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Morpho-histological characterization and direct shoot organogenesis in two types of explants from Bacopa monnieri on unsupplemented basal medium

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Abstract

In the present study, high frequency regeneration has been obtained via de novo direct shoot organogenesis from leaf and internode explants in Murashige and Skoog (MS) basal medium without any phytohormone supplementation in Bacopa monnieri, an indigenous traditionally used medicinal herb. Leaves and internodes from different positions were excised from 4-weeks-old in vitro propagated B. monnieri plants and cultured on MS basal medium supplemented with 3% (w/v) sucrose and 0.75% (w/v) agar for 4 weeks. The induction of de novo shoot buds was observed at petiolar cut edges of leaf and both proximal and distal cut ends of internode explants within 10–15 days of culture. The first histological changes could be observed after 4–5 days, with meristematic activity of vascular bundles. Proliferation of epidermal cells gave rise to dome-shaped protuberances followed by shoot apical meristems formation and their vascular connections with explant tissues within 2 weeks of culture. However, a basipetal gradient of shoot regeneration from both types of explants collected along the branch axis was noticed after 4 weeks of culture. Leaf and internode explants near the basal region exhibited significantly higher number of shoot buds and micro shoots (8.8/leaf explant and 15/internode explant). Microshoots (7–12 micro shoots/leaf or internode explants) elongated (shoot length 8–9 cm) within 8 weeks on phytohormone free MS medium. Excised micro shoots rooted (100%) in hormone free MS medium within two weeks of culture. Rooted plants were then acclimatized and transferred to field with 95% survival. This protocol may be used for micropropagation, genetic transformation as well as a model system for evaluation of changes associated with acquisition of competence of differentiated cells in phytohormone free medium.

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References

  • Bairu MW, Aremu AO, Van Staden J (2011) Somaclonal variation in plants: causes and detection methods. Plant Growth Regul 63:147–173

    Article  CAS  Google Scholar 

  • Chen L, Tong J, Xiao L, Ruan Y, Liu J, Zeng M, Huang H, Wang JW, Xu L (2016) YUCCA-mediated auxin biogenesis is required for cell fate transition occurring during de novo root organogenesis in Arabidopsis. J Exp Bot 67:4273–4284

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Das S, Jha TB, Jha S (1996) Organogenesis and regeneration from pigmented callus in Camellia sinensis (L.) O. Kuntze cv. Nandadevi, an elite Darjeeling tea clone. Plant Sci 121:207–212

    Article  CAS  Google Scholar 

  • Grzyb M, Kalandyk A, Waligórski P, Mikuła A (2017) The content of endogenous hormones and sugars in the process of early somatic embryogenesis in the tree fern Cyathea delgadii Sternb. Plant Cell Tiss Organ Cult. doi:10.1007/s11240-017-1185-8

    Google Scholar 

  • Hicks GS (1994) Shoot induction and organogenesis in vitro: a developmental perspective. In vitro Cell Dev Biol 30:10–15

    Article  Google Scholar 

  • Karp A (1991) On the current understanding of somaclonal variation. In: Miflin BJ (ed) Oxford survey of plant molecular and cell biology. Oxford University Press, Oxford, pp 1–58

    Google Scholar 

  • Klu GYP, Raemakers CJJM, Jacobsen E, van Harten AM (2002) Direct organogenesis and somatic embryogenesis in mature cotyledon explants of winged bean (Psophocarpus tetragonolobus (L.) DC) using cytokinin-based media. Plant Genet Resour Newsl 131:63–69

    Google Scholar 

  • Kumar N, Reddy MP (2011) In vitro plant propagation: a review. J For Environ Sci 27:61–72

    Google Scholar 

  • Li R, Bruneau AH, Qu R (2010) Tissue culture-induced morphological somaclonal variation in St. Augustinegrass [Stenotaphrum secundatum (Walt.) Kuntze]. Plant Breed 129:96–99

    Article  Google Scholar 

  • Majumdar S, Basu A, Paul P, Halder M, Jha S (2013) Bacosides and Neuroprotection. In: Ramawat KG, Merillon JM (eds) Handbook of natural products—phytochemistry, botany, metabolism. Springer, Berlin, pp 3639–3660

    Google Scholar 

  • Marques NT, Nolasco GB, Leitão JP (2011) Factors affecting in vitro adventitious shoot formation on internode explants of Citrus aurantium L. cv. Brazilian. Sci Hort 129:176–182

    Article  CAS  Google Scholar 

  • Mukherjee DG, Dey CD (1966) Clinical trial on Brahmi. Int J Exp Med Sci 10:5–11

    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 

  • Nicholson KL, Tarlyn N, Armour T, Swanson ME, Dhingra A (2012) Effect of phyllotactic position and cultural treatments toward successful direct shoot organogenesis in dwarf ‘Pixie’ grapevine (Vitis vinifera L.). Plant Cell Tiss Organ Cult 111:123–129

    Article  Google Scholar 

  • Owens LD, Cohen JD, Seelke R (1988) Gene introduction to induce morphogenesis. Hort Sci 23:520–525

    Google Scholar 

  • Sang YL, Cheng ZJ, Zhang XS (2016) Endogenous auxin biosynthesis and de novo root organogenesis. J Exp Bot 67:4011–4013

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sato M, Hosokawa M, Doi M (2011) Somaclonal variation is induced de novo via the tissue culture process: a study quantifying mutated cells in Saintpaulia. PLos ONE 6:e23541

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sharma P, Rajam MV (1995) Genotype, explant and position effects on organogenesis and somatic embryogenesis in eggplant (Solanum melongena L.). ‎J Exp Bot 46:135–141

    Article  CAS  Google Scholar 

  • Shrivastava N, Rajani M (1999) Multiple shoot regeneration and tissue culture studies on Bacopa monnieri (L.) Pennell. Plant Cell Rep 18:919–923

    Article  CAS  Google Scholar 

  • Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissue cultured in vitro. Symp Soc Exp Biol 11:118–131

    CAS  PubMed  Google Scholar 

  • Sokal RR, Rohlf FJ (1987) Introduction to biostatistics. WH Freeman, New York

    Google Scholar 

  • Talbot MJ, White RG (2013) Methanol fixation of plant tissue for scanning electron microscopy improves preservation of tissue morphology and dimensions. Plant Methods 9:36–42

    Article  PubMed  PubMed Central  Google Scholar 

  • Thorpe TA (1980) Organogenesis in vitro: structural, physiological and biochemical aspects. In: Vasil IK (ed) International review of cytology suppl 11 A. Perspectives in plant cell and tissue culture. Academic Press, New York, pp 71–111

    Google Scholar 

  • Tiwari V, Singh BD, Tiwari KN (1998) Shoot regeneration and somatic embryogenesis from different explants of Brahmi [Bacopa monnieri (L.) Wettst.]. Plant Cell Rep 17:538–543

    Article  CAS  Google Scholar 

  • Tiwari V, Tiwari KN, Singh BD (2001) Comparative studies of cytokinins on in vitro propagation of Bacopa monniera. Plant Cell Tiss Org Cult 66:9–16

    Article  CAS  Google Scholar 

  • Tran Than Van K (1980) Control of morphogenesis by inherent and exogenously applied factors in thin cell layers. Int Rev Cytol Suppl. 11A:175–194

    Google Scholar 

  • Wabiko H, Minemura M (1996) Exogenous phytohormone-independent growth and regeneration of tobacco plants transgenic for the 6b gene of Agrobacterium tumefaciens AKE10. Plant Physiol 112:939–951

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yamuna P, Bose I, Jha TB, Jha S (1993) Direct organogenesis from internodal segments of in vitro grown shoots of Ipecac, Cephaelis ipecacuanha Rich. Ind J Expt Biol 31:926–929

    Google Scholar 

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Acknowledgements

The authors thank the Head, Department of Botany, Calcutta University and the Programme Coordinator, CAS in Botany, CU, for facilities provided.

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Authors and Affiliations

  1. Center of Advanced Study, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata, West Bengal, 700 019, India

    Sayantika Sarkar & Sumita Jha

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  1. Sayantika Sarkar
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  2. Sumita Jha
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Correspondence to Sumita Jha.

Additional information

Communicated by Sergei Krasnyanski.

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11240_2017_1231_MOESM1_ESM.tif

Supplementary material 1: A 4-week-old in vitro grown plant of B. monnieri illustrating positions of leaf and internode explants excised for in vitro culture (TIF 243 KB)

11240_2017_1231_MOESM2_ESM.tif

Supplementary material 2: Scanning electron micrographs of leaf explants showing different stages of shoot organogenesis. a Proximal cut end of leaf explant showing no distinct changeat day 5, b Portion of leaf in transverse section showing formation of shoot apical meristem (SAM) with leaf primordial (LP) at day 10, c Development of shoot buds (SB enlarged view in inset) and micro shoots (MS) from proximal cut endat day 15, d Elongation of micro shoots with expansion of leaves at day 21 (TIF 1646 KB)

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Sarkar, S., Jha, S. Morpho-histological characterization and direct shoot organogenesis in two types of explants from Bacopa monnieri on unsupplemented basal medium. Plant Cell Tiss Organ Cult 130, 435–441 (2017). https://doi.org/10.1007/s11240-017-1231-6

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  • DOI: https://doi.org/10.1007/s11240-017-1231-6

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