Back
 AJPS  Vol.5 No.5 , March 2014
Highly Efficient in Vitro Regeneration Method of Endangered Medicinal Plant Heliotropium kotschyi (Ramram) in the Kingdom of Bahrain
Abstract: Heliotropium kotschyi (Ramram), is a very important plant in relation to environment protection and for its medicinal value in Bahrain. It has been detected as one of the endangered plants in Bahrain. Ex situ conservation of this plant applying tissue culture method was used in the present study. In vitro plant regeneration has been established via shoot organogenesis using nodal meristems. The effect of various plant growth regulators was evaluated to regenerate the whole plants by in vitro micropropagation in modified MS media supplemented with different plant growth regulators (PGRs). In vitro regeneration of plants was involved in three steps by shoot initiation, multiplication and rooting of microshoots. Initially 100% explants of Heliotropium kotschyi responded to initiate shoot in MS medium supplemented with 8.88 μM BAP with 5.71 μM IAA after 4 weeks of culture. Multiplication of shoots and ultimate plant regeneration capacity of Heliotropium kotschyi were compared and the highest performance of 578 shoots/explant was observed in presence of 8.88 μM BAP and 5.71 μM IAA after second transfer. Rooted plantlets were transferred to soil pots and 60% plants were survived after one and a half months. The highly efficient plant regeneration protocol of the desert plant is of great value in rapid plant propagation program for the purpose of conservation of biodiversity.
Cite this paper: Sadeq, M. , Pathak, M. , Salih, A. , Abido, M. and Abahussain, A. (2014) Highly Efficient in Vitro Regeneration Method of Endangered Medicinal Plant Heliotropium kotschyi (Ramram) in the Kingdom of Bahrain. American Journal of Plant Sciences, 5, 736-747. doi: 10.4236/ajps.2014.55088.
References

[1]   Jameel, A., Al-Laith, A. and Qaher, M. (2010) Knowledge, Attitudes, and Practices Regarding Medicinal Plants Used by the Indigenous People of Bahrain. Arab Gulf Journal of Scientific Research, 28, 105-117.

[2]   El-Oqlah, A. and Abbas, J. (1994) A Checklist of Vascular Plants of Bahrain. Dirasat Journal, 21, 95-118.

[3]   Bennett, R.N. and Wallsgrove, R.M. (1994) Secondary Metabolites in Plant Defense Mechanisms. New Phytology, 127, 617-633. http://dx.doi.org/10.1111/j.1469-8137.1994.tb02968.x

[4]   Grayer, R.J. and Harborne, J.J. (1994) A survey of Antifungal Compounds from Higher Plants 1982-1993. Phytochemistry, 37, 19-42. http://dx.doi.org/10.1016/0031-9422(94)85005-4

[5]   Vicente, M.J., Segura, F., Aguado, M., Migliaro, D., Franco, J.A. and Martinez-Sanchez, J.J. (2011) Genetic Diversity of Astragalus nitidiflorus, a Critically Endangered Endemic of SE Spain, and Implications for Its Conservation. Biochemical Systematics and Ecology, 39, 175-182. http://dx.doi.org/10.1016/j.bse.2011.03.005

[6]   Paunescu, A. and Vantu, S. (2002) Micropropagation of the Endemic Species Andryala levitomentosa (E.Nyar.) Sell. Review Roumaine de Biologie Series. Biologie Vegetation, 47, 9-11.

[7]   Khanna, A. (2012) Implementation of UNCCD in the GCC Countries. In: Khanna, A., Eds., Global Environmental Governance and Desertification: A Study of Gulf Corporation Council Countries, Concept Publishing Company Pvt. Ltd., New Delhi, 272.

[8]   Khan, S., Al-Qurainy, F. and Mohammad, N. (2012) Biotechnological Approaches for Conservation and Improvement of Rare and Endangered Plants of Saudi Arabia. Saudi Journal of Biotechnological Services, 19, 1-11.

[9]   Paunescu, A. (2009) Biotechnology for Endangered Plant Conservation: A Critical Overview. Romanian Biotechnological Letters, 14, 4095-4103.

[10]   Al-Eisawi, D. (2003) Effect of Biodiversity Conservation on Arid Ecosystem with a Special Emphasis on Bahrain. Journal of Arid Environments, 54, 81-90. http://dx.doi.org/10.1006/jare.2001.0888

[11]   Abbas, J.A., El-Oqlah, A.A. and Mahasneh, A.M. (1992) Herbal Plants in the Traditional Medicine of Bahrain. Economic Botany, 46, 158-163. http://dx.doi.org/10.1007/BF02930630

[12]   Earth, Google (2013) http://earth.google.com

[13]   Pathak, M.R. and Hamzah, R.Y. (2008) An Effective Method of Sonication-Assisted Agrobacterium-Mediated Transformation of Chickpea. Plant Cell Tissue and Organ Culture, 93, 65-71. http://dx.doi.org/10.1007/s11240-008-9344-6

[14]   Murashige, T. and Skoog, F. (1962) A Revised Medium for Rapid Growth and Bioassay with Tobacco Tissue Culture. Physiologia Plantarum, 15, 473-497. http://dx.doi.org/10.1111/j.1399-3054.1962.tb08052.x

[15]   Yeoman, M.M. and Macleod, A.J. (1977) Tissue (Callus) Culture Technique. In: Street, H.E., Ed., Plant Tissue and Cell Culture, Blackwell Scientific Publications, Oxford, 31-59.

[16]   Chen, C.C., Chen, S.-J., Sagare A.P. and Tsay. H.-S. (2001) Adventtitious Shoot Regeneration from Stem Internode Explants of Adenophora triphylla (Thunb) A.C. (Campanulaceae)—An Important Medicinal Herb. Botanical Bulletin of Academia Sinica, 42, 1-7.

[17]   Turker, A.U., Yucesan, B. and Gurel, E. (2010) Adventitious Shoot Regeneration from Stem Internode Explants of Verbena officinalis L., a Medicinal Plant. Tubitak, 34, 297-304.

[18]   Sharon, M. and Marie, C.C. (2000) In Vitro Clonal Propagation of Bixa orellana L. Current Science, 78, 1532-2534.

[19]   Sharon, M., Marie, C.C. and Sharon, M. (2012) In Vitro Culture Studies of Bixa orellana L: III Plant Regeneration from Roots through Direct and Indirect Somatic Embryogenesis. European Journal of Experimental Biology, 2, 156-162.

[20]   Makunga, N. and Staden, J. (2008) An Efficient System for the Production of Clonal Plantlets of the Medicinally Important Aromatic Plant: Salvia Africana-lutea L. Plant Cell Tissue and Organ Culture, 92, 63-72.
http://dx.doi.org/10.1007/s11240-007-9305-5

[21]   Bagadekar, A. and Jayaraj, M. (2011) In Vitro Flowering of Heliotropium indicum L.—An Important Medicinal Herb. Asian Journal of Experimental Biological Science, 2, 90-95.

[22]   Gokhale, M. and Bansal, Y.K. (2009) Direct in Vitro Regeneration of a Medicinal Tree Oroxylum indicum (L.) Vent. through Tissue Culture. African Journal of Biotechnology, 8, 3777-3781.

[23]   Tavares, A.C., Pimenta, M.C. and Gonalves, M.T. (1996) Micropropagation of Melissa officinalis L. through Proliferation of Axillary Shoots. Plant Cell Report, 15, 441-444. http://dx.doi.org/10.1007/BF00232072

[24]   Meszaros, A., Bellon, A., Pinter, E. and Horvath, G. (1999) Micropropagation of Lemon Balm. Plant Cell Tissue and Organ Culture, 57, 149-152. http://dx.doi.org/10.1023/A:1006359825024

[25]   Da Silva, S., Sato, A., Luiz, C., Lage, S., Azevedo, A. and Espuibel, M.A. (2005) Essential oil Composition of Melissa officinalis L. in Vitro Produced under the Influence of Growth Regulators. Journal of . Brazelian Chemical Society, 16, 1387-1390. http://dx.doi.org/10.1590/S0103-50532005000800014

[26]   Ghiorghita, G.I., Mafteli, D.E.S. and Nicuta, D.N. (2005) Investigations on the in Vitro Morphgenetic Reaction of Melissa officinalis L. Species. Genetics and Molecular Biology, 5, 119-125.

[27]   Yucesan, B., Turker, A.U. and Gurel, E. (2007) TDZ-Induced High Frequency Plant Regeneration through Multiple Shoot Formation in Witloof Chicory (Cichorium intybus L.). Plant Cell Tissue and Organ Culture, 91, 243-250.
http://dx.doi.org/10.1007/s11240-007-9290-8

[28]   Kohlenbach, H.W. and Wernicke, W. (1978) Investigations as Inhibitory Effect of Agar and Functions of Active Carbon in Anther Culture. Zeitschrift fur Pflanzenphysiologie, 86, 463-472.
http://dx.doi.org/10.1016/S0044-328X(78)80292-X

[29]   Wang, P.J. and Haung, L.C. (1976) Beneficial Effects of Activated Charcoal on Plant Tissue and Organ Culture. In Vitro Plant Cellular and Developmental Biology, 12, 260-262.

[30]   Fridberg, G., Pedersen, M., Landstrom, L.E. and Erikson, T. (1978) The Effect of Activated Charcoal on Tissue Cultures: Absorption of Metabolites Inhibiting Morphogenesis. Physiologia Plantarum, 43, 104-106.
http://dx.doi.org/10.1111/j.1399-3054.1978.tb01575.x

[31]   Renert, J. and Bajaj, Y.P.S. (1977) Anther Culture, Haploid Production and Its Significance in Applied and Fundamental Aspects of Plant Cell. In: Bajaj, Y.P.S., Ed., Biotechnology in Agriculture and Forestry 12, Haploids in Crop Improvement I, Springer-Verlag, New York, 48-58.

[32]   Mikulík, J. (1999) Propagation of Endangered Plant Species by Tissue Cultures. Acta Universitatis Palackianae Olomucensis. Gymnica, 37, 27-33.

[33]   Rout, G.R., Mahapatra, A. and Jain, S.M. (2006) Tissue Culture of Ornamental Pot Plant: A Critical Review on Present Scenario and Future Prospects. Biotechnology Advance, 24, 531-560.
http://dx.doi.org/10.1016/j.biotechadv.2006.05.001

[34]   Grando, F.M., Franklin, C.I. and Shatters, R.G. (2002) Optimizing Embryogenic Callus Production and Plant Regeneration from “Tifton 9” Bahia Grass Seed Explants for Genetic Manipulation. Plant Cell Tissue and Organ Culture, 71, 213-222. http://dx.doi.org/10.1023/A:1020303522530

[35]   Cui, S.X., Wang, W. and Zhang, C.L. (2002) Plant Regeneration from Callus Cultures in Two Ecotypes of reed grass (Phragmites communis trinius). In Vitro Cellular and Developmental Biology Plant, 38, 325-329.

[36]   Sarma, K.S. and Rogers, S. (2004) Plant Regeneration and Multiplication of the Emergent Wetland Monocot Juncus accuminatus. Plant Cell Reports, 17, 656-660. http://dx.doi.org/10.1007/s002990050460

[37]   Sharma, N., Chandel, K.P.S. and Paul, A. (1993) In Vitro Propagation of Gentiana Kurroo: An Indigenous Threatened Plant of Medicinal Importance. Plant Cell, Tissue and Organ Culture, 34, 307-309.
http://dx.doi.org/10.1007/BF00029722

[38]   Adelberg, J.W., Zhang, X.P. and Rhodes, B.B. (1997) Micropropagation of Citrullus lanatus (Thumb.) Matsum. and Nakai (Watermelon). In: Bajaj, Y.P.S., Ed., Biotechnology in Agriculture and Forestry. High-Tech and Micropropagation V, Vol. 39, Springer-Verlag, Berlin, 60-76.

[39]   Baksha, R.M., Jahan, A.A., Khatum, R. and Munshi, J.L. (2005) Micropropagation of Aloe barbadensis Mill. Through in Vitro Culture of Shoot Tip Explants. Plant Tissue Culture and Biotechnology, 15, 121-126.

[40]   Fraternale, O., Giamperi, L., Ricci, D. and Rocchi, M. (2002) Micropropagation of Bupleurum Fruticosum: The Effect of Triacontanol. Plant Cell, Tissue and Organ Culture, 69, 135-140. http://dx.doi.org/10.1023/A:1015245810053

[41]   Roy, A. and Bhattacharyya, S. (2008) An Improved Micropropagation of Eclipta alba by in Vitro Priming with Chlorocholine Chloride. Plant Cell, Tissue and Organ Culture, 92, 315-319. http://dx.doi.org/10.1007/s11240-007-9328-y

[42]   Pathak, M.R. and Hamzah, R.Y. (2010) In Vitro Plant Regeneration and Genetic Transformation of Chickpea (Cicer arietinum L.). In: Sivakumar, P., Gnanam, R., Khawar, K.M. and Thangadurai, D., Eds., Grain Legume Research: Tissue Culture, Biotechnology and Genetic Engineering, Vol. 2, Bioscience Publications, Tamil Nadu 635201, 11-19.

[43]   Sharma, U. and Mohan, J.S.S. (2006) Reduction of Vertification in in Vitro Raised Shoots of Chlorophytum borivillianum Sant and Fernand: A Rare Potent Medicinal Herb. Indian Journal of Experimental Biology, 44, 499-505.

[44]   Sen, A., Sharma, M.M., Grover, D. and Batra, A. (2009) In Vitro Regeneration of of Phyllanthus amarus Schum. and Thonn: An Important Medicinal Plant. Our Nature, 7, 110-115.

[45]   Mohebalipour, N., Aharizad, S., Mohammadi, S., Motallebiazar, A. and Arefi, H. (2012) Effect of Plant Growth Regulators BAP and IAA on Micropropagation of Iranian Lemon Balm (Melissa officinalis L.) Landraces. Journal of Food, Agriculture and Environment, 10, 280-286.

[46]   Varshney, V., Kant, T., Sharma, V.K., Rao, A. and Kothari, S.L. (1996) High Frequency Plant Regeneration from Immature Embryo Cultures of Triticum aestivum and T. durum. Cereal Research Communications, 24, 409-416.

[47]   Torregrosa, L. and Bouquet, A. (1996) Adventitious Bud Formation and Shoot Development from in Vitro Leaves of Vitis x Muscadinia Hybrids. Plant Cell Tissue and Organ Culture, 45, 245-252. http://dx.doi.org/10.1007/BF00043637

[48]   Saini, R., Sharma, A., Sharma, M.M. and Batra, A. (2011) Impact of Phytohormones in Micropropagation of Medicinally Potent Plant: Boerhavia Diffusa L. International Journal of Pharmaceutical Sciences Review and Research, 8, 85-89.

[49]   Ujjwala, J.S. (2007) In Vitro Regeneration of Aloe barbadensis. Biotechnology, 6, 601-603.
http://dx.doi.org/10.3923/biotech.2007.601.603

[50]   Sharma, M.M. and Batra, A. (2006) High Frequency Plantlet Regeneration in Indian Ginsing: Withania samnifera L. (Dunal). Physiology and Molecular Biology of Plants, 12, 289-293.

[51]   Ghimire, B.K., Seong, E.S., Goh, E.J., Kim, N.Y., Kang, W.H., Kim, E.H., Yu, C.V. and Chung, I.M. (2010) High Frequency Direct Shoot Regeneration from Drymaria cordata Wild, Leaves. Plant Cell, Tissue and Organ Culture, 100, 209-217. http://dx.doi.org/10.1007/s11240-009-9627-6

 
 
Top