JP2748348B2 - Propagation method of seeds and tubers of Araceae plants - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a method for growing a large amount of potato seeds or tubers by tissue-culturing a genus Araceae using a liquid medium by a specific method.

[Conventional technology]

Taro is a very important crop in the world, but the problem of virus yield reduction and quality deterioration is a major problem. For this reason, in recent years, a method of mass-producing seeds or tubers by tissue culture using shoot tips not contaminated with virus has been attempted. However, the culture method reported to date is agar culture, liquid shaking culture, or air aeration culture, and in agar culture, the culturing operation requires a lot of manpower, and it is difficult to use the incubator three-dimensionally and mass culture is performed. Not efficient to do. In addition, in liquid shaking culture and air aeration culture, air was dissolved in the culture medium, so it was difficult for the dissolved oxygen concentration in the culture medium to exceed 9 ppm at normal culture temperatures, and this was not always the optimal culture condition for the cultured plant. .

[Problems to be solved by the invention]

The present inventors have recognized the problems of the conventional tissue culture method of Araceae plants, and studied a culture method for efficiently growing high-quality potato seeds or tubers excellent in both growth and properties.

[Means for solving the problem]

As a result of the study, the present inventors have found that the dissolved oxygen concentration in the liquid medium at a normal culture temperature is 9 ppm by means such as aerating a gas having a higher oxygen content than normal air in the liquid medium.
Through the above, it has been found that the growth of a cultured plant of the genus Araceae can be promoted, and that seeds and tubers having excellent properties can be proliferated, and the present invention has been completed.

The plant used in the present invention is a genus Araceae,
Specific examples include taro and yam.

The tissue or cells to be cultured in the present invention include tissue fragments of the above-mentioned plants, cultured cells, and the like. Further, for seedlings or tubers of insufficient size obtained by culturing, the plant which is this cultured object The whole body can be used as a material for culture for tuber formation. Examples of tissue pieces include cut tissues such as shoot tips, stems, leaves, flowers, seeds, tubers, and roots. The cultured cells are undifferentiated atypical cells obtained by subjecting the above-mentioned tissue piece to tissue culture by a known method.

The liquid medium used in the present invention is a medium containing an inorganic component and a carbon source as essential components, a plant growth regulator, vitamins, and, if necessary, amino acids. As the inorganic component of the medium, inorganic components that are usually used for tissue culture of plants can be used, and as the carbon source, sucrose, glucose, fructose, maltose, and the like can be given, and the amount of addition is generally 5 g / l-. 150 g / l is good, preferably 10 g / l-100 g / l.

In the present invention, known plant growth regulators can be used to promote the growth and differentiation of the cultured plant. Examples of the plant growth regulator include benzyladenine, kinetin, zeatin, isopentenyladenine, indoleacetic acid and the like. , Indolebutyric acid, naphthaleneacetic acid, 2,4-dichlorophenoxyacetic acid, chlorocholine chloride and the like.

As the liquid medium used in the present invention, specifically, Murashige-Skoog medium, white medium, niche and niche medium, medium used for normal tissue culture such as Rinsmeyer-Skoog medium, or modified these mediums Examples of the medium include a medium in which the carbon source and the plant growth regulator are added to the prepared medium, and vitamins and amino acids are further added as necessary. Medium
The pH is preferably from 4.0 to 8.0. Although light is not necessarily required during the culture, better results may be obtained when the culture is performed under illumination with a light amount of 100 to 10,000 lux. The temperature of the culture is preferably 15-35 ° C.

The present invention is characterized by culturing under the condition that the oxygen content in the liquid medium is 9 ppm or more. In this case, the concentration of dissolved oxygen in the medium is preferably 9 to 20 ppm. For this reason, in the present invention, the culture is carried out by aerating a gas containing oxygen at a higher concentration than ordinary air into the liquid medium. In this case, as the oxygen-containing gas, oxygen can be used alone, or a gas obtained by mixing two or more kinds of gases such as oxygen, air, nitrogen, and carbon dioxide can be used. In the present invention, the oxygen content in these gases is 22-100 vol%, preferably 25-100 vol%.
% Is desirable. The aeration rate of the gas into the liquid medium varies depending on the shape of the incubator, but is preferably an aeration rate that is generally about 0.1 to 30 as expressed by an oxygen transfer capacity coefficient (KLa).

〔Example〕

Example A lump of taro (variety name: Ishikawa Sayo) obtained from a culture plant derived from a growing point was cut so that one section weighed 0.2 g on average, and sucrose 4% and benzyl adenine 1 ppm were added at pH 5.
The sections were added to an incubator (capacity 150 ml) containing 50 ml of a sterile Murashige-Skoog liquid medium of No. 7. Attach a gas vent tube with a glass filter to the incubator, and add 0.22μ
The gas containing 30% of oxygen and 70% of nitrogen passed through a sterilizing filter of m m was blown into the liquid medium through a gas vent tube at a rate of 3 ml / min (oxygen transfer capacity coefficient (KLa) was 5.0). The cells were cultured at 3,000 lux, at 3,000 lux for 16 hours and for 60 days. The average dissolved oxygen concentration in the medium during the culture was 12.3 ppm. A bud was formed on the tuber surface, and the base of the bud was enlarged to form a small tuber. Table 1 shows the number of tubers formed per section and the average diameter of the tubers. When the tubers were transferred to soil and cultivated, they showed healthy growth.

Comparative Example The culture was performed in the same manner as in the example except that air was blown. At this time, the average dissolved oxygen concentration in the medium was 7.9 ppm. Table 1 shows the measured values of the obtained small tubers.

[Effect of the Invention] If a culture method using a liquid medium having a dissolved oxygen concentration of 9 ppm or more of the present invention is employed, a high-quality seedling and tuber can be produced in a large amount more efficiently than the conventional method by tissue culture of Araceae plants. Can proliferate.

Claims (1)

(57) [Claims] 1. A method for cultivating a seed, a tuber or a tuber of a genus Araceae by culturing a tissue, cell or plant of the genus Araceae, wherein the cultivation is performed using a liquid medium having a dissolved oxygen concentration (DO) of 9 ppm or more. A method for growing seedlings or tubers of Araceae plants, characterized by the following.