JP2591477B2 - How to Induce Kenaf Callus - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a kenaf (hibiscus)
Cannabis, Hibiscus cannabinu
s) The method for inducing callus, more specifically, by culturing kenaf seeds under a specific culture condition using a specific callus-inducing solid medium,
The present invention relates to a method for stably and efficiently inducing kenaf callus in a short time.

[0002]

2. Description of the Related Art Induction of plant callus is necessary in various biochemical treatments on plants, such as cell fusion and introduction of foreign genes. The callus is induced by various plants including tobacco and rice, and the callus induction method is disclosed in JP-A-4-53495 and JP-A-2-53.
No. 1,861,631.

[0003]

Problems to be Solved by the Invention
The publication discloses a method for inducing the callus of hibiscus, but the callus induction conditions are completely different for different plant species.Therefore, with this conventional method, it is possible to induce kenaf callus with high efficiency in a short period of time. could not.

Japanese Patent Application Laid-Open No. 2-128631 discloses a method for inducing kenaf callus. After germinating kenaf seeds on an agar medium, a piece of the stem of the shoot is planted with a plant hormone. Since the callus is induced by placing the medium on another medium containing the medium, the operation is complicated, and 40
There is a problem that it takes days and a long time.

An object of the present invention is to solve the above problems and to provide a method for guiding kenaf callus stably, with high efficiency and easily in a short period of time.

[0006]

Means for Solving the Problems A first invention of the present invention is:
After sterilizing kenaf seeds, the seeds are placed on a pH-adjusted synthetic solid medium containing a carbon source and a plant hormone consisting of auxins and cytocanins, and the culture is aseptically cultured. A method for inducing kenaf callus, wherein the synthetic solid medium contains 3% by weight of sucrose as a carbon source.
And a plant hormone consisting of 300 μg / l of 2,4-dichlorophenoxyacetic acid as an auxin and 300 μg / l of kaitinene as cytokinins;
7 is a synthetic solid medium adjusted to 7 and 1 at 25 ° C.
It is a method for inducing kenaf callus, which comprises aseptically culturing under a culture condition of one hour photoperiod.

[0007]

[0008]

[0009]

Next, an embodiment of the present invention will be described. Callus induction experiments were carried out using kenaf (Hibiscus cannabinus), such as Chinese blue skin No.3.

[0010] Sterilization was performed by immersing kenaf seeds in a hypochlorous acid solution having an effective chlorine concentration of 1% for 20 minutes.
Since the seeds are wrapped in thick shells, the sterilization operation can be performed without destroying the cells necessary for the induction of callus inside the shell, which is advantageous in sterilization operation as compared with the case where the callus is induced from leaves or stems.

As a medium, an MS medium (Murasige-Skoog medium) is used as a basic medium, and 2,4-dichlorophenoxyacetic acid (300 μg / l) and kaitinene (300 μg /) are added at this concentration ratio, and a sodium hydroxide solution is added. The pH was adjusted to 7 with. 1% by weight of agar was added to this medium, and the medium was sterilized by pressurized steam in a conventional manner, and 20 ml of the medium was dispensed into test tubes for plant culture having a diameter of 40 mm.
According to the method of the present invention for adjusting the pH of a medium, the medium can be solidified, and its pH is preferably near neutral. The medium and the MS medium used in the conventional method have an acidic pH of 5.6 to 5.8, are not adjusted in pH, and cannot be solidified in this pH range. Was.

Next, the kenaf seeds are placed on the synthetic solid medium in the test tube, and the illuminated light is 1200 L at 25 ° C.
When cultivated under the irradiation conditions of 11 hours of photoperiod with a fluorescent light for plant growth of ux, kenaf seeds germinate after about 24 hours. Since the growth point of the germinated seeds has a high growth rate, the cell mutation starts immediately after germination, and the callus is induced, unlike the case where the callus is induced using leaves or stems. According to this method, the time required for callus cell clusters to be observed and callus induction to be confirmed was 3 days.
Callus could be induced stably with an induction efficiency of 00%. This induction period is considerably shorter than the time required for the conventional callus induction. FIG. 1 shows a photograph of kenaf callus induced according to the present invention.

[0013]

According to the method of the present invention, the callus of kenaf can be induced stably, with high efficiency and easily in a short period of time.

[Brief description of the drawings]

FIG. 1 is a photograph of kenaf callus induced according to the present invention.

[Explanation of symbols]

 None

Claims (1)

(57) [Claims] 1. After sterilizing kenaf seeds, the seeds are placed on a synthetic solid medium having a pH adjusted and containing a carbon source and a plant hormone consisting of auxins and cytocanins, Induction of kenaf callus cultured aseptically
The method, wherein the synthetic solid medium comprises 3% by weight of sucrose as the carbon source.
And 2,4-dichlorophenoxy as the auxins
300 μg / l of acetic acid and calcium as the cytokinins
Contains plant hormone consisting of 300 µg / l of itinene
And a synthetic solid medium whose pH has been adjusted to near neutrality,
Aseptically cultured at a temperature of 25 ° C under a culture condition of 11 hours photoperiod
A method for inducing kenaf callus, comprising: