STANDARDIZATION OF PROTOCOL FOR IN VITRO MULTIPLICATION OF ORCHIDS (original) (raw)
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Role of Organic Growth Supplement In vitro Multiplication of Orchid Species- A Review
Journal of Pharmaceutical Research International
The main purpose of this article is to review role of several organic growth additives such Apple juice, coconut water (CW), maize extract, banana homogenate (BH), peptone and protocorms etc which stimulate the multiplication rate of various orchid species in in vitro multiplication. These organic growth supplements help to increase the number of shoots, root and leaf in culture medium. In many orchid tissue culture, organic growth supplements, which are the most essential medium aspect to stimulate tissue growth, production and facilitate the regeneration of shoot. The banana homogenate (BH) had the highest rate of regeneration and root developments. The use of organic growth supplements resulted in increased regeneration, the creation of more shoots and the development of fresh plantlets. Amino acids, proteins, vitamins, carbohydrates and various types of organic compounds are present in these growth supplements. These components have the potential to play a significant role in th...
Plant Tissue Culture and Biotechnology, 2011
High frequency direct shoot proliferation was induced from the shoot tip explants derived from the in vitro grown seedlings of a critically endangered and horticulturally important ground orchid Phaius tancarvilleae (L'Her) Blume. Shoot tip explants cultured on solidified MS with alone or combination of various concentrations of NAA and BAP produced shoots and multiple shoots. The maximum number of healthy shoots was observed on MS with BAP (1.0 mg/l) with an average of 13.3 shoots per culture in 20 weeks; where shoot multiplication was initiated after 4 weeks of culture. Regenerated shoots rooted on MS with various concentrations of NAA, IAA, IBA. MS with NAA (0.5 mg/l) was the most appropriate condition for rooting. The well developed in vitro rooted plantlets were hardened successfully in the potting mixture containing cocopeat and sphagnum moss in the ratio of 2 : 1. Key words: Mass propagation, Phaius tancarvilleae, shoot multiplication D. O. I. 10.3329/ptcb.v21i2.10241...
Journal article, 2022
Orchids are one of the most important flowers grown in Sri Lanka for exports and local market. One of the majors constrains in expanding the orchid cultivation is a limited supply of quality planting materials. Micropropagation techniques can be successfully applied for the mass propagation of orchids. But micropropagation is expensive, thus increasing the cost of production. Therefore, this research was undertaken to replacement of expensive agar for in vitro subculturing of Cattaleya seedlings. Two-month-old Cattleya seedlings were subcultures on Murashige and Skoog media (MS) and Knudson C media (KNC) as basal media. As gelling agents' sago, semolina, corn flour, semolina + agar, corn flour + agar were used. Control treatment was carried out with agar. The cultures were maintained inside the culture room. The number of leaves, number of roots, leaf length, leaf width and percentage of contamination were recorded at fourweek intervals. Sago and semolina + agar showed higher performance and low contamination percentage as alternative gelling agents in the MS media. Sago, corn flour, sago + agar, semolina + agar and corn flour + agar showed higher performances and low contamination percentages in KNC media. The results of this study propose that sago, semolina, and corn flour can be used as alternative gelling agents in MS and KNC media for subculture practices of orchids.
Low-cost tissue culture technology for the orchid regeneration
Peruvian journal of agronomy, 2022
Orchids are one of the most important flowers grown in Sri Lanka for exports and local market. One of the majors constrains in expanding the orchid cultivation is a limited supply of quality planting materials. Micropropagation techniques can be successfully applied for the mass propagation of orchids. But micropropagation is expensive, thus increasing the cost of production. Therefore, this research was undertaken to replacement of expensive agar for in vitro subculturing of Cattaleya seedlings. Two-month-old Cattleya seedlings were subcultures on Murashige and Skoog media (MS) and Knudson C media (KNC) as basal media. As gelling agents' sago, semolina, corn flour, semolina + agar, corn flour + agar were used. Control treatment was carried out with agar. The cultures were maintained inside the culture room. The number of leaves, number of roots, leaf length, leaf width and percentage of contamination were recorded at fourweek intervals. Sago and semolina + agar showed higher performance and low contamination percentage as alternative gelling agents in the MS media. Sago, corn flour, sago + agar, semolina + agar and corn flour + agar showed higher performances and low contamination percentages in KNC media. The results of this study propose that sago, semolina, and corn flour can be used as alternative gelling agents in MS and KNC media for subculture practices of orchids.
Plant Tissue Culture and Biotechnology, 2016
In vitro propagation of a rare orchid, Bulbophyllum nipondhii was carried out. Five different media were tested to find the suitable medium for seed germination and seedling development. The tangible results were obtained on VW medium. To assess the effect of pollination types on seed germination and seedlings development, seeds derived from different self?, cross? and open?pollination were examined. Open?pollinated seeds produced the best germination and the highest seedling development, followed by cross? and self? pollination. VW medium was supplemented with 0, 25, 50 or 75 g/l potato extract (PE) and 0, 50, 100, 150 or 200 ml/l coconut water (CW) to identify their most suitable concentration. PE (75 g/l) with 100 ml/l CW was found to be best combination.Plant Tissue Cult. & Biotech. 26(1): 37-46, 2016 (June)
In Vitro Propagation Of Nepalese Orchids: A Review
Journal of Horticultural Research, 2014
Nepalese orchids are made up of 458 taxa. Despite a ban on the collection and trade of all orchid species in Nepal, numerous anthropogenic factors are leading to the rapid loss of natural stands of germplasm. Biotechnology, specifically in vitro propagation, may be the only viable solution for preserving and reintroducing endangered germplasm back into the wild. Despite the large germplasm base, only tissue culture studies have been conducted, and most have focused almost exclusively on in vitro seed germination, the bulk of which have been conducted in the past few years. No other biotechnological advances have yet been made. This brief review provides a short synopsis of the advances made thus far in the in vitro propagation of Nepalese orchids.
In VitroPropagation ofPhaius luridus Thwaites - A Terrestrial and Endemic Orchid of Western Ghats
Family Orchidaceae constitutes one of the largest families of flowering plants, having around 20,000 species. They are unique in forms, colors and flower structure. The genus Phaius luridus Thwaites is the terrestrial orchid, endemic to the Western Ghats and is an endangered species. A rapid in vitro seed germination technique is described here. MS, VW, B5 and KC media supplemented with various concentrations of auxins and cytokinins were used in combination for asymbiotic seed germination and plantlet formation. In the evaluation of the media MS medium supplemented with 2 mg BAP/L-1+ 5mg NAA/L-1 was found to be suitable with both liquid and solid. Even B5 solid and liquid medium supplemented with 2 mg BAP/L-1+1mg IAA/L-1 was found to be suitable. Further, hormonal concentrations of auxins and cytokinins were evaluated for minimal and optimal levels in the medium. Hardened plants were transferred to green house after ex vitro rooting technique. Significance of the present work is discussed here.
International Journal of Science and Research (IJSR), 2016
An efficient, protocol for in vitro germination seeds of and micropropagation of two the valuable indigenous medicine Orchids namely; Bulbophyllum lilacinum Ridl. and Cymbidium aloifoloum (L) Sw. were developed by enhanced shoot multiplication, nodal and leaf segments was designed. The medium type and various carbon sources, plant growth regulators markedly influenced in vitro propagation of B. lilacinum Ridl. and C. aloifoloum (L) Sw. The in vitro protocorms production system was investigated on three 0.8% (w/v) agar solidified basal media namely; Phytomax Sigma Chemical Co. (PM), Modified Vacin and Went (MVW), Murashige and Shook,1962 (MS) medium with the synergistic combination of Auxin (IAA 1.0-2.0 mgl-1 , IBA 2 mgl-1 , NAA 1.0-2.0mgl-1 , 2,4-D 1.0mgl-1) , Cytokinins (BAP 0.5-2.0mgl-1 , Kn 0.5-2mgl-1) and 2%-3% sucrose which induced maximum number of shoots as well as beneficial shoot length. Sub culturing of nodal segments and leaf segments on similar medium enabled continuous production of healthy shoots with similar frequency. Rooting was highest (2.96 cm) on half strength MS medium containing 1.5% sucrose. The in vitro growing seedling were then transferred to outside natural environment through successive phases of acclimatization.
International Journal of Current Microbiology and Applied Sciences, 2017
Phaius tankervilliae (L’Her.) Blume is one of the most attractive and horticulturally valuable ground orchids of the world. Due to illegal collection for trade and consumption and loss of habitat, the population of this species has decreased rapidly at an alarming rate in natural habitats. In vitro seed germination and seedling development of Phaius tankervilliae technique was successfully established for rapid multiplication using 0.8% (w/v) agar solidified MS medium supplemented with different concentrations and combinations of Kinetin (Kin) and NAA. MS medium supplemented with 1.0 mg L-1 Kin + 1.0 mg L-1 NAA was the most ideal condition for early seed germination (2.87 weeks), fastest protocorm formation (5.07 weeks), tallest shoots (6.85 cm) and maximum number of roots per plant (6.20). However, maximum number of shoots (52.27) per culture and maximum number of shoots with roots (50.27) were produced when the plantlets were transferred to MS basal medium supplemented with 1.5 mg L-1 Kin + 1.0 mg L-1 NAA. The well developed in vitro rooted plantlets were hardened successfully in the potting mixture containing only vermiculite (86.00 %) followed by vermiculite + vermicompost (71.00 %) at 1:1 ratio.
African Journal of …, 2010
Paphiopedilum rothschildianum is an endangered orchid species endemic to Mount Kinabalu, Sabah, and Malaysia. The vegetative propagation of this plant has always been restricted due to its slow growth and maturation rates. Thus, an in vitro tissue culture technique was explored in order to overcome this limitation. In this study, clonal propagation of P. rothschildianum was achieved through in vitro formation of multiple shoots from stem nodal and single shoot explants cultured onto halfstrength Murashige and Skoog medium. The responses of the explants to the presence of different types of organic nitrogen additives viz. casein hydrolysate, peptone and tryptone-peptone (in amount of 0.5, 1.0 and 2.0 g/l) in the culture medium were also evaluated. The addition of these organic nitrogen additives into the basal medium slightly enhanced the number of multiple shoots formed on both types of explants when compared to additive-free MS medium. After 16 weeks of culture, an average of 2.9 shoots per stem nodal explant and 2.8 shoots per single shoot explant were obtained on half-strength MS medium supplemented with 1.0 g/l peptone and 2.0 g/l tryptone-peptone, respectively. All the newly-formed shoots were divided into single plantlets and subcultured onto similar respective medium. After an additional 12 weeks of culture on the same medium, plantlets with 3 -4 roots were acclimatized and transferred to a glass house where they showed 90% survival rate. Thus, the method presented in this study had provided a promising strategy for the production of large numbers of phenotypically stable P. rothschildianum.