JPS62259513A - Promotion of organ differentiation in plant tissue/cell culture - 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 [Industrial Application Field] The present invention relates to a method for promoting differentiation of plant tissues into organs in cell culture.

[Conventional technology]

``Biotechnology'', also known as ``biological engineering'' or ``technology that utilizes the functions of living organisms,'' has attracted particular attention in recent years, and is contributing to a wide range of areas of human life, and further development is expected as we head into the 21st century. There is.

Plant tissue/cell culture is particularly closely related to agriculture in the technical field of biotechnology, and its applications are expected to be high and some of it has been put into practical use.

Application to breeding: 1) It has the potential to become a useful weapon as a method for rapidly propagating crops in large quantities, and 1 its importance is increasing.

It is based on the de-differentiation and re-differentiation of cells through the total forming ability of plant tissues and cell cultures. Regarding organ differentiation through culture, Skoog and Mil.
ler (1957) cannot be ignored.

This study was the first to examine in detail the control of organ differentiation through culture using tobacco stem parenchyma, and the differentiation and formation of adventitious roots or adventitious buds in the cultured tissue was determined by the amount of auxin and cytokinin added in the medium. It has been shown that it depends on the equilibrium.

Since then, similar tests have been carried out on many plant materials, but the trends do not apply to plants in general;
Rather, it came to be considered an exceptional material.

Even after that, organ redifferentiation remains the biggest problem when applying m-tissue and cell culture to breeding, mass proliferation, etc. Therefore, there are expectations for the development of ways to induce and promote redifferentiation of plant cells.

The compound of the present invention represented by formula 3 (1) below (hereinafter referred to as the compound of the present invention).

It's simply called brassinolide. ) is described in Japanese Patent Application Laid-Open No. 1983-1997 (1983).
It is disclosed in Japanese Patent No. 28471.

However, the description in this publication relates to the growth-promoting effect of brassinolides in culturing plant tissues or cells, and there is no statement that brassinolides promote organ differentiation.

[Aspects of the invention]

The present invention relates to plant tissue/cell culture.

(2α, 3α, 22R, 23R)-tetrahydroxy-243-methyl-B-homo=7-oxa- represented by the following formula (1)
5α-cholestan-6-one (hereinafter referred to as

It's simply called brassinolide. This invention relates to a method for promoting differentiation of tissues into organs in plant tissue/cell culture, which comprises adding an effective amount of the following:

The present inventors investigated various methods for promoting organ differentiation in plant tissue/cell culture, and discovered that brassinolide has a promoting effect on the process of stably inducing organ differentiation in an extremely wide concentration range. It was completed.

Plant tissue/cell culture in the present invention includes any tissue/cell taken out from a plant body.

Tissues derived from the shoot apical cambium, young hypocotyls, etc. are particularly desirable.

The method for treating brassinolide in the present invention is carried out in plant tissue culture using a medium such as Murashige-Skoog (Mura
Shige-Skoog) basic medium containing inorganic salts, trace elements and vitamins (generally referred to as MS-medium).

), and furthermore, growth promoters such as coconut milk or differentiation promoters such as synthetic plant hormones, which are used in normal culture, may be added to this medium depending on the purpose.

The amount of brassinolide added to the medium is 10-6 to 1
102 ppm, preferably 101 to ioppm.

In the present invention, the plants to which the present invention can be applied are not particularly limited, but the present invention can be applied to plants such as the following.

Cruciferous plants (Chinese cabbage, radish, wasabi).

turnips, cabbages, etc.), Rosaceae plants (strawberries, roses, etc.), Umbelliferae plants, carrots, celery, parsley, honeysuckle, etc.), Convolvulaceae plants (sweet potatoes, etc.), Araceae plants (konnyaku, taro, etc.), Chenopodiaceae plants plants (such as spinach).

Poaceae plants (corn, rice, wheat, etc.).

Liliaceae plants (salts, onions, green onions, garlic,
asparagus), nightshade plants (tomatoes, etc.).

peppers, eggplants, potatoes, etc.), Asteraceae plants (
Examples include burdock, Chinese chrysanthemum, lettuce, butterbur, etc.) and leguminous plants (soybeans, fava beans, peas, etc.).

The organ differentiation-promoting effect of brassinolide in plant tissue culture can be extremely effectively utilized for variety improvement and seed production in many plants and crops.

Hereinafter, the present invention will be explained in more detail with reference to examples.

The present invention is not limited to these.

Example 1 Using lettuce (variety: Wirehead) as an experimental material, 0.5% antiformin solution (seed sterilization solution) was added as usual.
After sterilizing the surface of the seeds with
% agar medium and grown in the dark at 25°C for 9 days.

After that, a 61IllIl long tissue piece was taken from the hypocotyl,
This was used as a culture material.

MS-medium was used as the basal medium. This medium contains 2.
Sodium 4-dichlorophenoxyacetate 0.1 mg/
1. , kinetin was added in a concentration of 0.1 mg/I, and brassinolide was added in a range of θ to 0.5 ppm.

After culturing in the dark for 6 days, the cells were transplanted to Tungdoku medium containing kinetin (0.1 mg/l) and cultured in the light. The adventitious bud formation rate was investigated on the 35th day of culture. The experiment was conducted at 20°C and 25°C.
It was conducted under two conditions. Four hypocotyl tissue pieces were placed per 1 colben, and 20 individuals were used in each group. The results are shown in Table 1. Incidentally, the above-mentioned kinetin is a common name of a plant hormone agent, and its compound name is 6-furfurylaminopurine.

Table 1 0 15 55 0.3 0.71.0
-4 20 65 0.4 1.01.
0-3 45 90 0.5 2.01
．． 0-2 55 95 1. ,O2,71
0-' 65 100 1゜02.85
X10-'-1003,7 As is clear from the results in Table 1, brassinolide concentrations 10-' to 5X10-
'ppm showed a high adventitious bud formation rate in each section, and the number of adventitious buds per individual in the cultured tissue also increased significantly.

In particular, the effect of promoting adventitious bud differentiation under poor conditions at 20°C was remarkable.

Claims (1)

[Claims] (1) In plant tissue/cell culture, the following formula (I): ▲
There are mathematical formulas, chemical formulas, tables, etc. ▼(I) (2α, 3α, 22R, 23R)-tetrahydroxy-24S-methyl-B-homo-7-oxa-
1. A method for promoting differentiation of tissues into organs in plant tissue/cell culture, which comprises adding an effective amount of 5α-cholestan-6-one to a medium.