In Vitro Shoot and Callus Induction and Alkaloid Contents of Ephedra intermedia (Schrenket) of Iran
Journal of Plant Sciences
Volume 3, Issue 1, February 2015, Pages: 1-8
Received: Dec. 12, 2014; Accepted: Dec. 18, 2014; Published: Jan. 19, 2015
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Azra Ataei Azimi, Associate professorof plant cell and developmental biology, Department of Biology, Saveh Branch, Islamic Azad University, Saveh, Iran
Babak Delnavaz Hashemloian, Associate professorof plant cell and developmental biology, Department of Biology, Saveh Branch, Islamic Azad University, Saveh, Iran
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Ephedra intermedia(Schrenket) isan evergreen shrub belongs to family Ephedraceae.The present study deals with Invitro callus induction, multiple shoots regeneration and alkaloid production of E.intermedia. Callus andmultiple shoots formation were initiated on MS medium supplementedwith different concentrations of 1-Naphthalene Acetic Acid (NAA)and Kinetin (Kn)hormonescombination. Alkaloid analysis was done by spectrophotometry, TLC, HPLC methods.Alkaloid was found in all tissue cultures and E. intermedia organs. Total alkaloids were higher in roots and some shoots cultures than of ephedra shoots and seeds, cultures shoots and callus. The ephedrine and pseudoephedrine were higher in ephedra roots than in ephedra shoots and seeds, all callusculturesand some shoots cultures but they balancedin roots with shoots cultures of 15 mgl-1 NAA supplemented with 10 mgl-1 Kn.This study can be utilized to initiate multiple shoots culture of Ephedrathat may provide a good source of pharmacologically active constituents.
Ma-Haung, Multiple Shoots, Ephedrine, Pseudoephedrine
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Azra Ataei Azimi, Babak Delnavaz Hashemloian, In Vitro Shoot and Callus Induction and Alkaloid Contents of Ephedra intermedia (Schrenket) of Iran, Journal of Plant Sciences. Vol. 3, No. 1, 2015, pp. 1-8. doi: 10.11648/j.jps.20150301.11
Friedman WE (1998) The evolution of double fertilization and endosperm: an historical perspective. Plant Reprod 11:6–16.
Stevens J (2003) The Ephedra Story. Application Note 215:1-6.
Caveney S., Charlet DA., Freitag H, Maier-Stolte M, Strarratt AN (2001) New observations of the secondary chemistry of world Ephedra (Ephedraceae). Am J Bot 88: 1199- 1280.
Pearson HHW (1929) Gnetales. Cambridge University Press, London, UK.
Freitag H, Maier-Stolte M (1989) The Ephedra-species of P. Forsskal: identity and typification. Taxon 38: 545-556.
Zhu YP (1998) Chinese MateriaMedica: chemistry, pharmacology and applications. Harwood Academic, Amsterdam, Netherlands.
White LM, Gardner SF, Gurley BJ, Marx MA, Wang PL, Estes M (1997) Pharmacokinetics and cardiovascular effects of Ma huang (Ephedra sinica) in normotensive adults. J ClinPharmacol 37: 116–122.
Qazilbach N A (1971) Pakistan Ephedra. PharmacologischeWeekblad 106: 345-349.
Parsaeimehr A, Sargsyan E,Javidnia K ( 2010) A Comparative Study of the Antibacterial, Antifungal and Antioxidant Activity and Total Content of Phenolic Compounds of Cellcultures and Wild Plants of Three Endemic Species of Ephedra. Molecules 15: 1668-1678.
Bagheri G, Bigdeli M (2009) Inhibitory Effects of Ephedra major Host on Aspergillusparasiticus Growth and Aflatoxin Production. Mycopathology 168: 249-255
Hollander JL, Wall SBV (2009) Dispersal syndromes in North American Ephedra. Inter J Plant Sci 170(3): 323-330.
Lee CH, Lee HS (2009) Growth Inhibiting Activity of Quinaldic Acid Isolated from Ephedra pachycladaagainst Intestinal Bacteria. Korean SocApplBiol 52(4): 331-335.
Singh D, Singh D, Choi SM, Han SS (2013) Enhanced Proliferation and Growth of Human Lung Epithelial Cells on Gelatin Microparticle Loaded with Ephedra Extracts. J Nanomaterials 2013:1-8.
Nawwar MAM,Barakat HH, Buddrust J, Linscheidt M(1985) Alkaloid, lignan and phenolic constituents of Ephedra alata. Phytochemistry 24: 878 – 879.
Ganzera M, Lanser C, Stuppner H (2005) Simultaneous determination of Ephedra sinicaand Citrus aurantium var. amara alkaloids by ionpair chromatography. Talanta 66(4): 889-894.
Inoko A, Kakiuchi N, Yoshimitsu M, Cai SQ, Mikage M (2007) Ephedra Resource in Sichuan and Yunnan Provinces. Biol Pharm Bull 32(9): 1621-1623.
Kitani Y, Zhu S, Omote T, Tanaka K, Batkhuu J, Sanchir C, Fushimi H, Mikage M, Komatsu K (2009) Molecular Analysis and Chemical Evaluation of Ephedra Plants in Mongolia. Biol Pharm Bull 32(7): 1235-1243. 907-916.
Liu YM, Tian W, Jia YX, Yue HY (2009) Simultaneous determination of methylephedrine and pseudoephedrine in human urine by CE with electrochemiluminescence detection and itsapplication to pharmacokeinetics Biomed. Chromatogra 23(11): 1138-1144.
Fujita Y, Tabata M, Geen CE, Somers DA, Hackett WP, Biesboer DD (1987) Secondary metabolites from plant cell- Pharmaceutical applications and progress in commercial production, Plant tissue and cell Culture. pp. 169-185.
Kim KW, De Hertogh AA (1997) Tissue culture of Ornamental flowering bulbs (Geophytes). Horticult Rev 18: 87-169.
Laetsch WM, Boasson R (1972) Effect of growth regulators on organelle development. Horm. Regul. Plant Growth devProcAdv Study Inst H Kaldewy, ed. VerlagChemie, Weinheim/Bergest, Germany, pp. 453-465.
Uddin A (1977) Production of amino acids in Ephedra foliata suspension cultures. CurrSci 46: 825‐826.
O'Dowd N A, Richardson DHS (1994) Production of tumours and roots by Ephedra following Agrobacterium rhizogenes infection, Can J Bot (72): 2 203-207.
O’Dowd NA, Richardson DHS (1993) In vitro regeneration and multiplication of shoots from Ephedra. Phytomorphology 43: 19‐24.
Bhatnagar SP and Singh MN (1984) Organogenesis in the cultured female gametophytes of Ephedra foliata. J Exp Bot 35: 268‐278.
Sharma V, Gupta SK, Dhiman M (2012) Regeneration of Plants from Nodal and Internodal Segment Cultures of Ephedra gerardiana using Thidiazuron, Plant Tissue Cult & Biotech 22(2): 153-161.
Hegazy GAM, El-Lamey TM (2011) Callus induction and extraction of ephedrine from Ephedra alata. Ameri- Eurasi J Agric& Environ Sci 11(1):19-25.
Dixon R A (1987) Plant cell and tissue culture. IRI Press, Oxford, 1-67
Sheu SJ, Huang MH (2001) Determination of Ephedra Alkaloids by High PerformancedLiquiedChromatogaphy. Chromatoga 54: 117- 119.
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473-497.
Patel K and Thorpe TA (1984) In vitro differentiation of plantlets from embryonic explants of lodgepole pine (PinuscontortaDougl. ex Loud.). Plant Cell Tiss Org Cult 3: 131‐142.
Toivonen PMA, Kartha KK (1988) Regeneration of plantlets from in vitro cultured cotyledons of white spruce (Piceaglauca(Moench) Voss.). Plant Cell Rep 7: 318‐ 321.
Mott RL and Amerson HV (1981) A tissue culture process for the clonal production of loblolly pine plantlets. Tech Bull 271:1‐14.
Kaul K (1987) Plant regeneration from cotyledon hypocotyls explants of PinusstrobusL. Plant Cell Rep 6: 5‐7.
Dhiman M, Sharma V, Moitra S ( 2010) Somatic Embryogenesis and Plant Regeneration in Ephedra foliata(Boiss.); A non-Coniferous Gymnosperm. Plant Tissue Cult & Biotech. 20(2): 133-143.
Banerjee M (1991) In vitro studies on two Himalayan gymnosperms: PinusgerardianaWall. and Ephedra gerardianaWall. Ph.D. Thesis: University of Delhi, Delhi, India.
Singh MN and Konar RN (1981) In vitro induction of haploid roots and shoots from female gametophyte of Ephedra foliataBoiss. BeitrBiolPflanzen 55: 169‐177.
Singh MN, Konar RN and Bhatnagar SP (1981) Haploid plantlet formation from female gametophyte of Ephedra foliataBoiss. In vitro Ann Bot 48: 215‐220.
Al-Gendy AA, Bakr RO, El-gindi OD (2013) Somatic embryogenesis and plant regeneration from callus and suspension culture of Iphionamucrona.Euro Scien J 9(27):37-49.
Mousavi B, ParsaeimehrA, Irvani N (2011) Influences of growth regulators on callus induction, ephedrine and pseudoephedrine contents and chemical analysis of mature embryo of Ephedra strobilacea Bunge. AdvaAgri& Bot 3 (1): 39-45.
Mungole AJ, Doifode VD, Kamble RB, Chaturvedi A and Zanwar P (2011)In vitro regeneration of Physalis minima. Anna of Biol Res, 2 (2): 79-85.
Cao Y, Xu X, Zhao J, Chen F (2000) Cell suspension culture of Ephedra intermediaSchrenket. Chin JAppli and Environ Biol 6 (1) 36-38.
O'Dowd NA, McCauley G, Wilson JAN, Parnell TAK, Kavanaugh D (1998) In vitro culture, micropropagation and the production of ephedrine and other alkaloids. In Biotechnology in Agriculture and Forestry; Bajaj, YPS, Ed, Springer: Berlin, Germany. pp.41.
Garla M, Pratush A, Kumar S, Singh MN,ShivaniS (2011) In-vitro callus induction and shoot regeneration in Ephedra:A medicinal Plant. AnnaBiol Res 2 (6):645-651.
Long C,Kakiuchi N, Zhong G,Mikage M (2005) Survey on Resources of Ephedra Plants in Xinjiang.Biol Pharm Bull 28(2)285-288.
Walker RB, Dholakia, VN, Brasfield KL, Bakhitiar R (1998) Effect of hydroxypropyl-b-cyclodextrin on the central stimulant activity of ephedrine and oxazolidineprodrug in rats. Gen Pharmacol 30: 725–731.
Parsaeimehr A, Sargsyan E, Javidnia K (2010) Influence of plant growth regulators on callus induction, growth, chlorophyll, ephedrine and pseudoephedrine contents in Ephedra procera. J Med Plants Res 4(13): 1308-1317.
Lodha D, Rathore N, KatariaV,Shekhavat NS (2014a)In vitro propagation of female Ephedra foliataBoiss. &Kotschy ex Boiss.: an endemic and threatened Gymnosperm of the Thar Desert. Physiology and Molecular Biology of plants. 20(3):375–383.
Lodha D, Kumar Patel A, RaiMK,Shekhavat NS (2014b)In vitro plantlet regeneration and assessment of alkaloid contents from callus cultures of Ephedra foliata (Unthphog), a source of anti-asthmatic drugs.Actaphysiol 36(11):1667 -1677.
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