KR102310362B1 - 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-sown microtuber potato seeds is presented. According to the method of the present invention, by mixing a red and blue LED light source and irradiating at 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. can produce

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 microtube potato seeds, and more particularly, to a method for producing large-sized microtube potato seeds in large quantities.

Potatoes are one of the world's four largest food crops, after rice, wheat, and maize, 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 or mainly used to make various processed products, but consumption of potatoes in Korea is on the rise.

Potatoes can be propagated by true seeds or vegetative propagation, and in general, vegetative propagation is the main method. When producing potatoes by vegetative propagation, it is important to select disease-free, high-quality seed potatoes, and seed potatoes supplied to farms must be disease-free and healthy, free from diseases such as viruses.

Seed potatoes usually produce small seed potatoes by growing tissue cultured potato seeds (sterile artificial seed potatoes) in the house or hydroponically, and after 3 years of cultivation in the house, they are cultivated twice in open air in high altitudes and supplied to farms. have. According to this system, it takes about 5-7 years to produce seed potatoes for supply to farmers from tissue-cultured potato seedlings. The reason why such a long time is required is that it takes a lot of money and time to produce the first tissue-cultured potato seed, and several proliferation processes are required because a system for mass production is not established. Therefore, as it takes a long time, there is a risk that the rate of virus infection coming from the soil or the environment will increase.

On the other hand, the mass production method of disease-free seed potatoes using a bioreactor incubator has an advantage in mass production of cultured bodies by putting a liquid medium of several hundred liters in the incubator. However, this culture method is complicated in culture technology and expensive to install, and an additional planting process is required to secure tubers of mini-tube size. In addition, if contamination occurs during the process of mass production of the culture, there is a risk of contamination of the entire culture body in the incubator.

The seed potato production method through in-flight microtube culture without using a bioreactor has the advantage of being able to produce disease-free seed potatoes year-round even in a small space at any time. However, this method has not yet been industrialized like the method for producing seed potatoes through nutrient solution small tubers or thinly shoveled tubers, because the size of the microtubers produced in-flight is too small or the quality is poor, so direct seeding like commercialized seed potatoes is impossible. It was. In addition, even if a rather large-sized microtuber is produced, there is a problem in that it is difficult to mass-produce it due to a low yield.

The present invention has been devised to solve this problem, and by using low-intensity red and blue LEDs (Light-Emiting Diodes) as light sources to increase the growth of potato plants, and ultimately to increase the productivity of large-sized microtuber potato seeds. related to ways to increase it.

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 photostress.

In order to achieve the above task,

According to one aspect of the present invention,

(a) based on MS medium, ammonium nitrate content of 800 to 1200 mg / L and potassium nitrate content of 3000 to 4000 mg / L, the carbon source is adjusted to 2 to 4% of the sterile potato plant stems in a medium 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 by dentition;} and

(b) the lower 1-3 nodes remaining after cutting the propagation 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 based on MS medium A method for producing micro-tube potato seeds comprising the step of culturing for 70 to 100 days under dark conditions in a medium adjusted to % can be presented.

In the method for producing a potato microtube of the present invention, the LED light source uses a red and a blue light source, preferably a 1:1 mixture of the two, and a light amount of 20 to 48 umol/m ² /sec, preferably It is recommended to maintain 35 to 40 umol/m ^{2 /sec.}

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

The potato microtuber production method of the present invention is not particularly limited to the potato varieties applied, such as Jijung, Daeseo, Sumi, Goun, Haryeong, Jayeong, Daeji, and Saebong, but is preferably applied underground.

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

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

According to the method for producing microtuber potato seeds of the present invention, by using a mixture of low-intensity red and blue LED light sources as light sources during tissue culture of potato stems, the growth of potato stems is improved and the welfare formation rate is increased, and ultimately A large amount of microtuber potato seeds can be produced.

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

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

Content not described herein will be omitted because it can be technically inferred sufficiently by those skilled in the art to which the present invention pertains.

the present invention

(a) based on the MS medium, ammonium nitrate content of 800 to 1200 mg / L and potassium nitrate content of 3000 to 4000 mg / L, the carbon source is adjusted to 2 to 4% by denting sterile potato stems 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) the lower 1-3 nodes remaining after cutting the propagation 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 based on MS medium 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 %.

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 proliferation stem, and cutting the proliferation stem The remaining nodes are produced by a series of cancer-culturing processes.

MS medium (Murashige & Skoog, 1962) was originally developed for tissue culture of tobacco plants, but is a plant medium that is widely used for culturing other plants. MS medium is most commonly used for plant cell/tissue culture, demonstrating effective growth in both dicot and monocot plants.

In the method of the present invention, the proliferation 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 include 2 to 4% sucrose.

In addition, the cancer 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, preferably in the MS medium composition. was used by adjusting 2600 to 3000 mg/L, and may contain 7 to 10% of sucrose as a carbon source.

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

The present applicant built light conditions suitable for the characteristics of the variety by optimizing the light source and the amount of light in the proliferative stem culturing stage. As a result, optimal light conditions were established, and the stem growth was improved by removing the stress phenomenon caused by insufficient or excessive light, and the quality of the produced microtubers was also improved.

In general, the light condition for growing potato plants to produce microtubers in-flight is about 50 ~ 60 umol/m ² /sec, but this light level is not the optimal light condition for all potato varieties. In some varieties, anthocyanin accumulation, callus formation, leaf curling, etc., which are found under high light stress in this light intensity range, are observed. If such light stress is continued in the growth of potato plants, it causes severe growth deterioration. Ultimately, potato plants grown under light stress significantly reduce the production rate of microtubers, and even if microtubers are formed, there is a problem in that the size thereof is reduced.

The present invention uses LED (Light-Emiting Diodes) as a light source, which has a small amount of electricity consumption and heat emission, and can selectively emit light in a specific wavelength band, for growing potato plants and furthermore for the production of potato microtubers according to the conditions thereof. It is the result of finding out the impact.

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

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

When a red and blue LED is mixed as a light source and used in the method of the present invention, proliferation of stem growth is improved, well-being is formed during cancer culture, and the production of large-sized microtubers can be increased.

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

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

As described above, by hardening the surface through greening, the vitality of the microtuber can be increased to improve storage properties. After that, it is stored at a low temperature, and if necessary, it is germinated and finally released and sown.

The method of the present invention showed excellent results in all measured items such as live weight, number of nodes, number of leaves, shoot length, stem diameter, and leaf area of the stem compared to the case produced under other light conditions when culturing potato proliferating stems (Table 1).

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

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

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

According to the method of the present application, disease-free microtuber potato seeds can be easily produced in large quantities. Since the method of the present invention produces potato seeds that can be directly sown in the field, it is possible to shorten the time it takes to produce conventional seed potatoes for replenishment, and it can help farmers by lowering the production cost by that much.

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 and cannot be construed as limiting the present invention.

Example 1: Production of microtubers

After culturing at the growth point of potato plants (underground), virus-free individuals were selected through a virus assay. 30 stems were placed in a 20 * 30 * 15 cm culture vessel containing a culture support plate (nonwoven fabric, PP 30g sm, Noja Co., Ltd.) divided into each compartment to obtain proliferated stems by densely culturing the obtained basic stems. did.

The medium for culturing the proliferation stem was adjusted to the nitrogen and potassium composition of 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. The culture conditions were 26 °C photoperiod of 16h/8h for 14 days, and the amount of light was irradiated at ^{38 umol/m 2 /s using an LED light source in which red and blue light sources were mixed 1:1.}

The three nodes of the lower part remaining after cutting the propagation stem were replaced with microtuber production medium (300ml per 20 * 30 * 10 cm container). This medium is a liquid medium adjusted to nitrogen and potassium content in the MS medium composition (NH ₄ NO ₃ 500 mg/L, KNO ₃ 2,800 mg/L) and adjusted to 8% sucrose. It was cultured for 90 days on a fixed culture shelf under 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 the LED light source in the proliferation stem culture.

Comparative Example 2: Production of microtubers

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

Comparative Example 3: Production of microtubers

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

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

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

division live weight number of bars number of leaves Shincho 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 of 3 replicates.

As shown in the table above, the proliferation stems cultured by mixing and irradiating red and blue light sources in a 1:1 ratio were used alone or other light sources were used in all measurement items such as live 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, overall leaf development was relatively normal under red and blue mixed light, blue monochromatic light, and fluorescent light, whereas under red monochromatic light, plant length was long and the number of nodes was small, so the internode spacing was longer. On the other hand, under blue monochromatic light, stem plant length was short and the number of nodes increased, indicating that internode elongation was suppressed. The growth difference 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 the interaction of different types of cryptochrome or phytochrome (blue and red photoreceptors).

Test Example 2: Welfare formation rate evaluation

After going through the growth stem culturing step according to the light source of Examples and Comparative Examples, the rate of formation of welfare (匐枝) according to time during cancer culture was examined. The welfare of potatoes in tissue culture means a stem with horizontally developed lateral buds in the culture vessel in dark culture, and the number of plants that have developed welfare is counted and expressed as a percentage, defined as the welfare formation rate. Microtubers are derived and produced from this welfare.

incubation 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 of 3 replicates.

As a result, in the case of irradiating 1:1 mixed light of red and blue and culturing, the welfare formation rate reached 100% in 2 weeks. Comparative Example 3, which was cultured using a fluorescent lamp, also showed similar results, but in the early stage of dark culture, Example 3 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 formation of wells, the faster the formation rate of microtubers, the larger the average size of microtubers, and the larger the size of microtubers, the higher the germination rate after storage.

Test Example 3: Microtuber productivity evaluation

Microtubers produced in Examples and Comparative Examples were harvested and the size was analyzed.

output Seeds over 8mm 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 table above, it is confirmed that in the method of Example 1, in which the red and blue LED light sources are mixed and irradiated at a 1:1 ratio, it is possible to effectively produce a large amount of seeds having a size of 8 mm or more. When the size of the microtuber is 7.9 mm or less, the storage rate (the storage rate refers to the ratio of seeds that can be used as final seeds capable of normal growth before sowing after storage for 3 months or more in a low-temperature storage at 4°C) is about 89%. On the other hand, the storage rate was increased to 97% in the case of 8mm or more. It can be considered that the larger the size of the microtuber, the harder the epidermis and the higher the storage rate because the starch content increases.

Test Example 4: Cultivation experiment

The microtuber potato seeds produced in Example were applied to the surface of the microtuber for 10 days with ^{an amount of light of 70 umol/m 2} /s at intervals of 2 days for 18 hours/6 hours, 16 hours/8 hours, 14 hours/10 hours and After irradiating and recording in a 12-hour/12-hour cycle, they were stored in a low-temperature storage at 4°C for about 3 months, then germinated and sown directly into the field. Most appeared 17-20 days after sowing, and G1 was normally harvested 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 a variety of different forms, and those of ordinary skill in the art to which the present invention pertains will appreciate the technical spirit of the present invention. However, it will be understood that the invention may be embodied in other specific forms without changing essential features. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (8)

(a) based on the MS medium, ammonium nitrate content of 800 to 1200 mg / L and potassium nitrate content of 3000 to 4000 mg / L, the carbon source is adjusted to 2 to 4% by denting sterile potato stems Culturing the proliferation stem by culturing for 10 to 30 days while irradiating at ^{a light amount of 20 to 48 umol/m 2} /sec by mixing a red and blue LED light source in a 1:1 ratio; and
(b) the lower 1-3 nodes remaining after cutting the propagation 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 based on MS medium A method for producing microtuber potato seeds, comprising the step of culturing for 70 to 100 days under dark conditions in a medium adjusted to %. The method of claim 1, wherein the red LED light source has a wavelength of 660 to 700 nm, and the blue LED light source has a wavelength of 450 to 480 nm. According to claim 1, wherein the amount of light is 35 to 40 umol / m ² /sec, Microtuber potato seed production method. The method according to claim 1, wherein the potato variety is underground. delete delete delete delete

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