JPH02283223A - Tissue culture of hiba arborvitae plant - Google Patents

Abstract

PURPOSE:To accomplish the title culture in such a manner that during the culture of the callus of a hiba arborvitae plant, an auxin contained in a culture medium ls replaced to easily induce adventitious roots from said callus. CONSTITUTION:The callus of a hiba arborvitae plant (e.g. hinoki hiba arborvitae) is subjected to culture in a medium containing 10<-6>-10-4mol/l of e.g. alpha-naphthalene acetate and <=10<-5>mol/l of a cytokinin. During the culture, said callus is transplanted to a second medium containing 10<-6>-10<-4> mol/l of 4- dichlorophenoxyacetic acid to effect stable induction of adventitious roots, thus accomplishing the objective culture.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to a method for efficiently inducing adventitious roots from callus tissue of plants of the genus Asunaro by tissue culture. Plants of the genus Asunaro contain trobolone compounds such as hinokitiol, phenolic compounds such as carvacrol, and aromatic components such as thujapsen, and the adventitious roots can be used as raw materials for producing these compounds.

(Prior Art) To date, there is no prior art for inducing adventitious roots from the callus tissue of plants of the genus Asunaro.

(Problems to be Solved by the Invention) An object of the present invention is to provide a tissue culture method for inducing adventitious roots from the callus of a plant of the genus Asunaro.

(Means for Solving the Problems) In order to achieve the above object, the present invention provides a method in which the concentration of auxins is 10~10~4 mol/! To induce adventitious roots from callus by substituting the type of auxin used during culture in the callus culture of plants of the genus Asunaro using a medium in which the concentration of cytokinins is 10-' mol/l or less within the range of The gist of the invention is a method for culturing tissues of plants of the genus Asunaro, which is characterized by the following.

(Function) In the present invention, the concentration of auxins is 10-6 to 10"'
In the range of mol/l, the concentration of cytokinins is 10-'
By culturing the callus tissue of a plant of the genus Asunaro in a medium containing mol/l or less, and culturing the callus by transplanting it to a medium containing a different type of auxin from that used in the culture medium until then, Adventitious roots can be easily induced from the callus tissue of plants of the genus Asunaro.

The medium used contains inorganic components, carbon sources, and plant hormones as essential components, and vitamins and amino acids are added as necessary. Components of the medium include inorganic salts containing elements such as nitrogen, phosphorus, potassium, sodium, calcium, magnesium, iron, manganese, zinc, molybdenum, copper, cobalt, sulfur, boron, chlorine, and iodine. . Specifically, potassium nitrate, sodium nitrate, ammonium nitrate, potassium chloride, calcium chloride, potassium dihydrogen phosphate, sodium dihydrogen phosphate, magnesium sulfate, ammonium sulfate, sodium sulfate, ferrous sulfate, ferric sulfate, manganese sulfate, Examples include compounds such as copper sulfate, zinc sulfate, sodium molybdate, potassium iodide, boric acid, and cobalt chloride.

Examples of carbon sources for the medium include carbohydrates such as sila sugar and their derivatives, organic acids such as fatty acids, and primary alcohols such as ethanol.

The plant hormones in the medium include α-naphthalene acetic acid (NAA), indole acetic acid (IAA), 2.4
Examples include auxins such as dichlorophenoxyacetic acid (2,4-D), indolebutyric acid (18^), and derivatives thereof, and cytokinins such as benzyladenine (BA), kinetin, and zeatin. In the present invention, auxin The concentration of
1. The method is characterized in that the concentration of cytokinins is used at a concentration of 10-s mol/l or less.

Examples of the vitamins in the medium include biotin, thiamine, pyridoxine, pyridoxal, pyridoxamine, calcium pantothenate, ascorbic acid, inositol, nicotinic acid, and nicotinamide.

Examples of amino acids in the medium include glycine, alanine, glutamic acid, cysteine, and phenylalanine.

The medium usually contains about 10-6 mol/l to about 0.1 mol/l of the inorganic component, about 10 g/l to about 50 g/l of the carbon source, and about 0.1 g/l of the vitamins. lff1
g/e to about 150 mg # of said amino acids, O to about 1 g/I! and the above-mentioned plant hormones are used at the above-mentioned concentration, but the components other than the above-mentioned plant hormones are not particularly limited.

Specifically, the medium used for culturing the Asunaro genus plant of the present invention includes a conventionally known culture medium used for tissue culture of plants, such as Murashige-Skoog medium, Linsmeyer-Skoog medium. , White's medium, Gamporg's B, -5 medium, H-Ech's medium, etc., by adding the above-mentioned carbon source and plant hormones, and further adding the above-mentioned vitamins and amino acids as necessary. For example, a medium prepared using a Murashige-Skoog medium is particularly preferred in the present invention.

The medium used in the present invention contains poly(III) such as agar at 0.
It is used as a solid medium containing 1 to 1%.

The present invention makes it possible to induce adventitious roots from callus by replacing the types of auxins contained in the medium during the cultivation of callus tissues of plants of the genus Asunaro in the medium. The callus was cultured in a medium containing α-naphthalene acetic acid at the above concentration.
- It is desirable to transplant and culture in a medium containing dichlorophenoxyacetic acid at the above concentration.

The present invention is a &IIwt culture method capable of stably inducing adventitious root induction from callus tissue of a plant of the genus Asunaro, which has not been previously reported.

(Example) Next, examples of the present invention will be described.

It should be noted that the embodiments are merely illustrative, and it goes without saying that various changes and improvements may be made without departing from the spirit of the present invention.

Example 1 In a 100 ml culture test tube, 10 α-naphthalene acetic acid
Murashige-Skoog's medium containing 4 mol/L of kinetin (1 mol/l) (3% by weight of sucrose, 0 vegetable gums)
．． A solid medium containing 3% of Thujopsis dolabrataSie
bold et Zuccarini var, tl
Place the callus (andai Makino) on the bed and hold it at 25°.
The cells were cultured under C1 dark conditions for 21 days. The callus is the same
Add 2,4-dichlorophenoxyacetic acid and kinetin at the concentrations listed in Table 1 to a Murashige-Skoog medium (containing 3% sucrose and 0.3% vegetable gum) prepared in a 100ml culture test tube. Transfer to solid medium and incubate at 25°C.
Table 1 shows the induction rate of adventitious roots from e-calli cultured for 3 months in the dark (n = IO).

(Effects of the Invention) As mentioned above, the present invention easily induces adventitious roots from the callus of a plant of the genus Asunaro by replacing the type of auxin contained in the medium during the cultivation of the callus of a plant of the genus Asunaro. There are effects that can be achieved.

(exterior stone)

Claims (3)

[Claims] (1) The concentration of auxins is 10^-^6~10^-^
In the range of 4 mol/l, the concentration of cytokinins is 10^
- A method of culturing the callus of a plant of the genus Asunaro using a medium with a concentration of 5 mol/l or less, and inducing adventitious roots from the callus by replacing the type of auxin used during the culture. Tissue culture methods. (2) A tissue culture method for a plant of the genus Asunaro, characterized in that adventitious roots are induced from callus by replacing the auxin according to claim 1 with α-naphthaleneacetic acid to 2,4-dichlorophenoxyacetic acid. (3) The method for tissue culture of a plant of the genus Asuna according to claim 1 or 2, wherein the plant of the genus Asuna is Cypress Asuna.