Plant regeneration from protocorm-derived callus of Cypripedium formosanum

In Vitro Cellular & Developmental Biology - Plant (Impact Factor: 1.16). 09/2003; 39(5):475-479. DOI: 10.1079/IVP2003450

ABSTRACT Totipotent callus of Cypripedium formosanum, an endangered slipper orchid species, was induced from seed-derived protocorm segments on a quarter-strength Murashige and
Skoog medium containing 4.52 μM 2,4-dichlorophenoxyacetic acid and 4.54 μM 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (thidiazuron). This callus proliferated well and was maintained by subculturing on
the same medium. On average, 13 protocorm-like bodies could be obtained from a piece of 4 mm callus after being transferred
to the medium with 4.44 μM N6-benzyladenine after 8 wk of culture. The regenerated protocorm-like bodies formed shoots and roots on medium containing 1
g l−1 activated charcoal and 20 g l−1 potato homogenate. After 24 wk of culture on this medium, well-developed plantlets ready for potting were established.

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Available from: Yung-I Lee, Jul 30, 2015
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    • "Rev. Ceres, Viçosa, v. 59, n.2, p. 185-191, mar/abr, 2012 Diferentes respostas foram observadas no meio de cultura e da concentração de carvão ativado empregada , dependendo da espécie utilizada. Lee & Lee (2003) obtiveram resultados positivos na regeneração de brotos e raízes a partir de protocormoides de Cypripedium formosanum Hayata adicionando 1 g L -1 de carvão ativado ao meio MS com ¼ da concentração de sais. Ket et al. (2004) observaram que a adição de 1 g L -1 ao meio Hyponex suplementado de 0,1 g L -1 de benziladenina e 1-2 mg L -1 de thidiazuron promovia a formação múltipla de brotos em segmentos nodais de Anoectochilus formosanus Hayata, entretanto o crescimento era lento e os caules não se alongavam. "
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    ABSTRACT: Orchids are over-exploited in their natural environment due to their ornamental importance, which is leading some species to extinction. In vitro culture is an alternative for ex-situ conservation. Our objective was to determine an efficient medium for in vitro seed germination and initial seedling growth of Cattleya forbesii and Cattleya harrisoniana. Seeds of Cattleya forbesii were incubated in Murashige & Skoog (MS) medium (T1) and MS added with 2,5 g L-1 of activated charcoal (T2). For the last, plants with 1 ± 0,2 cm of height were submitted to the following treatments: T1, T2, MS reduced to the half strenght (T3) and MS reduced to the half strenght supplemented with 1,25 g L-1 of activated charcoal (T4). Thirty days after seed inoculation of C. forbesii, it was verified 45% and 90% of germination in T1 and T2, respectively. The addition of activated charcoal also promoted an increase in the height of the C. forbesii seedlings after 180 days of in vitro cultivation. After 240 days of culture of C. harrisoniana shoot height, total fresh mass, number of roots and leaves, length of the largest root and diameter of the shoots were significantly stimulated in T2 compared to the other treatments. We suggest the use of MS medium added with 2,5 g L-1 of activated charcoal because is significantly favorable for seed germination and initial growth for both species.
    04/2012; 59(2):185-191. DOI:10.1590/S0034-737X2012000200006
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    ABSTRACT: Kaur Saranjeet., Bhutani K.K., 2012. Organic growth supplement stimulants for in vitro multiplication of Cymbidium pendulum (Roxb.) Sw. Hort. Sci. (Prague), 39: 47–52. The aim of this study was to establish protocol for in vitro regeneration and multiplication of Cymbidium pendulum through protocorms, as explants using organic growth supplements such as banana homogenate, coconut water, and peptone. The effect of growth supplements was tested on multiplication (neo-formation of secondary protocorms from primary protocorms) of protocorms, development of maximum number of shoots and early plantlets in M medium (Mitra medium). Though the explants regenerated in the absence or presence of growth adjuncts in the medium; the regeneration frequency was found significantly higher in organic growth supplement-enriched medium than control. The protocorm segments (primary) regenerated into protocorm-like bodies (secondary protocorms) which eventually differentiated into shoots; no intervening callus stage was observed. Among the treatments, the highest regeneration frequency, robust shoots and root formation was recorded in banana homogenate (50 g/l); the roots were lengthy, thick, and covered all over the surface by root hairs. Higher concentration of banana homogenate (75 g/l) proved detrimental for the survival of cultures; the protocorm-like bodies showed necrosis and they perished soon after. Coconut water (10%) and peptone (2 g/l) supplemented medium proved optimum for profuse multiplication of protocorm-like bodies (PLBs) their early growth into plantlets. The treatments with organic growth supplements gave better and early plantlets as compared to control. Among these tested organic growth supplements, peptone (2 g/l) and coconut water (10%) proved beneficial for multiplication of regenerants, maximum number of shoots formation and early plantlet develop-ment; whereas banana homogenate (50 g/l) favoured the highest regeneration frequency and healthy growth of plantlets.
  • Voenno-medit͡sinskiĭ zhurnal 04/1961; 3:62-5.
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