Volume: 02, October 2013, Pages: 268-272

International Journal of Computing Algorithm

IN VITRO PROPAGATION A POTENTIAL METHOD FOR PLANT CONSERVATION

A.Vinoth, R.Ravindhran Department of Plant Biology and Biotechnology, Loyola College, Chennai -600034, India raviloyola1998@gmail.com Abstract Conservation of plant biodiversity, a potential resource for health care, is essential for the very survival of the human race. Increasing importance to traditional medicine in the recent years has threatened the survival of rare species. Conventional plant propagation methods are affected by variable environmental factors and time consuming. In vitro propagation, an ex situ conservation strategy, provides new means for conservation and mass propagation of economically important plants. In addition, in vitro plant systems serve as an alternative approach for the production of bioactive compounds from medicinal plants. The present review is to focus on application of in vitro propagation via organogenesis, somatic embryogenesis and cell suspension culture for plant conservation. Keywords: Micropropagation, Synthetic seeds, Secondary metabolites, Germplasm conservation 1. Introduction Plant genetic resources are a vital part of the worlds biological diversity and an essential source for the human well being. Plants play a key role in maintaining the environmental balance and ecosystem stability and provide an important component of the habitats for the world's animal life. In addition to the crop plants that provide us food, many thousands of wild plants have great economic and cultural importance and potential for use in traditional medicines throughout the world. However, the worlds biodiversity is declining at an unprecedented rate due to the alarming increase in the world population and many plants are in danger of extinction, threatened by habitat transformation, over-exploitation, unsustainable agriculture and forestry practices, alien invasive species, pollution and climate change. Hence the loss of biodiversity in large scale in the recent years has set one of the greatest challenges for the world community, that is, to halt the destruction of the plant diversity to meet the present and future needs of humankind. The Global Strategy for Plant Conservation has been adopted unanimously at the sixth meeting of the Conference of the Parties to the Convention on Biological Diverstiy (CBD) held at Hague in April, 2002 to address the challenge of biodiversity conservation. This strategy aims to improve long-term conservation, management and restoration of plant diversity, plant communities, and the associated habitats and ecosystems both in situ (in natural habitats) and ex situ. Although species conservation is achieved most effectively through the management of wild populations and natural habitats (in situ conservation), ex situ conservation methods have been favourably used to complement in situ measures and, in some instances, may be the only option for some species [1-2]. The importance of ex situ conservation has gained international recognition with its inclusion in Article 9 of the Convention on Biological Diversity (CBD) [3] and in Target 8 of the Global Strategy for Plant Conservation (www.bgci.org.uk/files/7/0/global_strategy.pd f). Approaches to ex situ conservation include methods like seed storage, field genebanks and botanical gardens. DNA and pollen storage also contribute indirectly to ex situ conservation of plant genetic resources. Among the various ex situ conservation methods, seed storage is the most convenient for long-term conservation of plant genetic

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Volume: 02, October 2013, Pages: 268-272 resources. However, conventional seed storage strategies are not possible for a large number of important tropical and sub-tropical tree species which produce recalcitrant seeds that quickly lose viability and do not survive desiccation [4]. Field genebanks provide easy access to conserved material but they run the risk of destruction by natural calamities, pests and diseases. Hence, in vitro conservation options through tissue culture techniques have been the safest alternative with several distinct advantages for plant conservation. In vitro techniques have been found to be useful in the propagation of a large number of threatened plants [5-6] and the Micropropagation Unit at Royal Botanical Gardens, Kew has been involved in the propagation and maintenance of more than 3000 plant taxa, from all over the world, for over 30 years. In addition, cell suspension culture systems could be used for large scale culturing of plant cells from which secondary metabolites could be extracted. This method ultimately provides a continuous and reliable source of natural products. Thus the present review aims to highlight the importance of in vitro propagation methods in plant conservation. Development of techniques for conservation, with special emphasis on micropropagation, somatic embryogenesis and cell suspension cultures using both threatened and non-threatened plants are discussed. 2. In vitro propagation techniques 2.1. Micropropagation Micropropagation involves the production of plants using shoot tip or nodal culture. In this technique, newly formed shoots or shoot bases serve as explants for repeated proliferation of plants. Micropropagation plays a distinctive role in the conservation of species, particularly those having pharmacologic value [7]. It offers cost-effective protocols for large scale propagation of medicinal plants collected from the wild without affecting the survival of the species in their natural habitats [8]. The laboratory at Kings Park & Botanic Garden, Perth, Australia (http://www.kpbg.wa.gov.au) specialized in the conservation of threatened Integrated Intelligent Research (IIR)

International Journal of Computing Algorithm flora of Western Australia has developed techniques for the micropropagation of over 200 species representing 33 families of Australian plants. The Tropical Botanic Gardens and Research Institute in Kerala, India has studied the in vitro conservation of 46 medicinal plants and 30 orchids from the conservation hotspot of Western Ghats (www.tbgri.org). Micropropagation helps to improve medicinal plants through the selection of high-yield lines and their efficient cloning. Genetically similar clones of Talinum triangulare, a medicinal herb, were obtained through micropropagation [9]. The most significant advantage offered by this aseptic method of clonal propagation over the conventional methods is that in a relatively short time and space, a large number of plants can be produced starting from a single individual. For orchids, micropropagation is the only commercially viable method of clonal propagation. Over 250 orchid genera have been successfully propagated at Royal Botanical Gardens, Kew including species from Angraecum, Paphiopedilum, Phragmipedium, and Cypripedium, many of which are threatened in the wild [10]. Micropropagation allows for the creation and dissemination of large numbers of nursery plants without spreading pathogens across continents. It has been of great help in largescale production of disease-free planting materials of elite Vanilla planifolia genotypes [11].In the past decade, autotrophic micropropagation, an in vitro culture without a carbon source has gained importance in the propagation of economically important plants. In comparison with plantlets produced by conventional micropropagation systems, those produced by photoautotrophic systems with a sugar-free medium showed better growth, higher quality, lower contamination rate in vitro, and higher percentage survival ex vitro [12-13]. This method is still under laboratory trials and if good multiplication rates are achieved, this method should be explored to generate quality plants for reintroduction and long-term conservation of threatened species. 269

Volume: 02, October 2013, Pages: 268-272 2.2. Somatic embryogenesis Somatic embryogenesis is the technique involving the formation of embryo-like structures in plant cultures without the act of fertilization. Embryos thus formed in cultures and variously designated as accessory embryos, adventive embryos, embryoids or supernumerary embryos have the potential to develop into whole plants without the application of plant growth regulators in the culture medium. The first observations of in vitro somatic embryogenesis were made as early as 1950s in Daucus carota [14-16]. Although somatic embryos serve as a model system in embryological studies, the greatest interest lies in the practical application for large-scale vegetative propagation, especially through scale up of the propagation using bioreactors [17]. It has also been suggested that regeneration via embryogenesis has better chances of obtaining genetically uniform plants than through organogenic differentiation [18]. Another approach to the plant regeneration via somatic embryogenesis is the production of synthetic seeds or syn seeds. In this method, naked embryos are encapsulated in a protective covering which not only protects the embryo but also provides nutrients for germination thus resembling the true seeds. Thus, somatic embryogenesis is potentially the most amenable system to automation, not only at the production stage but also for mechanized planting in the field as synthetic seeds [19-20]. In most cases, somatic embryos or embryogenic cultures can be cryopreserved, which makes it possible to establish gene banks. 2.3. Cell suspension cultures Cell suspension culture is the process of culturing of cells or cell aggregates in an agitated liquid medium. Plant cell bioreactor technology promises exploitation of the plant kingdom as a rich resource of important and unique specialty chemicals [21]. Plant cell bioreactors also provide favorable conditions for improved secondary metabolite formation. Vast numbers of secondary metabolites have Integrated Intelligent Research (IIR)

International Journal of Computing Algorithm been produced using cell suspension cultures in the recent years. Calli and cell suspensions of different species of Linum, Callitris and Podophyllum [22-28] have been reported as sources of aryltetralin lignans. Suspension culture systems have been established for the production of caffeic acid derivatives (CAD) from Echinacea purpurea [29-30], flavonoid from Saussurea medusa [31-32] and gymnemic acid from Gymnema sylvestre [33]. Protoplasts isolated from embryogenic suspensions, may give rise to somatic embryos directly, without any intervening callus phase [34-35]. Cryopreservation for high-frequency plant regeneration via somatic embryogenesis has been achieved from zygotic embryoderived cell suspension cultures of Ranunculus kazusensis [36]. Though this technique is not sufficiently reliable for plant propagation, it is the best method available for the production of medicinally important compounds from plant species, thereby facilitating their sustainable utilization. 3. Conclusion In vitro propagation has a great potential for germplasm conservation of plant genetic resources. Ex situ conservation using in vitro methods provides a safe repository for plant populations derived from severely at risk locations. In vitro regeneration systems favour rapid production of large numbers of plantlets, without seasonal dependency, for ex situ commercial cultivation purposes and reduces the risk of endangered plants being sampled from wild habitats thus safeguarding the existing natural populations. In vitro propagated plants may also be acclimatized to natural environments and utilized in controlled breeding programmes for seed production. Hence in vitro propagation methods are not only an alternative means for mass proliferation of plants but also help in the conservation of endangered plant species. 4. References [1]Maunder M, Higgens S and Culham A. Neither common nor garden: the garden as a 270

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