Skip to main content
SpringerLink
Log in

Rapid in vitro propagation by liquid culture system and genetic homogeneity assessment of Plectranthus bourneae Gamble, an endemic plant species to South India

  • Original Article
  • Published:
Indian Journal of Plant Physiology Aims and scope Submit manuscript

Abstract

The present investigation describes the in vitro propagation of Plectranthus bourneae via liquid culture system from axillary bud and shoot tip explants. Multiple shoots were initiated from axillary bud and shoot tip explants on Murashige and Skoog (MS) medium supplemented with different concentrations (0.1–2.0 mg/l) and combinations of cytokinins [BA (6-benzyladenine), KN (kinetin), and TDZ (thidizauron)] along with (0.5–2.0 mg/l) auxins [NAA (α-naphthalene acetic acid), IAA (indole-3 acetic acid), IBA (indole-3-butyric acid)]. Maximum shoot multiplication (12.14) was achieved from axillary bud explant followed by shoot tip explant (8.85) at 0.5 mg/l TDZ with 1.0 mg/l NAA. A maximum of 9.71 roots/shoot was observed at 0.7 mg/l IBA. The rooted plantlets successfully hardened and transferred to greenhouse condition with 83% survival. Inter-simple sequence repeat markers exhibited genetic fidelity with 100% monomorphism in regenerants. The authenticated protocol supports rapid production of true-to-type plants by liquid culture in vitro and therefore could supply a valuable target material for genetic transformation study.

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

References

  • Akdemir, H., Suzerer, V., Tilkat, E., Onay, A., & Çiftçi, Y. O. (2016). Detection of variation in long-term micropropagated mature Pistachio via DNA-based molecular markers. Applied Biochemistry and Biotechnology, 180(7), 1301–1312.

    Article  PubMed  CAS  Google Scholar 

  • Ascough, G. D., & Fennell, C. W. (2004). The regulation of plant growth and development in liquid culture. South African Journal of Botany, 70, 181–190.

    Article  CAS  Google Scholar 

  • Ay, N. V., Duy, M. V., Baatartsogt, O., Altantsetseg, Kh, & Enkhchimeg, V. (2016). Plant regeneration of Kamchatic Plantain (Plantago camtschatica Link) through callus induction. Mongolian Journal of Agricultural Sciences, 19(3), 41–48.

    Google Scholar 

  • Bhatia, R., Singh, K. P., Sharma, T. R., & Jhang, T. (2010). Evaluation of the genetic fidelity of in vitro-propagated gerbera (Gerbera jamesonii Bolus) using DNA-based markers. Plant Cell, Tissue and Organ Culture. https://doi.org/10.1007/s11240-010-9806-5.

    Article  Google Scholar 

  • Bianchetti, R. E., Resende, C. F., Pacheco, V. S., Dornellas, F. F., Oliveira, A. M. S., Freitas, J. C. E., et al. (2017). An improved protocol for in vitro propagation of the medicinal plant Mimosa pudica L. African Journal of Biotechnology, 16(9), 418–428.

    Article  CAS  Google Scholar 

  • Carović-StanKo, K., Petek, M., Martina, G., Pintar, J., Bedeković, D., Ćustić, M. H., et al. (2016). Medicinal plants of the family Lamiaceae as functional foods–A review. Czech Journal of Food Sciences, 34(5), 377–390.

    Article  Google Scholar 

  • Chaudhary, Z., & Rashid, A. H. (2007). Effect of variety and plant growth regulators on callus proliferation and regeneration response of three tomato cultivars (Lycopersicon esculentum). Pakistan Journal of Botany, 39(3), 857–869.

    Google Scholar 

  • Chavan, J. J., Nalawade, A. S., Gaikwad, N. B., Gurav, R. V., Dixit, G. B., & Yadav, S. R. (2014). An efficient in vitro regeneration of Ceropegia noorjahaniae: An endemic and critically endangered medicinal herb of the Western Ghats. Physiology and Molecular Biology of Plants, 20(3), 405–410.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Chen, S. L., Yu, H., Luo, H. M., Wu, Q., Li, C. F., & Steinmetz, A. (2016). Conservation and sustainable use of medicinal plants: Problems, progress, and prospects. Chinese Medicine, 11(1), 37. https://doi.org/10.1186/s13020-016-0108-7.

    Article  PubMed  PubMed Central  Google Scholar 

  • Dhital, S. P., Lim, H., & Manandhar, H. K. (2010). Direct and Efficient plant regeneration from different explants sources of potato cultivars as influenced by plant growth regulators. Nepal Journal of Science and Technology, 12, 1–6.

    Google Scholar 

  • Doyle, J. J., & Doyle, J. L. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19(1), 11–15

    Google Scholar 

  • Fatima, N., & Anis, M. (2012). Role of growth regulators on in vitro regeneration and histological analysis in Indian ginseng (Withania somnifera L.) Dunal. Physiology and Molecular Biology of Plants, 18(1), 59–67.

    Article  PubMed  CAS  Google Scholar 

  • Gambhir, G., & Srivastava, D. K. (2015). Thidiazuron induces high frequency shoot regeneration in leaf and petiole explants of cabbage (Brassica Oleracea L. Var. Capitata). Journal of Biotechnology & Biomaterials, 5, 172.

    Google Scholar 

  • Gashi, B., Abdullai, K., Sota, V., & Kongjika, E. (2015). Micropropagation and in vitro conservation of the rare and threatened plants Ramonda serbica and Ramonda nathaliae. Physiology and Molecular Biology of Plants, 21(1), 123–136.

    Article  PubMed  Google Scholar 

  • Ghanbar, T., Hosseini, B., Jabbarzadeh, Z., Farokhzad, A., & Sharafi, A. (2016). High-frequency in vitro direct shoots regeneration from axillary nodal and shoot tip explants of clary sage (Salvia sclarea L.). Bulgarian Journal of Agricultural Science, 22, 73–78.

    Google Scholar 

  • Goel, M. K., Mehrotra, S., Kukreja, A. K., Shanker, K., & Khanuja, S. P. (2009). In vitro propagation of Rauwolfia serpentina using liquid medium, assessment of genetic fidelity of micropropagated plants, and simultaneous quantitation of reserpine, ajmaline, and ajmalicine. Methods in Molecular Biology, 547, 17–33.

    Article  PubMed  CAS  Google Scholar 

  • Grzegorczyk, I., & Wysokinska, H. (2008). Liquid shoot culture of Salvinia officinalis L. for micropropagation and production of antioxidant compounds; effect of triacontanol. Acta Societa Botanicorum Poloniae, 77(2), 99–104.

    Article  CAS  Google Scholar 

  • Kalpana, M., Anbazhagan, M., & Natarajan, V. (2009). Utilization of liquid medium for rapid micropropagation of Stevia rebaudiana Bertoni. Journal of Ecobiotechnology, 1(1), 016–020.

    Google Scholar 

  • Kim, D. H., & Sivanessan, I. (2016). Influence of benzyladenine and thidiazuron on shoot regeneration from leaf and shoot tip explants of Sedum sarmentosum Bunge. Brazilian Archives of Biology and Technology, 59(e16150), 717.

    Google Scholar 

  • Klerk, D. G. J., & Brugge, J. (2011). Micropropagation of dahlia in static liquid medium using slow-release tools of medium ingredients. Scientia Horticulturae, 127, 542–547.

    Article  CAS  Google Scholar 

  • Lata, H., Chandra, S., Techen, N., Khan, I. A., & ElSohly, M. A. (2016). In vitro mass propagation of Cannabis sativa L.: A protocol refinement using novel aromatic cytokinin meta-topolin and the assessment of eco-physiological, biochemical and genetic fidelity of micropropagated plants. Journal of Applied Research on Medicinal and Aromatic Plants, 3(1), 18–26.

    Article  Google Scholar 

  • Lata, H., Chandra, S., Techen, N., Yan-Hong, W., & Khan, I. A. (2013). Molecular analysis of genetic fidelity in micropropagated plants of Stevia rebaudiana Bert. Using ISSR marker. American Journal of Plant Sciences, 4, 964–971.

    Article  CAS  Google Scholar 

  • Lukhoba, C. W., Simmonds, M. S. J., & Paton, A. J. J. (2006). Plectranthus: A review of ethnobotanical uses. Ethnopharmacology, 103, 1–24.

    Article  PubMed  Google Scholar 

  • Mandal, J., & Laxminarayana, U. (2014). Indirect shoot organogenesis from leaf explants of Adhatoda vasica Nees. SpringerPlus. https://doi.org/10.1186/2193-1801-3-648.

    Article  PubMed  PubMed Central  Google Scholar 

  • Matthew, K. M. (1993). Precursory notes on the flora of the palni hills (pulney) hills, south India: II. Kew Bulletin, 48, 757–765.

    Article  Google Scholar 

  • Matthew, K. M. (1999). A report on the conservation status of south Indian plants. Biodiversity Conservation, 8, 779–796.

    Article  Google Scholar 

  • Medrado, H. H., Santos, E. O., Ribeiro, E. M. O., David, J. M., David, J. P., Araújo, J. F., et al. (2017). Rosmarinic and cinnamic acid derivatives of in vitro tissue culture of Plectranthus ornatus: Over production and correlation with antioxidant activities. Journal of the Brazilian Chemical Society, 28(3), 505–511.

    CAS  Google Scholar 

  • Minocha, S. C. (1987). Plant growth regulators and morphogenesis in cell and tissue culture of forest trees. In J. M. Banga & D. J. Durzan (Eds.), Cell and tissue culture in forestry (Vol. 1, pp. 50–66). Dordrecht: Martinus Nijhoff.

    Chapter  Google Scholar 

  • Muraseva, D. S., Novikova, T. I., & Erst, A. A. (2015). In vitro propagation and conservation of rare species Fritillaria meleagris L. from floral explants. Contemporary Problems of Ecology, 8, 6.

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nasir, N. A. N. M., Anuar, N., Zainuddin, Z., & Yaakob, Z. (2017). Evaluation of genetic homogeneity of Jatropha curcas L. hybrid at an early stage of shoot bud formation from petioles using ISSR marker. Biotechnology and Biotechnological Equipment, 31(1), 45–50.

    Article  CAS  Google Scholar 

  • Ngezahayo, F., & Liu, B. (2014). Axillary bud proliferation approach for plant biodiversity conservation and restoration. International Journal of Biodiversity. https://doi.org/10.1155/2014/727025.

    Article  Google Scholar 

  • Opabode, J., & Akinyemiju, O. (2016). In vitro propagation of Solanecio biafrae and determination of genetic stability of plantlets using rapid and ISSR markers. Journal of Horticultural Research, 24(1), 29–36.

    Article  CAS  Google Scholar 

  • Prakash, L., Middha, S. K., Mohanty, S. K., & Swamy, M. K. (2016). Micropropagation and validation of genetic and biochemical fidelity among regenerants of Nothapodytes nimmoniana (Graham) Mabb. employing ISSR markers and HPLC. 3. Biotech, 6, 171.

    Google Scholar 

  • Prameela, J., Ramakrishnaiah, H., Krishna, V., Deepalakshmi, A. P., Naveen Kumar, N., & Radhika, R. N. (2015). Micropropagation and assessment of genetic fidelity of Henckelia incana: An endemic and medicinal Gesneriad of South India. Physiology and Molecular Biology of Plants, 21(3), 441–446.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Raja, R. R. (2012). Medicinally potential plants of Labiatae (Lamiaceae) family: An overview. Research Journal of Medicinal Plants, 6, 203–213.

    Article  Google Scholar 

  • Razaq, M., Heikrujam, M., Chetri, S. K., & Agrawal, V. (2013). In vitro clonal propagation and genetic fidelity of the regenerants of Spilanthes calva DC. using RAPD and ISSR marker. Physiology and Molecular Biology of Plants, 19(2), 251–260.

    Article  PubMed  CAS  Google Scholar 

  • Resende, C. F., Bianchetti, R. E., Oliveira, A. M. S., Braga, V. F., & Peixoto, P. H. P. (2015). In vitro propagation and acclimatization of Lippia rotundifolia, an endemic species of Brazilian Campos Rupestres. The Revista Ciência Agronômica, 46(3), 582–589.

    Article  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 

  • San-Jose, C. M., Ballester, A., & Vieite, A. (2015). Effect of thidiazuron on multiple shoot induction and plant regeneration from cotyledonary nodes of chestnut. The Journal of Horticultural Science & Biotechnology, 5, 588–595.

    Google Scholar 

  • Sencie-Tarazona, A., Rojas-Idrogo, C., Kato, M. J., & Delgado-Paredes, G. E. (2015). In vitro rapid clonal propagation in static liquid medium and acclimatization of Piper solmsianum and P. tuberculatum. International Journal of Pure & Applied. BioScience, 3(1), 1–10.

    Google Scholar 

  • Sharma, V., Belwal, N., Kamal, B., Dobriyal, A. K., & Jadon, V. S. (2016). Assessment of genetic fidelity of in vitro raised plants in Swertia chirayita through ISSR, RAPD analysis and peroxidase profiling during organogenesis. Brazilian Archives of Biology and Technology. https://doi.org/10.1590/1678-4324-2016160389.

    Article  Google Scholar 

  • Shekhawat, M. S., & Manokari, M. (2016a). In vitro regeneration frequency, micro-morphological studies and ex vitro rooting of Hemidesmus indicus (L.) R. Br.: A multi-potent endangered climber. Indian Journal of Plant Physiology. https://doi.org/10.1007/s40502-016-0216-5.

    Article  Google Scholar 

  • Shekhawat, M. S., & Manokari, M. (2016b). In vitro propagation, micromorphological studies and ex vitro rooting of cannon ball tree (Couroupita guianensis aubl.): A multipurpose threatened species. Physiology and Molecular Biology of Plants, 22(1), 131–142.

    Article  PubMed  PubMed Central  Google Scholar 

  • Siddique, I., Bukhari, N. A. W., Perveen, K., & Siddiqui, I. (2015). Influence of plant growth regulators on in vitro shoot multiplication and plantlet formation in Cassia angustifolia Vahl. Brazilian Archives of Biology and Technology, 58(5), 686–691.

    Article  CAS  Google Scholar 

  • Siva, R., Mayes, S., Behera, S. K., & Rajasekaran, C. (2012). Anthraquinones dye production using root cultures of Oldenlandia umbellata L. Industrial Crops and Products, 37, 415–419.

    Article  CAS  Google Scholar 

  • Siva, R., Rajasekaran, C., & Mudgal, G. (2009). Induction of somatic embryogenesis and organogenesis in Oldenlandia umbellata L., a dye-yielding medicinal plant. Plant Cell, Tissue and Organ Culture, 98, 205–211.

    Article  CAS  Google Scholar 

  • Sivanandhan, G., Selvaraj, N., Ganapathi, A., & Manickavasagam, M. (2015). Effect of nitrogen and carbon sources on in vitro shoot multiplication, root induction and withanolides content in Withania somnifera (L.) Dunal. Acta Physiologiae Plantarum, 37(2), 12.

    Article  CAS  Google Scholar 

  • Solomou, A. D., Martinos, K., Skoufogianni, E., & Danalatos, N. G. (2016). Medicinal and aromatic plants diversity in Greece and their future prospects: A review. Agricultural Science, 1, 9–21.

    Article  Google Scholar 

  • Teshome, I., Teshome, S., Soromessa, T., & Feyissa, T. (2016). Development of an efficient in vitro propagation protocol for Satureja punctata—A rare aromatic and medicinal plant. Taiwania, 61(1), 41–48.

    Google Scholar 

  • Thaniarasu, R., Kumar, T. S., & Rao, M. V. (2015). In vitro propagation of plectranthus bourneae Gamble—An endemic red listed plant. Plant Tissue Culture & Biotechnology., 25(2), 273–284.

    Article  Google Scholar 

  • Thaniarasu, R., Kumar, T. S., & Rao, M. V. (2016). Mass propagation of Plectranthus bourneae Gamble through indirect organogenesis from leaf and internode explants. Physiology and Molecular Biology of Plants, 22(1), 143–151.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Tuan, P. N., Meier-Dinkel, A., & Höltken, A. M. (2016). Paving the way for large-scale micropropagation of Juglans × intermedia using genetically identified hybrid seed. Plant Cell, Tissue and Organ Culture. https://doi.org/10.1007/s11240-016-0986-5.

    Article  Google Scholar 

  • Wangdi, K., & Sarethy, I. P. (2016). Evaluation of micropropagation system of Bacopa monnieri L. in liquid culture and its effect on antioxidant properties. Journal of Herbs, Spices & Medicinal Plants, 22, 69–80.

    Article  CAS  Google Scholar 

  • Werner, E. T., Soares, T. C. B., Gontijo, A. B. P. L., Neto, J. D. S., & Amaral, J. A. T. (2015). Genetic stability of micropropagated plants of Crambe abyssinica Hochst using ISSR markers. Genetics and Molecular Research, 14(4), 16450–16460.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The authors express their sincere thanks to the University Grants Commission, New Delhi for financial support in the form of major research project (UGC REF. NO. 40-325/2011(SR) dated 30.06.2011), and Mr. R. W. Stewart and Mrs. Tanya Balcar of Vattakanal Conservation Trust, Kodaikanal, Tamil Nadu, India for the help rendered during the field survey. One of the authors (M. V. Rao) also thank the UGC for providing Emeritus fellowship.

Author information

Authors and Affiliations

  1. Department of Botany, Bharathidasan University, Tiruchirappalli, Tamil Nadu, 620024, India

    R. Thaniarasu, T. Senthil Kumar & M. V. Rao

Authors
  1. R. Thaniarasu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. Senthil Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. V. Rao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T. Senthil Kumar.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Thaniarasu, R., Senthil Kumar, T. & Rao, M.V. Rapid in vitro propagation by liquid culture system and genetic homogeneity assessment of Plectranthus bourneae Gamble, an endemic plant species to South India. Ind J Plant Physiol. 23, 376–384 (2018). https://doi.org/10.1007/s40502-018-0369-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40502-018-0369-5

Keywords

Search

Navigation