KR20130119186A - Plant tissues culture tabletting medium and manufacturing method thereof - Google Patents

Abstract

PURPOSE: A production method of a refined medium for the plant tissue culturing is provided to control the generation of dust, and to remove a measuring process before using. CONSTITUTION: An MS medium composition in a powder form is mixed with a diluting agent. The MS medium composition and the diluting agent are stirred to obtain medium paste. The medium paste is molded into tablets. 100 parts by weight of MS medium composition contains 105-148 parts by weight of sucrose. 100 parts by weight of MS medium composition additionally contains 0.097-0.114 parts by weight of 2,4-dichlorophenoxyacetic acid, 0.076-0.093 parts by weight of 1-naphthaleneacetic acid, and 0.076-0.093 parts by weight of 6-benzyladenine.

Description

Purifying medium for plant tissue culture and its manufacturing method {Plant tissues culture tabletting medium and manufacturing method

The present invention relates to a purified medium for plant tissue culture and a method of manufacturing the same, and more particularly, to a purified medium containing nutrients required for in-flight cultivation of plants as a main component. It is about.

Plant tissue culture refers to the propagation of plant tissues, organs, cells, etc. by giving nutrients, plant hormones, etc. necessary in glass containers or culture vessels. Plant tissue culture technology has been used as an effective technique in various fields such as mass production of individuals, molecular breeding, and plant raw material production through mass culture. In order to cultivate plant tissue, nutrients indispensable for survival and development require large elements and trace elements. All of them should be supplied to the medium as inorganic or organic compounds except those obtained in the gas phase. The medium that supplies the nutrients needed to raise plants is called a medium. Synthetic medium is called when only inorganic salts or a structure with a certain carbon source and nitrogen source are added. Liquid medium is suitable for mass cultivation, and solid medium with agar, gelatin, etc. is used for preservation or separation of liquor.

Recently, the importance of plant tissue culture technology has been recognized as one of the indispensable technologies in the use of biotechnology in plants beyond the initial simple mass growth or breeding technology. Botany, plant physiology, biochemistry, genetics, etc. Although it is widely used and researched in basic research, applied fields such as agriculture, breeding, etc., the conventional plant tissue culture medium is made in powder form, so there is a problem that dust occurs when used and flows into the respiratory system. There is an inconvenience to use. In particular, when culturing in large quantities, there was a problem of quantifying a large amount of medium each time.

In addition, using plant tissue culture technology, saponins, like the bellflower and ginseng, contain a lot of medicinal effects, and can be expected to develop low-calorie foods. In mass production, the conventional medium used for incubation in Deodeok had a low re-differentiation rate and a low rooting rate, making it difficult to efficiently infuse in-flight culture. Also, a large amount of medium must be quantified every time for mass culture of Deodeok. Hassle was a problem.

 The present invention has been made to solve the above problems, the first problem to be solved of the present invention provides a purified plant tissue culture medium that provides the convenience of suppressing dust generation, and does not have to be quantified every use. It is.

 The second problem to be solved of the present invention is to improve the regeneration rate and rooting rate of deodeok and to provide a tablet medium for deodorizing large-scale cultivation that can facilitate the ease of quantification.

 The third problem to be solved of the present invention is to provide a method for producing a tablet medium for deuldeul mass culture.

 In order to solve the above problems, the present invention,

In the medium for culturing plant tissue, it provides a tablet medium (tablet) for plant tissue culture, characterized in that the tableting (tabletting), including the MS medium composition.

According to a preferred embodiment of the present invention, the plant may be a tablet (tablet, 錠 劑) medium for plant tissue culture, characterized in that the deodeok.

According to another preferred embodiment of the present invention, the sucrose 105 to 148 parts by weight based on 100 parts by weight of the MS medium composition may be further included.

According to another preferred embodiment of the present invention, 2,4D (2,4-Dichlorophenoxyacetic acid) may further comprise 0.097 to 0.114 parts by weight based on 100 parts by weight of the MS medium composition.

According to another embodiment of the present invention, the NA medium (1-naphthaleneacetic acid) 0.076 to 0.093 parts by weight and BA (6-benzyladenine) 0.076 to 0.093 parts by weight based on 100 parts by weight of the MS medium composition may be further included.

In addition, the present invention comprises the steps of (1) adding an excipient to the MS medium composition in powder form; (2) stirring the MS medium composition and the excipient to form a medium dough; (3) provides a method for producing a plant tissue culture tablet (tablet, 錠 劑) medium comprising the step of purifying the medium dough (tablet, 錠 劑).

According to a preferred embodiment of the present invention, the sucrose 105 to 148 parts by weight based on 100 parts by weight of the MS medium composition of step (1) may be further included.

According to another preferred embodiment of the present invention, 2,4D (2,4-Dichlorophenoxyacetic acid) 0.097 to 0.114 parts by weight based on 100 parts by weight of the MS medium composition may be further included.

According to another preferred embodiment of the present invention, it may further comprise 0.076 to 0.093 parts by weight of NAA (1-naphthaleneacetic acid) and 0.076 to 0.093 parts by weight of BA (6-benzyladenine) based on 100 parts by weight of the MS medium composition. .

According to another preferred embodiment of the present invention, the excipient of step (1) is water,

It may include 4.22 to 5.06 parts by weight based on 100 parts by weight of the MS medium composition.

In addition, the present invention is a culture medium for culturing deodeok, plant tissue culture, characterized in that it comprises 38,000 to 42,500 parts by weight of water based on 100 parts by weight of the tablet (tablet) medium prepared according to the present invention described above Provide a culture solution.

 Purifying medium for plant tissue culture of the present invention and its manufacturing method can prevent the occurrence of fine dust through the purification of the medium, and reduce the hassle that must be quantified every time to facilitate the convenience in various aspects of the experiment applied to plant tissue culture technology can do.

In particular, the purified medium for plant tissue culture of the present invention and a method of manufacturing the same for reducing the inconvenience of having to quantitate a large amount of medium every time for mass cultivation of Deodeok used in a variety of medicinal and edible, more convenient mass culture At the same time, it improves the re-differentiation rate of Deodeok by about 30 to 40%, and also increases the rooting rate significantly, enabling mass cultivation of Deodeok in a short time with less labor in a limited space.

Hereinafter, the present invention will be described in more detail.

As described above, the conventional powder-type medium used for cultivating plant tissues had a problem in that dust was generated and introduced into the respiratory tract. There was a problem to quantify the medium every time.

In the present invention, in the medium for culturing plant tissue, the tablet medium (tablet, 錠 劑) for plant tissue culture, characterized in that the tableting (tabletting, 錠 劑 化) comprising the MS medium composition described above I tried to solve the problem.

 The MS (Murashige and Skoog) medium may use a conventional MS medium containing the elements necessary for plant growth, NO compared to other medium₃-N, NH₄The content of -N and K is very high. The tableting method is not particularly limited, but preferably water is used as an excipient, and 4.22 to 5.06 parts of water is stirred with respect to 100 parts by weight of the MS medium composition to form a medium dough, and then using a tablet machine. It can be tableted.

In the plant tissue culture tablet medium, the plant tissue may be a tablet (tablet, 錠 劑) medium for plant tissue culture, characterized in that the deodeok. In order to improve the re-differentiation rate and rooting rate of the deodeok it is preferable to further include sucrose 105 to 148 parts by weight based on 100 parts by weight of the MS medium composition, more preferably contains 125 to 128 parts by weight sucrose (sucrose) can do. If less than 105 parts by weight of the carbohydrate necessary for the culture of the deodeok is not supplied enough, the re-differentiation rate and rooting rate is lowered. If it exceeds 148 parts by weight, the carbohydrate is excessively supplied, the re-differentiation rate and rooting rate may fall. (See Examples 9, 10 and 11)

In addition, 2,4D (2,4-Dichlorophenoxyacetic acid) may further include 0.097 to 0.114 parts by weight based on 100 parts by weight of the MS medium composition, and more preferably 0.102 to 0.109 parts by weight. 2,4D plays a role in controlling or inhibiting plant physiological processes in a small amount, and may be most effective at a certain concentration, depending on species and form. When 2,4D is further included in 0.097 to 0.114 parts by weight based on 100 parts by weight of the MS medium, the re-differentiation rate and rooting rate may be further improved than when only sucrose is included. (See Examples 6, 7, and 8)

NAA (0.076 to 0.093 parts by weight and BA (6-benzyladenine) 0.076 to 0.093 parts by weight based on 100 parts by weight of the MS medium composition may be further included, more preferably 0.083 to 0.086 parts by weight NAA and 0.083 to 0.086 parts by weight BA) It may include parts by weight. If the NAA or BA is less than 0.076 parts by weight or more than 0.093 parts by weight, the regeneration rate and rooting rate may drop. If the NAA or BA is not mixed and contains only NAA or BA only, the regeneration rate and rooting of Deodeok The effect of rate improvement may be even more noticeable. (See Examples 2 to 5)

In addition, the present invention provides a method for producing a tablet medium for plant tissue culture, comprising the steps of: (1) adding an excipient to the MS medium composition in powder form; (2) stirring the MS medium composition and the excipient to form a medium dough; (3) provides a method for producing a plant tissue culture tablet (tablet, 錠 劑) medium comprising the step of purifying the medium dough (tablet, 錠 劑).

Sucrose 105 to 148 parts by weight based on 100 parts by weight of the MS medium composition of step (1) is preferably further included, more preferably may comprise 125 to 128 parts by weight of sucrose (sucrose). If less than 105 parts by weight of the carbohydrate necessary for the culture of the deodeok is not supplied enough, the re-differentiation rate is lowered, if it exceeds 148 parts by weight of the carbohydrate is excessively supplied, the re-differentiation rate and rooting rate may fall.

In addition, 2,4D (2,4-Dichlorophenoxyacetic acid) may further include 0.097 to 0.114 parts by weight based on 100 parts by weight of the MS medium composition, and more preferably 0.102 to 0.109 parts by weight. When the MS medium composition contains 0.097 parts by weight to 0.114 parts by weight of 2,4D, the redistribution rate and rooting rate may be further improved than when only sucrose is included.

NAA (0.076 to 0.093 parts by weight and BA (6-benzyladenine) 0.076 to 0.093 parts by weight based on 100 parts by weight of the MS medium composition may be further included, more preferably 0.083 to 0.086 parts by weight NAA and 0.083 to 0.086 parts by weight BA) It may include parts by weight. If NAA or BA is less than 0.076 parts by weight or more than 0.093 parts by weight, the root regeneration rate may be reduced due to a decrease in rooting rate. If NAA or BA are not mixed, only NAA or BA may be used. The effect of redifferentiation rate and rooting rate improvement may be significantly less.

Excipients included to purify the medium of step (1) are preferably water, and when using other excipients may affect the composition of the medium may not exhibit the effect of expected regeneration rate and rooting rate when a specific composition. The water may comprise 4.22 to 5.06 parts by weight based on 100 parts by weight of the MS medium composition. If the water is less than 4.22 parts by weight, tabletable medium dough is not formed. When the water content exceeds 5.06 parts by weight, the medium becomes liquid without forming the dough. (See Example 12)

In addition, the present invention is a medium for culturing deodeok, water based on 100 parts by weight of a tablet medium for plant tissue culture (tablet) medium prepared according to the present invention described above

It provides a culture medium for deodeok mass culture comprising 38,000 parts by weight to 42,500 parts by weight. In culturing the plant in-flight, the concentration of the medium composition greatly affects the plant culture, so when the water is included in an amount exceeding 38,000 parts by weight to 42,500 parts by weight, the re-differentiation rate and improvement of rooting rate may not occur effectively.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, and it is to be understood by those skilled in the art that the present invention is not limited thereto It will be obvious.

≪ Example 1 >

To 2368.22 mg of 1/2 MS medium in powder form, 3 g of sucrose and 2.5 mg of 2,4D were mixed, and 0.1 mL of water was added to a medium composition of NAA 2.0 mg and BA 2.0 mg. After stirring to make a constant mixing of water and the medium composition using the to make a medium dough, using a tablet machine of 2.4g deodeok culture medium for the culture medium.

 The composition of the used MS medium is shown in Table 1.

Medium composition Concentration (mg / L) Constituent MS medium 1/2 MS medium KNO ₃ 1900 950 NH ₄ NO ₃ 1650 825 MgSO ₄ .7H ₂ O 370 187 CaCl ₂ .2H ₂ O 440 220 KH ₂ PO ₄ 170 85 MnSO ₄ .4H ₂ O 22.3 11.2 Kl 0.83 0.42 H ₃ BO ₃ 6.2 3.1 ZnSO ₄ .7H ₂ O 8.6 4.3 CuSO ₄ .5H ₂ O 0.025 0.013 Na ₂ MoO ₄ .2H ₂ O 0.25 0.13 CoCl ₂ .6H ₂ O 0.025 0.013 FeSO ₄ .7H ₂ O 27.8 13.9 Na ₂ EDTA 37.3 18.7 Nicotinic acid 0.5 0.25 Pyridoxine-HCl 0.5 0.25 Thiamine-HCl 0.1 0.05 myo-lnositol 100 50 Glycine 2.0 1.0 Gross weight 4736.43 2368.33

After washing the deodeok washed with distilled water in 70% ethanol for 5 minutes, the deodeok washed three times with distilled water was cut out to 0.5mm. 1 liter of purified medium (2.4 g) was added to 1 liter of distilled water, followed by stirring using a stirrer for 5 minutes, followed by autoclave, and cultured for 4 weeks.

≪ Example 2 >

The deodeok was cultured in the same manner as in Example 1 except that BA was not included.

≪ Example 3 >

Except for not containing the NAA was carried out in the same manner as in Example 1 was cultured deodeok.

<Example 4>

NAA and BA were prepared in the same manner as in Example 1 except for including 1.5 mg.

&Lt; Example 5 >

NAA and BA were prepared in the same manner as in Example 1 except for including 2.5 mg.

&Lt; Example 6 >

It was prepared in the same manner as in Example 1 except that 2,4D was not included.

&Lt; Example 7 >

2,4D was prepared in the same manner as in Example 1 except for including 2 mg.

Example 8

2,4D was prepared in the same manner as in Example 1 except for including 3 mg.

Example 9

It was prepared in the same manner as in Example 1 except for including 2 g of sucrose and not including 2,4D, NAA, and BA.

Example 10

It was prepared in the same manner as in Example 1 except that 4 g of sucrose was included and 2,4D, NAA and BA were not included.

Example 11

It was prepared in the same manner as in Example 1 except that sucrose, 2,4D, NAA, and BA were not included.

Example 12

It was prepared in the same manner as in Example 1 except that the tablet medium was prepared by using starch instead of water as an excipient.

&Lt; Comparative Example 1 &

It was prepared in the same manner as in Example 1 except that the quantitative use of the powder form medium, not the purified medium.

<Experimental Example>

Table 2 shows the re-differentiation rate and rooting rate when 30 deodeok sections were cultured according to Examples 1 to 9 and Comparative Examples 1 to 4.

Sucrose
(g) 2,4D
(mg) NAA
(mg) BA
(mg) Deodeok subdivision () Root Count () Tablet badge Original badge
(powder) Refined medium Existing medium
(powder) water Starch water Starch 3 2.5 2 2 14.0 ± 1.05 10.2 ± 1.15 15.3 ± 2.1 27.4 ± 1.15 17 ± 2.03 28.3 ± 3.2 2 0 2.1 ± 0.53 1.3 ± 0.58 2.0 ± 1.3 20.7 ± 1.73 15 ± 2.08 22.3 ± 3.0 0 2 4.5 ± 0.3 6.7 ± 0.8 4.2 ± 1.9 20.1 ± 0.9 23.1 ± 1.3 21.2 ± 2.7 1.5 1.5 8.7 ± 0.73 5.3 ± 0.65 9.8 ± 1.4 19.2 ± 1.53 8.7 ± 1.53 24.3 ± 2.0 2.5 2.5 9.0 ± 0.53 13.2 ± 0.54 10.3 ± 1.2 23.3 ± 1.15 17.6 ± 1.53 25.5 ± 1.7 3 2 2 12.5 ± 0.58 11.7 ± 0.58 12.7 ± 1.6 22.4 ± 1.53 11.5 ± 1.08 24.3 ± 1.9 2 2 2 10.5 ± 1.02 4.7 ± 1.03 11.3 ± 1.2 25.7 ± 1.28 18.8 ± 1.34 26.4 ± 2.0 0 2 2 6.9 ± 0.4 6.2 ± 1.0 18.3 ± 1.1 20.3 ± 2.4 0 0 0 1.0 ± 0.02 1.1 ± 0.4 4.8 ± 1.2 5.1 ± 2.1 2 0 0 0 0.3 ± 0.08 0.6 ± 0.1 7.5 ± 0.8 7.3 ± 1.5 2.5 2 2 11.4 ± 0.6 12.3 ± 1.6 7.4 ± 1.3 9.3 ± 1.5 4 0 0 0 0.5 ± 0.1 0.5 ± 0.1 6.8 ± 0.8 6.6 ± 1.2 2.5 2 2 10.4 ± 0.71 12.7 ± 1.2 25.0 ± 1.15 26.0 ± 1.5 0 0 0 0 0 0 0 0 2.5 2 2 0 0 0 0

As can be seen in Table 2, when using a purified medium and quantitatively used in the form of a conventional powder form, the re-differentiation rate and rooting rate of the deodeok showed almost the same value. This is a result that can be confirmed that did not affect the composition of the medium during the purification of the medium. However, unlike refined media using water as an excipient, refined media using starch showed a different tendency for regrowth and rooting rate of Deodeok. Therefore, when using non-water excipients, it may affect the composition of the medium during the tableting process. I can see. In addition, when the culture medium is formed using the purified medium, the deviation value is generally small, and there is no fear of fine dust, so that the respiratory inflow is not introduced, and the trouble of having to quantify a large amount of medium for mass culture every time can be reduced. In addition, it can be seen that an efficient medium composition for cultivating a specific plant prevents an error from occurring in the quantification process so that the plant can be cultured more efficiently.

In the purification medium composition effective for the re-differentiation rate of deodeok, the re-differentiation rate and rooting rate of deodeok were much higher when 2368.22 mg of MS medium was composed of 3 g of sucrose, 2.5 mg of 2,4D, and 2.0 mg of NAA and BA. . When the NAA and BA are included in the range of 0.076 to 0.093 with respect to 100 parts by weight of the MS medium composition, the re-differentiation rate and rooting rate decrease, and it can be seen that NAA and BA have a great influence on the re-differentiation rate even with a small amount. In addition, when only one of the NAA and BA was used without mixing, the regeneration rate and rooting rate were significantly reduced than when the NAA and BA were mixed with less than 0.076 or more than 0.093. It can be seen that the use has a great effect in improving the regeneration rate and rooting rate of Deodeok.

When 2,4D is included in the range of 0.097 to 0.114 parts by weight with respect to 100 parts by weight of the MS medium composition, the re-differentiation rate and rooting rate of Deodeok are also shown to be low. You can see the crazy.

Sucrose showed better regeneration rate of deodeok when 3g was included than when 2g or 4g was included. However, it was found that sucrose had less effect on the re-differentiation rate of deodeok than 2,4D or NAA and BA. Can be. However, when sucrose is not included, even if other growth regulators are included, it can be seen that rooting and re-differentiation of deodeok do not occur.

Claims (11)

In the medium for culturing plant tissue,
Tablet medium for plant tissue culture, characterized in that the tableting (tabletting, 錠 劑 化) containing MS medium composition. The method of claim 1,
The plant is characterized in that the deodeok plant tissue culture tablets (tablet, 錠 劑) medium. 3. The method of claim 2,
Sucrose 105 to 148 parts by weight based on 100 parts by weight of the MS medium composition, characterized in that it further comprises a tablet medium (tablet, 錠 劑) medium for plant tissue culture. The method of claim 3,
2,4D (2,4-Dichlorophenoxyacetic acid) 0.097 to 0.114 parts by weight based on 100 parts by weight of the MS medium composition for plant tissue culture tablets (tablet, 錠 劑) medium. 5. The method of claim 4,
Tablets for plant tissue culture (tablet, 錠 劑) characterized in that it further comprises 0.076 to 0.093 parts by weight of NAA (1-naphthaleneacetic acid) and 0.076 to 0.093 parts by weight of BA (6-benzyladenine) based on 100 parts by weight of the MS medium composition ) Badge. (1) adding an excipient to the MS medium composition in powder form;
(2) stirring the MS medium composition and the excipient to form a medium dough; And
(3) The method for producing a plant tissue culture tablet (tablet, 錠 劑) medium comprising the step of purifying the medium dough (tablet, 錠 劑). The method according to claim 6,
Method for producing a plant tissue culture tablet (tablet, 錠 劑) medium, characterized in that it further comprises sucrose 105 to 148 parts by weight based on 100 parts by weight of the MS medium composition of step (1). The method of claim 7, wherein
Method for producing a plant tissue culture tablet (tablet, 錠 劑) medium characterized in that it further comprises 2,4D 0.097 to 0.114 parts by weight based on 100 parts by weight of the MS medium composition. 9. The method of claim 8,
NAA 0.076 to 0.093 parts by weight and BA 0.076 to 0.093 parts by weight based on 100 parts by weight of the MS medium composition method for producing a plant tissue culture tablet (tablet, 錠 劑) medium characterized in that it further comprises. The method according to claim 6,
The excipient of step (1) is water,
Method for producing a plant tissue culture tablet (tablet, 錠 劑) medium comprising 4.22 to 5.06 parts by weight based on 100 parts by weight of the MS medium composition. In the medium for culturing deodeok,
A culture medium for plant tissue culture, comprising 38,000 parts by weight to 42,500 parts by weight of water based on 100 parts by weight of the deodeok cultured tablet medium of any one of claims 1 to 4.