Chemistry: molecular biology and microbiology – Plant cell or cell line – per se ; composition thereof;... – Culture – maintenance – or preservation techniques – per se
Reexamination Certificate
2002-01-29
2004-04-27
Campell, Bruce R. (Department: 1661)
Chemistry: molecular biology and microbiology
Plant cell or cell line, per se ; composition thereof;...
Culture, maintenance, or preservation techniques, per se
C435S410000, C435S430100
Reexamination Certificate
active
06727094
ABSTRACT:
BACKGROUND OF THE INVENTION
(1) Field of the Invention
The present invention relates to cultured cells and a tissue-culturing method, and particularly the invention relates to cultured cells of
Rhaphiolepis umbellata
Thunb. and a method for culturing tissues of the
Rhaphiolepis umbellata
Thunb. by using said cultured cells.
(2) Related Art Statement
Recently, investigations have been vigorously carried out on the tissue culturing of the higher plants, and techniques for mass culturing plant tissues removed from leaves, stems, roots, etc. have public attracted attentions. When a plant piece cut off is place on agar or in a liquid containing a nutriment, a mass of cells swell up from a cut edge of the plant piece. This is called callus.
In general, cells of different tissues, for example, tissues of a root and a leaf, have different shapes and functions, respectively. This results from gradual differentiation during a time period in which the cells grow. However, the callus continue to grow without differentiation.
Generally, seeding or cutting-planting must be done so as to propagate plants. Further, the soil and the environment largely influence the growth of the plants. However, the culturing of the callus is not influenced by changes in these matters. In addition, the callus grows faster than ordinary plant bodies. When a hormone or a chemical substance that promotes germination or rooting is added to the callus, a complete plant body is obtained.
As such a callus tissue-culturing method, methods for culturing tissues of trees such as poplar and eucalyptus are known.
However, what has been established as the tissue-culturing methods are limited to the kinds of trees such as poplar and eucalyptus only. One of reasons for this is that it is difficult to grow or root other breeds unless appropriate hormones or the like are used.
The
Rhaphiolepis umbellata
Thunb., which is a plant belonging to the Rosaceae, is a roadside tree planted in a median strip of a road, etc. The
Rhaphiolepis umbellata
Thunb. is strong against the public pollution, and has saline resistance. In order to assuredly and swiftly provide such plants having strong public pollution resistance, the tissue-culturing method is effective. However, a method for stably mass propagating the
Rhaphiolepis umbellata
Thunb. has not been established.
SUMMARY OF THE INVENTION
It is an object of the present invention to provide cultured cells and a tissue-culturing method, which enable the establishment of stable mass propagation and transformation system of the
Rhaphiolepis umbellata
Thunb.
In order to accomplish the above object, the present inventors had repeatedly made strenuous investigations on conditions suitable for inducing the formation of callus and the redifferentiation of the
Rhaphiolepis umbellata
Thunb., and consequently have come to discover the cultured cells and the tissue-culturing method according to the present invention.
The cultured cell according to the present invention has high differentiating power, and is obtained by culturing a part of a tissue of the
Rhaphiolepis umbellata
Thunb. in a culture medium containing benzyladenine and naphthaleneacetic acid (NAA) in an amount effective to induce callus formation of the
Rhaphiolepis umbellata
Thunb.
In a preferred embodiment of the cultured cell of the
Rhaphiolepis umbellata
Thunb. according to the present invention, the tissue of the
Rhaphiolepis umbellata
Thunb. is a tissue selected from the group consisting of a shoot apex, a stem, a leaf, an embryonic cell and a root.
In another preferred embodiment of the cultured cell of the
Rhaphiolepis umbellata
Thunb. according to the present invention, the tissue of the
Rhaphiolepis umbellata
Thunb. is a tissue originated from a plant body raised from a seedling obtained by germinating a sterilized seed of the
Rhaphiolepis umbellata
Thunb.
In a further preferred embodiment of the cultured cell of the
Rhaphiolepis umbellata
Thunb. according to the present invention, the tissue of the
Rhaphiolepis umbellata
Thunb. is a tissue less than 2 weeks after seeding the
Rhaphiolepis umbellata
Thunb.
In a still further preferred embodiment of the cultured cell of the
Rhaphiolepis umbellata
Thunb. according to the present invention, the tissue of the
Rhaphiolepis umbellata
Thunb. is a tissue obtained by aseptically growing a seed of the
Rhaphiolepis umbellata
Thunb.
In a still further preferred embodiment of the cultured cell of the
Rhaphiolepis umbellata
Thunb. according to the present invention, the culture medium is a WP culture medium or an MS culture medium.
The method for culturing a tissue of a
Rhaphiolepis umbellata
Thunb. according to the present invention comprises the steps of subculturing any of the above cultured cells in a culture medium containing benzyladenine and naphthaleneacetic acid (NAA) in an amount effective to induce callus formation of the
Rhaphiolepis umbellata
Thunb., and thereby obtaining plantlet of the
Rhaphiolepis umbellata
Thunb.
In a preferred embodiment of the
Rhaphiolepis umbellata
Thunb. tissue-culturing method according to the present invention, the culture medium is a WP culture medium or an MS culture medium.
The cultured cell according to the present invention is a cultured cell of the
Rhaphiolepis umbellata
Thunb. having high differentiating power, said cultured cell being obtained by culturing a part of a tissue of the
Rhaphiolepis umbellata
Thunb. in a culture medium containing benzyladenine and naphthaleneacetic acid (NAA) in an amount effective to induce callus formation of the
Rhaphiolepis umbellata
Thunb. In general, the plantlet is formed according to the tissue-culturing method in the course of (1) formation of the callus, (2) formation of a polyblast, (3) redifferentiation of the polyblast and (4) formation of the plantlet. The tissue culturing of the
Rhaphiolepis umbellata
Thunb. takes this formation course. Here, the term “polyblast” means a mass of numerous shoots in which cells are differentiated.
In the following, the cultured cells of
Rhaphiolepis umbellata
Thunb. and the tissue-culturing method therefor according to the present invention will be explained.
Although the tissues of the
Rhaphiolepis umbellata
Thunb. are not particularly limited, tissues from stems, leaves, roots, stem apexes, root apexes, embryonic cells, etc. may be recited. Stems, leaves, roots, stem apexes, embryonic cells and roots are preferred.
The tissues of the
Rhaphiolepis umbellata
Thunb. may be ones taken from a matured tree of the
Rhaphiolepis umbellata
Thunb. Preferably, the tissue is a tissue originated from a relatively young cell tissue obtained after seeding the
Rhaphiolepis umbellata
Thunb. For example, the tissue obtained 2 weeks, more preferably less than 9 to 10 days, after seeding the
Rhaphiolepis umbellata
Thunb.
Seeds of the
Rhaphiolepis umbellata
Thunb. can be aseptically germinated and grown on the culture medium. The aseptic treatment for the cutting is not particularly limited, for example, such a treatment can be effected by treating the tissue with ethanol or the like for a few or several hours and with sodium hypochlorite for a few or several hours and is washed with sterilized water. It is preferable to use a seed fed with water at 4 to 10° C. in dark for not less than 1 to 3 nights.
The cultured cells of the present invention can be obtained by culturing a part of the tissue as mentioned above, in a culture medium containing benzyladenine and naphthaleneacetic acid (NAA) in an amount effective to induce callus formation of the
Rhaphiolepis umbellata
Thunb. The amount of benzyladenine depends upon the culture medium used and the culturing condition, and is preferably 0.5 to 10.0 &mgr;M, more preferably 0.5 to 5.0 &mgr;M. The reason for this range is to enhance the redifferentiation percentage. The amount of naphthaleneacetic acid (NAA) is not particularly limited, but is preferably 0.1 to 1 &mgr;M. The reason for this range is also to enhance the redifferentiation percentage.
As the culture medium,
Morikawa Hiromichi
Takahashi Misa
Campell Bruce R.
Hiroshima University
McCormick Susan B.
LandOfFree
Cultured cells of Rhaphiolepis umbellata Thunb. and a method... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Cultured cells of Rhaphiolepis umbellata Thunb. and a method..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Cultured cells of Rhaphiolepis umbellata Thunb. and a method... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3276642