In Vitro Callus Induction and Shoot Regeneration from Leaf Explants of Glinus lotoides (L.)—An Important Medicinal Plant

Abstract

G. lotoides L. is a threatened plant that is frequently harvested for medicinal purpose. However, its distribution in the world is limited because of short period of seed viability and poor seed germination. The objective of this study was to develop in vitro propagation protocol for G. lotoides through callus induction. For callus induction, different concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), NAA (α-naphthalene acetic acid) and BAP (6-benzyl amino purine) were used. Seeds were sown on growth regulator-free MS medium and shoots from the in vitro germinated seedlings were excised and cultured on MS medium containing 0.5 mg/l BAP. Young leaves from these shoots were used as explant for callus induction and shoot regeneration. Maximum callus induction (100%) was observed on medium containing 2,4-D (0.5, 2.0, 3.5 mg/l) or NAA (2.0, 2.5 mg/l) in combination with 0.5 mg/l BAP. However, 2,4-D was the best in overall callus induction. The highest regeneration (20%) frequency was achieved on the medium containing 0.5 mg/l BAP. Highest number of shoot (2.83 ± 1.22) and shoot length (2.16 ± 0.87 cm) per explant were obtained in the presence of 0.25 mg/l BAP + 0.5 mg/l KIN (Kinetin). In shoot multiplication media, maximum mean (6.43 ± 0.87) of shoot was observed on MS medium containing 0.5 mg/l BAP. The best shoot length (1.70 ± 0.14 cm) was recorded on control medium. The highest (95%), maximum root number (14.10 ± 1.47) and root length (1.01 ± 0.10 cm) were obtained on a medium supplemented with 1.5 mg/l Indole-3-butyric acid (IBA). All the plants (100%) were survived after acclimatization in greenhouse. The present study can be useful for callus induction and indirect shoot regeneration form G. lotoides.

Share and Cite:

Teshome, S. and Feyissa, T. (2015) In Vitro Callus Induction and Shoot Regeneration from Leaf Explants of Glinus lotoides (L.)—An Important Medicinal Plant. American Journal of Plant Sciences, 6, 1329-1340. doi: 10.4236/ajps.2015.69132.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] Dequan, L. and Hartmann, H.E.K. (2003) Molluginaceae. Flora of China, 5, 437-439.
[2] Gilbert, M.G. (2000) Magnoliaceae to Flacourtiaceae. In: Edwards, S., Tadesse, M., Demissew, S. and Hedberg, I., Eds., Flora of Ethiopia and Eritrea, Vol. 2, The National Herbarium, Addis Ababa University, Ethiopia, 234.
[3] Kloos, S. (1976) Preliminary Studies of Medicinal Plants and Plant Products in Ethiopian Markets. Journal of Ethiopian Pharmaceutical Association, 2, 18-28.
[4] Mulatu, D. (1978) Taenicidal Activity of Glinus lotoides (Aizoaceae). Journal of Ethiopian Pharmaceutical Association, 3, 9-11.
[5] Gidey, Y., Mekonen, T. and Mezgebe, K. (2011) Survey of Medicinal Plants Used to Treat Human Ailments in Hawzen District, Northern Ethiopia. International Journal of Biodiversity and Conservation, 3, 709-714.
http://www.academicjournals.org/IJBC
[6] Mesfin, T. and Obsa, T. (1994) Ethiopian Traditional Veterinary Practices and Their Possible Contribution to Animal Production and Management. Revue Scientifique et Technique (International Office of Epizootics), 13, 417-424.
[7] Abebe, E. and Youan, B.C. (2010) Anticancer Activity and Nutritional Value of Extracts of the Seed of Glinus lotoides. Journal of Nutritional Science and Vitaminology, 56, 311-318.
http://dx.doi.org/10.3177/jnsv.56.311
[8] Kumar, G., Banu, G.S., Rajarajan, T. and Sathishkumar, G. (2011) Medicinal Flora of Palayapalayam, Namakkal District, Tamilnadu. Indian Journal of Applied and Pure Biology, 26, 135-158.
[9] Shanmugam, S., Rajendran, K. and Suresh, K. (2012) Traditional Uses of Medicinal Plants among the Rural People in Sivagangai District of Tamil Nadu, Southern India. Asian Pacific Journal of Tropical Biomedicine, 2, 429-434. www.elsevier.com/locate/apjtb
http://dx.doi.org/10.1016/S2221-1691(12)60201-9
[10] Qureshi, R. and Bhatti, G.R. (2008) Ethno Botany of Plants Used by the Thari People of Nara Desert, Pakistan. Fitoterapia, 79, 468-473. www.elsevier.com/locate/fitote
http://dx.doi.org/10.1016/j.fitote.2008.03.010
[11] Qureshi, R., Bhatti, G.R. and Memon, R.A. (2010) Ethno Medicinal Uses of Herbs from Northern Part of Nara Desert, Pakistan. Pakistan Journal of Botany, 42, 839-851.
[12] Chopra, R.N., Nayar, S.I. and Chopra, I.C. (1956) Glossary of Indian Medicinal Plants. CSIR, New Delhi.
[13] Berhanu, A. and Berhane, T. (1980) A New Triterpenoid Glycoside from the Seeds of Glinus lotoides. Phytochemistry, 19, 1553-1554.
http://dx.doi.org/10.1016/0031-9422(80)80221-4
[14] Endale, A., Getachew, M. and Gebre-Mariam, T. (1997) In Vitro Taenicidal Activity of the Seeds of Glinus lotoides on Hymenolepis nana Worms. Ethiopian Pharmaceutical Journal, 15, 46-51.
[15] Kavimani, S., Manisenthilkumar, K.T., Ilango, R. and Krishnamoorthy, G. (1999) Effect of the Methanolic Extract of Glinus lotoides on Dalton’s Ascitic Lymphoma. Biological & Pharmaceutical Bulletin, 22, 1251-1252.
[16] Balcha, A. (2009) Studies on Propagation Biology of Four and Phenology of One Medicinal plant. Ph.D. Dissertation, Addis Ababa University, Addis Ababa, Ethiopia.
[17] Endale, A., Kassa, M. and Gebre-Mariam, T. (1998) In Vivo Anthelminthic Activity of the Extract of the Seeds of Glinus lotoides in Albino Mice Infested with Hymenolepis nana Worms. Ethiopian Pharmaceutical Journal, 16, 34-41.
[18] Biftu, T., Berhanu, A. and Teffera, S. (1979) Fatty Acid Composition of “Mettere” (Glinus lotoides Linne) Seeds by Gas Liquid Chromatography (GLC). SINET: Ethiopian Journal of Science, 2, 19-22.
[19] Endale, A., Schmidt, P.C. and Gebre-Mariam, T. (2004) Standardisation and Physicochemical Characterisation of the Extracts of Seeds of Glinus lotoides. Die Pharmazie, 59, 34-38.
[20] El-Zayat, S.A. (2008) Preliminary Studies on Laccase Production by Chaetomium globosum an Endophytic Fungus in Glinus lotoides. American-Eurasian Journal of Agricultural & Environmental Sciences, 3, 86-90.
[21] Abebe, E. (2005) Isolation, Structural Elucidation, Quantification and Formulation of the Saponins and Flavonoids of the Seeds of Glinus lotoides. PhD Dissertation, Eberhard Karls University, Tubingen, Germany.
[22] Endale, A., Gebre-Mariam, T. and Schmidt, P.C. (2008) Granulation by Roller Compaction and Enteric Coated Tablet Formulation of the Extract of the Seeds of Glinus lotoides Loaded on Aeroperl® 300 Pharma. AAPS PharmSciTech, 9, 31-38.
http://dx.doi.org/10.1208/s12249-007-9027-3
[23] Demma, J., Gebre-Mariam, T., Asres, K., Ergetie, W. and Engidawork, E. (2007) Toxicological Study on Glinus lotoides: A Traditionally Used Taenicidal Herb in Ethiopia. Journal of Ethno Pharmacology, 111, 451-457. www.elsevier.com/locate/jethpharm
http://dx.doi.org/10.1016/j.jep.2006.12.017
[24] Ould Ahmedou, M.L., Bouaichi, A. and Idrissi Hassani, L.M. (2001) Evidence of Deterrent and Toxic Effects of Glinus lotoides (Aizoacées) on Nymphs of the Desert Locust Schistocerca gregaria Forskål (Orthoptera, Acrididae). Zoologica Baetica, 12, 109-117.
[25] Hamed, A.I., Piacente, S., Autore, G., Marzocco, S., Pizza, C. and Oleszek, W. (2005) Antiproliferative Hopane and Oleanane Glycosides from Roots of Glinus lotoides. Planta Medica, 71, 554-560.
http://dx.doi.org/10.1055/s-2005-864158
[26] UNEP (United Nations Environment Programme) (2012) Capacity Building for Access and Benefit Sharing and Conservation and Sustainable Use of Medicinal Plants (Ethiopia ABS CSUMP). Project document.
[27] Murashige, T. and Skoog, F. (1962) A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Cultures. Physiologia Plantarum, 15, 473-97.
http://dx.doi.org/10.1111/j.1399-3054.1962.tb08052.x
[28] Priya, G. and Chellaram, C. (2014) In Vitro Derived Callus and Shoot of a Medicinal Herb Solanum trilobatum and Their Effect on Hepatocellular Carcinoma. International Journal of Pharmacy and Pharmaceutical Sciences, 6, 634-637.
[29] Tanveer, H., Ali, S. and Asi, M.R. (2012) Appraisal of an Important Flavonoid, Quercetin, in Callus Cultures of Citrullus colocynthis. International Journal of Agriculture and Biology, 14, 528-532.
http://www.fspublishers.org
[30] Kaladhar, D.S.V.G.K. (2012) An in Vitro Callus Induction and Isolation, Identification, Virtual Screening and Docking of Drug from Convolvulus alsinoides Linn against Aging Diseases. International Journal of Pharma Medicine and Biological Sciences, 1, 92-103.
http://www.ijlbpr.com/currentissue.php
[31] Sen, M.K., Nasrin, S., Rahman, S. and Jamal, A.H.M. (2014) In Vitro Callus Induction and Plantlet Regeneration of Achyranthes aspera L., a High Value Medicinal Plant. Asian Pacific Journal of Tropical Biomedicine, 4, 40-46. www.elsevier.com/locate/apjtb
http://dx.doi.org/10.1016/S2221-1691(14)60206-9
[32] Lim, Z.X., Ling, A.P.K. and Hussein, S. (2009) Callus Induction of Ocimum sanctum and Estimation of Its Total Flavonoids Content. Asian Journal of Agricultural Sciences, 1, 55-61.
[33] Arunkumar, B.S. and Jayaraj, M. (2011) Effect of Auxins and Cytokinins on in Vitro Propagation of Ionidium suffruticosum Ging—A Seasonal Multipotent Medicinal Herb. International Journal of Research in Ayurveda and Pharmacy, 2, 198-203. www.ijrap.net
[34] Wani, M., Pande, S. and More, N. (2010) Callus Induction Studies in Tridax procumbens L. International Journal of Biotechnology Applications, 2, 11-14.
http://dx.doi.org/10.9735/0975-2943.2.1.11-14
[35] Acemi, A., Ozen, F. and Kiran, R. (2012) Development of an Efficient Callus Production Protocol for Amsonia orientalis: A Critically Endangered Medicinal Plant. EurAsian Journal of BioSciences, 6, 105-112.
http://dx.doi.org/10.5053/ejobios.2012.6.0.13
[36] Kumar, A., Kaushal, S., Sharma, S. and Sood, H. (2014) In Vitro Callus Induction and Plantlet Regeneration of Swertia chirayita: A Critically Endangered Medicinal Plant. International Journal of Pharmaceutical Sciences Review and Research, 27, 94-96. www.globalresearchonline.net
[37] Padmavathy, S. (2014) Callus Production Studies in Strobilanthus foliosus and Smilax wightii. International Journal of Innovative Drug Discovery, 4, 41-45. www.ijidd.com
[38] Guruchandran, V. and Sasikumar, C. (2013) Organogenic Plant Regeneration via Callus Induction in Stevia rebaudiana Bert. International Journal of Current Microbiology and Applied Sciences, 2, 56-61.
http://www.ijcmas.com
[39] Lalabadi, M.A., Jelodar, N.B. and Bagheri, N. (2013) Effect of Plant Hormones on Callus Induction of Explant Types in Endangered Medicinal Herb Tashnedari. International Journal of Biosciences, 3, 39-43.
http://www.innspub.net
http://dx.doi.org/10.12692/ijb/3.12.39-43
[40] Sridhar, T.M. and Naidu, C.V. (2011) An Efficient Callus Induction and Plant Regeneration of Solanum nigrum (L.)— An Important Antiulcer Medicinal Plant. Journal of Phytology, 3, 23-28. www.scholarjournals.org
[41] Reshi, N.A., Sudarshana, M.S. and Rajashekar, N. (2013) Callus Induction and Plantlet Regeneration in Orthosiphon aristatus (Blume) Miq.—A Potent Medicinal Herb. IOSR Journal of Pharmacy and Biological Sciences, 6, 52-55. www.iosrjournals.org
http://dx.doi.org/10.9790/3008-0635255
[42] Zare, A.R., Solouki, M., Omidi, M., Irvani, N., Nezad, N.M. and Rezazadeh, Sh. (2010) Callus Induction and Plant Regeneration in Ferula assafoetida L. (Asafetida), an Endangered Medicinal Plant. Trakia Journal of Sciences, 8, 11-18.
http://www.uni-sz.bg
[43] Martin, K.P. (2004) Efficacy of Different Growth Regulators at Different Stages of Somatic Embryogenesis in Eryngium foetidum L.—A Rare Medicinal Plant. In Vitro Cellular & Developmental Biology-Plant, 40, 459-463.
http://dx.doi.org/10.1079/IVP2004543
[44] Jain, A.K. and Bashir, M. (2010) In-Vitro Propagation of a Medicinal Plant Portulaca grandiflora. Hook. World Journal of Agricultural Sciences, 6, 327-330.
[45] Aicha, N., Rachida, T.C. and El Abdelmalek, M. (2013) Micropropagation of Thymus satureioides Coss. an Endangered Medicinal Plant of Morocco. Journal of Agricultural Technology, 9, 487-501.
http://www.ijat-aatsea.com
[46] Oluk, E.A. and Çakyr, A. (2009) Micropropagation of Origanum sipyleum L., an Endemic Medicinal Herb of Turkey. African Journal of Biotechnology, 8, 5769-5772.
http://www.academicjournals.org/AJB
[47] Echeverrigaray, S., Carrer, R.P. and Andrade, L.B. (2010) Micropropagation of Salvia guaranitica Benth. through Axillary Shoot Proliferation. Brazilian Archives of Biology and Technology, 53, 883-888.
http://dx.doi.org/10.1590/S1516-89132010000400018
[48] Tejavathi, D.H. and Padma, A.V. (2012) In Vitro Multiplication of Majorana hortensis Moench—An Aromatic Medicinal Herb. Indian Journal of Plant Sciences, 1, 48-56.
http://www.cibtech.org/jps.htm
[49] Balaraju, K., Agastian, P., Preetamraj, J.P., Arokiyaraj, S. and Ignacimuthu, S. (2008) Micropropagation of Vitex agnus-castus, (Verbenaceae)—A Valuable Medicinal Plant. In Vitro Cellular & Developmental Biology-Plant, 44, 436- 441.
http://dx.doi.org/10.1007/s11627-008-9155-9
[50] Rasool, R., Kamili, A.N., Ganai, B.A. and Akbar, S. (2009) Effect of BAP and NAA on Shoot Regeneration in Prunella vulgaris. Journal of Natural Sciences and Mathematics, 3, 21-26.
[51] Pandey, P., Mehta, R. and Upadhyay, R. (2013) In-Vitro Propagation of an Endangered Medicinal Plant Psoralea corylifolia Linn. Asian Journal of Pharmaceutical and Clinical Research, 6, 115-118.
[52] Bera, T., Haque, R. and Saha, S. (2009) Micropropagation of an Important Medicinal Plant Chlorophytum borivilianum. International Journal of Physical Sciences, 1, 153-163. www.ijps.info
[53] Jitendra, M., Monika, S., Ratan, S.D., Priyanka, G., Priyanka, S. and Kiran, D.J. (2012) Micropropagation of an Anti Diabetic Plant—Stevia rebaudiana Bertoni, (Natural sweetener) in Hadoti Region of South-East Rajasthan, India. ISCA Journal of Biological Sciences, 1, 37-42. www.isca.in
[54] Rathore, V. and Gupta, D. (2013) In Vitro Clonal Propagation of Solanum nigrum. Research Journal of Chemical and Environmental Sciences, 1, 32-33.
http://www.aelsindia.com/rjces.htm
[55] Fracaro, F. and Echeverrigaray, S. (2001) Micropropagation of Cunila galioides, a Popular Medicinal Plant of South Brazil. Plant Cell, Tissue and Organ Culture, 64, 1-4.
http://dx.doi.org/10.1023/A:1010626200045
[56] Sebastinraj, J., Britto, S.J. and Kumar, S.R.S. (2013) Micropropagation of Operculina turpethum (Linn.) Silva Manso. Using Cotyledonary Node Explants. Academic Journal of Plant Sciences, 6, 77-81.
[57] Rout, G.R. (2004) Effect of Cytokinins and Auxins on Micropropagation of Clitoria ternatea L. Biological Letters, 41, 21-26.
http://www.biollett.amu.edu.pl
[58] Rahman, Md.M., Amin, M.N., Islam, Md.Z. and Sultana, R.S. (2011) Mass Propagation of Solanum surattense Bum. Using Direct Adventitious Shoot Organogenesis from Internode. Acta Agriculturae Slovenica, 97, 11-17.
http://dx.doi.org/10.2478/v10014-011-0002-z

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.