Chemistry: molecular biology and microbiology – Plant cell or cell line – per se ; composition thereof;... – Culture – maintenance – or preservation techniques – per se
Patent
1997-06-10
2000-05-09
Lankford, Jr., Leon B.
Chemistry: molecular biology and microbiology
Plant cell or cell line, per se ; composition thereof;...
Culture, maintenance, or preservation techniques, per se
47 581, C12N 500
Patent
active
060603135
DESCRIPTION:
BRIEF SUMMARY
FIELD OF TECHNOLOGY
The present invention relates to a method for producing the clone plants of Paphiopedilums in a large scale.
BACKGROUND TECHNOLOGY
Paphiopedilum is one of the most attractive garden plant in orchids. Studies in the prior art for establishing a clone multiplication technology of Paphiopedilums include a scape terminal bud culture method (K. Kawase, Multiplication of Paphiopedilums by tissue cultures, J. J. Soc. Hort. Sci. 63(Suppl.1), 1988), an axillary shoot culture method (L. C. Huang, A procedure for asexual for multiplication of Paphiopedilums in vitro, American Orchid Society Bulletin, 57: 274-278, 1988), a protocorm culture method (B. F. Mark, Tissue culture method for the genus Paphiopedilum. Australian Orchid Review, Feb. 4-10, 1991) and a shoot apex culture method (S. Yasuki et al., Meri-clone seedling formation of Paphiopedilums by a shoot apex culture method, Engeigakkaishi, 64 Extra number 1: 516-517, 1995).
However, if the multiplication of Paphiopedilum clone is carried out by such methods, there may be provided with low multiplication rate or no multiplication. Accordingly, such methods have suffered from difficulty in being employed as a practical technology for clone multiplication of Paphiopedilum.
The difficulty may be attributable to the plant death due to browning of tissues during culture as well as the missing of the combination of culture materials and culture conditions suitable for clone multiplication.
DISCLOSURE OF THE INVENTION
An objective of the present invention is to provide a method for producing a clone plants of a Paphiopedilum on a practical basis.
The first aspect of the present invention is a method for clone-multiplication of a Paphiopedilum wherein a scape terminal bud or an axillary shoot, whose surface has been sterilized, or an aseptic shoot obtained by an aseptic cultivation, are cut into cross-sectional pieces which are then cultivated in a medium containing a plant hormone.
The second aspect of the present invention is a method for clone-multiplication of a Paphiopedilum according to claim 1 wherein said medium in which said cross-sectional pieces are cultivated is a liquid medium containing a solid support capable of dispersing or adsorbing polymeric growth inhibiting substances, such as polyphenols, produced in said culture.
BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 shows the preparation of the cross sectional pieces from a viable plant being cultured.
PREFERABLE EMBODIMENTS OF THE INVENTION
In a method according to the present invention, a starting material is a scape terminal bud derived from a blooming stock or a bud before blooming or a young axillary shoot. Cultivation of the scape terminal bud of a Paphiopedilum stock may be conducted, according to an ordinal condition, at an atmospheric temperature (usually 23 to 27.degree. C., preferably at about 25.degree. C.) under a continuous lighting or a 16-hour photoperiod. Such scape terminal bud cultivation is conducted preferably by an aseptic method. A scape terminal bud or an axillary bud obtained by a non-aseptic method should be employed after sterilizing its surface.
A scape terminal bud enclosed in a floral bud before blooming is preferably selected as a starting material, since it exhibits a higher survival rate after sterilization of the surface. In such case, a floral bud is taken from a Paphiopedilum and washed several times with an aqueous ethanol, preferably 70% ethanol in water, and then immersed 5 to 10 minutes, preferably for about 10 minutes, in an aqueous ethanol, preferably 70% ethanol in water, and subsequently shaken for about 10 minutes in a chlorinated sterilizing agent such as Wilson solution containing a surfactant such as Tween 20 at about 0.1%, whereby accomplishing sterilization of the surface, and then further washed several times with a sterilized water, resulting in a minimum contamination with environmental bacteria.
A scape terminal bud or an axillary shoot obtained by a non-aseptic method as mentioned above is employed as a starting material after
REFERENCES:
K. Kawase, "Multiplication of Paphiopedilums by Tissue Cultures", J. J. Soc. Hort. Sci, 63(Suppl. 1), 1988 with Abridged Translation.
Li-Chun Huang, "A Procedure for Asexual Multiplication of Paphiopedilums in Vitro", American Orchid Society Bulletin, vol. 57, pp. 274-278, 1988.
Prakash Lakshmanan et al., "An In Vitro Method for Rapid Regeneration of a Monopodial Orchid Hybrid Aranda Deborah Using Thin Section Culture", Plant Cell Reports, vol. 14, pp. 510-514, (1995).
Database WPI, Section Ch, Week 8939, Derwent Publications Ltd., London, GB, AN 89-282442, XP002084843 & JP 01 206991 A, Aug. 21, 1989.
Shoji Kazuhiko, "Tissue Culture of Plant of Family Orchidaceae", Patent Abstracts of Japan, vol. 095, No. 001, Feb. 28, 1995.
K. Zimmer "Phalaenopsis--Zur Vegetativen Vermehrung", vol. 79, No. 11, pp. 258-260, 1979. (English Abstract Provided).
Tanaka Michio
Zhou Tian Su
Lankford , Jr. Leon B.
Sapporo Breweries Limited
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