JPH0731308A - Plant reproduction method and liquid medium therefor - Google Patents

Abstract

PURPOSE:To enhance reproduction rate and keep a regenerated plant in a state for the easy division, by performing reproduction of the regenerated plant on a supporting stand using a liquid medium. CONSTITUTION:A regenerated plant 1 derived from a plant tissue is placed on a supporting stand 2, in a sealed container 3. In the said sealed container a liquid medium 4 which is in contact with the plant, is placed and the container is fixed on a shaker to be shaken throughout the period of cultivation. The supporting stand has a net 2a and supporting legs 2b made of a stainless steel. As the liquid medium, it is favorable to use a medium prepared by diluting the MS medium by 4 fold and by adding 1weight% of sucrose and up to the concentration of 0.01mg/l of benzyladenine.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a method for growing plants and a liquid medium used for the method.

[0002]

2. Description of the Related Art Conventionally, in the case of growing a plant, a method of producing a regenerated plant body by utilizing the tissue of the plant and culturing the plant body on a solid medium and growing the same (hereinafter referred to as a stationary culture method). And a method of producing a regenerated plant body by utilizing the tissue of a plant and culturing the plant while immersing the regenerated plant body in the liquid medium while agitating the liquid medium. (Hereinafter referred to as the tank culture method).

[0003]

According to the conventional plant growth method described above, in the static culture method, the nutrients dispersed in the solid medium are not effectively supplied to the plant, so that the plant is It has a drawback that it is difficult to grow and its growth rate is low. Further, in the tank culture method, since the plant grows while being stirred in a liquid medium while being stirred, the direction in which the plant grows is not determined, and for example, the leaf part of the plant is the petiole part and root part of the plant. It grows so as to wrap around the part corresponding to the boundary (hereinafter referred to as the crown part) so as to wrap it, block the irradiation light, and consume nutrients for the growth of the roots that grew and the plant grows. It had the drawback of disappearing.

Furthermore, when it is desired to divide the proliferated plant into further strains and to further proliferate them, the plant can be easily divided into strains because the growth direction of the plant is determined by the static culture method. In the tank culture method, the growth direction of the plant body was not determined, so that the leaves and roots interfered with each other and the plants could not be easily separated.

In view of the above-mentioned drawbacks, an object of the present invention is to grow a plant with a high growth rate while keeping the grown plant in a state where it can be easily divided.

[0006]

[Means for Solving the Problems] A feature of the method for growing a plant according to the first aspect of the present invention to achieve this object is to produce a regenerated plant body using a plant tissue and to support the regenerated plant body. The plant is placed on a stand and allowed to grow while being kept in contact with a liquid medium. Further, the characteristic means of the method for growing a plant of the second invention is to cultivate a tissue at a growing point of the plant. To produce a regenerated plant body, place the regenerated plant body on a support, and keep the plant body in contact with a liquid medium to grow the plant body. The characteristic means of the method is to cut the leaf part, petiole part, and root part of the plant to obtain a part corresponding to the boundary between the petiole part and the root part, and to place the part corresponding to the boundary on a support stand, Proliferation by maintaining the state of contact with liquid medium There also characterizing feature of the liquid medium of the fourth invention, 0.01 to 0.5 benzyladenine
The amount of sucrose is 0.5 to 1%, benzyladenine is 0.01 to 0.5 mg / liter, and kinetin is added to MS medium diluted 1/10 to 1/2.
0.2 to 1 mg / liter is added, and
The characteristic composition of the liquid medium of the fifth invention is that benzyladenine is 0.05 mg / liter or less and kinetin is 0.2 mg.
/ Liter or less, and their effects are as follows.

[0007]

That is, according to the first aspect of the present invention, since the plant is placed on the support, the growth direction is constant even when the plant is cultivated and proliferated using a liquid medium. In addition to allowing the leaves and roots to be easily separated without disturbing, by using a liquid medium, by maintaining a state in which the liquid medium is contacted with a plant that is effective for growth, It has become possible to effectively supply nutrients to the plant body and to obtain a high growth rate.

According to the second aspect of the present invention, the growth point tissue of a plant is generally not infected with a virus because the tissue at the growth point of a plant is proliferated using a plant obtained by regenerating the tissue. Since it is known, it is possible to cultivate a high quality plant which is not infected with a virus.

Further, according to the third invention, a leaf portion of a plant,
By cutting the petiole part and the root part and obtaining a part corresponding to the boundary between the petiole part and the root part, the crown part containing the tissue necessary for the growth of the plant body is secured, and further, nutrients for growth are further provided. Since the leaf portion and the root portion consuming the rice are cut and removed, the nutrient is not consumed for the growth of the propagated leaf portion or the root portion, and the used nutrient effectively acts on the growth of the plant body.

[0010] Further, MS diluted to 1/10 to 1/2
0.5 to 1% of sucrose, 0.5 mg / liter or less of benzyladenine, and 1 mg of kinetin in the medium.
It was experimentally proved that the liquid medium added at 1 / liter or less can significantly improve the growth rate of strawberries. That is, according to the liquid culture medium of the fourth aspect of the present invention, the strawberry can be significantly grown as compared with the conventional method.

Further, when plant plants are grown by adding plant hormones such as benzyladenine and kinetin to a liquid medium which is generally used for a plant growing method, malformed leaves and flowers of the grown plant can be seen and flowers It is known that although it blooms a lot, a mutation that makes it impossible to produce a large fruit is likely to occur, but the present inventors have found that 0.05 mg of benzyladenine was added to the liquid medium used for the above-described plant growth method.
It was confirmed that the above mutation did not occur in the plant if the amount of kinetin contained was 0.2 mg / liter or less and kinetin was 0.2 mg / liter or less.

That is, according to the liquid culture medium of the fifth aspect of the present invention, the above-mentioned new findings have made it possible to improve the growth rate in a state where no mutant species is generated in the plant body.

[0013]

[Effects of the Invention] Therefore, since the plants can be propagated at a high growth rate while the strains can be easily divided, it becomes possible to mass-produce the plants and further to infect the virus. It has become possible to produce high-quality plants by supplying plants that have not been produced, and to provide them in large quantities.

[0014]

Embodiments of the present invention will be described below with reference to the drawings. First, a growth point at the tip of a strawberry runner is obtained according to a conventional method, and is cultured according to a conventional method to form one plant 1 (hereinafter, the plant at this stage is referred to as a seedling).
This seedling is placed in a sealed container 3 while being placed on a support base 2 composed of a stainless steel net 2a and support legs 2b for supporting the net. In addition, a medium A (described later) as a liquid medium 4 is placed in the sealed container 3 so as to come into contact with the plant body. Then, the hermetically sealed container was fixed on a shaking table and continued to be shaken during the culturing period, and the medium A was constantly stirred to continuously supply nutrients to the crown portion, and the housed plant was Culture and grow for a day. (See Figure 1)

When the strawberry seedlings were grown in this manner, the average number increased 98 times. The further grown strawberry seedlings were divided into strains, and as shown in FIG. 3, 1-2 cm from the petiole side of the crown part.
After cutting at the root and immediately below the crown, the root 1a not containing the crown 1d, the leaf 1b, and the petiole 1c are removed, and the cut strawberry seedlings containing this crown are regenerated as a plant body. It is further cultured and propagated. (See Figure 2)

In the shaking table used here, the sealed container 3 fixed on the shaking table is rotated around the vertical axis so as to draw a circle on a horizontal plane, and the medium 4 in the sealed container 3 is removed. It can be stirred. Further, the hermetically-sealed container used herein means a container that allows gas to flow in and out and blocks invasion of various bacteria.

The medium A is an MS medium (described later) diluted 4-fold (hereinafter referred to as 1/4 MS) (hereinafter 1 / n).
The term "MS" refers to an MS medium diluted n-fold) with 1% by weight of sucrose and benzyladenine (hereinafter referred to as "BA").
Abbreviated as “)” to 0.01 mg / liter. The composition of the MS medium is shown below in terms of weight per liter (unit: mg). Ammonium nitrate (NH ₄ NO ₃ ) ………… 1,650 Potassium nitrate (KNO ₃ ) ……………… 1,900 Calcium chloride dihydrate (CaCl ₂ · 2H ₂ O) 440 Magnesium sulfate 7 Hydrate (MgSO ₄ / 7H ₂ O) 370 Potassium dihydrogen phosphate (KH ₂ PO ₄ ) ………… 170 Iron sulphate heptahydrate (FeSO ₄ / 7H ₂ O) ………… 27.8 Ethylenediaminetetraacetic acid disodium (Na ₂ -EDTA) 37.3 Manganese sulfate tetrahydrate (MnSO ₄ .4H ₂ O) 22.3 Zinc sulfate heptahydrate (ZnSO ₄ 7H ₂ O) 8.6 Copper sulfate pentahydrate (CuSO ₄ .5H ₂ O) 0.025 Sodium molybdate dihydrate (Na ₂ MoO ₄ 2H ₂ O) 25 Potassium iodide (KI) ………… ......... 0.83 Boric acid (H ₃ BO ₃₎ ........................ 6.2 cobalt chloride hexahydrate _{(CoCl 2 · 6H 2 O)} ... 0.025 nicotinic acid .................. …… 0.5 thiamine hydrochloride …………………… 0.1 pyridoxine hydrochloride …………………… 0.5 m-inositol …………………… 100 glycine ………… ………… 2

The following are test examples in which the strawberry seedling 1 was grown under various conditions.

[Test Example 1]: BA concentration dependency of strawberry seedling growth number 3 strawberry seedlings 1 obtained by cultivating growing points according to a conventional method, 50 ml of culture medium A, B, C, D, E The strawberry seedlings were cultivated for 35 days under the condition that the liquid medium 4 was constantly stirred using the shaking table, and the number of grown seedlings was examined. The mediums B to E are obtained by changing the BA concentration of the medium A in various ways, as shown in Table 1. The result is shown in Table 2.

[0020]

[Table 1]

[0021]

[Table 2]

That is, it was found that when the strawberry seedlings 1 were grown in the liquid medium 4 containing BA and grown, the growth was about 7 to 16 times in 35 days although there were variations due to experimental errors.

[Test Example 2]: Dependency of the number of strawberries grown on the concentration of MS medium Instead of the liquid medium 4 used in Test Example 1, BA was not contained, but kinetin, which was a component used by adding to the normal medium, was contained. The strawberry seedlings 1 were cultured for 32 days in the same manner as in Test Example 1 using the following culture media F to I, and the growth number of the strawberry seedlings 1 was examined. The components of the liquid medium used are shown in Table 3 below, and the culture results are shown in Table 4.

[0024]

[Table 3]

[0025]

[Table 4]

As a result, when the liquid medium 4 containing kinetin added to 1/4 MS had a lower growth rate as compared with the case of using BA, the growth rate was comparable to that using BA. The growth rate was shown. That is, 1 / 2MS
A high growth rate can be obtained by using the liquid medium 4 in which kinetin is added to ˜1 / 10 MS. Kinetin is
It has been empirically confirmed that, when used in the range of 0.2 to 1 mg / liter, the same or higher effect is achieved as compared with a medium containing benzyladenine at 0.5 mg / liter.

[Test Example 3]: Dependence of strawberry growth number on sucrose concentration In place of the liquid medium 4 used in Test Example 1, the sucrose concentration of the liquid medium 4 containing kinetin without BA was changed variously. 4 using the culture media J to M prepared in the same manner as in Test Example 1.
Cultivation of strawberry seedling 1 was carried out for 2 days, and the growth number of strawberry seedling 1 was examined.
The components of the liquid medium 4 used are shown below in Table 5, and the culture results are shown in Table 6.

[0028]

[Table 5]

[0029]

[Table 6]

As a result, when the liquid medium 4 in which kinetin was added to 1/4 MS was used, the amount of sucrose was 0.5% to 0.5%.
It was found that if it was set to 1%, the proliferation rate was comparable to that when BA was used. In other words, the sucrose concentration is 0.5% ~
If it is 1%, a high proliferation rate can be obtained.

[Comparative Example 1]: Conventional culture test example [0031] The liquid medium 4 used in Test Example 1 was replaced with the conventionally used agar medium N under the same conditions as in Test Example 1 for 62 days. The static growth rate of Strawberry seedlings 1 was examined by performing static culture.

The agar medium N is prepared by adding 3% by weight of sucrose, 0.5 mg / liter of BA, 1.0 mg / liter of kinetin and 0.8% by weight of agar to LS medium (described later). It is a certain medium. Below is the composition of the LS medium
It is shown by weight per liter (unit: mg). Potassium nitrate (KNO ₃₎ ........................ 2,830 ammonium sulfate _{((NH 4) 2 SO 4} ) ............ 463 Calcium chloride dihydrate _{(CaCl 2 · 2H 2 O)} 440 Magnesium sulphate Nanamizu hydrate _{(MgSO 4 · 7H 2 O)} 370 potassium dihydrogen phosphate (KH ₂ PO ₄₎ ............... 170 iron sulfate heptahydrate _{(FeSO 4 · 7H 2 O)} ............ 27.8 ethylenediamine Disodium tetraacetate (Na ₂ -EDTA) 37.3 Manganese sulfate tetrahydrate (MnSO ₄ .4H ₂ O) 22.3 Zinc sulfate heptahydrate (ZnSO ₄ 7H) ₂ O) ... 8.6 Copper sulfate pentahydrate (CuSO ₄ .5H ₂ O) ... 0.025 Sodium molybdate dihydrate (Na ₂ MoO ₄ .2H ₂ O) 0.25 Potassium iodide (KI) ……………… ...... 0.83 Boric acid (H ₃ BO ₃₎ ........................ 6.2 cobalt chloride hexahydrate _{(CoCl 2 · 6H 2 O)} ... 0.025 Thiamine hydrochloride .................. …… 0.4 m-inositol …………………… 100

[Test Example 4]: Examination of Strawberry Seedling Growth Rate Strawberry seedlings were cultured for 59 days in the same manner as in Test Example 1 using the medium O instead of the liquid medium 4 used in Test Example 1 The growth rate of 1 was investigated. The medium O is a liquid medium 4 in which 1 mg of kinetin and 0.2 mg / liter of sucrose are added to 1/4 MS.

The results of Comparative Example 1 and Test Example 4 are shown below in Table 7.
Shown in.

[0035]

[Table 7]

As a result, it can be seen that when the strawberry seedlings are cultured and grown using the liquid medium 4 of the present invention, the growth rate is remarkably improved as compared with the case where the conventional agar medium is used.

[Test Example 5]: Examination of the effect on the strawberry seedling growth rate by cutting and removing the root part, petiole part and using a support base. Instead of the liquid medium 4 used in Test Example 1, the medium P was used and further growth was performed. 1 to 2 on the petiole side of the crown 1d
cm, and the root part immediately below the crown part, respectively, the part containing the crown part 1d is taken as the strawberry seedling 1, and the strawberry seedling 1 is placed on the support base 2 and the sealed container 3 is placed.
The cells were placed in the cells and cultured for 64 days in the same manner as in Test Example 1 to examine the proliferation rate. The medium P is a medium prepared by adding 0.2 mg / liter of kinetin and 1% by weight of sucrose to 1/4 MS.

Table 8 shows the results of Test Example 5 and the test results obtained by growing the same liquid medium 4 in Test Examples 2 and 3 for comparison.

[0039]

[Table 8]

As a result, it can be seen that, when the strawberry seedlings are cultured and propagated by using the plant growth method of the third invention, the growth rate is remarkably improved as compared with the case where the conventional growth method is used.

[Test Example 6]: Examination of a medium containing both BA and kinetin, and examination of the effect on the growth rate of strawberry seedlings by the use of a support stand Instead of the liquid medium 4 used in Test Example 1, the medium Q to After the strawberry seedling 1 grown in Test Example 1 was further divided into strains using U, the crown part 1d is included among those cut from the crown part 1d at the petiole part of 1-2 cm and the root part immediately below the crown part. The portion was designated as strawberry seedling 1, and the strawberry seedling 1 was cultured for 62 days in the same manner as in Test Example 1 to examine the growth rate. Moreover, after replacing the liquid medium 4 used in Test Example 1 with the mediums V and W, and further dividing the strawberry seedlings 1 grown in Test Example 1 into strains,
Location of petiole of 1-2 cm from crown 1d, and
Among the pieces cut with the root portion directly below the crown portion 1d, the portion containing the crown portion 1d is called strawberry seedling 1, and this strawberry seedling 1 is placed in a sealed container in a state of being placed on a support 2 and in a sealed container. Each of the culture mediums V and W is housed in contact with the plant, and the hermetically sealed container is fixed on a shaking table and shaken continuously during the culturing period.
The strawberry seedlings 1 were cultivated while constantly stirring (see FIG. 1).
The mediums Q to W have the following compositions.

[0042]

[Table 9]

As a result, the result is shown in Table 10.

[0044]

[Table 10]

From Table 10, it can be seen that in any case, a higher proliferation rate than the conventional method was obtained. Further, in the example using the shaking table, it was found that the growth rate was 98 times higher in the medium V, which was significantly higher than that in the medium Q, and also in the medium W, as compared with the case of Test Example 5. It was found that a high growth rate was obtained. That is,
It can be seen that the growth rate is further improved under the condition that the liquid medium 4 is intermittently in contact with the crown portion using the shaking table.

It should be noted that reference numerals are given in the claims for convenience of comparison with the drawings, but the present invention is not limited to the configurations of the accompanying drawings by the entry.

[Brief description of drawings]

FIG. 1 is a schematic view showing a method for growing a plant of the present invention.

FIG. 2 is a process diagram showing a method for growing a plant of the present invention.

[Figure 3] Overall view of the plant

[Explanation of symbols]

 1 plant 2 support 4 liquid medium

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Fumiaki Oda 1-1-1, Hama, Amagasaki City, Hyogo Prefecture Kubota Technology Development Laboratory Co., Ltd.

Claims (5)

[Claims] 1. A regenerated plant body (1) is produced by using plant tissue, the regenerated plant body (1) is placed on a support (2), and the regenerated plant body (1) is placed in a liquid medium ( 4) A method for growing a plant, which grows while maintaining contact with 2. A plant (1) regenerated by culturing a tissue of a plant growing point, the regenerated plant (1) is placed on a support (2), and the plant (1) is A method for growing a plant, which grows while maintaining contact with a liquid medium (4). 3. A leaf part, a petiole part, and a plant (1),
The root part is cut to obtain a part corresponding to the boundary between the petiole part and the root part, and the part corresponding to the boundary is placed on the support base (2),
A method for growing a plant, wherein the plant (1) is maintained in contact with a liquid medium (4) and grown. 4. A liquid medium used in the method for growing a plant according to any one of claims 1 to 3, wherein MS is diluted 1/10 to 1/2.
Liquid medium obtained by adding sucrose 0.5 to 1% benzyladenine 0.5 mg / liter or less kinetin ............ 1 mg / liter or less to the medium. 5. A liquid medium used in the method for growing a plant according to claim 1, wherein benzyladenine is 0.05
A liquid medium containing not more than mg / liter and not more than 0.2 mg / liter of kinetin.