Abstract
An efficient micropropagation protocol was developed for the medicinal plant Phyllanthus caroliniensis (Euphorbiaceae) using nodal segments for axillary shoot proliferation. Maximum multiplication (21–23 shoots per explant) was achieved on MS or AR media supplemented with either 5.0 μM BA, 1.25–5.0 μM kinetin or 2.5–5.0 μM 2iP. Rooting was achieved with 80–100% of the microshoots on MS medium without growth regulators, although 1.25 μM NAA and 1.25–5.0 μM IAA promoted significant increases in the number of roots per explant. Regenerated plants were successfully acclimatized and about 88% of plantlets survived under ex vitro conditions. Flowering was observed on in vitro grown plantlets and after 3–4 weeks of acclimatization. High frequency callus initiation and growth was achieved when nodal segment explants were inoculated in the vertical position on MS medium supplemented with 5.0 μM 2,4-D. Root cultures were successfully established on MS medium containing 1.1 μM NAA. The optimized micropropagation, callus and root culture protocols offer the possibility to use cell/root culture techniques for vegetative propagation and secondary metabolism studies.
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References
Alizadeh SA, Mantell SH & Viana AM (1998) In vitro shoot culture and microtuber induction in the steroid yam Dioscorea composita Hemsl. Plant Cell Tiss. Org. Cult. 53(2): 107–112
Bollmark M & Eliasson L (1986) Effects of exogenous cytokinins on root formation in pea cuttings. Physiol. Plant. 68: 662–666
Calixto JB, Santos ARS, Cechinel Filho V & Yunes RA (1998) A review of the plants of the genus Phyllanthus: Their chemistry, pharmacology and therapeutic potential. Res. Med. Rev. 4: 225–258
Catapan E (1999) In vitro culture and phytochemical analysis of Phyllanthus spp. MSc. Thesis, Federal University of Santa Catarina, Florianopolis, SC, Brazil (in Brazilian Portuguese with English summary)
Cechinel Filho V, Santos ARS, Campos ROP, Miguel OG, Yunes RA, Ferrari F, Messana J & Calixto JB (1996) Chemical and pharmacological studies of Phyllanthus caroliniensis in mice. J. Pharm. Pharmacol. 48: 1231–1236
Cho DY (1985) Distribution and quantitative determination on IAA by high performance liquid chromatography on adventitious root formation in Azukia epicotyl cuttings. Korean J. Plant Tissue Cult. 12: 79–87
Fabijan D, Taylor JS & Reid DM (1981) Adventitious rooting in hypocotyls of sunflower (Helianthus annuus) seedlings II. Action of gibberellins, cytokinins, auxins and ethylene. Physiol. Plant. 53: 589–597
Haicour R, Rossignol L, Rossignol M & Gaisne C (1994) Patterns of diversification and evolution in Phyllanthus-Odontadenius (Euphorbiaceae). Annals of the Missouri Botanical Garden 81(2): 289–301
Ishimaru K, Yoshimatsu K, Yamakawa T, Kamada H & Shimomura K (1992) Phenolic constituents in tissue cultures of Phyllanthus niruri. Phytochemistry 34: 2015–2016
Ivanov P, Encheva J & Ivanova I (1998) A protocol to avoid pecocious flowering of sunflower plantlets in vitro. Plant Breeding 117(6): 582–584
Kintzios S & Michaelakis A (1999) Induction of somatic embryogenesis and in vitro flowering from inflorescences of chamomile (Chamomilla recutita L.). Plant Cell Rep. 18(7-8): 684–690
Krogh R, Kroth R, Berti C, Madeira AO, Souza MM, Cechinel Filho V, Delle-Monache F & Yunes RA (1999) Isolation and identification of compounds with antinociceptive action from Ipomoea pes-capre (L.). R. Br. Pharmazie 54: 1–6
Lomax TL, Muday GK & Rubery PH (1995) Auxin transport. In: Davies PJ (ed) Plant Hormones: Physiology, Biochemistry and Molecular Biology (pp 509–530). Kluwer Academic Publishers, Dordrecht
McCartan SA & Van Staden J (1998) Micropropagation of the medicinal plant, Scilla natalensis Planch. Plant Growth Regul. 25: 177–180
Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497
Ogata T, Higuchi H, Mochida S, Matsumoto H, Kato A, Endo T, Kaji A & Kaji H (1992) HIV-1 reverse transcriptase inhibitor from Phyllanthus niruri. AIDS Res. Hum Retroviruses 8: 1937–1944.
Pierik RLM (1989) In Vitro Culture of Higher Plants (pp 203–209). Martinus Nijhoff Publishers, Dordrecht
Rao CS, Eganathan P, Anand A, Balacrishna P & Reddy TP (1998) Protocol for in vitro propagation of Excoecaria agallocha L., a medicinally important mangrove species. Plant Cell Rep. 17(11): 861–865
Remeshree AB, Hariharan M & Unnikrishnan K (1994) Micropropagation and callus induction of Aristolochia bracteolata Lam. - a medicinal plant. Phytomorphology 44: 247–252
Santos ARS, Cechinel Filho V, Viana AM, Moreno FN, Campos MM, Yunes RA & Calixto JB (1994) Analgesic effects of callus culture extracts from selected species of Phyllanthus in mice. J. Pharm. Pharmacol. 46: 755–759
Taha RM (1997) In vitro flowering of Murraya paniculata (Jack) Linn. Asia Pacific J. Molec. Biol. Biotechnol. 5: 68–71
Ueno H, Horie S, Nishi Y, Shogawa H, Kawasakim M, Suzuki S, Hayashi T, Arisawa M, Shimizu M, Yoshizakim M, Morita N, Berganza LH, Ferro E & Basualdo I (1988) Chemical and pharmaceutical studies on medicinal plants in Paraguay. Geraniin, an angiotensin-converting enzyme inhibitor from ‘papaparai mi’, Phyllanthus niruri. J. Nat. Products 51: 357–359
Unander DW (1991) Callus induction in Phyllanthus species and inhibition of viral DNA polymerase and reverse transcriptase by callus extracts. Plant Cell Rep. 10: 461–466
Unander DW (1996) Phyllanthus species: in vitro culture and production of secondary metabolites. In: Bajaj YPS (ed) Biotechnology in Agriculture and Forestry, Vol 37 (pp 304–318). Springer-Verlag, Berlin
Unander DW(1998) Phyllanthus as a chemopreventive: a case study of why it is hard to translate ethnobotany to the clinic. Cancer Prevention International 3: 143–152
Unander DW, Bryan HH, Lance CJ & McMillan RT (1995) Factors affecting germination and stand establishment of Phyllanthus amarus (Euphorbiaceae). Econ. Bot. 49(1): 49–55
Vicent K, Mathew KM & Hariharan M (1992) Micropropagation of Kaempferia galanga L., a medicinal plant. Plant Cell Tiss. Org. Cult. 28: 229–230
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Catapan, E., Otuki, M.F. & Viana, A.M. In vitro culture of Phyllanthus caroliniensis (Euphorbiaceae). Plant Cell, Tissue and Organ Culture 62, 195–202 (2000). https://doi.org/10.1023/A:1006406806839
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DOI: https://doi.org/10.1023/A:1006406806839