JPH09233962A - Method for proliferating shoot of orchid plant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for proliferating shoot of orchid plant with which many shoots can be stably proliferated. SOLUTION: A shoot 2 is formed by using the inflorescence of orchid plant as an explant and cultivating the tissue of the explant. The provided shoot 2 is proliferated by a solid culture medium 5 containing phytohormone. The phytohormone is composed of NAA, BA and TDZ. The orchid plant is Paphiopedilum or the like. It is desirable for the content of TDZ is to be from 10mg to 30mg per liter of solid culture media.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for growing shoots of orchid plants.

[0002]

2. Description of the Related Art Among flowers, high-quality orchids, which are high-quality flowers, are uniformly and mass-proliferated by tissue culture using a part of plant tissue as an explant. For example, Phalaenopsis orchid and Cymbidium are inexpensively and stably supplied to the general market because the technology for mass-propagation by tissue culture has been established.

On the other hand, Paphiopedilum, which is a kind of western orchid, is one of the types in which tissue culture is difficult, and the tissue culture technology has not been established. Therefore, it is currently being increased by the stock split method and the seed method. The stock split method only increases the number of stocks to one or two. Also,
In the method by species, there are variations in individuals. for that reason,
With these methods, it is not possible to stably secure a uniform strain.

[0004] Although few in number, several methods for proliferating Paphiopedilum by tissue culture have been disclosed.
First, the prior literature (Tissue Cultures Studies in Paphi
opedilum, Joyce Stewart et. al., American Orchid S
ociety, pp.591-599, 1975) and prior literature (The Ef
fect of Benzyl Adenine on the Development of Later
al Buds of Paphiopedilum, Joyce Stewart et. al., A
The American Orchid Society, pp.415-418, 1977) describes a breeding method using shoot tips as explants. However, this has not been put to practical use because it was prone to contamination by miscellaneous bacteria because the shoot was taken out after decomposing the strain.

Next, the prior literature (Propagation of Paphiopedilum by Tissue Culture, Koshiro Kawase, Horticultural Society Abstracts No. 3
76-377, 1988), and the prior literature (Propagation of Paphiopedilum by Tissue Culture (Part 2), Koshiro Kawase, Abstracts of the Horticultural Society, 464-465, 198).
9) describes a method of using inflorescences as explants. However, although this method can significantly suppress contamination by various bacteria and obtain shoots, on the other hand, the number of shoots formed per explant varies from individual to individual. Moreover, the number of shoots obtained is only a small number from one individual to several tens.

In view of such conventional problems, the present invention aims to provide a method for multiplying shoots of Orchidaceae plants, which enables stable and large-scale growth of shoots.

[0007]

The invention according to claim 1 uses inflorescences of the Orchidaceae plant as explants, and the explants are tissue-cultured to form shoots, and the resulting shoots contain plant hormones. In the method for growing in a medium described above, the plant hormone comprises NAA, BA, and TDZ, and is a method for growing shoots of Orchidaceae plants.

What is most noticeable in the above invention is that
Culturing shoots of orchid plants using a solid medium containing TDZ. As shown in FIG. 1, inflorescence 1 in the above invention means bud 1 in an unflowered Orchidaceae plant.
1, the ovary 12, and the flower stem 13 are collectively referred to.

The NAA means 1-naphthalene acetic acid (sodium aceto acid). The above BA is 6-
Benzyladenine (6-benzyladenine)
Say. The above TDZ is Thidiazon (1-
PHENYL-3- (1,2,3-THIADIAZO
L-5-YL) UREA. The chemical formula of TDZ is shown in FIG.
It was shown to.

Next, the function and effect of the above invention will be described. Shoots of orchid plants are NAA, BA, and TD
It is grown on a solid medium supplemented with a plant hormone consisting of Z. Therefore, shoots can be propagated with high efficiency. In particular, by adding TDZ, it is possible to differentiate and proliferate into a plurality of individuals with higher efficiency than ever before. Therefore, it becomes possible to grow orchidaceae plants such as Paphiopedilum, and it is possible to stably supply them to the general market in large quantities at low cost. Therefore, it is possible to establish a technique for mass-propagating tissue culture of orchid plants.

Next, as described in claim 2, the TD
The Z content is 10 to 3 per liter of the solid medium.
It is preferably 0 mg. If it is less than 10 mg or more than 30 mg, a high shoot growth rate may not be obtained. Next, as described in claim 3, the orchidaceous plant is, for example, Paphiopedilum, but is not limited thereto.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION A method for multiplying shoots of orchidaceous plants according to an embodiment of the present invention will be described with reference to FIGS. First, as shown in FIG. 1, the proliferation method of this example is as follows: unflowered Paphiopedilum (Clairde).
Inflorescence 1 of Lune AM-RHS) was cut off. This inflorescence 1 consists of a bud 11, an ovary 12, and a flower stem 13.

Then, various bacteria were removed from this inflorescence 1 with alcohol and antiformin. Then, inflorescence 1
Cut into round slices, ovary 12 shown in FIG. 2 (a), or FIG.
The flower stem 13 and the ovary 12 shown in (b) were obtained. Next, these were placed in a test tube and cultured at a temperature of 25 ° C. in an aerated environment. Then, as shown in FIG. 2, the ovary 12 and the flower stem 13 were continuously cultured until one to several shoots 2 were formed. The medium is Vacin & Men
t medium and coconut milk (10%) was added to it.

Next, one to several shoots formed were cut out and tissue culture was performed in a solid medium. As the solid medium, the seedling N3f medium shown in Table 1 was used. To the N3f medium for seedlings, 1% by weight of glucose and 1% by weight of fructose as sugar sources and 2% of gellan gum were added to the above N3f medium for seedlings. Furthermore, in this seedling N3f medium, 10 mg / L NAA, 5 mg / L BA,
A plant hormone consisting of TDZ was added. As shown in Table 2, the addition amount of TDZ was changed within the range of 0 to 40 mg / L. Solid medium containing the same concentration of TDZ
The tubes were sorted into test tubes.

As shown in FIG. 3, each test tube 8
1 to 5 shoots 2 were placed as an outgrowth on the solid medium 5 in (diameter 25 mm) and covered with a plastic stopper 6. The test tube 8, the solid medium 5 and the plastic stopper 6 have been sterilized in advance.

Tissue culture of shoots in solid medium
I went for 50 days. The conditions for tissue culture are that the light-dark cycle is 12 hours light and 12 hours dark. A fluorescent lamp was used as a light source in the light period, and the illuminance was 2000 to 4000 lux. The temperature was constant at 25 ° C. The solid medium was subcultured every two weeks. Count the number of shoots after a certain period of time,
Cultured for 8 months. The results are shown in Table 2. In Table 2, the average number of shoots refers to the average number of shoots formed in 5 solid culture media (5 test tubes) containing the same concentration of TDZ.

From the table, when TDZ is added, T
The proliferation rate was higher than that in the case where DZ was not added. In particular, 10 to 30 mg of TDZ per liter of solid medium
In the case of adding, the growth rate was more than 3 times, showing the highest growth rate. However, at 40 mg or more, shoot growth was suppressed.

The reason why shoot growth is promoted in the presence of TDZ is not clear, but TD
It is considered that Z is to induce the growth and change of side buds.

[0019]

[Table 1]

[0020]

[Table 2]

[0021]

EFFECTS OF THE INVENTION According to the present invention, it is possible to provide a method for growing shoots of Orchidaceae, which enables stable and large-scale growth of shoots.

[Brief description of drawings]

FIG. 1 is an explanatory view showing the inflorescence of an unflowered Orchidaceae plant according to the present invention.

FIG. 2 is an explanatory diagram of shoot-forming inflorescences in the embodiment.

FIG. 3 is an explanatory view showing a method of growing shoots with a solid medium in the embodiment.

FIG. 4 is an explanatory diagram showing a chemical formula of TDZ in the embodiment.

[Explanation of symbols]

 1. . . Inflorescence, 11. . . Bud, 12. . . Kobo, 13. . . Flower stem, 2. . . Shoot, 5. . . Solid medium,

Claims (3)

[Claims] 1. An inflorescence of an Orchidaceae plant is used as an explant,
In the method of culturing the explants in tissue to form shoots and growing the shoots in a medium containing plant hormones, the plant hormones are NAA, BA and T.
A method for multiplying shoots of an Orchidaceae plant, which comprises DZ. 2. The method for growing shoots of Orchidaceae according to claim 1, wherein the TDZ content is 10 to 30 mg per liter of the solid medium. 3. The method for growing shoots of Orchidaceae according to claim 1, wherein the Orchidaceae is Paphiopedilum.