KR20210036433A - Method of producing seed potatoe using red and blue LED of low light intensity as light source - Google Patents

Abstract

A method for increasing the productivity of direct planting microtuber potato seeds is presented. According to the method of the present invention, by mixing red and blue LED light sources and irradiating with a low light intensity during tissue culture of potato stems, the growth of potato stems is improved, and the welfare formation rate is increased, so that a large amount of microtuber potato seeds are finally produced.

Description

{Method of producing seed potatoe using red and blue LED of low light intensity as light source}

The present invention relates to a method for producing microtuber potato seeds, and more particularly, to a method for producing large amounts of microtuber potato seeds having a large size.

Potatoes are one of the world's four largest food crops, followed by rice, wheat, and corn because of their short growing period, relatively high production per unit area, and strong environmental adaptability. Potatoes are used as a staple food in each country and mainly used to make various processed products, but the consumption of potatoes in Korea is on the rise.

Potatoes can be truly propagated by seeds or vegetative propagation, but in general, the method by vegetative propagation is dominant. When producing potatoes by nutrient breeding, it is important to select disease-free and superior seed potatoes, and the seed potatoes supplied to farms must be disease-free, disease-free, such as viruses.

Seed potatoes usually produce small-sized seed potatoes by cultivating tissue-cultivated potato seeds (sterile artificial seed potatoes) on top soil or hydroponics in a house. have. According to this system, it takes about 5-7 years to produce seed potatoes for supply to farmers from tissue-cultivated potato seeds. The reason why a long time is necessary is that it takes a lot of cost and time to produce the first tissue cultured potato seeds, and a system for mass production is not established, requiring several proliferation processes. Therefore, as it takes a long time, there is a concern that the rate of virus infection coming from the soil or environment increases.

On the other hand, the method for mass production of disease-free seed potatoes using a bioreactor incubator has an advantage in mass production of a culture medium by putting a liquid medium of several hundred liters in the incubator. However, this cultivation method requires a complex cultivation technique, high facility cost, and requires an additional planting process to secure tubers the size of a mini tuber. In addition, if contamination occurs during the process of mass production of the culture medium, there is a concern that the entire culture medium in the incubator may be contaminated.

The method of producing seed potatoes through in-flight microtuber cultivation without using a bioreactor has the advantage of being able to produce disease-free seed potatoes year-round when desired even in a small space. However, this method has not yet been industrialized like the method of producing seed potatoes through nutrient solution small tubers or small small tubers. This is because the size of the microtubers produced in-flight is too small or the quality is poor, so direct selling like commercialized seed potatoes is impossible. Was. In addition, even if a rather large-sized microtuber is produced, there is a problem that it is difficult to mass-produce because the yield is low.

The present invention has been devised to solve such a problem, and increases the growth of potato plants by using low-intensity red and blue LEDs (Light-Emiting Diodes) as light sources, and ultimately increases the productivity of microtuber potato seeds with large sizes. It has to do with ways to improve.

An object of the present invention is to provide a method for increasing the productivity of microtuber potato seeds having a large size.

Another object of the present invention is to provide a method capable of inducing good potato plant growth by reducing light stress.

To achieve the above task,

According to an aspect of the present invention,

(a) Based on the MS medium, the stem of a sterile potato plant in a medium having an ammonium nitrate content of 800 to 1200 mg/L and a potassium nitrate content of 3000 to 4000 mg/L and a carbon source of 2 to 4% by weight Culturing the proliferation stem by culturing for 10 to 30 days while irradiating the LED light source with a light amount of 20 to 48 umol/m ^{2 /sec.} And

(b) Cutting the proliferation stem and cutting the remaining 1 to 3 nodes on the basis of the MS medium, the ammonium nitrate content is 200 to 800 mg/L, the potassium nitrate content is 2500 to 3000 mg/L, and the carbon source is 7 to 10 It is possible to present a method for producing micro-tuber potato seeds comprising the step of culturing for 70 to 100 days under dark conditions in a medium adjusted to% by weight.

In the production method of the potato microtuber of the present invention, the LED light source uses a red and blue light source, and preferably, a light quantity of 20 to 48 umol/m ² /sec by mixing the two at 1:1, preferably It is good to maintain it at 35 to 40 umol/m ^{2 /sec.}

The red LED light source may have a wavelength of 660 to 700 nm, and the blue LED light source may have a wavelength of 450 to 480 nm.

The production method of the potato microtuber of the present invention is not largely limited to the potato varieties applied, such as underground, Daeseo, Sumi, Goun, Haeryeong, Jayeong, Earth, and Saebong, but it is preferably applied to the underground.

The method for producing micro-tuber potato seeds of the present invention may further include the step of recording the micro-tuber potato seeds that have undergone step (b). ^{The recording process includes 60 to 80 umol/m 2} /s of light on the surface of the microtuber for 7 to 10 days at intervals of 2 days, 6 hours/18 hours, 8 hours/16 hours, 10 hours/14 hours, 12 hours It can be carried out by irradiation with a total of 4 photoperiod changes with a light/dark cycle of /12 hours.

In addition, according to another aspect of the present invention, it is possible to present microtuber potato seeds produced by the above method.

According to the method for producing microtuber potato seeds of the present invention, by mixing and using a low-intensity red and blue LED light source as a light source for tissue culture of the potato stem, it improves the growth of the potato stem and increases the welfare formation rate. Large microtuber potato seeds can be produced in large quantities.

The left drawing of FIG. 1 is a photograph showing a microtuber (MCT) produced according to an embodiment of the present invention, and the right drawing is an enlarged photograph of a germinated microtuber.
The left drawing of FIG. 2 is a photograph showing the growth of the microtuber (G0) produced according to an embodiment of the present invention directly in the field, and the growth after 60 days, and the right drawing is a photograph showing the G1 harvested after 100 days of sowing. to be.

Hereinafter, the configuration and operation of the present invention will be described in more detail through preferred embodiments of the present invention. However, this has been presented as a preferred example of the present invention, and for any reason cannot be construed as limiting the present invention.

Contents not described herein can be sufficiently technically inferred by those skilled in the technical field to which the present invention pertains, and the description thereof will be omitted.

The present invention

(a) Based on the MS medium, the ammonium nitrate content is 800 to 1200 mg/L and the potassium nitrate content is 3000 to 4000 mg/L, and the carbon source is 2 to 4% by weight. The step of culturing the proliferation stem by culturing for 10 to 30 days while maintaining the LED light source at a light amount of 20 to 48 umol/m ^{2 /sec;} And

(b) Based on the MS medium, 1 to 3 nodes at the bottom left after cutting the proliferation stem have an ammonium nitrate content of 200 to 800 mg/L, a potassium nitrate content of 2500 to 3000 mg/L, and a carbon source of 7 to 10 It provides a method for mass production of microtuber potato seeds, comprising the step of culturing for 70 to 100 days under dark conditions in a medium adjusted to% by weight.

The microtuber potato seed of the present invention is based on a tissue culture method, by collecting and culturing the growth point of a potato plant to obtain a basic stem, cutting/culturing the upper part of the stem to obtain a proliferating stem, and cutting the proliferating stem It is produced by a series of processes in which the remaining nodes are cultured in the dark.

MS medium (Murashige & Skoog, 1962) was originally developed for tissue cultivation of tobacco plants, but is a plant medium widely used for cultivation of other plants. MS medium has proven effective growth in both dicotyledonous and monocotyledonous plants and is most commonly used for plant cell/tissue culture.

In the method of the present invention, the growth stem medium contains ammonium nitrate (NH ₄ NO ₃ ) 800 to 1200 mg/L, preferably 900 to 1100 mg/L _{, based on the MS medium, and potassium nitrate (KNO 3} ). It may contain 3000 to 4000 mg/L, preferably 3200 to 3800 mg/L. The carbon source may contain 2 to 4% by weight of sucrose.

In addition, the dark culture medium for inducing and producing microtubers contains 200 to 800 mg/L of ammonium nitrate, preferably 400 to 600 mg/L, and 2500 to 3000 mg/L of potassium nitrate in the MS medium composition. Was used by adjusting to 2600 to 3000 mg/L, and may contain 7 to 10% by weight of sucrose as a carbon source.

The tissue culture medium used in the present invention does not contain artificial growth regulators or hormones.

The Applicant established light conditions suitable for the characteristics of the variety by optimizing the light source and the amount of light in the step of culturing the proliferation stem. As a result, the optimal light conditions were established to remove the stress phenomenon caused by insufficient or large light, thereby improving stem growth and improving the quality of the produced microtubers.

In general, the light intensity condition for growing potato plants to produce microtubers on board is about 50 to 60 umol/m ² /sec, which is not the optimal light intensity condition for all potato varieties. Some varieties have anthocyanin accumulation, callus formation, and leaf curl, which are found in high light stress in this light range. In the growth of potato plants, if this light stress persists, it causes serious growth degradation. Ultimately, potato plants grown under light stress conditions have a problem in that the production rate of microtubers is significantly lowered, and the size of the microtubers is reduced even if the microtubers are formed.

The present invention uses LEDs (Light-Emiting Diodes) that can selectively emit light in a specific wavelength range with a small amount of electricity and heat emission as a light source, and is suitable for the growth of potato plants and further production of potato microtubers according to the conditions thereof. This is the result of examining the impact.

Specifically, in the production method of the potato microtuber of the present invention, the LED light source is a mixture of red and blue light sources at a ratio of 1 to 1 to obtain a light quantity of 20 to 48 umol/m ² /sec, preferably 35 to 40 umol. It is recommended to keep it in the range /m ^{2 /sec} Here, the light cycle may be 12 to 18 hours: 12 to 6 hours (L:D).

The red LED light source may have a wavelength of 660 to 700 nm, and the blue LED light source may have a wavelength of 450 to 480 nm.

When red and blue LEDs are mixed as a light source and used in the method of the present invention, growth of proliferating stems is improved, welfare formation is well formed during dark culture, and the production of large-sized microtubers can be increased.

In the method of the present invention, the proliferation stem culture may be cultured at 24 to 28°C for 10 to 30 days, and the cancer culture may be cultured at 18 to 22°C for 70 to 100 days to grow microtubers. The number of cultivation days may be adjusted within the above range in consideration of the growth rate, the size of the vessel, and the efficiency of the operation.

The microtuber production method of the present invention may further include the step of recording the microtuber that has undergone cancer culture. The recording process is, for example, 6 hours / 18 hours, 8 hours / 16 hours, 10 hours / light ^{of 60 to 80 umol / m 2} / s luminous intensity on the surface of the microtuber for 7 to 10 days at intervals of 2 days. It can be conducted by irradiating a total of 4 times in a 14-hour, 12-hour/12-hour (L/D) cycle.

As the surface is hardened through greening in this way, it is possible to increase the vitality of the microtuber and increase storage properties. After that, it is stored at a low temperature, germinated if necessary, and finally shipped and sown.

The method of the present invention showed excellent results in all measured items such as the fresh weight of the stem, the number of nodes, the number of leaves, shoot length, stem diameter, and leaf area compared to the case where the potato growth stem was grown under different light conditions (Table 1).

In addition, the welfare formation rate was excellent in subsequent cancer culture (Table 2), and the final produced microtubers were also produced in large quantities with a size of 8 mm or more (Table 3).

The applicant of the present invention has been studying a method for increasing the productivity of microtuber seeds for several varieties of potatoes for many years. In one aspect, efforts have been made to produce large-sized microtuber seeds, and the emergence rate of large-sized microtubers when they are directly sown on the field (ratio of sprouts appearing on the soil surface after 20 days of sowing compared to the sown seeds) This is because there was a correlation that was high, and the production quantity per week and production weight also increased.

The microtuber produced according to the method of the present invention showed normal growth and yield even when directly sown in the packaging (FIGS. 1 and 2).

According to the method of the present application, it is possible to conveniently produce a large amount of disease-free microtuber potato seeds. Since the method of the present invention produces potato seeds that can be directly stalked on a package, it is possible to shorten the time taken to produce the conventional seed potatoes for replenishment, and to help farmers by lowering the production cost accordingly.

Hereinafter, the present invention will be described in more detail through preferred embodiments of the present invention. This is for illustration of the present invention, whereby the present invention cannot be interpreted as being limited.

Example 1: Production of microtubers

Virus-free individuals were selected through virus assay after culturing the growth point of potato plants (underground). Placing 30 stems in a 20 * 30 * 15 cm culture vessel containing a culture support plate (non-woven fabric, PP 30g sm, Laoja Co., Ltd.) divided into each compartment to obtain a proliferated stem by densely culturing the basic stem obtained therefrom. I did.

The medium for culturing the proliferation stem was adjusted to the composition of nitrogen and potassium in the MS medium composition (NH ₄ NO ₃ 1,000 mg/L, KNO ₃ 3,600 mg/L), and a liquid medium containing 3% sucrose (200 mL per container) Was used. Culture conditions were cultivated for 14 days at 26°C photoperiod 16h/8h, and the amount of light was irradiated at ^{38 umol/m 2 /s using an LED light source in which a red and blue light source was mixed 1:1.}

After cutting the proliferation stem, the remaining 3 nodes of the lower part were replaced with microtuber production medium (300ml per 20 * 30 * 10 cm container). This medium is a liquid medium in which the contents of nitrogen and potassium in the composition of the MS medium were adjusted (500 mg/L of _{NH 4} NO _{3 and 2,800 mg/L of KNO 3} ) and was adjusted with 8% sucrose. This was incubated for 90 days on a fixed culture shelf in dark conditions at 20°C.

Comparative Example 1: Production of microtubers

Microtubers were produced under the same conditions as in Example 1, except that 100% of a red light source (660-700 nm, Seoul Semiconductor) was used as an LED light source in the growth stem culture.

Comparative Example 2: Production of microtubers

Microtubers were produced under the same conditions as in Example 1, except that 100% of a blue light source (450-480 nm, Seoul Semiconductor) was used as an LED light source in the growth stem culture.

Comparative Example 3: Production of microtubers

Microtubers were produced under the same conditions as in Example 1, except that a fluorescent lamp (40W) was used as a light source in the growth stem culture.

Test Example 1: Comparison of proliferation stem growth according to various light sources

The fresh weight, number of nodes, number of leaves, shoot length, stem diameter, and leaf area of potato plants obtained in the proliferation stem culture process of Examples and Comparative Examples were measured. The results are shown in the table below.

division Live weight Number of nodes Number of leaves Shoot length
(cm) Stem diameter
(mm) Leaf area
(cm ² ) Example 1.59 5.8 7.5 7.6 2.7 4.5 Comparative Example 1 1.01 3.7 5.1 7.9 1.5 3.1 Comparative Example 2 0.88 4.3 6.4 4.5 1.8 3.9 Comparative Example 3 1.37 5.4 6.9 6.8 2.3 4.2

* Average value of 3 iteration tests.

As shown in the table above, the proliferation stems cultured by mixing and irradiating red and blue light sources in a ratio of 1:1 use these alone or use other light sources in all measurement items such as fresh weight, number of nodes, number of leaves, shoot length, stem diameter, and leaf area. It can be seen that the growth is excellent compared to the case of use.

On the other hand, under red and blue mixed light, blue monochromatic light, and fluorescent light, overall leaf development was relatively normal, whereas under red monochromatic light, the plant length was long and the number of nodes was small, resulting in an overgrowth phenomenon in which the interval between nodes became longer. On the other hand, under blue monochromatic light, stem length was short and the number of nodes increased, indicating that internode elongation was suppressed. The difference in growth in the mixed light, blue monochromatic light, and red monochromatic light is thought to be due to the promotion or inhibition of stem elongation due to different kinds of interactions of cryptochrome or phytochrome (blue and red photoreceptors).

Test Example 2: Welfare formation rate evaluation

After going through the step of culturing the proliferation stem according to the light sources of Examples and Comparative Examples, the rate of formation of welfare according to time during cancer culture was examined. In tissue culture, the welfare of potatoes refers to the stems with horizontally developed side buds of the base in the culture vessel in the culture of cancer, and the number of plants with welfare is counted and expressed as a percentage was defined as the welfare formation rate. Microtubers are derived and produced from this welfare.

Cultivation period Welfare formation rate (%) Example Comparative Example 1 Comparative Example 2 Comparative Example 3 2 days 25.6 11.5 13.3 11.9 4 days 80.3 30.1 46.8 44.3 1 week 98.9 44.2 60.1 98.3 2 weeks 100 89.7 85.5 100 3 weeks - 98.5 96.2 - 4 weeks - 100 100 -

* Average value of 3 iteration tests.

As a result, when cultured by irradiating 1:1 mixed light of red and blue, the welfare formation rate reached 100% in 2 weeks. Comparative Example 3 cultured using a fluorescent lamp showed similar results, but in the early stage of cancer culture, Example had a slightly higher welfare formation rate than Comparative Example 3. Under red or blue monochromatic light, it took 4 weeks for the welfare formation rate to reach 100%. The faster the welfare formation, the faster the formation rate of microtubers, the larger the average size of the microtubers, and the larger the microtubers, the higher the germination rate after storage.

Test Example 3: Evaluation of microtuber productivity

Microtubers produced in Examples and Comparative Examples were harvested and analyzed for size.

output 8mm or more seeds Seeds less than 7.9 mm Example 1 1,208 587 Comparative Example 1 780 813 Comparative Example 2 874 799 Comparative Example 3 501 1,235

* This is the result of culture in 20 containers.

As shown in the above table, it was confirmed that in the method of Example 1 in which a red and blue LED light source was mixed and irradiated at 1:1 and cultured, a large amount of seeds having a size of 8 mm or more can be effectively produced. If the size of the microtuber is less than 7.9mm, the storage rate (the storage rate refers to the ratio of seeds that can be used as final seeds that can be normally grown before sowing after storage for 3 months or more in a low temperature storage at 4℃) is about 89%. On the other hand, in the case of more than 8mm, the storage rate increased to 97%. It can be considered that the larger the size of the microtuber, the harder the epidermis and the higher the starch content, the higher the storage rate.

Test Example 4: Cultivation experiment

The microtuber potato seeds produced in the examples were subjected to ^{a light amount of 70 umol/m 2} /s on the surface of the microtuber for 10 days at intervals of 2 days for 18 hours/6 hours, 16 hours/8 hours, 14 hours/10 hours, and After irradiation at 12/12 hour cycles and greening, it was germinated after a storage period of about 3 months in a cold storage at 4°C, and then sprouted to the packaging. Most appeared 17 to 20 days after sowing, and G1 was harvested normally after 100 days of sowing (FIGS. 1 and 2).

Although the embodiments of the present invention have been described above, the present invention is not limited to the above embodiments, but may be implemented in various different forms, and those of ordinary skill in the technical field to which the present invention pertains to the technical idea of the present invention. It will be appreciated that it can be implemented in other specific forms without changing any or essential features. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not limiting.

Claims (8)

(a) Based on the MS medium, the ammonium nitrate content is 800 to 1200 mg/L and the potassium nitrate content is 3000 to 4000 mg/L, and the carbon source is 2 to 4% by weight. Culturing the growth stem by culturing for 10 to 30 days while irradiating the LED light source with a light amount of 20 to 48 umol/m ^{2 /sec;} And
(b) Based on the MS medium, 1 to 3 nodes at the bottom left after cutting the proliferation stem have an ammonium nitrate content of 200 to 800 mg/L, a potassium nitrate content of 2500 to 3000 mg/L, and a carbon source of 7 to 10 A method for producing microtuber potato seeds comprising the step of culturing 70 to 100 days under dark conditions in a medium adjusted to% by weight.
The method of claim 1, wherein the LED light source is a 1:1 mixture of red and blue light sources.
The method of claim 2, wherein the red light source has a wavelength of 660 to 700 nm, and the blue light source has a wavelength of 450 to 480 nm.
The method of claim 1, wherein the amount of light is 35 to 40 umol/m ² /sec.
The method according to claim 1, wherein the variety of the potato is cultivated.
The method of claim 1, further comprising the step of recording the microtuber that has been subjected to step (b).
The method of claim 6, wherein the recording is performed by applying a light amount of 60 to 80 umol/m ² /s to the surface of the microtuber for 7 to 10 days at intervals of 2 days, 18 hours/6 hours, 16 hours/8 hours, 14 hours/10 Time and 12 hours / 12 hours will be carried out by irradiation, the production method of microtuber potato seeds.
Microtuber potato seeds produced by the method of claim 1.

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