KR100392514B1 - The method for plant culture using polyester petri dish - Google Patents

Abstract

The present invention relates to a plant culture method using a plant culture vessel of a polyester (PET) material, and more particularly composed of a polyester (PET) material and to provide a ventilation number by forming a ventilation hole on the top or both sides of the lid The medium is prepared by dispensing the medium in a plant culture container attached with a micropore tape as a material for providing adequate ventilation, and then the photon flux density (PPF) of 50 to 300 μmol · m ⁻² · s ⁻¹ and the ventilation frequency of 0.8 to 4.4 The present invention relates to a plant cultivation method using a plant culture vessel made of polyester (PET), characterized in that the plant is cultivated under the condition of / hour.

According to such a method, the material of the culture vessel is high light transmittance, light weight, and economical, so that the conditions suitable for plant growth can be more effectively formed.

Description

The method for plant culture using polyester petri dish}

The cell or tissue culture of a plant is a tissue in which organs, tissues, or cells of a multi-cell plant are extracted and separated from the plant, cultured in a nutrient-containing aircraft, and the callus is actively formed and the organ is not formed. Often referred to as a series of manipulations that abandon an amorphous mass formed from a wound) or a single cell population or directly differentiate a plant or organ from a callus, cell or protoplasm.

In vitro culture of plants means that the seeds, embryos, organs, tissue cells and protoplasts of the plant are grown under sterile conditions on a medium containing nutrients. There is a big difference in technology from culture.

Plant tissue culture, which began in 1950 with the cultivation of easy-to-renewable meristems in simple media, has undergone many advances and can now be reproduced in almost any plant or plant. In addition, dealing with cells and genes to improve varieties and study the basic physiology of plants is becoming more common.

Reproduction by tissue culture of such plants is carried out on a small scale in relatively narrow containers (triangular flasks, petri dishes, glass bottles), all bacteria and fungi that are reproduced in all microorganisms and other higher plants are removed, Is more suitable for physical growth as well as the nutrients and plant growth regulators needed for tissue growth, and it is possible to use a number of prototyping methods such as those previously impossible using protoplasts or cells to grow independently. It has several features.

In general, the plant tissue culture was cultured by dispensing artificial medium containing various nutrients in a glass container, or by dispensing in a container made of heat-resistant polypropylene (PP) material of light transmittance lower than glass but light and easy to handle.

However, glass containers are heavy and fragile and require careful attention to handling and are expensive. Polypropylene (PP) is light and easy to handle, but has low light transmittance and is expensive like glass containers. The use of propylene (PP) containers has increased the price of seedling production.

In addition, these containers are difficult to provide more effective ventilation times and economical photon flux density (PPF), so clarification occurs frequently in cultured plants, and the development of coliform cells in the in-flight plants and photosynthesis during in-building and in-flight growth. The development of other organs, such as chlorophyll concentration, affects the development of incomplete conditions, resulting in a very low rate of use in direct seedling production due to high mortality from outside purifying.

The growth of in-flight plants varies greatly depending on not only the light intensity in the culture chamber, but also the amount of light transmitted to the culture bottle, and the number of ventilations in the incubator. It's working.

The present invention has been made in consideration of the above matters, an object of the present invention is made of a polyester (PET) material and forming a ventilation hole on one side or both sides of the upper surface of the lid to attach the micropore tape as a material for providing the number of ventilation By developing a method of cultivating plants using culture vessels, plant culture vessels made of polyester (PET) can efficiently provide conditions suitable for in-flight plant growth and achieve desirable effects in terms of convenience and economics. It is to provide a plant culture method using.

In order to achieve the above object, the plant culture method using the plant culture vessel of the polyester (PET) material according to the present invention is composed of a polyester (PET) material and the ventilation number is formed by forming a ventilation hole on one side or both sides of the upper surface of the lid. A medium is prepared by dispensing a medium in a plant culture container that provides a suitable ventilation number by attaching micropore tape as a material for supply, and then the photon flux density (PPF) of 50 to 300 μmol · m ⁻² · s ⁻¹ and the number of ventilation of 0.8 to 4.4 The present invention relates to a plant cultivation method characterized by cultivating a plant at a condition of / hour, and thus, it is possible to provide a plant cultivation method which can more effectively form conditions suitable for plant growth because of its high light transmittance, light weight, and economical efficiency. .

Hereinafter, the present invention will be described in detail.

Plant cultivation method using a plant culture vessel of the polyester (PET) material according to the present invention is added to the medium and the inorganic and organic salts required for plant growth in the large volume dispensing vessel 40 to 50 minutes at 120 ~ 125 ℃ After sterilization and put in a clean bench (cool bench) and cooled to 60 ~ 70 ℃,

Dispensing the sterilized culture vessel, characterized in that the cooled medium is made of polyester (PET) material and forms a ventilation hole on one side or both sides of the upper surface of the lid to attach a micropore tape as a material for providing the number of ventilation times;

Inoculating the plant in the culture vessel in which the medium was dispensed, and culturing the plant under conditions of 50 to 300 μmol · m ⁻² · s ⁻¹ and a ventilation frequency of 0.8 to 4.4 / hour It features.

The polyester container used in the present invention is very weak to heat, and the shape of the container is broken when the medium is dispensed into the culture vessel as in the conventional method, and then sterilized at high temperature and high pressure. Therefore, the medium preparation method of the plant culture method using the culture vessel of the polyester (PET) material according to the present invention is preferably carried out in the same manner as the medium preparation method described above.

Hereinafter, the present invention will be described in detail with reference to the following examples.

Example 1

Luminous intensity is one of the important plant growth factors in creating a suitable culture environment in the in-flight culture. In-flight cultivation is to grow plants depending on the artificial light source using a fluorescent lamp and each fluorescent lamp is combined to provide a suitable amount of light for plant growth.

Therefore, in this experiment, in order to find out the growth of in-flight plants according to photosynthetic photon flux density (PPFD), the standard medicinal plant Rehmannia glutinosa and Korea's representative cut flowers The results of observing the growth state by cultivating Chrysanthemum are shown in Table 1 and Table 2.

As shown in Table 1 and Table 2, the higher the culture PPF, the more effective the plant growth, and the higher the dry matter and chlorophyll concentration was observed.

Of chrysanthemums and Rehmannia have showed me the PPF 100~140μ㏖ · m ^-2 · s ^-1 was good in most plant growth PPF 200μ㏖ · m ^-2 · s ^-1 over the size of the plants tend to be smaller in total construction And the chlorophyll content is increased, such that it is growing more firm and healthy.

Therefore, it was judged to show the most preferable result in the range of 50-300 micromol * m <^-2> s <^-1> PPF.

Example 2

As in the result of Example 1, in order to have the most economical and effective plant growth in in-flight culture, it should be possible to provide the maximum amount of light as possible. However, the increase in the amount of light should be considered in the most economical aspect, since the amount of light must be artificially increased by using power in in-flight culture.

Therefore, in this experiment, a 40W fluorescent lamp was installed in a culture bed having a width of 60 cm and a height of 40 cm, and the photon flux density in the culture table was measured according to the number of fluorescent lamps.

As can be seen from the results in Table 3, when using fluorescent lamps as artificial light sources in the culture chamber, the PPF of the culture table when four fluorescent lamps were installed was 115.5 μmol · m ⁻² · s ⁻¹ and 94.4 when three fluorescent lamps were installed. only μ㏖ · m ^-2 · 81.7% of the photon flux density of 115.5μ㏖ · m ^-2 · s ^-1 s ^-1 when to turn on the fluorescent lamp 4 shown to penetrate.

Example 3

It is very important to develop a container capable of transmitting the amount of light emitted from a fluorescent lamp in a culture chamber directly to plants without filtration. Therefore, in this experiment, the amount of light in the container according to the material of the culture vessel was investigated and the results are shown in Table 4.

As shown in Table 4, when the PPF in the culture chamber was 115.5 μmol · m ⁻² · s ⁻¹ , the glass container was 100.7 μmol · m ⁻² · s ^−1, and only 87.2% of the light amount was transmitted into the container. When cultivated using a container made of propylene (PP) material, it was not possible to pass as much light as one fluorescent lamp, which was judged to be economically unfavorable.

The material that transmits the most light is polycarbonate (PC) material, but its price is very expensive (185% of the glass culture vessel). It is not economical to use as a culture vessel, and it has a light transmittance above glass. This cheapest (11.14% of glass culture vessel) and light weight polyester (PET) material was considered to be the most suitable for use as in-flight culture vessels or as commercial bulk breeding culture vessels.

Example 4

Photosynthesis of plants is facilitated by adequate light and supply of CO ₂ in the presence of basic nutrients, providing adequate PPF and ventilation times in the incubation, and increasing CO ₂ concentrations in carnations, potatoes, symbidium It has been reported that photosynthesis is promoted in cultured plants such as strawberry and strawberry, and the growth of sections, shoots and plant seedlings in the incubator has been reported. (Professor Kozai 1900, 1991, Kozai, 1990, 1991, Environ. Biol. 28 (4): 147-154 (JE), Environ.Control Biol. 28 (1): 21-21. (JE), Environ.Control Biol. 28 (2): 31-39 (JE), Plant Cell, Tissue and Organ cult. 25: 107-115)

Therefore, in this experiment, using sulfuric acid and chrysanthemum as the test material, PPF was fixed at 70 μmol · m ⁻² · s ⁻¹ at a culture temperature of 25 ± 1 ° C., and CO ₂ was 1,000 μmol · m ⁻² · s ^-1. After increasing the cultivation effect according to the number of ventilation was shown in Table 5 and Table 6.

As shown in Rehmanning glutinosa of Table 5 and chrysanthemum morifolium Ramat. Plant growth also increased rapidly by providing the number of times, and the most effective in-flight cultured plant was obtained when the number of ventilations of 1.5 to 1.9 was provided. In addition, as the number of ventilation increased, the number of pores of the in-vehicle plants increased, the pore size decreased, and the development of cotyledon cells and sub-cells was good, which resulted in an increase in purifying rate during in vitro transplantation.

Therefore, it was judged that providing a ventilation number in the range of 0.8 to 4.4 / hour is preferable for in-flight plant culture.

Example 5

As can be seen from the results of Example 4, it is essential to provide adequate number of ventilation to plant growth, but a culture vessel provided with a suitable number of ventilation for plant growth has not been developed at present, and thus a conventional culture vessel made of polypropylene (PP) There is a difficulty in investing a lot of effort and time in the preparation of the culture vessel as a situation that the membrane filter (membrane filter) is attached to the culture by manual work.

Currently, the membrane filter used to provide the number of ventilations is imported in its entirety, and the price is also very expensive as shown in Table 7 below, so Milliseal uses Milliseal because it contains 375 discs at 100,000 won per box. If you provide 1.5 ~ 1.9 / hour ventilation times, it costs 266.7 won per culture bottle, and even if you use a slightly cheaper SAN filter, it costs 160 won per culture bottle. Even though the number of times is effective for plant growth, it is not possible to apply the number of ventilation economically in commercial culture at such a price.

Therefore, in this experiment, in order to select the filter that can provide the most economical number of ventilation, a hole of 10mm inside diameter (ø) is made in the lid of 500ml PET culture vessel and the filter is attached to it. The results of the measurement are shown in Table 8.

As shown in Table 8, Milliseal showed the best number of ventilations at 1.928 / hour, and the San- io filter had 1.536 / hour and Micropore tape had 1.295 / hour.

However, as shown in Table 7, it is determined that the use of the micropore tape is most preferable because the difference in the number of ventilation is not large when considering economic aspects together.

Example 6

In this experiment, essential ingredients necessary for plant growth, such as organic and inorganic substances, are added to the medium, and dissolved by adding Agar, then dispensing into a container, sterilizing at high temperature and high pressure (about 121 ℃, 20 to 40 minutes), cooling, and then curing Agar. After the completion of the conventional medium preparation method (hereinafter referred to as Method 1) consisting of plant-healing and the addition of inorganic and organic salts necessary for plant growth, 3,000 to 3,500 ml are dispensed in a large capacity container of about 7,000 ml to about 121 ° C. The present invention consists of sterilizing at 40-50 minutes in a clean bench, cooling for 20-25 minutes, cooling to 60-70 ° C., and dispensing at a Gamma-ray sterilized polyester (PET) culture vessel. Table 9 shows the results of comparing the contamination rates by preparing the medium according to the method of preparing the medium (hereinafter Method 2).

As shown in Table 9, the medium prepared by the conventional medium preparation method (Method 1) showed a contamination rate of 5.3%, whereas the medium preparation method (Method 2) according to the present invention showed a contamination rate of 3.9%. In addition to simplifying the medium preparation process, the medium can be prepared quickly, and shows a lower contamination rate than the conventional method (Method 1).

As described above, a container made of polyester (PET) was judged to be the most preferable in conditions of light transmittance, economical efficiency, weight, and the like, and a micropore tape was most preferred as a material for providing an inexpensive and effective ventilation number. Plant cultivation method using an ester (PET) plant culture vessel was found to be effective and economically used for mass propagation of in-flight plants.

According to the present invention made as described above, by using a container of high transparency material can produce a good seeding more economically, by using a micropore tape as a material providing the number of ventilation ventilation times suitable for plant growth at a lower price It can be provided, and the material of the plant culture container is light, can save labor in the process of moving the medium, tooth, shipping.

In addition, by using a plant culture vessel that attaches micropore tape to polyester (PET) material to provide ventilation times, it is possible to provide a simple and efficient plant culture method by developing a medium preparation operation that is simpler and has a lower contamination rate.

Claims (3)

In the in-flight plant culture method, A medium preparation step of adding inorganic and organic salts necessary for plant growth to the medium, dispensing in a large volume dispensing container, and then sterilizing at 120 to 125 ° C. for 40 to 50 minutes, Dispensing the prepared medium into a sterile culture vessel made of a polyester material (PET), Inoculating the plant in the culture vessel in which the medium was dispensed, and culturing the plant under conditions of 50 to 300 μmol · m ⁻² · s ⁻¹ and a ventilation frequency of 0.8 to 4.4 / hour Plant culture method using a plant culture vessel of the polyester (PET) material characterized in that. The method of claim 1, The plant culture vessel using a plant culture vessel of the polyester (PET) material, characterized in that the micropore tape is attached as a material for providing the number of ventilation by forming a ventilation hole on one side or both sides of the upper surface of the lid. The method of claim 1, The medium preparation step is a plant culture vessel of the polyester (PET) material characterized in that it further comprises the step of putting the sterilized medium in a clean bench (cool bench) and cooled to 60 ~ 70 ℃ and then used as a medium Plant culture method used.