KR100723665B1 - Method for producing virus-free potato minitubers for rapid multiplication of seed potato - Google Patents

Abstract

The present invention relates to a method of producing excellent disease free bird seed potatoes for mass production of seed potato, and more specifically, seed seed potato (potato) grown in three weeks by potato stem culture which is directly cut and fertilized after cultivation of potato stem. It relates to a rapid production method of minituber) or a mass production method of potato seedlings (potato minituber) using the same after increasing the size and quantity of potato microtuber.

The mass production method of bird seed potato of the present invention is a stem position, stem growth stage, leaf area, type and concentration of plant growth regulators, the type and concentration of culture soil, culture medium composition added during tissue culture By carrying out the addition time under optimum conditions, it is possible to mass-produce disease-free soybean seed potatoes and new seed potatoes, which are superior in number, size and weight of the produced seed potatoes.

Bean seed potato, bird seed potato, stem culture, culture composition, disease-free seed potato, valley potato

Description

METHOD FOR PRODUCING VIRUS-FREE POTATO MINITUBERS FOR RAPID MULTIPLICATION OF SEED POTATO}

1 is a flow chart of cutting the potato stem of the present invention,

Figure 2a is the size and area of the initial leaf of a single node according to the direct fertilization cutting method of potato stem of the present invention,

Figure 2b shows the size and area of the leaves after three weeks of culture of the cut single node,

Figure 2c shows the size of the seedlings harvested after three weeks culture of the cut single node,

Figure 3 shows the size of the new grain seed potato harvested according to the cultivated potato stem of the present invention sterile culture soil,

4 is a size classification diagram of soybean seed potatoes according to the addition of the plant growth regulator to the MS liquid medium in the mass production method of bird seed potato of the present invention,

FIG. 5 is a size classification diagram of bird seed potatoes according to the addition time of MS liquid medium in MS solid medium in the method of mass production of seed grains of the present invention.

The present invention relates to a production method of excellent disease-free bird seed potatoes for mass production of seed potatoes, and more particularly, rapid mass production of potato seedlings (potato minituber) which is cultured by directly cutting and cutting the nodes at various positions of potato stems. Method and mass production of potato minituber after mass production of potato seed potato (potato microtuber), stem position, stem growth time, leaf area, type of plant growth regulator when cutting potato stem And it relates to a mass production method of the new seedlings to carry out the concentration, the culture solution composition added during the cultivation and the addition time of the optimum conditions.

Potatoes are a member of the eggplant family, and are a nutritive breeding crop by tubers. They are not only a product of crop cultivation, but also a seed of cultivation. However, one of the most serious problems in the breeding of crops is the reduction of the yield due to viral infections. Therefore, it is important to ensure that potato seed is harvested every year, which is free of virus infection. In other words, securing the seed potatoes of a large number of large-sized, excellent disease-free potatoes produced by sprouting sprouts is a top priority.

Therefore, among the recently developed tissue culture technology, after collecting the growth point area with little virus infection density in the plant, go through the tissue culture proliferation step, the stem or tubers undergoing the culture step 6-8 times in the soil and soil harvesting process After being used as a seed paper. In addition, a method of mass-proliferating disease-free potato plants that are not infected with virus has been developed in parallel with the growth point culture technology and high temperature (38 ° C.) heat treatment technology. However, even if a large amount of shoots are produced in-flight by the above method, it is very difficult to transplant the plants grown on the plane to the soil in order to produce the seed seeds that are actually needed. However, because of the fact that a large number of plants died due to the low unit yield, it was necessary to go through several stages of cultivation.

Therefore, two methods are proposed as a solution to this problem. The first method is to produce young plants through viru-free potato stem culture in a container medium ( in vitro ), Since this is transferred to sterile soil (clean soil) to produce a large number of seed seed potatoes.

The second method is to produce potato microtuber in a container and use the bean seed potato as a starting seed seed in a minituber production process. In particular, in the latter case, soybean seed potatoes can be produced and stored for a long time, so that they can be used in large quantities in a timely manner. The soybean seed potato (potato microtuber) is also called in-flight tuber, and the stem of potato is artificially formed by giving appropriate culture environment conditions while cultivating it on an artificial medium. It is a tuber of about the size of soybeans that are extremely reduced in size (0.5-1.0 cm in diameter and about 200-1000 mg in weight). Hereinafter, in the present specification, "bean seed potatoes".

Regarding the formation of soybean seed potato (potato microtuber), a method of mass production of superior disease-free seeds of potato by the tissue culture method has been reported [Wang PJ and CY Hu. 1982. In vitro mass tuberization and virus-free seed potato production in Taiwan. Amer. Potato Jour . 59: 33-39] Since potatoes require so many seed potatoes every year, even if they can be used as seed potatoes, a huge number of soybean seed potatoes can be produced in large quantities in a small area and supplied at low prices. That's still not enough.

In addition, the potato microtuber is about the size of soybeans and is so small compared to conventional fist-sized seed potatoes, which not only gives psychological anxiety to farmers as end-users, but also gives rise to fragile shoots at the beginning of growth. It is disadvantageous in that it is very disadvantageous in the damage caused by natural disasters and pests, and in the competition for growth with the surrounding weeds.

Therefore, in recent years, such a microtuber by inserting a tissue culture seedling in the container or pot cultivation of soybean seed-sized potatoes, soybean seed potatoes that are about 10 times larger in size while maintaining the quality of the disease-free soybean seed potatoes Trying to produce. Hereinafter, in the present specification, seed potatoes having a weight of 1 to 15 g of potatoes are referred to as potato minitubers.

On the other hand, as a method for shortening the various stages of the cultivation process due to the low unit yield, which is a conventional problem, it is proposed a method for producing small tubers with high yield using nutrient solution cultivation [Korea Patent No. 0298475, 0298476. And 0298477. However, nutrient cultivation is very likely to be contaminated with various soil bottles, and once contaminated, it is contaminated through nutrient medium at once, so the risk is very high. For this reason, developed countries in potato countries prefer a system for producing seed grains from small pots using sterile clay rather than nutrient farming.

Therefore, the present inventors endeavored to mass-produce a fine disease-free potato seed, as a result of the rapid mass production method of potato seedlings (minituber) to cultivate by directly cutting the shoot (shoot) of potato stem and cultivated in sterile culture soil Stem location, growth time, leaf area, type and concentration of plant growth regulators, culture composition to be added during cultivation, and the timing of its addition are optimized to maximize the efficiency of bird seed potatoes after mass production. The present invention has been completed by providing a method for mass production of higher seed grain potatoes.

It is an object of the present invention to provide a method for rapid mass production of minituber potatoes, which are directly cut and fertilized with potato stems, transferred to sterile culture soil, and cultured in three weeks.

Another object of the present invention is to provide a mass production method of microtuber (microtuber) to produce a large seed and microtuber using the same to transfer to cultivated in sterile culture soil (minituber).

The present invention to sterilize potato tubers and then cultured in-flight tissue to obtain potato stems, by directly cutting the fertilization of the potato stems, two or more species comprising a single or a top soil selected from the group consisting of top soil, vermiclite and perlite Transfer to a sterile culture soil in mixed form, supplying the culture composition and providing a rapid mass production method of minituber to cultivate by irradiating for 20 days to 40 days with a photoperiod of 16 hours / day at a temperature of 24 ± 2 ℃. do.

The mass production method of the new seed seed potato is proportional to the area and size of the leaves in the stem cut during the cutting of the potato stem.

According to the mass production method of the new seed potatoes, the new seed potatoes produced average weight is 1.0 ~ 1.6g, 30 ~ 50% of the range of the new seed potatoes tuber size 5g or less, 40 ~ 60% of the tuber size 5 ~ 15g range and the residual ratio It can be obtained that the tuber size is 30g or more.

In addition, the culture composition further comprises any one plant growth regulator selected from the group consisting of gibberellic acid, paclobutrazole, 6-benzylaminopurine, kinetin, succinic acid 2,2-dimethylhydrazide and chlorochlorine chloride It may contain.

배지 또는 MS 배지에 배양용 조성물을 함유하는 MS 액체배지를 첨가하여, 2차 배양하고, 암 상태에서 유지하여 재배하는 콩알씨감자(potato microtuber)의 대량생산 공정 및 상기 콩알씨감자의 무균 배양토에 이양하고 다양한 배양조건과 배양조성액 하에서 효율적인 새알씨감자(minituber)의 대량생산 방법을 제공한다. In addition, the present invention is the primary culture of potato seedlings cultured in-flight in MS solid medium containing 6% sucrose, the MS solid medium containing 8% sucrose

MS liquid medium containing the composition for cultivation is added to the medium or MS medium, followed by secondary culture, and then transferred to a mass production process of soybean seed potato (potato microtuber) grown and maintained in a dark state, and transferred to the sterile culture soil of the bean seed potato. The present invention provides an efficient method for mass production of minituber under various culture conditions and culture compositions.

In addition, the culture composition further comprises any one plant growth regulator selected from the group consisting of gibberellic acid, paclobutrazole, 6-benzylaminopurine, kinetin, succinic acid 2,2-dimethylhydrazide and chlorochlorine chloride It may contain.

MS liquid medium containing the culture composition is preferably added one week before the culture in MS solid medium to remain in the dark state.

The soybean seed potato obtained according to the mass production method of the new seed seed potato has an average weight of 0.3-0.6g, 50-70% of the range of seed potato tuber size is 0.01g or less, 20-35% of the tuber size is 0.5g or less, and the remaining ratio This tuber has a size of 1.0 g or more.

Hereinafter, the present invention will be described in detail.

The present invention provides a mass production method of minituber.

The first embodiment for the preferred embodiment of the present invention, sterilized potato tubers and then cultured in-flight tissue to obtain potato stems, the cutting of the potato stems directly, selected from the group consisting of topsoil, vermiclite and perlite Transfer to aseptic culture soil in two or more mixed forms, including single or top soil.

1 is a flow chart for cutting the fertilization of tissue cultured potato stem of the present invention, the mass production method of bird seed potato (minituber) is proportional to the area and size of the leaf in the stem cut in the process of cutting the fertilization of the potato stem do. That is, the larger and larger the area and size of the leaves in the stem, the higher the number, size, and weight of the produced new seed potatoes can be mass-produced superior seed potatoes. From this, it is preferable to cut the fourth to sixth potato stems in Fig. 1, which is advantageous for mass production of seed grains ( Figs. 2A to 2C and Fig. 3 ).

According to the mass production method of the new seed potatoes, the new seed potatoes produced average weight is 1.0 ~ 1.6g, 30 ~ 50% of the range of the new seed potatoes tuber size 5g or less, 40 ~ 60% of the tuber size 5 ~ 15g range and the residual ratio It can be obtained that the tuber size is 30g or more.

A second embodiment for the preferred embodiment of the present invention provides a method for mass production of new seed seed potatoes grown in sterile culture soil using mass production of soybean seed potatoes.

배지 또는 MS 배지에 무기양액의 배양용 조성물을 함유하는 MS 액체배지를 첨가하여, 2차 배양하고, 암 상태에서 유지하여 재배하는 콩알씨감자(potato microtuber)의 대량생산 공정 및 상기 콩알씨감자의 괴경을 무균 배양토에 이양하고 줄기배양 직접 배양을 통한 새알씨감자 생산 방법으로 새알씨감자(minituber)의 대량생산 방법을 제공한다.More specifically, the present invention is the primary culture of potato seedlings cultured in-flight in the MS solid medium containing 6% sucrose, the MS solid medium containing 8% sucrose

Mass production process of soybean seed potatoes and tubers of soybean seed potatoes, which are secondary cultured and cultured in a dark state by adding an MS liquid medium containing a composition for cultivating an inorganic nutrient solution to a medium or MS medium, are maintained in a dark state. This method provides a mass production method of bird seed potatoes (minituber) as a method of producing seed eggs by cultivating them in sterile culture soil and directly cultivating stem culture.

배지 또는 MS 배지에 배양용 조성물이 함유된다. 이때, 배양용 조성물은 KNO₃ 0.4∼2.0 g/L; Ca(NO₃)₂ㆍ4H₂O 0.1∼0.5 g/L; NH₄H₂PO₄ 0.1∼2.0 g/L; 및 MgSO₄ 0.2∼1.0 g/L;로 이루어진 수경재배용 배양용 조성물을 사용한다. 따라서, 통용되는 값싼 수경재배용 배양용 조성물을 함유하는 MS 액체배지를 사용함으로써, 종래 고가의 MS 고체배지만을 사용한 배양공정보다 경제적인 효과가 있다. The method is characterized by performing a two-step culturing process of primary culture in MS solid medium and secondary culture by adding MS liquid medium, wherein the MS liquid medium contains 8% sucrose.

The culture composition is contained in the medium or MS medium. At this time, the composition for culture is KNO ₃ 0.4 _~ 2.0 g / L; Ca (NO ₃ ) ₂ .4H ₂ O 0.1-0.5 g / L; NH ₄ H ₂ PO ₄ 0.1-2.0 g / L; And 0.2 to 1.0 g / L of MgSO ₄ ; Therefore, by using the MS liquid medium containing the commonly used inexpensive hydroponic culture composition, there is an economical effect than the conventional culturing process using only expensive MS solid medium.

In addition, the MS liquid medium of the present invention is more preferable by showing an even result of the tuber size of the produced potato seed (potato minituber) is added one week before the culture in MS solid medium to remain in the dark state ( Fig. 5 ).

The soybean seed potato obtained according to the mass production method of the new seed seed potato has an average weight of 0.3-0.6g, 50-70% of the range of seed potato tuber size is 0.01g or less, 20-35% of the tuber size is 0.5g or less, and the remaining ratio This tuber has a size of 1.0 g or more.

The culture composition of the present invention may further contain plant growth regulators (PGRs), the plant growth regulators Gibberellic acid (Gibberellic acid), Paclobutazol, 6-benzylaminopurine (BAP), kinetin , Any one selected from the group consisting of succinic acid 2,2-dimethylhydrazide (B 9) and chlorochlorine chloride is used.

Potato varieties that can be used in the present invention are Atlantic, Supermi, Winter Valley, NorValley, Early Valley and Golden Valley in the embodiment of the present invention. Although any one selected from the group consisting of) is described, but not particularly limited to all potato varieties can be applied.

Sterile culture soil that can be used in the present invention may be used in the form of a single soil or a mixture of two or more selected from the group consisting of top soil, vermiclite and perlite, more preferably the top soil and vermiclite in the same amount It is to use mixed culture soil.

Hereinafter, the present invention will be described in detail by way of examples.

The following examples are merely illustrative of the present invention, but the content of the present invention is not limited to the following examples.

I. Mass production of minituber

<Example 1>

A superior tuber was used, which was immersed in 70% ethanol for 5 minutes and continuously immersed in 10% chlorax (commercial finitex) for 10 minutes, and then washed 3-4 times with purified distilled water. Sterilized tubers were seeded in 21 cm diameter vessels containing top soil and incubated for 45 days. For fast and good growth, 2.4 g / L of fertilizer (20-20-20 / NPK) was fed for one month after sowing and kept in cancer for one week before the shoot cut. Correction cutting was performed just before flowering, and a total of eight samples were prepared by cutting from the end of the potato stem, that is, the youngest leaf, in the labeled order of FIG. 1 . The cut stems were handed over to a plastic tray in which topsoil and vermiculite were mixed 1: 1, and the plastic trays were sterilized with 10% Chlorox before use. Then, hydroponic cultivation consisting of KNO ₃ 404.0 mg / L, Ca (NO ₃ ) ₂ ㆍ 4H ₂ O 354.0mg / L, NH ₄ H ₂ PO ₄ 76.7mg / L, MgSO ₄ 246.5mg / L Up to a week ago, each plant was continuously fed through a pipe. After 3 days, 10 mg / L plant growth regulator 6-benzylaminopurine (BAP) was sprayed on the leaves, and cultured for 3 weeks by irradiating with a photoperiod of 16 hours / day at 25 ℃ temperature.

<Example 2>

The same procedure as in Example 1 was carried out except that Winter Valley was used instead of Sumi potato species.

Characteristics of Bird Seed Potatoes Prepared by Direct Fertilization Cutting of Potato Stalks Cutting order Example 1 Example 2 Leaf area (cm ² ) Germination Rate (%) Tuber seed potato weight / node (g) Leaf area (cm ² ) Germination Rate (%) Tuber seed potato weight / node (g) 1st single node 10.1 68.3 0.37 12.3 76.6 0.31 2nd single node 17.6 88.0 0.70 17.8 83.3 0.61 3rd single node 33.3 90.0 0.95 31.6 93.3 0.88 4th single node 41.8 93.3 1.11 40.6 100.0 1.07 5th single node 47.3 100.0 1.51 48.0 100.0 1.25 6th single node 45.3 91.6 1.25 46.3 96.6 1.20 7th single node 34.6 96.6 1.28 40.6 100.0 1.08 8th single node 24.3 90.0 1.01 25.0 83.3 1.07

Figure 2a shows the size and area of the initial leaf of a single node according to the direct fertilization cutting method of potato stem of the present invention, Figure 2b shows the size and area of the leaf after 3 weeks of culture of the cut single node 2C shows the size of the harvested minituber after 3 weeks of incubation of the cut single node. As shown in FIG. 2A to FIG. 2C, the larger the size and area of the initial leaf, the larger the harvested minituber and the higher the germination rate. This is because the larger the leaf area, the higher the photosynthetic rate and the faster the carbohydrate transfers from the leaf to the tuber.

In addition, as shown in Table 1, in the direct fertilization cutting method of potato stems, the fourth to sixth single node is preferred, the fifth single node having the largest leaf size and area is most preferred.

Therefore, when grown to a single node of potato stem with a sufficiently large and large leaf, and cultured in sterile culture soil, it is advantageous for the mass production of minituber which is superior in number, size and weight.

Conventional methods using growth points and stem nodes have been widely used, but the results of studies on the location of the cutting nodes have not been published. In addition, in the present invention, the production of new seed potatoes using a conventional plant takes about 60 to 90 days, but according to the technique of the present invention, it is possible to produce a new seed seed potato in 21 days, thereby enabling mass production within a predetermined time.

<Example 3>

Except for culturing soil mixed 1: 1 and vermiclite 1: 1, it was carried out in the same manner as in Example 1 except for culturing in a plastic tray containing only the clay.

<Example 4>

Winter Valley was used as potato species, and the same procedure as in Example 1 was performed except that the culture was carried out in a plastic tray containing only the clay, instead of the culture soil mixed with the clay and vermiclite 1: 1.

Example 5

Instead of cultured soil mixed with 1: 1 and vermiculite 1: 1, it was carried out in the same manner as in Example 1 except for culturing in a plastic tray containing vermiclite alone.

<Example 6>

Winter Valley was used as the potato species, and was cultured in the same manner as in Example 1 except for culturing in a plastic tray containing vermiclite alone, instead of cultured soil mixed with top soil and vermiclite 1: 1. .

Characteristics of Bird Seed Potato According to the Cultured Soil division Potato species process Leaf area (cm ² ) Germination Rate (%) Tuber seed potato weight / node (g) Example 1 Sumy Clay + Vermiclite 34.7 100.0 1.52 Example 3 Topsoil 37.0 92.5 1.30 Example 5 Vermiculite 32.7 87.5 1.21 Example 2 Winter Valley Clay + Vermiclite 37.0 100.0 1.55 Example 4 Topsoil 36.4 92.5 1.27 Example 6 Vermiculite 33.0 82.5 1.00

Figure 3 shows the size of the new seed (minituber) harvested according to the sterile culture soil, the tuber of the new seed potato (minituber) was observed in the culture soil mixed with the top soil and Vermiclite 1: 1. In addition, as shown in Table 2, in the case of Example 1 and 2 cultured in a culture soil mixed with the top soil and vermiclite 1: 1, the germination rate is 100.0%, the weight of minituber 1.52g respectively And 1.55 g.

From these results, the use of clay alone produces many roots and increases the leaf area, but delays the formation of minituber, making it difficult to improve tuber size. Since nutrition is not enough to obtain these, therefore, it is most preferable in the case of cultured soil mixed in a 1: 1 equivalent amount thereof.

<Example 7>

As a plant growth regulator, 50mg / L of kinine (kinetin) was sprayed on the leaves, except that it was grown in the same manner as in Example 1.

<Example 8>

As a plant growth regulator, it was carried out in the same manner as in Example 2, except that 50 mg / L of kinetin was sprayed onto the leaves and grown.

Characteristics of Bird Seed Potato According to Plant Growth Regulators division Potato species process Leaf area (cm ² ) Germination Rate (%) Tuber seed potato weight / node (g) Example 1 Sumy BAP injection 36.7 100.0 1.35 Example 7 kinetin sprayed 36.3 95.0 1.17 Comparative Example 1 No treatment 34.5 95.0 1.05 Example 2 Winter Valley BAP injection 31.2 100.0 1.31 Example 8 kinetin sprayed 31.7 96.0 1.20 Comparative Example 2 No treatment 29.7 93.0 1.10

As shown in Table 3 , the germination rate was promoted when the plant growth regulator was sprayed on the leaves, more preferably in Examples 1 and 2 using 6-benzylaminopurine (BAP), the germination rate is maximized The seeds of minituber weighed 1.35 g and 1.31 g, respectively.

Example 9

Except for using Atlantic as the potato species, the same culture as in Example 1 above, but as a sterile culture soil, using top soil and plant growth regulator, 50mg / L of Gibberellic acid solution leaves After spraying on, it was incubated for 90 days and then grown. At this time, the tubers were classified according to the size (<5g, 5-15g, 15-30g, and> 30g). Half of the cultivated potatoes were stored at 4 ° C. and the other half were observed for 4 months at room temperature and dark. Experiments were carried out in a vinyl house condition and the number of tubers was determined according to plant height, total tuber weight, weight and size. Data were analyzed using an analyzer and average separation was performed at a Duncan multi-domain test at 0.05 using a statistical analysis system (SAS, 1990) package.

<Example 10>

Except for the use of Atlantic as the potato species, the same culture as in Example 1 above, but using the soil as a sterile culture soil and spraying 100mg / L Gibberellic acid solution as a plant growth regulator to the leaves To produce minituber.

<Example 11>

Except for using Atlantic as the potato species, the same culture as in Example 1 above, but using the top soil as sterile culture soil and 50mg / L Paclobutazol solution as a plant growth regulator to the leaves Sprayed to produce minituber.

<Example 12>

Except for the use of Atlantic as the potato species, the same culture as in Example 1 above, but using the top soil as sterile culture soil and 100 mg / L Paclobutazol solution as a plant growth regulator to the leaves Sprayed to produce minituber.

Comparative Example 3

Except for using the Atlantic (Atlantic) as a potato species, but was cultured in the same manner as in Example 1, using a top soil as a sterile culture soil and untreated plant growth regulator was produced minituber.

division process Go Kyoung Soo / potato Tuber size (%) Storage days <5 g 5-15g 15-30 g > 30 g 80 days 95 days 110 days 125 days 4 ℃ RT 4 ℃ RT 4 ℃ RT 4 ℃ RT Example 9 Topsoil-GA ₃ ^a 50 10.3 32.6 58.1 9.3 - 0 40 0 100 0 100 92.5 100 Example 10 Topsoil-GA ₃ 100 10.0 42.7 44.3 4.6 8.4 0 90 0 100 0 100 100 100 Example 11 Topsoil-PBZ ^b 50 10.8 48.8 48.8 2.4 - 0 0 0 90 0 100 82.5 100 Example 12 Topsoil-PBZ 150 14.0 50.1 42.9 7.0 - 0 0 0 100 0 100 80 100 Comparative Example 3 Topsoil-Free Treatment 7.5 43.3 50.0 6.7 - 0 10 0 100 0 100 87.5 100

a: Gibberellic acid, and b: Paclobutazol

As shown in Table 4 , the tuber number of potatoes formed when the plant growth regulator was not treated in culture soil was 7.5, whereas when the plant growth regulator of gibberellic acid or paclobutrazole was added, the tuber number was 10 or more, High yields of minituber can be expected. Gibberellic acid promoted kidney growth of harvested potato stems, whereas paclobutrazole inhibited kidney growth of harvested potato stems (results not shown). From these results, as shown in Examples 11 and 12, when the plant growth regulator of paclobutrazole was sprayed, the tuber number of the potato was 10.8 and 14.0, and the even growth of the potato stem was suppressed by showing an even size distribution. On the other hand, it was confirmed that nutrients can move to the tuber part to produce enlarged tuber.

In addition, half of the cultivated potatoes were stored at 4 ° C. and the other half of the shoots were observed at room temperature and in the dark for 4 months. As a result, room temperature germination germinated one month earlier than cold storage (4 ° C.), Gibberellin Tubers sprayed with acid (GA ₃ ) solution showed a shorter germination time at 30-80% at room temperature and 5-12.5% at 4 ° C than untreated group. On the other hand, tubers sprayed with paclobutrazole (PBZ) solution were confirmed to have an effect of delaying germination by 0 to 10% at room temperature and 5 to 7.5% at 4 ° C than the untreated group.

From the above, gibberellic acid (GA ₃ ) is useful for mass production because it promotes dormancy breakdown of minituber at 4 ° C. and room temperature storage conditions. .

II. Mass production method of minituber using mass production of soybean seed potato (microtuber)

<Examples 13-19>

Step 1: Mass Production of Potato Microtuber

Select any one from the group consisting of disease-free, Superior, Early Valley, Golden Valley, and potato genotype F-9 99 varieties provided by KPGR. It can be, but this embodiment will be described by performing the rice bran tuber. In-flight tissue cultured potato seedlings for 60 days with a photoperiod of 16 hours / day at a temperature of 24 ± 2 ° C in Murushige and Skoog's solid medium (Murashige & Skoog, hereinafter referred to as "MS medium") containing 6% sucrose. It was cultured by light irradiation.

배지 및/또는 MS 배지 및 세가지 조성 중에서 선택되는 수경재배용 배양용 조성물을 포함하며, 이때의 바람직한 양은 pH 5.6를 만족시킬 수 있을 정도의 양으로 정의한다.Then, after as described below in the solid medium Table 5a and Table 5b culture was added to a liquid medium and inorganic nutrient solution and the growth transferred to a dark state were harvested pea seed potatoes (potato microtuber). At this time, the liquid medium containing 8% sucrose

Hydroponic culture culture composition selected from the medium and / or MS medium and three compositions, the preferred amount at this time is defined as an amount sufficient to satisfy the pH 5.6.

Step 2: Mass production of new seed potatoes using soybean seed potatoes

Tubers of soybean seed potatoes obtained in step 1 were sown in culture soil and cultured to produce a large amount of potato minituber. At this time, the production method was carried out in the same manner as in Example 1, the composition of the plant growth regulator and the culture soil may vary.

Production of Soybean Seed Potato and Its Characteristics division process Seed Potato / Sapling FW / seed potato (mg) Tuber size of seed potatoes (%) Count Weight (mg)

0.01

0.01

0.5

1.0 Example 13

, 8% Sucrose, Inorganic Nutrition1 1.20 394.8 329.0 69.2 25.0 5.8 Example 14

, 8% Sucrose, Inorganic Nutrition 2 1.40 564.9 406.4 50.0 33.2 16.8 Example 15

, 8% sucrose, inorganic nutrients3 1.23 511.7 416.0 67.5 21.9 10.6 Example 16 MS, 8% Sucrose, Inorganic Nutrition 1 1.27 546.7 430.5 52.7 25.9 21.4 Example 17 MS, 8% Sucrose, Inorganic Nutrition 2 1.36 602.2 442.8 51.9 28.3 20.1 Example 18 MS, 8% sucrose, inorganic nutrients3 1.33 512.7 358.5 50.9 29.1 20.0 Example 19 8% Sucrose, Inorganic Nutrition 2 1.13 391.0 346.0 62.0 29.2 8.8 Comparative Example 4 MS, 8% sucrose 1.07 424.7 396.9 56.1 31.8 12.1

Classification Inorganic nutrients (g / L) KNO ₃ NH ₄ H ₂ PO ₄ Ca (NO ₃ ) ₂ 4H ₂ O MgSO ₄ One 0.81 0.94 0.15 0.49 2 1.22 1.41 0.23 0.74 3 1.62 1.88 0.30 0.98

0.01g의 50.0%,

0.5g의 33.2% 및

1.0g의 16.8%를 얻었다. 가장 바람직하게는 실시예 14의 액체배지 조성인

, 8% 수크로오스 및 무기영양 2로 이루어진 배양액의 경우, 각각 1.4개수 및 564.9 mg을 나타내었다. As shown in Table 5a, in terms of the number and weight of soybean seed potato (microtuber), preferably in the case of the culture medium consisting of the liquid medium composition of Example 17, 8% sucrose and inorganic nutrition 2, 1.36 pieces and 602.2 mg, the distribution of its size

50.0% of 0.01g,

33.2% of 0.5 g and

1.0 g of 16.8% was obtained. Most preferably the liquid medium composition of Example 14

For cultures consisting of, 8% sucrose, and inorganic nutrients 2, 1.4 and 564.9 mg, respectively.

0.01g의 51.9%,

0.5g의 28.3% 및

1.0g의 20.1%를 얻음으로써, 350∼600mg의 무게 및 얻어진 콩알씨감자의 괴경크기가 0.01∼1.0g 범위 내에서 고른 분포를 나타내었다.In addition, the distribution of the size

51.9% of 0.01 g,

28.3% of 0.5 g and

By obtaining 20.1% of 1.0 g, the weight of 350-600 mg and the tuber size of the soybean seed potatoes obtained were evenly distributed within the range of 0.01-1.0 g.

<Examples 20-21>

, 8% 수크로오스 및 무기영양 2로 이루어진 배양액)에서, 하기 표 6에 기재된 바와 같이, 식물성장조절제 10mg/L의 6-벤질아미노퓨린(BAP) 및 200mg/L의 숙신산 2,2-디메틸하이드라지드(succcinic acid 2,2-dimethylhydrazide;B 9)를 첨가하는 배양액 조성물을 제조하였다.The liquid medium composition of Example 14 most preferred from the above results (

, A culture solution consisting of 8% sucrose and inorganic nutrients 2), as shown in Table 6 below, a plant growth regulator 10 mg / L 6-benzylaminopurine (BAP) and 200 mg / L succinic acid 2,2-dimethylhydra A culture composition was prepared to which zide (succcinic acid 2,2-dimethylhydrazide; B 9) was added.

division process Germination time (4 days) (%) Bean Seed Potato / Potato Seedling FW / Bean Seed Potato (mg) One 2 3 4 5 6 Count Weight (mg) Example 14

, 8% 수크로오스, 무기영양2

, 8% Sucrose, Inorganic Nutrition 2 28 50 65 79 86 95 1.28 602.5 470.7 Example 20 MS, 8% sucrose, 10 mg / L BAP, 200 mg / L B 9 46 74 92 92 100 100 1.08 814.0 755.7 Example 21

, 8% sucrose, 10 mg / L BAP, 200 mg / L B 9, inorganic nutrition 2 38 67 90 95 100 100 1.4 971.0 693.6 Comparative Example 4 MS, 8% sucrose 30 65 85 90 95 100 1.0 490.0 490.0

As shown in Table 6 , the germination time was shortened according to the addition of the plant growth regulator, it was confirmed that the bean seed potatoes produced better than Comparative Example 4. More preferably, in the case of Example 21, by showing the number 1.4 and the weight 971.0mg in terms of number and weight, a superior new seed potato was secured.

In addition, Figure 4 is a size classification diagram of soybean seed potatoes according to the treatment group of Table 6, preferably in the case of Examples 20 and 21, that is, when the plant growth regulator is further contained in the liquid medium of the present invention, The size of soybean seed potatoes showed even distribution in the range of> 0.0g and> 0.5g.

<Examples 22 to 25>

After culturing in MS solid medium in Example 13, to determine the desired time to add the MS liquid medium, it was carried out as described in Table 7 below depending on the time before or after the transfer to the dark state.

division process Sumy Seed Potato / Sapling FW / seed potato (mg) Count Weight (mg) Example 22 2 weeks ago 1.40 977.0 697.9 Example 23 1 week ago 1.60 1207.7 754.8 0 days 1.40 803.3 573.8 Example 24 In a week 1.50 777.7 518.5 Example 25 2 weeks later 0.97 516.7 532.7

From the above results, it can be seen that it is most preferable to add MS liquid medium after culturing in MS solid medium, and one week before transferring to the dark state.

In addition, Figure 5 is a size classification diagram of the seed grains produced according to the addition time of the MS liquid medium in the MS solid medium in the mass production method of the potato minituber of the present invention. Sumi confirmed the even distribution of the new seed potato produced when the liquid medium was added one week before growth in cancer after transplantation, that is, the> 0.5g size was 43%, and the largest distribution was> 0.01g. 36% in size and 21% in size> 1g.

As described above, the present invention

First, we provided a method for mass production of minituber, which can be rapidly grown in three weeks by directly cutting the fertilization of potato stem and handing it over to sterile culture soil.

배지 또는 MS 배지에 배양용 조성물을 함유하는 MS 액체배지를 첨가하여 2차 배양하고, 암 상태에서 유지하여 재배하는 콩알씨감자(potato microtuber)의 대량생산 공정을 제공하였고, Second, in-flight tissue cultured potato seedlings were first cultured in MS solid medium containing 6% sucrose, and the MS solid medium contained 8% sucrose.

To provide a mass production process of soybean seed potato (potato microtuber) to cultivate by secondary culture by adding the MS liquid medium containing the culture composition to the medium or MS medium, maintained in the dark state,

Third, the composition of the MS liquid medium used in the mass production process of the soybean seed potato (potato microtuber) and the optimum conditions of the addition time was provided,

Fourth, in the composition of the MS liquid medium, since the culture composition uses a conventional hydroponic culture medium composition, it is possible to avoid the use of conventionally expensive MS solid medium, so there is an economic effect.

Although the present invention has been described in detail only with respect to the embodiments described, it will be apparent to those skilled in the art that various modifications and variations are possible within the technical spirit of the present invention, and such modifications and variations belong to the appended claims. .

Claims (7)

Potato stems are obtained by sterilizing potato tubers and then incubating tissues in-flight, and cutting the fertilization of the potato stems directly, and in the form of two or more mixed forms including single or top soil selected from the group consisting of top soil, vermiclite and perlite. Transfer to sterile culture soil, and supplying the culture composition, the rapid mass production method of bird seed potatoes, characterized in that grown for 20 to 40 days light irradiation with a photoperiod of 16 hours / day at a temperature of 24 ± 2 ℃. delete The method of claim 1, wherein the culture composition is any one plant growth selected from the group consisting of gibberellic acid, paclobutrazole, 6-benzylaminopurine, kinetin, succinic acid 2,2-dimethylhydrazide and chlorochlorine chloride Rapid mass production method of bird seed potatoes, characterized in that it further comprises a regulator. Primary culture of potato cultured potato seedlings in MS solid medium containing 6% sucrose; The MS solid medium containing 8% sucrose

Secondary culture by adding MS liquid medium containing the composition for culturing the inorganic nutrient solution to the medium or MS medium; Maintaining the MS solid medium in the secondary culture in a dark state to produce soybean seed potatoes in large quantities; Sterilizing the tubers of the mass produced soybean seed potatoes and then incubating tissue to obtain potato stems; And Directly cutting the fertilization of the potato stem, handed over to sterile culture soil of two or more mixed forms, including single or top soil selected from the group consisting of top soil, vermiclite and perlite, and supply the culture composition to supply 24 ± 2 ℃ The method of mass production of bird seed potato, characterized in that consisting of; culturing by irradiating light for 20 days to 40 days with a photoperiod of 16 hours / day at a temperature. The method of claim 4, wherein the culture composition is KNO ₃ 0.4 _~ 2.0 g / L; Ca (NO ₃ ) ₂ .4H ₂ O 0.1-0.5 g / L; NH ₄ H ₂ PO ₄ 0.1-2.0 g / L; And MgSO ₄ 0.2 ~ 1.0 g / L; mass production method of the seed seedlings characterized in that the composition for cultivation for hydroponic cultivation. The method of claim 4, wherein the culture composition is any one plant growth selected from the group consisting of gibberellic acid, paclobutrazole, 6-benzylaminopurine, kinetin, succinic acid 2,2-dimethylhydrazide and chlorochlorine chloride The method of mass production of the new seed potato characterized in that it further comprises a regulator. [5] The method of claim 4, wherein the MS liquid medium is added one week before being cultured in MS solid medium and maintained in a dark state.

Images