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Somatic embryogenesis of Pelargonium sidoides DC.

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Abstract

Pelargonium sidoides is a high value medicinal plant endemic to South Africa and the Lesotho Highlands. Establishing an efficient regeneration system through somatic embryogenesis is necessary for its conservation. An efficient reproducible protocol for in vitro plant regeneration via somatic embryogenesis of Pelargonium sidoides, an important medicinal plant, is reported for the first time. Embryogenic callus was obtained on Murashige and Skoog (MS) basal medium supplemented with 2.0 mg L−1 picloram, 0.5 mg L−1 thidiazuron and 20 mg L−1 glutamine. Different developmental stages of somatic embryos (SEs: globular embryos, heart shaped embryos, torpedo shaped embryos and cotyledon embryo with radicle) were directly obtained and further matured from embryogenic callus by subsequent subculture on the same medium. The highest frequency of somatic embryos (25.89 ± 1.50) was recovered after 6 weeks. Scanning electron microscopic (SEM) analysis revealed the presence of embryogenic cell clusters that formed cotyledonary embryos. Mature somatic embryos germinated and developed into plantlets after 4 weeks on half-strength MS medium. High plant regeneration frequency (94.4 %) was achieved on half-strength MS medium supplemented with 1.0 mg L−1 gibberellic acid. Rooted plantlets were successfully acclimatized in the greenhouse with a survival rate of 90 %. The protocol developed would be helpful in reducing stress on natural habitats, provide a system for germplasm conservation, regeneration of large numbers of high value clonal plants for commercial production, the isolation of bioactive compounds and genetic transformational studies.

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Abbreviations

GA3 :

Gibberellic acid

MS:

Murashige and Skoog medium

PGRs:

Plant growth regulators

PPF:

Photosynthetic photon flux

SEs:

Somatic embryos

SEM:

Scanning electron microscopy

TDZ:

Thidiazuron

References

  • Aderkas P, Rohr R, Sundberg B, Gutmann M, Be Boux ND, Lelu MA (2002) Abscisic acid and its influence on development of the embryonal root cap, storage product and secondary metabolite accumulation in hybrid larch somatic embryos. Plant Cell Tissue Organ Cult 69:111–120

    Article  Google Scholar 

  • Baskaran P, Jayabalan N (2009) In vitro propagation of Psoralea corylifolia L. by somatic embryogenesis in cell suspension culture. Acta Physiol Plant 31:1119–1127

    Article  CAS  Google Scholar 

  • Baskaran P, Van Staden J (2012) Somatic embryogenesis of Merwilla plumbea (Lindl.) Speta. Plant Cell Tissue Organ Cult 109:517–524

    Article  CAS  Google Scholar 

  • Baskaran P, Van Staden J (2014) Plant regeneration via somatic embryogenesis in Drimia robusta. Plant Cell Tissue Organ Cult 109:517–524

    Article  Google Scholar 

  • Brendler T, van Wyk B-E (2008) A historical, scientific and commercial perspective on the medicinal use of Pelargonium sidoides (Geraniaceae). J Ethnopharmacol 119:420–433

    Article  CAS  PubMed  Google Scholar 

  • Cangahuala-Inocente GC, Dal Vesco LL, Steinmacher D, Torres AC, Guerra MP (2007) Improvements in somatic embryogenesis protocol in Feijoa (Acca sellowiana (Berg) Burret): induction, conversion and synthetic seeds. Sci Hortic 111:228–234

    Article  CAS  Google Scholar 

  • Colling J, Groenewald JH, Makunga NP (2010) Genetic alterations for increased coumarin production lead to metabolic changes in the medicinally important Pelargonium sidoides DC. (Geraniaceae). Metab Eng 12:561–572

    Article  CAS  PubMed  Google Scholar 

  • Deo PC, Taylor M, Harding RM, Tyagi AP, Becker DK (2010) Initiation of embryogenic cell suspensions of taro (Colocasia esculenta var. esculenta) and plant regeneration. Plant Cell Tissue Organ Cult 100:283–291

    Article  CAS  Google Scholar 

  • Dhir SK, Dhir S, Widholm JM (1991) Plantlet regeneration from immature cotyledon protoplasts of soybean (Glycine max L.). Plant Cell Rep 10:39–43

    CAS  PubMed  Google Scholar 

  • Gerdakaneh M, Mozafari AA, Mardah A, Sarabi B (2011) Effects of different amino acids on somatic embryogenesis of strawberry (Fragaria 3 ananassa Duch.). Acta Physiol Plant 33:1847–1852

    Article  CAS  Google Scholar 

  • Hua YM, Rong HN (2010) A simple cryopreservation protocol of Dioscorea bulbifera L. embryogenic calli by encapsulation-vitrification. Plant Cell Tissue Organ Cult 101:349–358

    Article  Google Scholar 

  • Jeong JH, Jung SJ, Murthy HN, Yu KW, Paek KY, Moon HK, Choi YE (2005) Production of eleutherosides in in vitro regenerated embryos and plantlets of Eleutherococcus chiisanensis. Biotechnol Lett 27:701–704

    Article  CAS  PubMed  Google Scholar 

  • Ke X, Zhang X, Shi H, Li B (1996) Electroporation of immature maize zygotic embryos and regeneration of transgenic plants. Transgenic Res 5:219–221

    Article  CAS  Google Scholar 

  • Khan MA, Abbasi BH, Ali H, Ali M, Adil M, Hussain I (2015) Temporal variations in metabolite profiles at different growth phases during somatic embryogenesis of Silybum marianum L. Plant Cell Tissue Organ Cult 120:127–139

  • Kumar V, Chandra S (2014) High frequency somatic embryogenesis and synthetic seed production of the endangered species Swertia chirayita. Biologia 69:186–192

    Article  CAS  Google Scholar 

  • Manjkhola S, Dhar U, Joshi M (2005) Organogenesis, embryogenesis, and synthetic seed production in Arnebia euchroma—a critically endangered medicinal plant of the Himalaya. In Vitro Cell Dev Biol Plant 41:244–248

    Article  Google Scholar 

  • Martin KP (2004) Plant regeneration through somatic embryogenesis in medicinally important Centella asiatica L. In Vitro Cell Dev Biol-Plant 40:586–591

    Article  CAS  Google Scholar 

  • Moyo M, Finnie JF, Van Staden J (2012) Topolins in Pelargonium sidoides micropropagation: do the new brooms really sweep cleaner? Plant Cell Tissue Organ Cult 110:319–327

    Article  CAS  Google Scholar 

  • Moyo M, Aremu AO, Gruz J, Šubrtova M, Szüčova L, Doležal K, Van Staden J (2013) Conservation strategy for Pelargonium sidoides DC: phenolic profile and pharmacological activity of acclimatized plants derived from tissue culture. J Ethnopharmacol 149:557–561

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Pathi KM, Tula S, Tuteja N (2013) High frequency regeneration via direct somatic embryogenesis and efficient Agrobacterium mediated genetic transformation of tobacco. Plant Signal Behav 8(6):e24354

    Article  PubMed Central  PubMed  Google Scholar 

  • Paul S, Dam A, Bhattacharyya A, Bandyopadhyay TK (2011) An efficient regeneration system via direct and indirect somatic embryogenesis for the medicinal tree Murraya koenigii. Plant Cell Tissue Organ Cult 105:271–283

    Article  Google Scholar 

  • Peña-Ramírez Y, García-Sheseña I, Herna´ndez-Espinoza Á, Domíguez-Herna´ndez A, Barredo-Pool F, González-Rodríguez J, Robert M (2011) Induction of somatic embryogenesis and plant regeneration in the tropical timber tree Spanish red cedar [Cedrela odorata L. (Meliaceae)]. Plant Cell Tissue Organ Cult 105:203–209

    Article  Google Scholar 

  • Raju CS, Kathiravan K, Aslam A, Shajahan A (2013) An efficient regeneration system via somatic embryogenesis in mango ginger (Curcuma amada Roxb.). Plant Cell Tissue Organ Cult 112:387–393

    Article  Google Scholar 

  • Raomai S, Kumaria S, Tandon P (2014) Plant regeneration through direct somatic embryogenesis from immature zygotic embryos of the medicinal plant, Paris polyphylla Sm. Plant Cell Tissue Organ Cult 118:445–455

    Article  CAS  Google Scholar 

  • Red Data List of South African plants version 2013.1, 2013 http://redlist.sanbi.org/. Accessed 09 Dec 2013

  • Robichaud RL, Lessard VC, Merkle SA (2004) Treatments affecting maturation and germination of American chestnut somatic embryos. J Plant Physiol 161:957–969

    Article  CAS  PubMed  Google Scholar 

  • Shrivastava S, Chawla HS (2001) Synergistic effect of growth regulators and glutamine on regeneration response in high yielding cultivars of wheat (Triticum aestivum L.). Indian J Genet 61:12–15

    CAS  Google Scholar 

  • Siddiqui Z, Mujib A, Maqsood M (2011) Liquid overlaying improves somatic embryogenesis in Catharanthus roseus. Plant Cell Tissue Organ Cult 104:247–256

    Article  Google Scholar 

  • Sivanesan I, Lim MY, Jeong BR (2011) Somatic embryogenesis and plant regeneration from leaf and petiole explants of Campanula punctata Lam. var. rubriflora Makino. Plant Cell Tissue Organ Cult 107:365–369

    Article  Google Scholar 

  • Tribulato A, Remotti PC, Loffler HJM (1997) Somatic embryogenesis and plant regeneration in Lilium longiflorum Thunb. Plant Cell Rep 17:113–118

    Article  CAS  Google Scholar 

  • Vani AKS, Reddy VD (1996) Morphogenesis from callus cultures of chickpea, Cicer arietinum L. Indian J Exp Biol 34:285–287

    Google Scholar 

  • Yin Y, Li S, Chen Y, Guo H, Tian W, Chen Y (1993) Fertile plants regenerated from suspension culture-derived protoplasts of an indica-type rice (Oryza sativa L.). Plant Cell Tissue Organ Cult 32:61–68

    Article  CAS  Google Scholar 

  • Zouine J, Hadrami IEI (2007) Effect of 2, 4-D, glutamine and BAP on embryogenic suspension culture of date palm (Phoenix dactylifera L.). Sci Hortic 112:221–226

    Article  CAS  Google Scholar 

Download references

Acknowledgments

V. Kumar and M. Moyo thank the National Research Foundation and the University of KwaZulu-Natal, South Africa for support in the form of postdoctoral fellowships. The authors are grateful to the Microscopy and Microanalysis Unit (MMU), UKZN, Pietermaritzburg for microscopic assistance.

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Authors and Affiliations

  1. Research Centre for Plant Growth and Development, School of Life Sciences, University of KwaZulu-Natal Pietermaritzburg, Private Bag X01, Scottsville, 3209, South Africa

    Vijay Kumar, Mack Moyo & Johannes Van Staden

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  1. Vijay Kumar
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  2. Mack Moyo
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  3. Johannes Van Staden
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Correspondence to Johannes Van Staden.

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Kumar, V., Moyo, M. & Van Staden, J. Somatic embryogenesis of Pelargonium sidoides DC.. Plant Cell Tiss Organ Cult 121, 571–577 (2015). https://doi.org/10.1007/s11240-015-0726-2

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  • DOI: https://doi.org/10.1007/s11240-015-0726-2

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