JPS5995816A - Culturing of plant tissue - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for culturing the entire proboscis tissue without inducing callus tissue, and a method for culturing in a specific organ rVpi body medium such as roots and stems and leaves separated from a young seedling L1li7. ◇ In the conventional 4iR tissue culture method, a tissue piece is aseptically collected from a transplanted body and planted in a specific medium, and amorphous cells (callus tissue) are generated from this tissue piece. However, even if all the callus tissue is induced and cultured in this way, the cultured callus tissue will not contain alkaloids such as those produced in the mother cells, fragrance pigments, pollination inducers, etc. It does not produce secondary metabolic substances such as sugar. Therefore, in order to obtain this kind of secondary metabolic $lJ quality, it is necessary to use callus tissue. It is necessary to redifferentiate into root, stem, and leaf tissues. The striations that induce redifferentiation differ depending on the species. Therefore, the only way to induce redifferentiation is through repeated trial and error over a long period of time. For example, vegetative growth hormones (auxins) are essential for callus culture.
Experiments have been carried out to drastically change the balance of plant growth promoting substances such as hormones (cytokinins) and hormones related to functional differentiation. Ikuriki who can synthesize hormones? have. Then, in accordance with the AJ rK of growth, these hormones are combined to produce bidifferentiated growth.Cultivating the entire callus tissue is an attempt to artificially inhibit such a plant's original differentiated growth system. suitable for

Moreover, inducing redifferentiation is an operation that takes extra time and effort.

The object of the present invention is to eliminate the need for callus formation and its cultivation. An object of the present invention is to provide a stacked tissue culture method capable of producing desired plant tissues by direct culturing.

The invention will be fully explained below.

In the present invention, first, plant seeds are sterilized by a method. Next, they are germinated on this persimmon purple solid medium or WIj main medium to obtain seedlings and plants. This solid medium for germination only needs to contain water τ necessary for germination, and there are no direct limitations on its type or composition. As an example.

Contain water, agar, sterilized wet absorbent cotton, or Murashige Starb medium with agar 7 added. There are no restrictions on the type or #A composition of the wave medium for germination.

For the medium, Murashige-Skoog's medium, White's medium, etc. can be used.

2 Germination is the culture of seedlings obtained by germination of seeds? If germination is carried out on a solid medium, it is preferable to transplant the seedling cuttings into a liquid medium and culture them from the viewpoint of the growth rate.0Or, if germination is carried out in a liquid medium, it may be possible to culture as is. Alternatively, the cells may be transplanted into a different type of liquid medium and cultured. There are no restrictions on the liquid medium at this time. As an example.

Murashige-Skoog medium and White medium can be purchased for $. 41a growth promoting substances such as auxins and cytokinins may be added to this inoculation culture medium as necessary. Its concentration varies depending on the plant species, but it is usually 20 ppm or less.

Preferably it is 5 pprn or less.

In the present invention, when the seedlings are propagated and the properly elongated roots, stems, and leaves are obtained, a d vessel is required.
It is preferable to separate the whole plant by cutting it off or the like, and then separate and culture it in a time-controlled liquid medium for whole plant culture. As the liquid medium at this time, the same medium as the liquid medium used for culturing the seedlings in the procedure #t+i may be used.

Of course, it is not limited to this. Plant growth promoting substances such as auxins and cytokinins may be added to this medium as appropriate depending on the safety.
Although it varies depending on the species, it is usually 20 ppm or less, preferably 5 PPm or less.

When performing isolation culture on the root organ, its elongation and storage should be investigated. Dark fir is used to speed up the process. When performing isolation culture on stems and leaves, do you use light? When hit, chloroplast production becomes active.

In the present invention, it goes without saying that the seedlings can be directly cultured in a liquid medium for plant culture without separating the seedlings from specific vessels.

Example Akamaru radish miko 770 Soaked in ethanol for 2 minutes (Peshi seed table 11k Well-humidified grains, 0.1% boiling water VC
Seed surface after soaking for 10 minutes? Sterilized. After sterilization, the seeds were washed twice with water to remove the rising water. Next, the seeds were placed on a solid medium consisting of water and agar. 15
After 2-3 days at ~25°C.

The germinated whole shoots were transplanted into 150 ytu of sterile Murashige-Skoog medium in 300 mtg Ersinmeyer flasks. Plant growth promoting substances as shown in the lower coat are added to this medium. This seedling #t25
The cells were photographed and cultured in a dark place using a reciprocating two-spinner at 90 strokes per minute. This transplanted young plant's proboscis grew quickly, differentiated into leaves, and extended filamentous roots. Two weeks later, the filamentous roots were separated from the leaves and the roots were vigorously cultured in liquid. 0 This liquid medium was the same as the liquid medium used for culturing the seedlings above.The amount of the plant growth promoting substance added was also 4 liters as in the liquid medium for seedlings above.
Through this separation culture, the roots produced branched roots, but only filamentous roots were elongated without differentiation into stems and leaves.

The stem and leaves extend, while the thread-like roots grow from the stems and leaves? In some cases, the cells are separated and cultured at appropriate times. The table below shows the effect of plant growth-promoting substances on the growth rate of each part when cultured separately for 2 weeks in the God's Plant Society. 0 growth rate is the amount of growth (g) after 2 weeks of culture. is expressed as a ratio to the inoculation m (g). What we can see from the table is that q'a+Va
As a growth-promoting substance, kinetin, which belongs to the cytokinin family, exerts a remarkable effect on radish and turnips, especially on the roots of turnips, at concentrations of o.ippm.

table that's all Agent: Patent Attorney Hidetaka Yamamoto 79-

Claims (1)

[Scope of Claims] 1. A step of sterilizing the citrus fruit of the plant, and culm of the citron being germinated in a silk culture medium to obtain seedlings, which are then transplanted and cultured for protection; A step of separating the holding organ of the proboscis as necessary, and a step of culturing the young plant or the young seedling (specific organ W separated from the larva for plant culture) in the main medium. Including the method of weaving and cultivating the mash. 2. Said proboscis, I + (1) Planted in at least one of the culture medium for tissue culture and the liquid medium for plant tissue culture, 1 cloth 0 self ↑ pregnant Af a In the 1st J Sada area of Qingsui,) 4th edition 41144 Mushito 144 love culture smell method.