

    Article  CAS  Google Scholar 

  • Nowakowska K, Pińkowska A, Siedlecka E, Pacholczak A (2022) The effect of cytokinins on shoot proliferation, biochemical changes and genetic stability of Rhododendron ‘Kazimierz Odnowiciel’in the in vitro cultures. Plant Cell Tiss Organ Cult 149:675–684

    Article  CAS  Google Scholar 

  • Razak UNAA, Ong CB, Yu TS, Lau LK (2014) In vitro micropropagation of Stevia rebaudiana Bertoni in Malaysia. Braz Arch Biol Technol 57(1):23–28

    Article  Google Scholar 

  • Rohilla P, Naik R, Acharya RN (2016) Single herbal remedies for Sandhivata (Osteoarthritis): a review on evidence based researches in Dravyaguna department of IPGT & RA, Jamnagar. Int J Herb Med 4(5):97–101

    Google Scholar 

  • Saha S, Adhikari S, Dey T, Ghosh P (2016) RAPD and ISSR based evaluation of genetic stability of micropropagated plantlets of Morus alba L. Variety S-1. Meta Gene 7:7–15

    Article  PubMed  Google Scholar 

  • Shekhawat MS, Manokari M (2015) Efficient in vitro propagation by ex vitro rooting methods of Artemisia absinthium L., an ethnobotanically important plant. Chin J Biol 273405:8–15

    Google Scholar 

  • Shekhawat MS, Priyadharshini S, Jogam P, Kumar V, Manokari M (2021) Meta-Topolin and liquid medium enhanced in vitro regeneration in Scaevola taccada (Gaertn.) Roxb. In Vitro Cell Dev Biol Plant 57:296–306

    Article  CAS  Google Scholar 

  • Shukla R, Chakravarty M, Gautam MP (2013) Ethno medicinal practices among the Binjhwar Tribe of Chhattisgarh, India. Glob J Res Med Plants Indig Med 2(7):525–531

    Google Scholar 

  • Singh P, Dwivedi P (2013) Two-stage culture procedure using thidiazuron for efficient micropropagation of Stevia rebaudiana, an anti-diabetic medicinal herb. 3 Biotech 4:431–437

    Article  PubMed  PubMed Central  Google Scholar 

  • Subrahmanyeswari T, Laha S, Kamble SN, Singh S, Bhattacharyya S, Gantait S (2023) Alginate encapsulation of shoot tips and their regeneration for enhanced mass propagation and germplasm exchange of genetically stable Stevia rebaudiana Bert. Sugar Tech 25(3):542–551

    Article  CAS  Google Scholar 

  • Sujatha G, Kumari BD (2007) Effect of phytohormones on micropropagation of Artemisia vulgaris L. Acta Physiol Plant 29:189–195

    Article  Google Scholar 

  • Ved DK, Kinhal GA, Ravikumar K, Sankar RV, Sumathi R, Mahapatra AK, Panda PC (2008) Conservation Assessment and Management Prioritisation for Medicinal Plants of Orissa. Regional Plant Resource Centre, Bhubaneswar and Foundation for Revitalisation of Local Health Traditions, Bangalore

  • Voicu D (2017) In vitro plant regeneration from nodal segments of the alpine wormwood Artemisia eriantha Ten. (Asteraceae). AAB Bioflux 9(2):90–96

    Google Scholar 

  • Yesmin S, Hashem A, Islam MS (2015) Micropropagation of an important medicinal herb Eclipta alba (L.) Hassk. J Biol Sci 4(1):61–69

    Google Scholar 

Download references

Acknowledgements

We acknowledge to the Department of Forest and Environment, Govt. of Odisha for allowing us to visit different forest areas and collect the plant materials for this study. SKN also gratefully acknowledged the funding in the form of ‘UGC-Research Award-2016’. Our thanks are also to Mr Biswa Nath Hota, Former Deputy Conservator of Forest for his help and enlightening discussion during this work.

Author information

Author notes

  1. Shashikanta Behera

    Present address: Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab, 143005, India

Authors and Affiliations

  1. Department of Botany, Ravenshaw University, Cuttack, Odisha, 753003, India

    Kumari Monalisa, Shashikanta Behera, Partha S. Nial & Soumendra K. Naik

  2. Department of Botany, Rayagada Autonomous College, Rayagada, Odisha, 765001, India

    Siba P. Pidika

Authors
  1. Kumari Monalisa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shashikanta Behera
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Siba P. Pidika
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Partha S. Nial
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Soumendra K. Naik
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

SKN conceptualized the study. KM carried out all experiment and prepared the first draft. SKN, SB, SPP visited different forest areas and collected plant samples. SB analyse data and corrected the manuscript. SKN critically reviewed and edited the final version of the manuscript. All authors read and finally approved the manuscript for publication.

Corresponding author

Correspondence to Soumendra K. Naik.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Monalisa, K., Behera, S., Pidika, S.P. et al. In vitro propagation and assessment of genetic fidelity of Blepharispermum subsessile DC.: an endangered medicinal plant of India. Vegetos (2024). https://doi.org/10.1007/s42535-024-00851-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s42535-024-00851-1

Keywords

Search

Navigation