JP3330365B2 - Culture fluid for Scutellaria plants - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to the orchid family in large quantities grown Cypripedium plants, about the Cypripedium plant for the culture medium for growing.

[0002]

BACKGROUND OF THE INVENTION In general, a plant of the genus Atsushi is a member of the orchid family, and is also referred to as the genus Cypripedum. . Cypripedium is said to be a phantom wild grass, and in particular, Cypripedium is registered as a rare wildlife by the Act on Regulations on the Transfer of Endangered and Endangered Wildlife and Plants due to Overfishing. Plant. In addition, the plants of the genus Atsushi are difficult to proliferate, and one of the reasons is that the germination rate is low. The germination rate in nature is 1 / 100,000
Also called. This is because ordinary plants have parts that grow on leaves and roots, and endosperm that contains nutrients necessary for germination. The germination rate is thought to be low from the structure. Therefore, in recent years, in order to preserve the species of the plant belonging to the genus Hevea, research has been conducted to increase the germination rate and to proliferate using biotechnology. At present, there are twelve research facilities in Japan that are working on cultivation of Cypripedium with the permission of the government.

Hitherto, as a method for cultivating a plant of the genus Hemlock, there has been known, for example, a method in which sterile water is poured on an agar medium by aseptic sowing to disperse seeds.
As the sterilized water, for example, an aqueous solution to which sodium hypochlorite is added is known (for example, see Japanese Patent Application Laid-Open No. 5-14 / 1993).
No. 8115).

[0004]

However, such a conventional method for cultivating a plant of the genus Hemlock has a problem that a satisfactory germination rate cannot always be achieved. The inventors of the present invention have been working on the research since 1995, and in the meantime, using about 100 or more kinds of mediums, spreading the seeds on these mediums using the above-mentioned sterilized water to which sodium hypochlorite was added. The test showed that the germination rate did not increase. In addition, there is also a problem that even if the plant of the genus Athleticum germinates, the plant is more likely to brown and die before it grows into a seedling. Browning death is a phenomenon in which a plant emits a phenolic substance that has turned black and browns and dies. Therefore, it is difficult to improve the germination rate by the conventional cultivation method, medium and culture solution, and even if germinated, the plants often brown and die during cultivation, which is a serious problem.

The present invention has been made in view of such problems, and improves the germination rate, suppresses the occurrence of browning death during cultivation after germination, increases the survival rate, and increases the number of seedlings at once. It is an object of the present invention to provide a culture solution for a plant of the genus Apex genus, which can produce Escherichia coli.

[0006]

Means for Solving the Problems In order to solve such problems, the present inventors have proposed a culture solution containing nutrients as a substitute for sterile water used for spreading seeds on a medium when sowing. We studied the use of cultivation and worked on its development, and developed the following culture fluids for Hempericum genus plants, culture media for genus Hemperial genus plants, and methods for cultivating Hempericum genus plants, which can suppress the occurrence of browning and improve the germination rate . The culture fluid for the Athens plant of the present invention is used for cultivating the Athens plant from seeds, and a culture solution for the Athens plant obtained by adding an additive to water,
In 1000 ml of the culture solution, saccharides 0 to 0 are added as additives.
10 g, wood vinegar 0.1-2.0 ml, and vitamins 8
４５45 mg and 0.1 to 1.0 ml of the plant active agent. Here, the plant active agent is a crude protein
0.05-0.15% by weight, crude fat 0.3-0.5%
%, 35-45 mg / L of Na, Ca30
~ 35mg / l, Fe1.5 ~ 2.0mg / l, Mg
3.0-4.0 mg / l, Si 7.0-8.0 mg /
1, N 90 to 100 mg / l. When this culture solution is used for aseptic seeding, the seeds can easily absorb nutrients due to the interaction of the wood vinegar solution, vitamins, and plant activators, promoting the formation of protocorms and greatly improving the germination rate. Also, by adding during the subculture after germination, the occurrence of browning of the plant is suppressed, the browning of the plant is suppressed, and the plant is well absorbed by nutrients and is grown smoothly. Is greatly improved.
In addition, the addition of a plant active agent, good nutritional balance,
It has the effect of promoting germination.

[0007] If necessary, the culture solution is adjusted to pH
It is effective to set it to 5.5 to 6.0. By adjusting the pH to a value close to the pH of the self-made cloth of snapper, the environment for germination and culture can be optimized, and the germination rate can be improved and the growth can be improved. If necessary, a saccharide is used as the additive. In this case, it is effective to contain 1 to 10 g of saccharides in 1000 ml of the culture solution. Then, if necessary, the saccharide was used as saccharose. And, if necessary, the above vitamins were configured to be thiamine hydrochloride. In addition, the above vitamins
Is effective to contain 1 to 10 mg of thiamine hydrochloride . Further, if necessary, the above-mentioned vitamins were configured to be nicotinic acid. In addition, the above vitamins,
It is effective to include 1 to 5 mg of nicotinic acid . If necessary, the above-mentioned vitamins were constituted by pyridoxine hydrochloride. In addition, the above vitamins, salt
It is effective to comprise 1 to 10 mg of pyridoxine acid . Furthermore, if necessary, the above-mentioned vitamins are configured to be myo-inositol. In addition,
Containing 5-20 mg of myo-inositol
It was it is effective. In the cultivation of orchid plants, vitamins often have the effect of promoting cultivation, and the selection is important. However, the addition of the above-mentioned vitamins allows the plant to be balanced during germination and subculture. Works well and effectively.

[0008]

[0009] Next, the culture for a plant of the genus Hemlock according to the present invention.
In the medium for the genus Athliana plants developed during the study of the liquid
I will mention it. The A Tsumorisou Genus medium is used in culturing Cypripedium plants from the seeds, an aqueous solution prepared by adding an additive to water at Cypripedium planting medium in gel form, as an additive in the aqueous solution 1000ml , Fertilizer containing nitrogen, phosphorus and potassium 1
3 g, peptone 1 to 3 g, sugar 10 to 30 g, plant active agent 0.2 to 1.0 ml, vitamins 16 to 70
mg. Here, the plant active agent is a crude tan.
Protein 0.05-0.15% by weight, crude fat 0.3-0.
5% by weight, 35-45 mg / L of Na as inorganic substance, C
a 30-35 mg / l, Fe 1.5-2.0 mg / l,
Mg 3.0-4.0 mg / l, Si 7.0-8.0 mg
/ L, N 90-100 mg / l. By using this medium for aseptic seeding, the formation of a protocomb is promoted, so that the germination rate can be improved. Also,
By adding a plant active agent, nutritional balance is good and germination
It has the effect of promoting. If necessary, potato cubes were added in an amount of 0.5 to 1.5 g in 50 ml of the medium instead of the plant active agent. And it was set as the structure which makes the said aqueous solution pH 5.5-6.0 as needed. By adjusting the pH to a value close to the pH of the self-made cloth of Hemlock, the environment for germination and culture can be optimized, and the germination rate can be improved and the growth can be improved.

[0010] If necessary, the above saccharides may be made into saccharose. And if necessary, the above vitamins were configured to be thiamine hydrochloride. In addition, the above Vita
Mins containing 5 to 20 mg of thiamine hydrochloride.
And it is effective. And if necessary, the above-mentioned vitamins are configured to be nicotinic acid. In addition,
Tamines containing 1 to 10 mg of nicotinic acid
And it is effective. Furthermore, if necessary, the above vitamins were constituted by pyridoxine hydrochloride. Also on
Contains the vitamins, pyridoxine hydrochloride 5-20 mg
It is effective to have the configuration . If necessary, the above-mentioned vitamins were configured to be myo-inositol. Ma
The above vitamins, myo-inositol 5-20m
It is effective that g is contained . In cultivation of orchid plants, vitamins often have the effect of promoting cultivation, and the selection is important. By adding the above-mentioned vitamins, vitamins are balanced in the plant during germination and subculture. Works well and effectively.

[0011] Then, if necessary, has a structure using gellan gum as the gelling agent. This has an effect on germination and growth.

[0012] If necessary, the above aqueous solution may be used in combination with activated carbon as an additive. By adding activated charcoal, the occurrence of browning of the plant body during subculture after germination is suppressed, so that the plant can be grown without browning death and the survival rate can be increased. In this case, it is effective to contain 0.5 to 3 g of the activated carbon.

Next, the present invention relates to the
Cultivation of the plant of the genus Hemlock developed during the study of the culture solution
Nourishment methods are listed. The method of culturing the A Tsumorisou genus plant is used in the culture method of Cypripedium plant culturing Cypripedium plants from seeds, and seed selection steps to obtain the seeds to select and use the resulting seeds culture sterile Aseptic seeding step of seeding solid medium, germination culturing step of seeding and culturing until germination, subculture step of subculturing germinated and organ differentiated by changing medium, and cultured seedling The plant was configured to include a low-temperature treatment step of exposing it to a low-temperature environment after growing to a certain size, and a pot-raising step of replanting the soil after the low-temperature treatment. Thereby, after sowing the seeds in the medium, a protocomb is formed, and the germination rate is remarkably improved. The seeds are sown in a medium, and aseptic sowing is performed under the conditions of nutrition, temperature, humidity, etc. under ideal conditions. The seedlings are resilient to environmental changes and acclimatization proceeds smoothly.

[0014] If necessary, the above-mentioned culture solution may be a culture solution having a composition different from that of the medium. With this, not only elements due to the components of the medium but also components of the culture solution are added, so that nutrients are abundant and germination can be promoted. If necessary, in the germination culture step, the seeded medium was cultured in the dark. Cypripedium plants are sensitive to light, and when exposed to light, browning occurs and browns to death. Also, if necessary,
In the germination culture step, the inoculated medium was
It was configured to culture in an environment of ° C. It is the optimal temperature for cultivation of a plant of the genus Athliana. If the temperature is higher than 25 ° C., browning occurs and browning ends. If the temperature is lower than 10 ° C., the growth is slow, browning occurs, and browning occurs.

[0015] If necessary, a culture solution is added to the medium in the subculturing step. There is a high risk of dying in the seeds and plants immediately after germination and young seedlings that cannot withstand drying, but do not dry by adding a culture solution, and easily absorb nutrients for the liquid, Furthermore, the occurrence of browning death during cultivation after germination can be suppressed, and the survival rate can be increased. Further, if necessary, in the above subculture step, the medium was cultured in an environment of 10 to 25 ° C. It is the optimal temperature for cultivation of a plant of the genus Athliana. If the temperature is higher than 25 ° C., browning occurs and browning ends. If the temperature is lower than 10 ° C., the growth is slow, browning occurs, and browning occurs.

[0016] If necessary, the low-temperature treatment step may have a low-temperature treatment period of 30 to 90 days. In this case, desirably, in the low-temperature treatment step, the low-temperature treatment period is set to 60 days ± 5 days. Vernalization treatment in which the seedlings grown by cultivation have been successfully grown by cultivation in a natural environment is required to be kept at a low temperature for 90 days or more in order to grow them in the natural environment. However, in the present culture method , the same effect as the vernalization treatment can be obtained in the low-temperature treatment step of about 60 days ± 5 days.

Further, if necessary, the low-temperature processing step is configured so that the low-temperature processing temperature is 0 to 10 ° C. When the temperature is lower than 0 ° C., the seedling freezes and necroses. On the other hand, when the temperature is higher than 10 ° C., the effect of the low-temperature treatment is not seen and the growth does not proceed smoothly. In this case, desirably, in the low-temperature processing step, the low-temperature processing temperature is 3 to 5
C is effective.

Further, a culture solution used in this culture method
And The medium, it is preferable to use a culture solution及 beauty culture medium described above. In this case, the germination rate becomes 80% or more, the survival rate after germination becomes 70% or more, the mass growth of genus Plants becomes possible, and seedlings after potting can be supplied.

[0019]

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective view of a culture solution for a snapper plant according to an embodiment of the present invention.
Will be explained. Cypripedium plant for culture, because used in the process of culture A Tsumori <br/> Saw genus plants are described in the description of the culture methods.

[0020] Scutellaria genus according to an embodiment of the present invention
As shown in FIGS. 1 to 3, a method for cultivating a snapper plant using a plant culture solution is a seed selection step (1) of obtaining seeds to be used by sorting, and aseptically collecting the obtained seeds in a solid medium. Aseptic sowing step (2), germination culture step (3) of culturing after seeding until germination, and subculture step (4) of subculture of germinated and differentiated organs by changing the medium. A low-temperature treatment step (5) of exposing the cultured seedlings to a certain size when exposed to a low-temperature environment; and a potting step (6) of replanting the medium after the low-temperature treatment. Hereinafter, each step will be described in detail.

(1) Seed Sorting Step First, excellent seeds of snapdragons suitable for culture used for aseptic sowing are obtained as follows. Artificial Crossing Within 30 days after flowering, preferably within one week after flowering, pollen is collected, pollen is pollinated at the fertilization destination of the strain to be mated, and mating is completed. Protection of pods After the end of artificial mating, if mating is confirmed, cover the entire pod with a bag to prevent damage from pests and insects. If not covered with a bag, germicide is regularly applied to the sheath. Pod collection time Pods of up to 40 to 120 days, preferably 60 to 80 days after completion of artificial mating are collected and used as aseptic seeding material. Here, in Hemlock, seeds around 60 days after mating, at which embryo formation begins, have the highest germination rate and are desirable. Embryos are not sufficiently formed until less than 40 days after mating, and the germination rate decreases after 80 days after mating. Seed sorting After the collected pods are sterilized, the seeds are removed from the pods, and the embryos inside the seeds are observed and selected using a microscope. The seeds are selected from those having high germination ability, having a rounded embryo shape, which is characteristic, and having a white or yellow embryo color. Embryos that are white or yellow in color are not susceptible to germination inhibitors and therefore are easy to germinate. Conversely, the shape of the embryo is thin, and the rice-like one has low germination ability. Embryos with brown or black color have extremely low germination ability.

(2) Aseptic Seeding Step The selected seeds are put into a culture solution and aseptically seeded on a germination medium used for germination. The seeds in the culture solution are poured into the medium and spread so that the seeds spread throughout the medium. At this time, there are about 300 to 1000 seeds in the flask.

The culture solution used here is the culture solution according to the embodiment of the present invention, and contains 0-10 g of saccharose, 0.1-2.0 ml of wood vinegar, 1-10 mg of thiamine hydrochloride as a vitamin, nicotine. 1-5 mg of acid, 1-10 mg of pyridoxine hydrochloride and 5-20 mg of myo-inositol
And 0.1 to 1.0 ml of a plant active agent, distilled water is added to these additives, and the pH is adjusted to 5.5 to 6.0 using 0.1 N hydrochloric acid or 0.1 N sodium hydroxide. 1000 ml of the adjusted culture solution was obtained. Then, high pressure sterilization by autoclaving, cool sterile room. Here, the plant active agent is composed of 0.05 to 0.15% by weight of crude protein, 0.3 to 0.5% by weight of crude fat, and 35 to 45% Na as an inorganic substance.
mg / l, Ca 30-35 mg / l, Fe 1.5-2.
0 mg / l, Mg 3.0-4.0 mg / l, Si 7.0
-8.0 mg / l, N90-100 mg / l. For example, HB101 (manufactured by Flora Co., Ltd.) or Menedale (manufactured by Menedale Chemical Laboratory Co., Ltd.), which are commercially available, are used as the plant activator.

[0024] In addition, Cypripedium for plants Ru used here
The medium contains 6.5 nitrogen, 6 phosphate and 19 potassium.
As fertilizers (commercially available under the trade name "Hyponex" (manufactured by Hyponex Co., Ltd.)) 1-3 g, peptone 1-3 g, saccharose 10-30 g, and vitamins Thiamine hydrochloride 5
Using 20 mg, 1 to 10 mg of nicotinic acid, 5 to 20 mg of pyridoxine hydrochloride and 5 to 20 mg of myo-inositol and 0.1 to 1.0 ml of a plant active agent, distilled water was added to these additives, and 0.1N hydrochloric acid was added. Or adjusted to pH 5.5 to 6.0 using 0.1N sodium hydroxide and
0 ml of an aqueous solution is obtained. Then, 3 to 4 g of gellan gum is added to this aqueous solution as a gelling agent, and then heated with stirring to dissolve gellan gum, and the flask is heated to about 50 m.
The mixture is dispensed in 1-liter increments, sterilized by autoclaving under high pressure, allowed to cool in a sterile room, and solidified. As the plant active agent, the same components as described above are used. For example, commercially available products such as HB101 (manufactured by Flora Co., Ltd.) and Menedale (manufactured by Menedale Chemical Laboratory Co., Ltd.) are used. The weight of the vitamins contained in the medium is twice that of the above-mentioned culture solution.

(3) Germination culture step After aseptic seeding, the room temperature is set to 15 to 20 ° C. and the culture is started in the dark. The culture is performed in the dark because:
This is because the seeds brown and die when exposed to light. About 2 to 3 months after sowing, PLB (protocomb)
Seeds and germinates. Then, until organ differentiation is seen,
Incubate. (4) Subculture Step The germinated and organ differentiated cells are subcultured by changing the medium.
The culture medium used here is a germination medium containing 0.5 to
This is a subculture medium having a composition to which 3.0 g is added. Transfer the tweezers to the subculture medium to which activated carbon has been added, add about 0.5 to 1.0 ml of the culture solution in 50 ml of the medium onto the medium, and place the medium in the dark, and bring the room temperature to 10 to 25 ° C. Set and start culture. Subculture is performed for 1 to 6 months. During this time, the medium is replaced one to five times in the same manner as described above, and at that time, a culture solution is also added. Here, when it is desired to promote the growth of the cultured seedlings, if generation of the foliage is observed, a very small amount of light (the light amount is 0 to 2000 lux (lux)) may be irradiated.

(5) Low temperature treatment step When the cultured seedlings have grown to about 10 mm or more, the temperature is raised to 3 to 5 ° C.
And place in a low temperature environment for 60 days or more. Vernalization treatment in which the seedlings grown by cultivation have been successfully grown by cultivation in a natural environment is required to be kept at a low temperature for 90 days or more in order to grow them in the natural environment. However, in the main culture method , the same effect as the vernalization treatment for 90 days or more can be obtained in the low-temperature treatment step for 60 days or more. (6) Pot-raising process After the low-temperature treatment is completed, the plants are replanted into a medium and then potted. To raise the pots, the cultured seedlings grown on the medium are taken out using tweezers, and immersed in water or water containing a bactericide to wet the seedlings sufficiently. Next, soil is added to the pot prepared in advance, and a hole having a depth of about 1 cm is made in the soil, planted so that the whole seedling is buried (so that the plant body is not taken out), and potted. . After the start of cultivation, the plant elongates from the soil and starts growing.

Therefore, according to the present culturing method , the germination rate is extremely high, and the cultivation can be carried out while suppressing the occurrence of browning during the cultivation. . In addition, when the cultivation is performed such that the potting step is performed in the spring, the growth cycle of the snapper and the season coincide with each other, so that the snapper can be smoothly grown even after the potting.

[0028]

Example (A) Culture solution (named "Tono culture solution") As shown in FIG. 4, a culture solution according to an example of the present invention comprises 10 g of saccharose, 1.0 to 2.0 ml of wood vinegar solution. Thiamine hydrochloride 5mg as vitamins, nicotinic acid 2.5
mg, pyridoxine hydrochloride 5 mg and myo-inositol 1
0 mg, distilled water was added to these additives, and 0.1 to 1.0 ml of HB101 (produced by Flora Co., Ltd.) as a plant activator was appropriately added dropwise.
The pH was adjusted to 5.65 using 0.1N hydrochloric acid or 0.1N sodium hydroxide to obtain 1000 ml of a culture solution. HB101 as a plant activator is composed of 0.1% crude protein, 0.4% crude fat, and Na41 as mineral.
mg / l, Ca 33 mg / l, Fe 1.8 mg / l, M
g 3.3 mg / l, Si 7.4 mg / l, N97 mg / l
1 is provided. The hydrogen ion concentration (stock solution) was pH 4.
0. Thereafter, the mixture was sterilized by high pressure in an autoclave and allowed to cool in a sterile room.

(B) Germination medium (named "Tono medium") The medium according to the main culture method is, as shown in FIG. 5, "Hyponex" (manufactured by Hyponex Co., Ltd.) as a fertilizer.
Was used. Here, "Hyponex" means that nitrogen is 6.
5, a fertilizer containing 6 phosphoric acid and 19 potassium (weight%). And 2 g of "Hyponex", 2 g of peptone, 20 g of saccharose,
Thiamine hydrochloride 10mg, nicotinic acid 5 as vitamins
mg, pyridoxine hydrochloride 10 mg and myo-inositol 20 mg, distilled water was added to these additives, and 0.1 to 1.0 ml of HB101 (produced by Flora Co., Ltd.) as a plant activator was appropriately added dropwise. Furthermore,
The pH was adjusted to 5.65 using 0.1 N hydrochloric acid and 0.1 N sodium hydroxide to obtain 1000 ml of an aqueous solution. Gellan gum 3 is added to the aqueous solution in this state as a gelling agent.
４4 g was dissolved by heating with stirring, dispensed into flasks by about 50 ml each, and sterilized by autoclaving under high pressure.
Allow to cool in a sterile room to solidify. (C) Subculture medium (named "Tono medium") The subculture medium according to the main culture method has a composition containing 0.5 to 3.0 g of activated carbon as an additive in the aqueous solution. Was used.

The culture method, and therefore were carried out to the above-described method. In the subculturing step (4), the subculturing is 1 to
At 6 months, the medium was replanted 1 to 5 times, and the cultured seedlings were grown to about 10 mm or more. The cells were cultured without irradiating light even if stems and leaves were generated. In the low-temperature treatment step (5), the temperature of the seedling was set at 3 to 5 ° C., and the seedling was placed in a low-temperature environment for 60 days in a dark place.

[0031]

[Experimental Examples] (II) Experiments on germination medium First, the following experiment was conducted on the medium (B) according to the main culture method together with Comparative Examples. As a comparative example of a germination medium, M
S medium, Hyponex medium, MT medium, Nadson B medium, Nadson C medium, Ringmeyer-Skoog medium,
White medium, Basin bent medium, Winver medium, Burgeft. H. Eg-1 medium, Curtis. J
Medium, Yamada medium, banana honey (Karazawa) medium, fish soluble cultivation medium, western orchid onion medium, GS medium,
A total of 20 kinds of culture mediums, a Kyoto solution medium, a Burgeft N ₃ F medium, a Gamborg B5 medium, and a Harvars medium, were used. Aspergillus seeds were aseptically seeded on these media. It is to be noted that only sterilized water was used for spreading the seeds at the time of sowing, and the other steps were the same as those in the culture method described above. FIG. 6 shows the results. As a result, the present culture
The medium according to the feeding method could be adjusted relatively easily, and both germination and growth were good.

(I-II) Experiment on germination medium and subculture medium In order to examine the difference between germination and growth in each medium ,
The following experiment was conducted with the comparative examples for the culture media (B) and (C) according to the method. As a comparative example of the germination medium and the subculture medium, a medium prepared by halving the components of the medium according to the main culture method (1/2 of the concentration of the medium according to the main culture method)
Medium), Havers medium, MT medium, MS medium
Aspergillus seeds were aseptically seeded on various media. still,
The culture was performed in the same manner as the above-described culture method, except for the seeding medium and the subculture medium. The height of the grown seedlings was measured two months, four months, and six months after sowing. The results are shown in FIGS. The final germination rate using the mediums (B) and (C) according to the main culture method was the highest at 82.6%, and the germination rate of a medium prepared by halving the components of the medium according to the main culture method. Was 71.0%, the germination rate of the Havers medium was 70.5%, the germination rate of the MT medium was 1.4%, and the germination rate of the MS medium was 0.6%. In FIG. 8,
The height of the grown seedlings two months, four months and six months after sowing is the average value of 20 to 30 seedlings. As a result,
Six months after seeding, the seedlings of snapdragon cultivated in the medium according to the main culture method became 50 mm on average, indicating that the seedlings cultured in other mediums grew.

(II) Experiments on Culture Solution In order to examine the difference between germination and growth of each culture solution, the following experiment was conducted on the culture solution according to the example of the present invention together with a comparative example. A solution obtained by adding HB101 to sterilized water and saccharose, a solution obtained by adding wood vinegar to sterilized water and saccharose, a solution obtained by adding HB101 and wood vinegar to sterilized water and saccharose, and a solution obtained by adding wood vinegar and vitamin to sterilized water and saccharose The culture was performed using the culture solution of (1). Further, a liquid obtained by changing the addition concentration of HB101 in the above liquid was also examined. In addition, it carried out similarly to the above-mentioned culture method except a culture solution. FIG. 9 shows the results. From these results, it was found that the culture solution of the present invention in which HB101, wood vinegar solution, and vitamins were added to sterilized water and saccharose suppressed the generation of browning most and showed good growth.

(III) Experiment of Culture Temperature In order to examine the influence of each culture temperature, the following experiment was performed as a comparative example in which the culture temperature was changed in the germination culture step (3) of the main culture method . Culture is performed at 10 ° C, 15 ° C, 20
C., 25.degree. C., and 30.degree. C., and the number of tests at each temperature was 50, and the rate at which browning occurred during culturing and did not germinate (browning death rate) was examined. In addition, it carried out similarly to the above-mentioned culture method except culture temperature. As shown in FIG. 10, the result was about 70% in the culture at 25 ° C. and about 9% in the culture at 30 ° C.
5% browned and died, and the culture temperature after seeding was 10
It has been found that performing at ~ 20 ° C is optimal.

(IV-I) Experiment for Examining the Effect of Low-Temperature Treatment In order to examine the effect of low-temperature treatment, the following experiment was performed as a comparative example in which the main culture method and the low-temperature treatment were not performed. The survival rates of the seedlings that had been potted for 60 days in a dark place at 20 ° C. without a low-temperature treatment and the seedlings that had been potted for 60 days in a light place at 20 ° C. were examined. In the examples, the low-temperature treatment is performed by heating the seedlings at 3 to 5 ° C.
Of the seedlings in the dark place for 60 days, but also in the low-temperature treatment step, the survival rate of the seedlings after being potted and the subsequent growth were examined. The conditions other than the low-temperature treatment were the same as in the culture method described above. As shown in FIG. 11, the survival rate of the seedling when the pot was raised without performing the low-temperature treatment was as follows.
% Low survival rate. In contrast, low-temperature treated
Both the dark place and the light place had a high survival rate of 80% or more, and the survival rate in the light place was slightly higher. Further, as shown in FIG. 12, the results of the subsequent growth of the dark place and the light place of the low-temperature treatment were as follows:
In the dark place, the growth was inferior to the light place but no browning occurred. In the light place, the color of the foliage became light but the growth proceeded. However, light browning occurred slightly in the light.

(IV-II) Experiment on Growth State of Hemlock Performing Low-Temperature Treatment Next, the following experiment was performed as a comparative example in which the main culture method and the low-temperature treatment were not carried out in order to examine the growth state of snapper. did. In Examples, the low-temperature treatment was performed in a state where the seedlings had grown to 10 mm or more. In the comparative example, the state after low-temperature treatment and the subsequent growth were examined with the protocomb and the seedling immediately after germination grown to about 5 mm. The conditions other than the low-temperature treatment were performed in the same manner as in the culture method described above. As shown in FIG. 12, as shown in FIG. 12, in the protocomb, growth stopped and browning died. In plants of about 5 mm, no change was observed in growth, and in plants of 10 mm or more (seedling), growth proceeded smoothly. It is.

[0037] (IV-III) of the period in which the low-temperature treatment experiment Next, in order to examine the period in which the low-temperature treatment, the culture lateral
The following experiment was performed as a comparative example in which the method and the low-temperature treatment were not performed. In the examples of the present invention, the low-temperature treatment was performed for 60 days. As a comparative example, plants (seedlings) of about 10 mm in each of 15 days, 30 days, 60 days, and 90 days were stored at 3 to 5 ° C. At low temperature. As shown in FIG.
Since the growth of the seedlings subjected to the low-temperature treatment for 90 days and 90 days was good, the effect of the low-temperature treatment is seen when the seedlings are treated for 60 days or more.

According to the experiments on the low-temperature treatment, it is optimal that the low-temperature treatment is carried out at a temperature of 3 to 5 ° C. for 60 days or more at a low temperature treatment, and the brightness may be either dark or light. I understood that.

In the above embodiment, a plant activator was added to the medium, but without adding the plant activator,
Potato cubes may be used.

[0040]

Industrial Applicability As described above, according to the culture solution for a snapper plant of the present invention, a culture solution for a snapper plant obtained by adding an additive to water, which is used when cultivating a snapper plant from a seed, is used. In the solution, the culture solution 1000
In ml, as an additive, wood vinegar liquid 0.1-2.0 ml
And vitamins 8 to 45 mg and plant active agent 0.1 to
Since the liquid contains 1.0 ml of liquid, the liquid easily absorbs nutrients to form a protocorm, thereby improving the germination rate, and is added during the subculture after germination. Thus, the occurrence of browning of the plant body is suppressed, so that the plant body can easily absorb nutrients without causing browning and death, so that the plant works effectively and grows smoothly, thereby increasing the survival rate.

When the pH of the culture solution is adjusted to 5.5 to 6.0, the environment for germination and culture can be optimized, the germination rate can be improved, and the growth can be improved. When saccharides are used in combination as additives, nutrients can be given to the seeds and the germination rate is improved. In this case, the culture solution 10
It is effective to contain 1 to 10 g of saccharide in 00 ml. In addition, when sugar is saccharose and vitamins contain 1 to 10 mg of thiamine hydrochloride, 1 to 5 mg of nicotinic acid, 1 to 10 mg of pyridoxine hydrochloride, and 5 to 20 mg of myo-inositol, germination of a snapper plant rate to improve, Ru can be made to promote growth.

[0042]

[0043]

[0044]

[0045]

[0046]

[0047]

[Brief description of the drawings]

FIG. 1 shows a plant of the genus Athliana according to an embodiment of the present invention.
FIG. 4 is a process chart showing a culture method using a culture solution for use .

FIG. 2 shows a plant of the genus Attachium according to the embodiment of the present invention.
FIG. 4 is a process chart showing a culture method using a culture solution for use .

FIG. 3 shows a plant of the genus Attachium according to the embodiment of the present invention.
FIG. 4 is a process chart showing a culture method using a culture solution for use .

FIG. 4 is a table showing the composition of a culture solution for a Scutellaria plant according to the embodiment of the present invention.

FIG. 5: Species of the genus Athliana according to an embodiment of the present invention
FIG. 2 is a table showing the composition of a culture medium for a plant of the genus Hevea in a culture method using a culture solution for culture .

FIG. 6 shows a plant of the genus Hemlock according to the embodiment of the present invention.
FIG. 4 is a table showing the results of comparison of germination in other culture media with respect to a culture media for the Athens plant in a culture method using a culture solution for culture .

FIG. 7 shows a plant of the genus Attachium according to the embodiment of the present invention.
FIG. 3 is a diagram showing the germination rate in a medium for a snapper plant in another culture medium in a culture method using a culture solution for culture .

FIG. 8 shows a plant of the genus Athliana according to an embodiment of the present invention.
It is a figure which shows the growth condition of the seedling cultured by the other culture medium concerning the culture medium for the genus Epimedium in the culture method using the culture solution for culture .

FIG. 9 is a diagram showing a growth state of a culture solution of a plant of the genus Hemperium according to the embodiment of the present invention, using another culture solution.

FIG. 10 shows a plant belonging to the genus Hemlock according to an embodiment of the present invention.
It is a figure which shows the generation | occurrence | production situation of browning by the culture temperature regarding the culture method using the culture liquid for products .

FIG. 11 is a plant of the genus Attachium according to the embodiment of the present invention.
It is a figure which shows the effect of low-temperature processing regarding the culture method using the culture solution for articles .

FIG. 12 shows a plant belonging to the genus Hemlock according to the embodiment of the present invention.
It is a figure which shows the growth situation at the time of changing the time which performs low-temperature processing regarding the culture method using the culture liquid for articles .

FIG. 13 shows a plant belonging to the genus Hemlock according to the embodiment of the present invention.
It is a figure which shows the growth situation at the time of changing the period which performs low-temperature processing regarding the cultivation method using the culture liquid for articles .

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Claims (13)

(57) [Claims] The present invention relates to a culture solution for a snapdragon plant which is used when cultivating a snapdragon plant from a seed and which is obtained by adding an additive to water. 2.0 ml, 8 to 45 mg of vitamins, and a plant active agent, crude protein 0.05 to 0.15
Weight%, crude fat 0.3-0.5 weight%, as minerals,
Na 35-45 mg / l, Ca 30-35 mg / l, F
e 1.5-2.0 mg / l, Mg 3.0-4.0 mg /
1, Si 7.0 to 8.0 mg / l, N 90 to 100 mg
/0.1 ml of a plant activator having 1 / l . 2. The culture solution for a plant of the genus Athleticum according to claim 1, wherein the culture solution has a pH of 5.5 to 6.0. 3. The culture solution for a plant of the genus Athliana according to claim 1, wherein a saccharide is used as the additive. 4. The method according to claim 1, wherein the saccharide is added in an amount of 1 to 1000 ml of the culture solution.
The culture solution for a plant of the genus Athliana according to claim 3, comprising 10 g. 5. The culture solution for a plant of the genus Athliana according to claim 3, wherein the saccharide is saccharose. 6. The culture solution for a plant of the genus Athletic plant according to claim 1, wherein the vitamins are thiamine hydrochloride. 7. The method according to claim 1, wherein the vitamins are thiamine hydrochloride in an amount of 1 to 3.
The composition according to claim 1, characterized in that the composition comprises 10 mg.
The culture solution for a plant of the genus Apex genus according to 2, 3, 4 or 5 . 8. The culture solution for a plant of the genus Athletic plant according to claim 1, wherein the vitamins are nicotinic acid. 9. The method according to claim 9, wherein the vitamins are nicotinic acid in an amount of from 1 to 5.
3. The composition according to claim 1, wherein
8. The culture solution for a plant of the genus Apex genus according to 3, 4, 5, or 7 . 10. The method of claim 1, 2, 3, 4 or 5 SL, characterized in that the vitamins and pyridoxine hydrochloride
The culture solution for the plants of the genus Atsuma. 11. The method according to claim 11, wherein the vitamins are pyridoxine hydrochloride.
Characterized by comprising 1 to 10 mg of
Item 1. A culture solution for a plant of the genus Athliana according to Item 1, 2, 3, 4, 5, 7, or 9 . 12. The method of claim 1, 2, 3, 4 or 5 SL, characterized in that the vitamins and myo-inositol
The culture solution for the plants of the genus Atsuma. 13. The method according to claim 12, wherein the vitamins are myo-inositol.
Characterized by comprising 5 to 20 mg of
Item 1. A culture solution for a plant of the genus Apex genus according to item 1, 2, 3, 4, 5, 7, 9, or 11 .