JPH0724519B2 - Potato tuber manufacturing method - Google Patents

Description

Detailed Description of the Invention a. INDUSTRIAL APPLICABILITY The present invention relates to a potato (Solanum tubero by tissue culture method).
sum L.) tuber production method. More specifically, potato plants are cultivated at a growing point, a multibud,
The present invention relates to a method for efficiently forming several tubers in a sterile plant body grown by a tissue culture technique such as node culture using a specific growth control substance. This tuber can be taken out of the culture container and stored for a long period of time, and the germination and subsequent growth are also good. By this method, it is possible to grow a large number of disease-free excellent seedlings of new varieties obtained by cross breeding of potatoes or varieties obtained by introduction from abroad in an inexpensive and genetically uniform state in a short period of time. Become.

B. Conventional technology Conventionally, much research has been conducted on the production method of potato tubers using the tissue culture method, and estrada and the like are produced in a medium containing 2-chloroethyl-trimethylammonium chloride (CCC), which is one of the growth regulators. Potato tubers were obtained by cultivating potato aseptic plants (Plant Cell, T
issue and Organ Culture, 7 , 3 ~ 10,1986). In this method, tuber formation is promoted, but its effect is not always sufficient, the manufacturing concentration is high because the concentration of the drug used is as high as 500 ppm, and CCC is a deleterious substance, so special handling is required. .

C. DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention The present invention is to solve the above-mentioned drawbacks of the prior art, and an object of the present invention is to improve efficiency by a plant tissue culture method in a short time regardless of climate, soil, season, etc. It is often the production of tubers for the rapid spread of new potato varieties and introduced varieties.

D. Means for Solving the Problems For the purpose of solving the above problems, the present inventors have conducted a detailed study on a method for producing potato tubers by tissue culture, and as a result, have identified a specific growth control substance, namely Dole (α-cyclopropyl-α- (p-methoxyphenyl)
-5-pyrimidine methyl alcohol), paclobutrazol ((2- RS , 3- RS ) -1- (4-chlorophenyl) -4--4-dimethyl-2- (1H-1,2,4-triazole- 1-yl) penten-3-ol) or uniconazole ((E) 1- (4-chlorophenyl) -4,4-dimethyl-2- (1,2,4-triazol-1-yl) -1-pentane- It was found that the above problems were remarkably improved by adding (3-ol) to the medium, and the present invention was completed.

The production method of the present invention is preferably carried out by a tissue culture method comprising the following steps A and B.

<Step A: Establishment of Aseptic Culture System> Aseptic plants of potato can be obtained by a known method.

For example, potato tubers, shoot tips, stems and the like or tissue sections obtained by cutting them are sterilized with ethyl alcohol, sodium hypochlorite and the like, and then washed thoroughly with sterile water. The plant body or tissue section thus surface-sterilized is placed in a sterilized solid medium or liquid medium at a rate of 1 per 1 to 100 ml of the medium. As the solid medium or liquid medium, any medium can be used as long as it is a medium usually used for tissue culture of plants. For example, Murashige and Skoog's medium, Rinsmeier's and Skoog's medium, etc., or various modifications of these as basic medium are used. Agar, agarose, gellan gum and the like are used to make them solid. It is also possible to add various concentrations of plant growth regulators such as auxins and cytokinins to the medium in combination. The amount of these plant growth regulators added is
Although it varies depending on the type of plant growth regulator, plant part, culture stage, etc., it is generally about 0.01 to 10 mg / l. The pH of the medium is preferably 4.0 to 7.0. Lighting is 100-100
It is desirable to do it with a luminous intensity of 000 lux. Also in the step B described below, the above medium and plant growth regulator are appropriately used. After placement, plants develop from the tissue slices planted by culturing at 10 to 30 ° C, preferably 21 to 25 ° C for 1 to 20 weeks. As described above, a sterile culture system for potato is established.

<Step B: Formation of tubers / promotion of growth by growth control substance> The sterile plant body established in step A is cut / divided at a ratio of 1 slice per 1 to 100 m1 of liquid medium containing the growth control substance of the present invention. After placing the plate, incubate at 10 to 30 ° C for 1 to 20 weeks. The culturing method is not particularly limited, and a known method, that is, static, shaking, swirling, aerating or stirring culturing can be performed. By culturing, the foliar shoots of the shoots present in the tissue section as the explant are suppressed, the shoots are efficiently converted into tubers, and the tubers are also enlarged. Growth regulator concentration is 0.01-10 mg
A range of / l is suitable. As the growth regulator, a specific growth regulator, that is, ancimidol, uniconazole, or paclobutrazol is used, and uniconazole and paclobutrazol are particularly effective. Irradiation of light may be omitted. By repeating these steps A and B, tuber production by tissue culture can be performed stably and rapidly.

In the method of the present invention, the apical bud of the explant is first enlarged and tuberized, and then the axillary bud is sequentially tuberized from the top,
If the apical buds are cut off, a large amount of axillary buds will be tuberized, and many tubers can be obtained.

E. Example Next, an example will be described.

Example 1 Tubers of the potato variety May Queen are laid down in vermiculite in a garden vat and grown in a greenhouse to germinate. The germinated buds are made to have a length of about 5 cm, soaked in a 10% sodium hypochlorite solution (effective chlorine amount: 1%) for 15 minutes to sterilize the surface, and then washed 3 times with sterile distilled water. Under a dissecting microscope, the young leaves are removed with tweezers to expose the growing points. Using a scalpel, cut the growth point into 0.5 mm high slices,
16mm inner diameter including 5m1 of agar medium for growing the composition shown in Table 1 below
Place one test tube in a 130 mm high test tube per test tube, stopper with aluminum foil, and incubate at 25 ° C under 5000 lux illumination.

Dissolve the above components in distilled water to make 1 liter of the medium, and adjust the pH.
Was adjusted to 5.8 and steam sterilized using an autoclave.

By this culture, the growing point developed into a young plant in about 1 month. When the seedlings have about 10 leaves, 1
Each node was cut off and replanted in the above medium. The raw material for tuber production was grown by repeating this section culture. For the production of tubers, the sucrose concentration of the medium composition in Table 1 was changed to 60 g / l, the agar was removed, and paclibutrazol was added in an amount of 0-100.0 mg / l. It was carried out by transplanting a young plant one month after the start of the node culture, and culturing for one week. The culture was performed at 20 ° C. in the dark. As shown in Table 2, 2 to 5 tubers grew on the plant cultivated in the medium containing 0.01 to 10 mg of paclobutrazol, whereas the medium containing no paclobutrazol at all. No tuber colonization was observed in the plants cultured in 1. The tuber thus obtained was taken out from the Erlenmeyer flask, washed with running water to remove the liquid medium with a liquid, and then stored in a refrigerator at 3 ° C. Three months later, tubers were planted in a greenhouse in which tubers were planted in a medium in which equal volumes of synthetic horticultural soil made by Kureha Chemical Co., Ltd. and peat moss were mixed. Sprouting was observed within 1 week and 100% germination within 2 weeks.

The above data are average values per 20 straight objects.

Example 2 In the same manner as in Example 1, seedlings of the barley barley of the potato variety were grown by tissue culture. Plants three weeks after the start of node culture were adjusted to a sucrose concentration of 60 g / l in the medium composition shown in Table 1, 0 to 10 mg / l of uniconazole was added, and transplanted to a liquid medium excluding agar. One week later, when cultured in a medium containing uniconazole, tuber formation was 2 to 4 per plant.
However, tuber colonization was not observed without uniconazole.

Example 3 In the same manner as in Example 1, seedlings of Baresque variety Baroness were grown by tissue culture. Plants 4 weeks after the start of node culture were adjusted to sucrose concentration of 60 g / l in the medium composition shown in Table 1, and 0.01 to 10 mg / l of paclobutrazol or CCC.
After adding 0.01 to 10 mg / l, the cells were transplanted to a liquid medium excluding agar.
One week later, when cultured in a medium containing paclobutrazol, 3 to 5 tubers were observed per plant, but in the case of containing CCC, tubers were not observed.

Example 4 In the same manner as in Example 1, potato varieties
Young plants of Norin No. 1, Tsunika, Benimaru, Hatsufuki, Yukijiro, Toyoshiro or Vaseshiro were grown by tissue culture. 4 weeks after the start of the section culture, the sucrose concentration in the medium composition in Table 1 was set to 60 g / l, and uniconazole was added to 0.01%.
-10 ml / l was added and the cells were transplanted to a liquid medium without agar. 1
After a week, 2 to 5 tubers were observed per plant in all the cultivars.

Example 5 In the same manner as in Example 1, seedlings of Baresque variety Baroness were grown by tissue culture. Four weeks after the start of knot culture, the plant was divided into sections containing four knots, grown in a liquid medium obtained by removing agar from the medium composition shown in Table 1 for 3 days at 25 ° C, and then grown. Of the medium composition in Table 1, the sucrose concentration was 60
The concentration was changed to g / l, 1.25 mg / l of ansimidol was added, and the medium was transplanted to a liquid medium excluding agar and a medium not containing ansimidol. Two weeks later, as shown in Table 3, when cultured in a medium containing ancimidol, 3 to 5 tubers were observed per plant, but in the plant in a medium containing no ancidol. Almost no tuber colonization was observed.

Three sections were seeded with 4 nodes per culture bottle. Data are average values per 3 culture bottles.

Comparative Test Example Potato tubers were produced in the same manner as in Example 1 using various concentrations of growth control substances, and the tuber formation rate was examined. The results are shown in Table 4.

From Table 4, the tuber formation rate of the growth regulator of the present invention is extremely higher than that of CCC, and especially paclobutrazol and uniconazole have excellent potency at an extremely low concentration of 0.1 to 1.0 mg / l. I understand.

F. EFFECTS OF THE INVENTION According to the present invention, tubers can be efficiently produced from an aseptic potato plant. Since the tissue culture method is used in the production method of the present invention, it is not affected by natural conditions such as season, weather, and soil, and fertilization, chemical spraying, water supply, and other cultivation management are not required, and a large area may be required. No, many potato tubers can be produced in a short period of time. The tuber obtained by the production method of the present invention can be taken out of the culture vessel and stored for a long period of time, and the germination and the subsequent growth are good.

Therefore, by using the production method of the present invention, it is possible to multiply a large number of disease-free excellent seedlings of a new variety obtained by cross breeding of potatoes in a genetically uniform state within a short period of time.

Claims (3)

[Claims] 1. A method for producing potato tubers, which comprises culturing a sterile potato plant obtained by tissue culture in a medium containing ansimidol, paclobutrazol or uniconazole. 2. The method according to claim 1, wherein the concentration of ansimidol, paclobutrazol or uniconazole in the medium is 0.01 to 10 mg / l. 3. The method according to claim 1, wherein the culture temperature is 10 to 30 ° C.