JP2963301B2 - Method for producing somatic embryos and propagation method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing and propagating somatic embryos of a stock, and provides a method for producing and propagating somatic embryos capable of directly regenerating a plant.

[0002]

2. Description of the Related Art Stock is a plant belonging to the genus Brassicaceae and the genus Araceae, which has grassy or slightly woody stalks. There are single-flowering and double-flowering strains. Is the actual situation.

[0003] For the stock for appreciation, the double-flowered strain is traded at a much higher price, but when the stock is cultivated by seedlings, the single-flowered strain and the double-flowered strain occur at a ratio of approximately 1: 1. . For this reason, a cultivation method capable of cultivating only a double-flowered strain is desired.

[0004] In recent years, research on a technique for regenerating a plant by culturing has been advanced, and a very small number of stocks have been reported. For example, in the Journal of the Horticultural Society, Vol. 55, separate volume 2, pages 332 to 333, there is a leaflet or glass of a mature strain cultivated in a house bed under the title of "Adventitious bud formation and histological observation by stock leaf fragment culture". A method of forming adventitious buds by culturing leaf pieces of young seedlings grown indoors in a medium supplemented with plant hormones is introduced.
On pages 521 to 521, the adventitious buds obtained by the above-mentioned leaf leaf culture of the stock were placed on a rooting medium to which plant hormones had been added, with the title "About rooting from shoots formed by the leaf leaf culture". And how to root them.

[0005]

SUMMARY OF THE INVENTION A method for growing a stock by culturing cotyledons, hypocotyls and mature leaves of the stock to regenerate a plant, as apparent from the above-mentioned report, resulted in the formation of shoots by adventitious buds. Since this is later transferred to a rooting medium for rooting, the process of proliferation is complicated. Further, since it takes a long time to regenerate a plant, it has disadvantages such as not only low regeneration efficiency but also high probability of bacterial contamination during culturing.

[0006] In contrast, the present invention relates to a method for producing a somatic embryo of a stock regenerated into a plant by culturing in a very simple step, and a method of growing the somatic embryo in the stock in a very simple step. It is possible to regenerate a plant in a short period of time by using the created somatic embryo or the grown somatic embryo, and also to enable mass propagation of a double-flowered strain and a strain having a useful trait.

[0007]

SUMMARY OF THE INVENTION The present invention comprises a step of extracting immature embryos from immature seeds that have passed 30 days or more after pollination and fertilization of a flowering stock; And culturing the plant in a medium containing plant hormones.

[0008] The present invention also includes a step of extracting immature embryos from immature seeds that have passed 30 days or more after pollination and fertilization of a flowering stock, and the step of extracting immature embryos extracted in the above step with a plant hormone. A method for propagating a somatic embryo, wherein the adventitious embryo obtained by the step of culturing by placing on a medium is subcultured to a medium containing a plant hormone.

In the method of the present invention for producing an adventitious embryo and propagating the same, the immature embryo extracted from an immature seed that has passed 30 days or more after pollination and fertilization of the flowering stock is performed. It is necessary to. Immature embryos less than 30 days after pollinating and fertilizing a flowering stock may die during cultivation, and more than 60 days after pollinating and fertilizing a flowering stock. From immature embryos, the rate of adventitious embryo formation is low.

For this reason, in the method for producing adventitious embryos and expanding the stock of the present invention, the immature seeds removed from the immature seeds between 30 and 60 days after pollination and fertilization of the flowering stock. Preferably, an embryo is used.

The phytohormone in the medium used in the method of producing and somatic embryos of the stock of the present invention is auxin such as indole acetic acid or naphthalene acetic acid, or auxin to which cytokinin such as benzyl adenine or kinetin is added. Is preferred. In particular, it has been confirmed that the use of indoleacetic acid alone or a mixture of indoleacetic acid and benzyladenine increases the somatic embryo formation rate of the stock.

It is preferable that the culture medium in which the immature embryo of the stock is placed and cultured is the same as the culture medium in which the adventitious embryo of the produced stock is subcultured and cultured.

[0013]

The method for producing somatic embryos of a stock according to the present invention comprises the steps of extracting immature embryos from immature seeds that have passed 30 days or more after pollination and fertilization of a flowering stock, A step of laying the embryo on a medium containing plant hormones and culturing the embryo, whereby an adventitious embryo having shoots and roots can be formed at once by a very simple operation.

The method for growing somatic embryos of a stock according to the present invention comprises the steps of: removing a immature embryo from an immature seed that has passed 30 days or more after pollination and fertilization of a flowering stock;
The step of placing the immature embryos removed in the above step on a medium containing plant hormones and culturing them, and subculture the stock somatic embryos produced in the medium containing plant hormones and culturing them. A large amount of somatic embryos can be propagated by a simple operation.

Thus, the desired plant can be regenerated by growing the somatic embryo obtained by the above-mentioned growth in a hormone-free medium.

[0016]

Furthermore, in the method for producing and growing a somatic embryo of a stock according to the present invention, the somatic embryo can be taken out of the medium extremely easily, and the handling operation is easy.

[0018]

EXAMPLES The specific construction of the method for producing somatic embryos and the method of propagation of the stock of the present invention will be described below based on examples.

Examples 1 to 6 Pollination and fertilization were performed on flowering stocks. Pods of immature seeds after 55 days were immersed in an antiformin solution containing 1% by weight of available chlorine for 15 minutes, and further sterilized water. And washed.

Subsequently, the immature seed embryos were taken out of the pods and placed on a solid medium (containing 0.3% by weight of gellan gum) to which the plant hormones described in the predetermined column of Table 1 were added. Cultivation in a constant temperature bath at a temperature of ℃ resulted in the formation of two somatic embryos at the base of the cotyledon on the surface of the immature embryos on the fifteenth day from the start of cultivation. The formation of was observed. Table 1 shows the somatic embryo formation rate on day 20 from the start of the culture.

Comparative Example 1 An immature embryo was cultured using a hormone-free solid medium (containing 0.3% by weight of gellan gum) in the same manner as in Examples 1 to 6 except for the corresponding procedures. The adventitious embryo formation rate on day 20 from the start of the culture is also shown in [Table 1].

[0022]

[Table 1] Examples 7 to 11 A solid medium (containing 0.3% by weight of gellan gum) to which a plant hormone described in a predetermined column of [Table 2] was added was used, and the other procedures were the corresponding procedures of Examples 1 to 6. The immature embryo was cultured in the same manner as described above. [Table 2] shows the somatic embryo formation rate on the 20th day from the start of culture.

[0023]

[Table 2] Example 12 Somatic embryos of the stock obtained in Examples 1 to 11 were
The somatic embryos were propagated by subculturing and culturing them in the same medium as the medium in which they were created.

When the somatic embryo obtained by this propagation was transferred to a hormone-free MS medium, it grew into a plant.

[0025]

According to the method for producing somatic embryos of the stock and the method of propagation of the present invention, the plant can be regenerated extremely easily and in a short period of time.

Further, according to the method for producing somatic embryos of a stock and the method of propagation according to the present invention, it is possible to mass-produce a double-flowered strain that is traded at a high price and to mass-produce useful traits.

Claims (5)

(57) [Claims] 1. A step of extracting an immature embryo from an immature seed that has passed 30 days or more after pollination and fertilization of a flowering stock, and placing the immature embryo extracted in the step on a medium containing a plant hormone. And producing the adventitious embryo from the stock. 2. The method for producing a somatic embryo of a stock according to claim 1, wherein the plant hormone comprises auxin or auxin and cytokinin. 3. The method according to claim 2, wherein the auxin is indoleacetic acid or naphthaleneacetic acid. 4. The method for producing a somatic embryo of a stock according to claim 2, wherein the cytokinin is benzyladenine or kinetin. 5. A method for propagating a somatic embryo, wherein the somatic embryo of the stock produced by the method according to claim 1 is subcultured into a medium containing a plant hormone.