JP2942101B2 - Method for growing plant and liquid medium used for the method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for growing a plant and a liquid medium used for the method.

[0002]

2. Description of the Related Art Conventionally, when a plant is grown, a method of producing a regenerated plant using plant tissue and culturing the plant on a solid medium and growing the plant (hereinafter referred to as a static culture method). A method of producing a regenerated plant using plant tissue and culturing and growing the plant while immersing the regenerated plant in the liquid medium while stirring the liquid medium. (Hereinafter referred to as a tank culture method).

[0003]

According to the above-mentioned conventional method for growing a plant, in a stationary culture method, nutrients dispersed in a solid medium are not effectively supplied to the plant, so that the plant is There is a drawback that it is difficult to grow and its growth rate is low. Further, in the tank culture method, the plant grows while being stirred while being immersed in the liquid medium, so that the growth direction of the plant is not determined, for example, the leaf of the plant is the petiole and the root of the plant. It grows to cover the area corresponding to the boundary (hereinafter referred to as the crown), blocks light for irradiation, consumes nutrients for the growth of the grown roots, etc., and grows, but the plant grows. There was a disadvantage that it disappeared.

[0004] Further, when it is desired to divide the grown plant and to further grow the plant, the stationary culturing method allows the plant to be strained easily because the growth direction of the plant is determined. In the tank culture method, since the growth direction of the plant body is not determined, the leaves and roots are in the way, and the plants cannot be easily divided.

[0005] In view of the above-mentioned drawbacks, an object of the present invention is to grow a plant at a high growth rate while maintaining the grown plant in a state where it is easy to divide the strain.

[0006]

Means for Solving the Problems To achieve this object, the first aspect of the method for growing a plant according to the present invention is to produce a regenerated plant using plant tissue and to support the regenerated plant. The method of growing a plant according to the second aspect of the present invention is to cultivate a plant growth point tissue by placing the plant on a table and maintaining the plant in contact with a liquid medium while maintaining the state. To produce a regenerated plant by placing the regenerated plant on a support, and growing the plant while maintaining the plant in contact with a liquid medium. The method is characterized in that the leaves, petiole, and root of the plant are cut to obtain a portion corresponding to the border between the petiole and the root, and the portion corresponding to the boundary is placed on a support, and the plant is cut. Growth while maintaining contact with liquid medium There also characterizing feature of the liquid medium of the fourth invention, 0.01 to 0.5 benzyladenine
Per liter, the sucrose 0.5-1%, benzyladenine 0.01-0.5 mg / liter, kinetin,
0.2 to 1 mg / liter.
The liquid medium according to the fifth aspect is characterized in that benzyladenine is 0.05 mg / liter or less and kinetin is 0.2 mg / liter.
Per liter or less, and their effects are as follows.

[0007]

In other words, according to the first aspect, since the plant is on the support, even if it is cultured and propagated using a liquid medium, its growth direction is constant, and it is difficult to separate strains. The leaves and roots can be easily separated without disturbing, and by using a liquid medium, maintaining the state where the liquid medium is in contact with plants that are effective for growth, Nutrients can be effectively supplied to plants, and a high growth rate can be obtained.

[0008]

[Function] Furthermore, the state where the liquid medium is brought into contact with the plant body is
Since this is realized by shaking the sealed container, the boundary phase
In this part, a state where intermittent contact with gas and liquid is obtained,
An even higher proliferation rate is easily obtained. Also of the plant
Regenerating and growing a plant that has regenerated the growing point tissue
In general, the growth point tissue of a plant is infected with a virus.
Is known to not be infected by a virus
High quality plants can be cultured.

According to a second aspect of the present invention, a leaf of a strawberry ,
By cutting the petiole and the root to obtain a portion corresponding to the border between the petiole and the root, securing the crown containing the tissue necessary for plant growth, and further consuming nutrients for growth Since the leaves and roots are cut and removed, nutrients are not consumed for the growth of the grown leaves and roots, and the nutrients used effectively affect the growth of plants.

In addition, MS diluted 1/10 to 1/2
In the medium, 0.5-1% of sucrose and 0.1% of benzyladenine.
A liquid medium containing 5 mg / liter or less and kinetin at 1 mg / liter or less employs the above-described culture method.
When you do, you can greatly improve the proliferation rate
Is experimentally known, that is, according to the third invention,
For example, it has become possible to greatly grow strawberries as compared with the conventional method.

Furthermore, when a plant hormone such as benzyladenine or kinetin is added to a liquid medium generally used for growing a plant, and the plant is grown, deformed leaves or fruits of the grown plant can be seen, or the flower may grow. Although it blooms a lot, it is known that mutations that make it impossible to produce large fruits are liable to occur.
It has been confirmed that the plant mutation does not occur when the kinetin content is 0.2 mg / liter or less and the kinetin content is 0.2 mg / liter or less.

[0012] In other words, by the upper Symbol new findings, in a state that does not generate a variant in plants, now it is possible to improve the growth rate.

[0013]

As described above, the plant can be grown at a high growth rate while the strains are easily maintained, so that a large amount of the plant can be produced, and furthermore, the virus can be infected. By producing plants that have not done so, high-quality plants can be produced and provided in large quantities.

[0014]

Embodiments of the present invention will be described below with reference to the drawings. First, a growing point at the tip of a strawberry runner is obtained according to a conventional method, and cultivation is performed according to a conventional method to obtain one plant 1 (hereinafter, the plant at this stage is referred to as a seedling).
The seedling is placed in a sealed container 3 while being placed on a support table 2 including a stainless steel net 2a and support legs 2b for supporting the net. Further, a medium A (described later) is placed as a liquid medium 4 in the sealed container 3 so as to be in contact with the plant. The hermetically sealed container is fixed on a shaking table and shaken continuously during the culture period, and the medium A is constantly stirred to continuously supply nutrients to the crown portion. Culture and grow for days. (See Fig. 1)

Propagating strawberry seedlings in this way increased the average 98-fold. The grown strawberry seedlings were further divided into strains and, as shown in FIG.
And the root immediately below the crown are cut off to remove the root 1a, leaf 1b, and petiole 1c that do not include the crown 1d. Then, the cut strawberry seedling containing the crown is regenerated as a regenerated plant. It is further cultured and expanded. (See Fig. 2)

The shaking table used here is such that the sealed container 3 fixed on the shaking table is rotated about a vertical axis so as to draw a circle on a horizontal plane, and the culture medium 4 in the sealed container 3 is removed. It can be stirred. Further, the sealed container used herein means a container that allows gas to enter and exit inside and outside, and that blocks invasion of various bacteria and the like.

The medium A is obtained by diluting an MS medium (described later) four-fold (hereinafter referred to as ４MS) (hereinafter 1 / n).
MS is an MS medium diluted n-fold with sucrose in 1% by weight and benzyladenine (hereinafter BA).
(Abbreviated as) is added at 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 Ferrous sulfate heptahydrate (FeSO ₄ .7H ₂ O) 27.8 Disodium ethylenediaminetetraacetate (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 nicotinic acid .................. 0.5 thiamine hydrochloride 0.1 pyridoxine hydrochloride 0.5 m-inositol 100 glycine 100 glycine ............ 2

The following are test examples in which strawberry seedlings 1 were grown under various conditions.

[Test Example 1]: Dependence of the number of growing strawberry seedlings on BA concentration The three strawberry seedlings 1 obtained by cultivating the growing point according to a conventional method were each 50 ml of mediums A, B, C, D and E. The strawberry seedlings were cultured for 35 days under conditions that the liquid medium 4 was constantly stirred using the above-mentioned shaking table, and the number of seedlings grown was examined. The mediums B to E were obtained by changing the BA concentration of the medium A in various ways, as shown in Table 1. The result is as shown in Table 2.

[0020]

[Table 1]

[0021]

[Table 2]

In other words, it was found that when the strawberry seedlings 1 were cultured in the liquid medium 4 containing BA and proliferated, they grew about 7 to 16 times in 35 days, although there were variations due to experimental errors.

Test Example 2 Dependence of Strawberry Proliferation Number on MS Medium Concentration Instead of liquid medium 4 used in Test Example 1, kinetin, which is a component added and used in a normal medium without BA, was used. The strawberry seedlings 1 were cultured for 32 days in the same manner as in Test Example 1 using the different culture media FI to I, and the number of the grown 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 to which kinetin was added to 1/4 MS was used, the growth rate was lower as compared with the case where BA was used, but it was comparable to the growth situation using BA. Growth rate. That is, 1 / 2MS
If the liquid medium 4 in which kinetin is added to １／1/10 MS is used, a high growth rate can be obtained. In addition, kinetin is
It has been empirically confirmed that when used in the range of 0.2 to 1 mg / liter, the same or more effect can be obtained as compared with a medium containing 0.5 mg / liter of benzyladenine.

Test Example 3 Dependence of Strawberry Proliferation Number on Sucrose Concentration Instead of the liquid medium 4 used in Test Example 1, the sucrose concentration of the liquid medium 4 containing BA and not containing kinetin was variously changed. In the same manner as in Test Example 1, 4
The strawberry seedlings 1 were cultured for 2 days, and the number of the grown strawberry seedlings 1 was examined.
Table 5 below shows the components of the liquid medium 4 used, and Table 6 shows the culture results.

[0028]

[Table 5]

[0029]

[Table 6]

As a result, when liquid medium 4 containing kinetin added to 1/4 MS was used, sucrose was reduced from 0.5% to 0.5%.
At 1%, it was found that the growth rate was comparable to that when BA was used. In other words, sucrose concentration is 0.5% ~
If it is 1%, a high growth rate can be obtained.

Comparative Example 1 Conventional Culture Test Example Strawberry seedlings 1 for 62 days under the same conditions as in Test Example 1 using the agar medium N conventionally used instead of the liquid medium 4 used in Test Example 1 Was subjected to static culture to examine the growth rate of strawberry seedlings 1.

The agar medium N is prepared by adding sucrose to 3% by weight, BA at 0.5 mg / l, kinetin at 1.0 mg / l, and agar at 0.8% by weight to an LS medium (described later). A medium. The composition of the LS medium is as follows.
Shown in 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 sulfate heptahydrate 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]: Investigation of strawberry seedling growth rate The strawberry seedling 1 was cultured for 59 days in the same manner as in Test Example 1 by using the medium O instead of the liquid medium 4 used in Test Example 1. 1 was examined. The medium O is a liquid medium 4 in which kinetin is added at 0.2 mg / liter and sucrose at 1% by weight to 1/4 MS.

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

[0035]

[Table 7]

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

[Test Example 5]: Examination of the effect on the growth rate of strawberry seedlings by cutting and removing roots and petiole parts and using a support stand In place of liquid medium 4 used in Test Example 1, medium P was further used. Strawberry seedlings are placed on the petiole side of crown 1d 1-2.
cm and the root immediately below the crown portion, the portion including the crown portion 1d is defined as a strawberry seedling 1, and the strawberry seedling 1 is placed on the support 2 in a sealed container 3.
And cultured for 64 days in the same manner as in Test Example 1 to examine the growth rate. The medium P is a medium in which kinetin is added at 0.2 mg / liter and sucrose at 1% by weight to 1/4 MS.

Table 8 shows the results of Test Example 5 and the test results obtained by using 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 were cultured and grown using the plant growth method of the third invention, the growth rate was remarkably improved as compared with the case where the conventional growth method was used.

[Test Example 6]: Examination of a medium containing both BA and kinetin, and examination of the effect on growth rate of strawberry seedlings by use of a support. In place of liquid medium 4 used in Test Example 1, mediums Q to U, the strawberry seedlings 1 grown in Test Example 1 were further strained, and the crown portion 1d was cut off at a location of the petiole portion of 1 to 2 cm and the root portion immediately below the crown portion, including the crown portion 1d. 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, and the growth rate was examined. Further, 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,
Place of petiole 1-2 cm from crown 1d, and
The portion including the crown portion 1d among the cut portions with the root portion immediately below the crown portion 1d was designated as a strawberry seedling 1, and the strawberry seedling 1 was placed on a support 2 in a sealed container while being placed in the sealed container. Contains the mediums V and W so as to be in contact with the plants, and the sealed container is fixed on a shaking table to continue shaking during the culturing period.
Was cultivated while constantly stirring (see FIG. 1).
The mediums Q to W have the following compositions.

[0042]

[Table 9]

The results are as shown in Table 10.

[0044]

[Table 10]

From Table 10, it can be seen that a higher growth rate was obtained in each case than in the conventional method. In addition, in the example using the shaking table, the medium V had a 98-fold increase in the medium V, indicating that the growth rate was greatly improved as compared to the medium Q. 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 intermittently contacts the crown using the shaking table.

Incidentally, reference numerals are written in the claims for convenience of comparison with the drawings, but the present invention is not limited to the configuration of the attached drawings by the entry.

[Brief description of the drawings]

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

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

FIG. 3 is an overall view of a plant.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Plant 2 Support 4 Liquid culture medium

──────────────────────────────────────────────────続 き Continuation of the front page (72) Inventor Fumiaki Oda 1-1-1 Hama, Amagasaki-shi, Hyogo Pref. Kubota Technology Development Laboratory Co., Ltd. (56) References JP-A-1-252278 (JP, A) Jiro Furukawa Author, Illustrated Introduction to Tissue Culture, Dec. 24, 1985, 4th print, Seibundo Shinkosha, p. 23 (58) Fields investigated (Int. Cl. ⁶ , DB name) A01H 4/00 C12M 3 / 00

Claims (3)

(57) [Claims] 1. A regenerated plant is produced using a plant tissue, and the leaves, petiole and root of the plant are cut off.
With obtaining the boundary substantial portion of the petiole and root portion, the boundary
The part corresponding to the field is placed on a support base and housed in a sealed container,
A method for growing a plant , wherein the hermetically sealed container is rotated so as to draw a circle on a horizontal plane, and the plant is grown while maintaining the state in contact with the liquid medium. 2. A regenerated plant is produced using strawberry tissue, and the leaves, petiole and root of the plant are cut off.
To obtain a portion corresponding to the border between the petiole and the root,
Place the part corresponding to the boundary on a support base , housed in a sealed container,
A method for growing a plant , wherein the hermetically sealed container is rotated so as to draw a circle on a plane, and the plant is grown while keeping the plant in contact with a liquid medium. 3. A liquid medium for growing strawberries for use in the method of growing a plant according to claim 2, wherein 0.5 to 1% sucrose is added to a 1/10 to 1/2 diluted MS medium. A liquid medium containing 0.5 mg / liter or less and kinetin 1 mg / liter or less.