JPS62275604A - Plant regenerated tissue, its production and artificial seed - 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 3. Detailed Description of the Invention Field of Industrial Application The present invention relates to a preserved form of meristem of a plant and to an artificial seed containing the same as a constituent.

Prior Art Recently, a method has been proposed that applies plant tissue culture technology and embeds produced somatic cell embryos in materials such as polymers to create artificial seeds (see Japanese Patent Application Laid-Open No. 102308/1982).
In this method, it is necessary to simply preserve the somatic cell embryo while it is still alive and in a growth-arrested state. Conventionally, this is done by adding a plant growth regulator or adding a sugar with a high <mW content. Methods have been proposed, but these methods have problems with storage, growth after sowing, and contamination by bacteria.

In connection with this, it has recently been reported that orchard grass somatic cell embryos have been dried (wheat) and regenerated (Am(!
riCan Journal of Botany
72816-817 (1985)). However, according to the research conducted by the present inventors, if the water content of somatic cell embryos is minimized, the period during which somatic embryos can be stored in a renewable state becomes shorter, and the regeneration rate also decreases. There was found. Moreover, the above-mentioned literature does not disclose any specific methods such as drying methods, or application to other plants or organs.

OBJECTS OF THE INVENTION An object of the present invention is to provide a regenerated plant tissue that does not have the above-mentioned problems and has excellent characteristics when used as artificial seeds, and a method for producing the same. Another object of the present invention is to provide artificial seeds containing the regenerated plant tissue as a steel component.

p' of 1J and Effects The present inventors have researched ways to improve the problems of the conventionally known techniques, and have found that it is easy to dry regenerated plant tissues, especially those dried at a specific moisture content and at a specific drying rate. saved,
They also discovered that regeneration is possible, and arrived at the present invention.

That is, the present invention relates to a regenerated plant tissue having a water content in the range of 5 to 15%, and a polymer that can resist external forces in the atmosphere and provides an environment in which water can grow through water permeation. The moisture content covered by the film is 5~
This is an artificial seed made of regenerated plant tissue in a state of 15% by weight.

The regenerated plant tissue of the present invention can be easily produced without using complicated procedures or expensive equipment, and can be easily stored. The tissue of the present invention can be widely applied regardless of plant species, is easy to regenerate, and has a high regeneration rate. In general, the tissue of the present invention is suitable as an artificial seed in terms of storage stability, germination properties, etc.

Summary of the invention The present invention will be explained in more detail below.

In the present invention, the term "plant regeneration tissue" refers to a tissue that can differentiate and regenerate the entire plant, and refers not only to meristems such as shoot tips, but also to part or the entire plant containing them. Specifically, somatic embryos, seedlings, shoot tips, etc. are used, and somatic embryos and seedlings are particularly preferred.

The somatic embryo referred to here is an embryo derived from somatic cells, also called a somatic embryo, and although it may be formed in a natural state in citrus fruits etc., it is often formed from cultured cells. For example, the manufacturing method is described by Dee Ni Evans (D., A.
``Handbook of Plant Cell Culture'' by Evans et al.
Hacmill Publishing Company
AnPublishing Co., ) published in 1983], etc. Somatic embryos are globular embryos depending on their growth stage.

They are distinguished from heart-shaped embryos, torpedo-shaped embryos, and mature embryos, but somatic embryos in the present invention are preferably those at a lower developmental stage.

Seedlings mean seedlings of seed plants that have sprouted from seeds, and in the present invention, seedlings are preferably seedlings that have a growth period as short as possible after germination.

Seedlings can be grown in a conventional manner, such as by keeping them in a bright place and at high humidity to avoid germination.

The plants targeted by the present invention can be applied to a wide variety of plant species in general, but are particularly suitable for plants such as Apiaceae, Fabaceae, and Poaceae.

In general, seeds can withstand dryness and can be stored for long periods of time while preserving their names, and it is known that the lower the moisture content in the seeds, the better their storage stability. However, it was thought that the other two organs would normally wither and die if they become dry. Some plants and organs, such as plants in the cactaceae family, tubers, and bulbs, can survive for long periods in a dry atmosphere, but this is because the cuticle of the epidermis prevents water evaporation. It is said that the interior maintains a high moisture content. However, according to research conducted by the present inventors on regenerated plant tissues other than seeds, it has been found that when these tissues are dried under specific conditions and the moisture content inside the tissues is within a specific range, they can survive for a long period of time. , was found to be reproducible.

It is necessary that the water content of the regenerated plant tissue is 5 to 15% (by weight), and in this range, a long survival period and high regeneration rate are exhibited. The water content referred to here is the water content (percentage) relative to the total weight of the regenerated plant tissue, and specifically, by the Karl Fischer method, for example, "Dehydrated Solvent FHr Mitsubishi" manufactured by Mitsubishi Kasei ■ is used. It can be calculated from the value measured as a solvent at 25°C.

There are no particular limitations on the drying method, as long as it is carried out at a temperature that does not cause tissue necrosis, such as 40°C or lower.
Generally, the following method is preferably used.

For example, a method in which the relative humidity in the air is lowered using a desiccant such as calcium chloride, sulfuric acid, pentaacid or phosphorus, zeolite, silica gel, or a dehumidifier, and the meristem tissue is allowed to stand in this atmosphere to dry it; A method may be adopted in which the meristematic tissue is allowed to stand still or is allowed to flow, and then air is circulated around it to dry it.

In terms of survival period and regeneration rate, it is necessary to set the drying rate such that the moisture content of the regenerated plant tissue falls within the range of 5 to 15% within 100 hours from the start of drying. Drying rate is the relative humidity of the surrounding air.

temperature, type, size, and shape of the regenerated plant tissue.

It is difficult to clearly state the conditions for the insulator because it changes depending on the conditions such as the amount, but for example, the thickness is about 0.5 mm.

Carrots with a length of 1 to 3 mm (DanCLISc)
Arota L.) somatic embryos, when arranged in a thin layer, reach a moisture content of about 15% in 48 hours when left standing in the atmosphere at a temperature of 25° C. and a relative humidity of 50%.

The regenerated plant tissue of the present invention can be suitably used as artificial seeds. A conventionally proposed method is to embed meristems such as somatic embryos in aqueous gel, but this method does not have good preservation properties because the meristems are alive and continue to grow. In addition, a method has been proposed to improve this method by adding abscisic acid or high-concentration sugar to the aqueous gel, but in the case of abscisic acid, it is removed by dissolving and diffusing in water after seeding. However, this takes time and germination is not good. On the other hand, when sugars are added, they encourage the proliferation of various bacteria and impede growth.

On the other hand, when the regenerated plant tissue of the present invention is used, it does not require any additives, remains dormant, and resumes growth when water is applied. It is easy to store and has a low moisture content, so it is light in weight.

In the present invention, the regenerated plant tissue is covered with a polymer film that can resist external forces in the atmosphere and provides an environment in which it can grow through water permeation, thereby forming an artificial seed. In this case, the regenerated plant tissue has a moisture content of 5 to 1.
It is necessary to keep it within 5%.

In order to handle and preserve artificial seeds, it is necessary to resist external forces and maintain the shape of the internal regenerated plant tissue. Especially when the water content of the regenerated plant tissue is low, it is necessary to resist external forces and maintain the shape of the regenerated plant tissue. This is necessary because it is relatively easy to destroy. Furthermore, the regenerated tissue of the plant resumes growth by providing water. Therefore, in order to germinate artificial seeds, the polymer needs to be water permeable.

Further, after water permeation, it is desirable that the film does not contain substances that inhibit the growth of plants, and that the film disintegrates as it stretches. Further, it is desirable that the inside can be kept sterile in the atmosphere and after water permeation.

Such polymers include polymers that are themselves water-absorbing, such as polyvinyl alcohol, carboxymethylcellulose, and methylcellulose.

Gelatin, metal alginate, carrageenan, agar,
Julan gum" (manufactured by Sanei Chemical Co., Ltd.) can be mentioned. In addition, the outer layer is made of a water-permeable polymer, such as polyamide or polyester, even if it itself is not water-absorbing.
A multilayer structure containing a water-absorbing substance, such as starch, may be adopted.

The method for producing the artificial seeds of the present invention can use many of the seed coating methods that have been disclosed in the past. For example, diatomaceous earth, calcium carbonate,
Inorganic powder such as talc, kaolin, clay, etc. and a binder such as polyvinyl alcohol, carboxymethyl cellulose, methyl cellulose, gelatin, etc. can be mixed with water and coated on the regenerated plant tissue. It is also possible to coat the meristem with -H with an aqueous gel such as sodium alginate or carrageenan and then dry it by the method described above. Furthermore, in the present invention, various additives may be added to the coating agent, such as sugars that serve as nutritional sources for the regenerated plant tissues, other substances, growth promoters, or drugs such as fungicides and herbicides.

The present invention will be briefly described below using Examples, but the present invention is not limited thereto.

Example 1 Seedlings of the carrot (Daucus carota L) cultivar "Kofuku Shisun Ginseng" were grown to a length of 0.5 cm according to the method described in "Plant Cell Tissue Culture" by Harada et al. (Rigakusha 1979), p. 94. ~5 mm somatic embryos were obtained. The somatic cell embryos were washed with water, and after absorbing adhering water on paper, they were placed in an atmosphere at each relative humidity listed in Table 1 and at 25°C to dry. After 180 days, the somatic embryos were taken out and planted into plants.
Hurashige-3kooy that does not contain L, -E
When cultured in a medium, regeneration rates as shown in Table 1 were obtained. Note that the regeneration rate referred to here is the ratio (%) of the number of regenerated somatic embryos to the number of dried somatic embryos.

Table 1 Example 2 Alfalfa () ledicaao 5ativa
Seeds of L.) were sown on hydrated Gavi and placed in the dark at 20'C for 2 days. Seedlings that have sprouted and grown to 5-10 mm are kept at 20% relative humidity.

It was left in an atmosphere at 25° C. for 6 days to dry. When the dried seedlings were placed on water-containing gauze again and left at 20℃, 8
0% were regenerated.

Procedural amendment δ month) 6th, 1985

Claims (5)

[Claims] (1) Regenerated plant tissue having a water content in the range of 5 to 15% by weight. (2) The regenerated plant tissue according to item 1, wherein the regenerated plant tissue is a somatic embryo. (3) The regenerated plant tissue according to item 1, wherein the regenerated plant tissue is a seedling. (4) A method for producing regenerated plant tissue, which comprises drying somatic embryos or seedlings in their natural state to a moisture content of 5 to 15% by weight within 100 hours. (5) From regenerated plant tissues with a water content of 5 to 15% by weight, covered with a polymer film that can withstand external forces in the atmosphere and provide an environment in which growth can occur through water permeation. Artificial seeds.