JP6316545B2 - Cutting seedling production method - Google Patents

Description

  The present invention relates to a method for producing cuttings.

  Cutting is a method of rooting in the root bed using artificially cut plant tissue and creating an independent plant body, which is an excellent method for mass propagation of genetically uniform seedlings It is.

  Propagation by cutting is simple and suitable for producing a large amount of seedlings at a time, and it is considered as a commercially advantageous method because individuals with excellent traits can be proliferated in large amounts. However, many plant species are difficult to root with cuttings.

  A medicinal plant is a plant used as a raw material of Chinese medicine and / or food. Plant secondary metabolites often act as active ingredients. Secondary metabolites often adversely affect the rooting of medicinal plants, and it was difficult to grow medicinal plants by cuttings. There are very few cases where medicinal plants are cultivated in Japan, and there is a current situation that depends on imports from overseas. Even overseas, cultivars are limited and many wild plants are collected. For this reason, in order to protect overfishing, China bans the collection of wild plants and restricts exports. Therefore, in order to stably supply medicinal plants in Japan, establishment of a method capable of growing a large amount of simple and uniform excellent seedlings by cutting is desired.

In Patent Document 1, in a method for promoting rooting by inserting an insertion head of a garden plant or the like on a support material in a hydroponic liquid, the light intensity of 100 to 500 μmol / m ² / sec and the repetition of the light-dark cycle of one day are repeated. Artificial light and carbon dioxide concentration set to 2 to 8 times, 500 to 3,000 ppm, air humidity 80 to 100% RH, air temperature 20 to 30 ° C., hydroponic solution and support material temperature 20 It is described that it is maintained at ˜30 ° C.

  In Patent Document 2, a rooting bed wetted with a liquid medium containing nitrogen, phosphorus, and potassium as essential elements and not containing a carbon source is prepared in a culture vessel, and a head is inserted and cultured. In addition, a method for producing cuttings seedlings characterized in that rooting from cuttings is performed while controlling the concentration of carbon dioxide in the culture vessel is described.

JP 2010-110225 A JP 2001-186814 A

  However, Patent Document 1 describes only application examples to foliage plants such as petunia and starches, and describes and suggests application to plants that accumulate secondary metabolites that are considered difficult to root. Absent. With the method described in Patent Document 2, it has been difficult to produce cuttings on a practical scale for plants that are difficult to root, such as medicinal plants.

  An object of the present invention is to provide an efficient method for producing cuttings that can increase the rooting rate of cuttings of plants that accumulate secondary metabolites.

The present invention provides the following [1] to [4].
[1] A method for producing cuttings and seedlings of a plant accumulating secondary metabolites, comprising a step of rooting from cuttings under conditions of light intensity of 120 to 270 μmol / m ² / sec and humidity of 60% Rh or higher.
[2] The production method according to [1], wherein the carbon dioxide concentration in the step of rooting from the cutting head is 500 to 1500 ppm.
[3] The production method according to [1] or [2] above, wherein the plant that accumulates secondary metabolites is a medicinal plant.
[4] The production method according to any one of [1] to [3], wherein the step of rooting from the cutting ear is performed in a culture vessel.

  ADVANTAGE OF THE INVENTION According to this invention, the rooting rate of the cutting head of the plant which accumulate | stores a secondary metabolite can be improved, and cutting production from a medicinal plant can be performed efficiently.

  The target of application of the present invention is a plant that accumulates secondary metabolites. The secondary metabolite (secondary metabolite) means a substance synthesized by metabolizing the primary metabolite. While primary metabolites are directly involved in plant cell growth, development, reproduction, and other life, secondary metabolites are not directly involved in life. It has functions such as protecting itself from external enemies (pathogenic bacteria, etc.). Examples of secondary metabolites include alkaloids (such as ephedrine), terpenoids, flavonoids, terpene glycosides (such as glycyrrhizin), glycosides, phenols, or derivatives thereof. Secondary metabolites are known to contain components useful for medicines, health aids, anesthetics, dyes and the like, and among these, components useful for medicines, health aids and the like are preferable.

  Medicinal plants are preferred as the plants that accumulate secondary metabolites. The medicinal plant means a plant in which at least a part of the plant is used for medicinal purposes. Medicines include herbal medicines (unrefined medicines, medicinal herbs), traditional Chinese medicines, folk medicines, etc., and include foods that are not classified as pharmaceuticals by the Pharmaceutical Affairs Law. Examples of medicinal plants used for medicinal purposes include whole plants, foliage, flowers, persimmons, seeds, fruits, pericarps, roots, rhizomes, rhizomes, xylem, bark, and plant parts containing two or more of these. Illustrated.

  Medicinal plants include citrus, apricot, irresen, nintendo, fennel, engosaku, ogi, ougon, oak, auren, onji, gayyo, kashi, kankon, casseki, carocon, caroten, licorice, kyouka, kikuka, pheasant , Kyonin, Kujin, Keigai, Keihi, Kouka, Koujin, Kouushi, Koubay, Kokuboku, Goshitsu, Goshuyu, Goboushi, Sesame, Garbage, Psycho, Saishin, Hawthorn, Sanshishi, Sanshuyu, Salamander, Sansonin, Sanyaku, Ganko, Zikoppi, Shikon, Shitsuri, Cinnamon, Peonies, Shazenshi, Zukujiou, Shukusha, Shokyo, Shobakaku, Shouma, Shinyi, Gypsum, Senkyu, Zenko, Senkotsu, Centai Senna, Sojutsu, Sakuhakuhi, Soboku, Soyo, Daio, Taiso, Takusha, Chikujo, Chikusetinjinjin, Chimo, Chaoyou, Clove, Butterflyfish, Chorei, Chinpi, Tennansho, Tenma, Tenmondo, Pepper, Toki, Tonin, Spruce , Tokon, Tochu, Dokatsu, Carrot, Nindo, Baimo, Bakuga, Bakumondo, Hakka, Hakubofuu, Hange, Byakugo, Byakushi, Byakujutsu, Biwayou, Binrouji, Bukkyou, Bushi, Fungatsuame, Bowie, Bofufu, Ukubo , Machinin, mokutsu, mokko, yokoinin, ryuganiku, ryukotsu, ryutan, ryokyou, forsythia, rennik, kyoko katsu, preferably kankyo, licorice Keihi, Goboushi, Sanshi, Chimpi, Biwayo, Carrot, Tonin, Chorei, Soyo, Daio, Taiso, Senna, Peonies, Salamander, Sunshu, Kojijin, Buttonpi, Onji, Ogi Ryutan. Even more preferred are licorice plants (Glycyrrhiza) and Ephedra plants. Examples of licorice plants include Russian licorice (G. echinata), Spanish licorice (G. glabra), American licorice (G. lepidota), Ural licorice (G. . Examples of the genus Maou include E. distachya, E. sinica, and E. equistina Bunge.

  In view of the object of the present invention, the application target of the present invention is preferably a medicinal plant of a line that accumulates a large amount of secondary metabolites. “Large amount” only needs to be larger than secondary metabolites accumulated in normal strains of the same species, and the amount varies from plant to plant and is difficult to define uniformly. If the amount of the active ingredient described in the above is exceeded, it can be said that the amount is large.

The present invention includes a step of rooting from the cuttings under conditions of light intensity of 120 to 270 μmol / m ² / sec and humidity of 60% Rh or more.

  The cuttings may be part or all of a medicinal plant. The cuttings may be at least part of the plant, branches such as green branches (current year branches), mature branches (branches extending before the previous year); buds such as top buds and buds; leaves, cotyledons; hypocotyls Etc. are exemplified. The cuttings in the case of woody plants are usually green branches or mature branches, and the cuttings in the case of herbaceous plants are usually leaves or buds, but are not limited thereto.

  On the other hand, shoots may be used as cuttings in that they are expected to form adventitious roots. Shoots refer to all tissues that have rooting ability. Examples of the tissue include branches, stems, apical buds, buds, adventitious buds, leaves, cotyledons, hypocotyls, adventitious embryos, shoot primordia, and the like. The origin of the shoot is not particularly limited, and may be a tissue obtained from an individual plant growing in a greenhouse or outdoors, or may be a cultured tissue obtained by a tissue culture method, or a part of a natural plant body It may be an organization. The shoot can be efficiently obtained from the main plant or multi-bud of the cutting ear. Among them, cuttings (cutting ears obtained from the mother plant), polyblasts obtained by aseptically culturing organs collected from the mother plant, or foliage obtained by aseptically growing the organs Preferably there is.

  A multi-bud can be derived by cutting out tissues such as apical buds and axillary buds from a plant to which a clone seedling is to be produced by applying the present invention, and culturing the tissue. In order to form and cultivate an organ collected from a mother plant aseptically, the polyblast can be formed according to the method and conditions described in JP-A-8-228621. The method and conditions are as follows. First, tissues such as apical buds and axillary buds are collected from a plant as a material, and about the collected tissues, an aqueous sodium hypochlorite solution having an effective chlorine content of about 0.5% to about 4% or an effective chlorine content of about 5% to Surface sterilization is performed by immersing in an aqueous solution of about 15% hydrogen peroxide for about 10 minutes to about 20 minutes. Next, this is washed with sterilized water, inserted into a solid medium, the buds are opened, and the elongated foliage is subcultured in a medium having the same composition to form a multi-bud. In the case of using Eucalyptus or Acacia tissues (for example, buds), the solid medium is 1 to 5% by weight of sucrose, and benzyladenine (hereinafter abbreviated as BA) as a plant hormone is about 0.02 mg / l or more. Murashige Sukug (hereinafter abbreviated as MS) medium containing 1 mg / l or less, gellan gum of about 0.2 wt% to about 0.3 wt% or agar of about 0.5 wt% to about 1 wt%, or MS It is preferable to use a modified MS medium in which the ammonium nitrate component and potassium nitrate component of the medium are halved. The shoots are actively extended from the multi-buds thus formed. The multiblasts themselves can be maintained and proliferated by appropriately dividing them and culturing them in a medium having the same composition as the medium used for the formation of the multibuds.

The light intensity (illuminance) of light applied to the cutting head in the step of rooting from the cutting head is expressed as a photosynthetic effective photon flux density, and is preferably 120 to 270 μmol / m ² / sec, preferably 130 to 270 μmol / m. ² / sec is more preferable, and 130-200 μmol / m ² / sec is more preferable. If it is less than 120 μmol / m ² / sec, the rooting rate of cuttings of plants that accumulate secondary metabolites cannot be improved, and if it is 270 μmol / m ² / sec or more, light such as leaf burns Failure occurs.

  The light applied to the cutting head may be red light, blue light, white light, and a combination of two or more selected from these, but is preferably white light.

  The repetition period of the light period and dark period of irradiation is not particularly limited, but it is preferably 14 to 18 hours of light period per 24 hours / 6 to 10 hours of dark period.

  The humidity of the cultivation environment in the process of rooting from the cuttings is 60% Rh or more. If the humidity is less than 60% Rh, the rooting rate of cuttings of plants that accumulate secondary metabolites cannot be improved. The humidity can be appropriately adjusted depending on the target plant, and is usually more preferably 80% Rh or more, and further preferably 90% Rh or more. When the humidity is in the above range, rooting from plants can be promoted. There is no restriction | limiting in particular about the upper limit of humidity, What is necessary is just 100% Rh or less. In the case of plants where mold tends to grow, it is more preferably 80% Rh or less, and even more preferably 70% Rh or more.

  The carbon dioxide concentration in the cultivation environment in the step of rooting from the cuttings is usually 300 ppm to 1500 ppm, preferably 500 ppm to 1500 ppm, more preferably 750 ppm to 1250 ppm, and further preferably 900 ppm to 1000 ppm. preferable. Control of the supply amount of carbon dioxide gas can be performed using equipment such as an artificial weather device, a culture vessel having a carbon dioxide permeable membrane in the opening, and the like.

  The temperature of the cultivation environment in the step of rooting from the cuttings is preferably about 23 ° C. or higher and about 28 ° C. or lower.

  In the process of rooting from the cuttings, the distribution of the light period and dark period is usually 1: 1 to 3: 1 and preferably 3: 2 to 5: 2.

  In the step of rooting from the cutting head, it is preferable to perform light shielding. The light shielding rate is preferably 30% or more and 70% or less, and more preferably 40% or more and 60% or less.

  The period of rooting from the cuttings varies depending on the cultivation conditions such as the type of medicinal plant, but is usually 2 weeks to 3 months, and preferably 4 weeks to 2 months. Continue until rooting is observed from the cuttings.

  In the step of rooting from cuttings, a rooting medium is usually used. In the present invention, the rooting medium means a medium used for rooting from cuttings.

The rooting medium preferably contains silver ions and / or antioxidants, and more preferably contains both silver ions and antioxidants. Silver ions may be added to the medium as a silver compound (silver ion source) such as silver thiosulfate (STS, AgS ₄ O ₆ ) or silver nitrate. Among them, STS is preferable as a silver ion source used in the present invention, since healthy roots and / or elongation is promoted when added to a medium and cultured for shoots. This is presumably because silver ions derived from STS take the form of silver thiosulfate ions in the medium and are negatively charged. The concentration of silver ions in the rooting medium depends on the type of silver ion source and other culture conditions, but the concentration of the silver ion source is preferably about 0.5 μM or more and about 6 μM or less, and about 2 μM or more and about 6 μM or less. Is more preferable.

  On the other hand, as the antioxidant, for example, known antioxidants such as oxidized glutathione (GSSG), ascorbic acid, sulfite and the like can be used, and oxidized glutathione is preferable. The concentration of oxidized glutathione in the rooting medium is preferably 1 mg / l to 100 mg / l, more preferably 5 mg / l to about 75 mg / l.

  The rooting medium may further contain components such as an inorganic component, a carbon source, vitamins, amino acids, and plant hormones.

  Inorganic components include elements such as nitrogen, phosphorus, potassium, sulfur, calcium, magnesium, iron, manganese, zinc, boron, molybdenum, chlorine, iodine, cobalt, and one or more elements selected from these elements Inorganic salts are exemplified. Examples of inorganic salts include potassium nitrate, ammonium nitrate, ammonium chloride, sodium nitrate, potassium hydrogen phosphate, sodium dihydrogen phosphate, potassium chloride, magnesium sulfate, ferrous sulfate, ferric sulfate, manganese sulfate, zinc sulfate, These hydrates include copper sulfate, sodium sulfate, calcium chloride, magnesium chloride, boric acid, molybdenum trioxide, sodium molybdate, potassium iodide, cobalt chloride and the like. 1 type may be sufficient as an inorganic component, and the combination of 2 or more types may be sufficient as it.

  The rooting medium preferably contains nitrogen, phosphorus and potassium as essential elements. Therefore, among the specific examples of the above-described inorganic components, nitrogen, phosphorus, potassium, an inorganic salt containing nitrogen, an inorganic salt containing phosphorus, and an inorganic salt containing potassium are preferable, and include nitrogen, phosphorus, potassium, and nitrogen. Inorganic salts are more preferred. The concentration of the inorganic component in the rooting medium is preferably 0.1 μM to 100 mM, and more preferably 1 μM to 100 mM, when there is only one inorganic component. When the inorganic component is a combination of two or more, it is preferably 0.1 μM to 100 mM, and more preferably 1 μM to 100 mM.

  Examples of the carbon source include carbohydrates such as sucrose and derivatives thereof; organic acids such as fatty acids; primary alcohols such as ethanol. 1 type may be sufficient as a carbon source, and 2 or more types may be sufficient as it. The concentration of the carbon source in the rooting medium is preferably 1 g / l to 100 g / l, and more preferably 10 g / l to 100 g / l. However, when culture is performed while supplying carbon dioxide gas, the culture medium does not need to contain a carbon source, and preferably does not contain it. Organic compounds that can serve as a carbon source such as sucrose can also serve as a carbon source for microorganisms. Therefore, when using a medium supplemented with these, it is necessary to culture in a sterile environment, but use a medium that does not contain a carbon source. This makes it possible to culture in a non-sterile environment.

  Examples of vitamins include biotin, thiamine (vitamin B1), pyridoxine (vitamin B4), pyridoxal, pyridoxamine, calcium pantothenate, inositol, nicotinic acid, nicotinamide and riboflavin (vitamin B2). 1 type of vitamin may be sufficient and the combination of 2 or more types may be sufficient. The concentration of vitamins in the rooting medium is preferably 0.01 mg / l to 200 mg / l, more preferably 0.02 mg / l to 100 mg / l when the number of vitamins is one. When the vitamin is a combination of two or more, it is preferably 0.01 mg / l to 150 mg / l, more preferably 0.02 mg / l to 100 mg / l.

  Examples of amino acids include glycine, alanine, glutamic acid, cysteine, phenylalanine and lysine. 1 type may be sufficient as an amino acid and 2 or more types may be sufficient as it. The concentration of the amino acid in the rooting medium is preferably about 0.1 mg / l or more and about 1000 mg / l or less when one amino acid is used, and is 0. It is preferable that it is 2 mg / l-1000 mg / l.

  Examples of plant hormones include auxin and cytokinin. Examples of auxins include naphthalene acetic acid (NAA), indole acetic acid (IAA), p-chlorophenoxyacetic acid, 2,4-dichlorophenoxyacetic acid (2,4D), indolebutyric acid (IBA), and derivatives thereof. . One auxin may be used or a combination of two or more auxins may be used. Examples of cytokinins include benzyladenine (BA), kinetin, zeatin, and derivatives thereof. Cytokinin may be one kind or a combination of two or more kinds. The plant hormone may be auxin alone, cytokinin alone, or a combination of auxin and cytokinin. In general, auxin acts on rooting, and cytokinin acts on growth. Therefore, after culturing the ear by adding only cytokinin, it can be rooted by adding auxin. The concentration of the plant hormone in the rooting medium is preferably 0.01 mg / l to 10 mg / l, more preferably 0.02 mg / l to 10 mg / l when one kind of plant hormone is used. preferable. When there are two or more kinds of plant hormones, it is preferably 0.01 mg / l to 10 mg / l, and more preferably 0.02 mg / l to 10 mg / l.

  As the rooting medium, a medium in which silver ions and / or an antioxidant, a carbon source, or plant hormones are appropriately added to a medium known as a plant tissue culture medium as necessary may be used. Examples of the medium known as a plant tissue culture medium include MS medium, Rinsmeier Skoog medium, white medium, Gamborg B-5 medium, and niche niche medium. Of these, MS medium and Gamborg B-5 medium are preferred. These media can be used by appropriately diluting them as necessary.

  The rooting medium may be either a liquid medium or a solid medium, but the liquid medium is preferable in terms of work efficiency and less damage to the roots during transplantation. In the case of a liquid medium, the medium composition may be mixed and prepared and used as it is. In the case of a solid medium, the medium composition can be mixed and prepared in the same manner as the liquid medium, or can be used after being adjusted and solidified with a solidifying agent such as agar or gellan gum. The amount of the solidifying agent added to the medium varies depending on the kind of the solidifying agent and the composition of the medium. When the solidifying agent is agar, it is preferably 0.5% by weight or more and 1% by weight or less. When the solidifying agent is gellan gum, the content is preferably 0.2% by weight or more and 0.3% by weight or less.

  What is necessary is just to select suitably the insertion method of the cutting head to the culture medium for rooting according to the kind of culture medium, culture conditions, etc. When the rooting medium is a solid medium, the rooting medium may be directly inserted into the rooting medium and cultured. On the other hand, when the rooting medium is a liquid medium, for example, the support described later may be wetted with the rooting medium, and then the base of the cutting ear may be inserted and cultured. In order to improve the rooting rate, it is also preferable to apply a physical stimulus such as damaging the base of the cutting head when inserted into the rooting medium. The base of the cutting head means a region (one side opposite to the end where the leaf is formed) which is one end of the cutting head and a root is formed. The base in the case of using a multi-bud as an insertion ear is a region having a cut surface when dividing the multi-bud. The size (size, shape, etc.) of the damage to the base of the cutting head is not particularly limited. For example, in the case where a multi-bud is used as the cutting head, it is preferable to scratch the base of the cutting head (the above-mentioned cut surface) so as to form a cross when viewed from the front. When scratching, instruments such as scissors and knives can be used.

  In the present invention, the support is a support for supporting the cutting head. When using a rooting medium (especially a solid medium), a support may be unnecessary. In other cases, a support is usually used.

  The support is preferably a support that can be held in a state where the cutting head is pointed during the cultivation period. When a liquid rooting medium is used for cultivation, it is usually used by infiltrating a support. Accordingly, the support is preferably a support that can be infiltrated with a liquid, and among them, a support that can be substantially uniformly wetted by a liquid medium is preferable. A conventional support may be used as the support, and is not particularly limited. Examples of the support include natural soils such as sand and red bean clay; artificial soils such as rice husk charcoal, coconut fiber, vermiculite, perlite, peat moss, and glass beads; and porous molded articles such as foamed phenol resin and rock wool. be able to. The root bed can be prepared by placing the support in a culture vessel. When the rooting medium is a solid medium, the rooting bed can be prepared by placing the solid medium directly into the culture vessel.

  In the present invention, it is preferable to use a culture vessel for storing a rooting medium or a support. As the culture vessel, a conventional culture vessel can be used, and is not particularly limited. For example, a seedling pot, a plug tray, etc. are illustrated. The culture vessel may be a closed type or an open type, but a closed type culture vessel is preferable because it facilitates maintaining the humidity of the environment surrounding the cuttings and cuttings and seedlings formed therefrom.

  When a branch is used as the cutting head, it is preferable to use a closed culture vessel as the culture vessel. This makes it easy to place the cutting head under high humidity, so that the transpiration action of the leaves on the branches is suppressed, and the conventional partial excision processing of the leaves can be omitted.

  The culture container is more preferably a container capable of supplying carbon dioxide gas into the container. As such a culture container, a container having an opening covered with a carbon dioxide permeable membrane is exemplified. By using a container having an opening covered with a carbon dioxide permeable membrane, the humidity of the culture environment can be easily adjusted. The shape of the opening is not particularly limited. The material for the carbon dioxide permeable membrane is not particularly limited, and examples thereof include polytetrafluoroethylene. The pore diameter of the membrane is not particularly limited, and examples thereof include membranes having a pore diameter of about 0.1 μm or more and about 1 μm or less.

  In the present invention, the temperature of the support and / or rooting medium is usually 20 ° C to 30 ° C, preferably 22 ° C to 26 ° C.

  In this invention, the process of growing a cutting seedling may be included. The process of growing the cuttings is usually performed after the process of rooting from the cuttings. In the step of growing the cutting seedling, the cutting seedling obtained in the step of rooting from the cutting ear can be transplanted and grown in a seedling container or a nursery. When the cuttings are shoots derived from cultured tissues such as adventitious shoots and shoot primordia, they usually undergo an acclimation step before transplantation to a seedling container or the like. Conditions suitable for cutting seedlings may be set as appropriate for conditions such as temperature, light intensity during cultivation, and the type and amount of soil to be stored in the seedling container. By going through the process of growing cuttings and seedlings, it is possible to obtain a plant that can be used as a raw material for medicines such as traditional Chinese medicines and foods, or secondary metabolites thereof.

Example 1
As a medicinal plant, licorice strain 1 (GuTS71-08IV2 strain) and strain 2 (GuTS71-08IV1 strain) [both strains refer to JP2012-115261A] were used. These are strains that accumulate a large amount of glycyrrhizin as a secondary metabolite. These above-ground parts were collected, branches were cut into about 3 cm to 10 cm so as to include one bud, and the ears were adjusted. A foamed phenol resin molded product (product name: Oasis) was used for the insertion floor. The root bed was infiltrated with 100 ml of 5-fold diluted B5 medium supplemented with 5 mg / ml of IBA. The cutting seedlings were produced by inserting the cuttings adjusted as described above into the root bed, temperature 25 ° C., light intensity (illuminance) 135 μmol / m ² / sec, carbon dioxide concentration 380 ppm, light period 8 hours. This was carried out by culturing in a culture vessel with a dark period of 16 hours. The number of cuttings in each test section was 100. The culture container is a 10 cm square polycarbonate container with holes with a diameter of 1 cm in the top, and this opening is covered with a carbon dioxide permeable membrane ("Milliseal" manufactured by Nihon Millipore). It was. Table 1 shows the rooting rate in each test section after 6 weeks.

Example 2
The procedure was the same as Example 1 except that the carbon dioxide concentration was 1,000 ppm.

Comparative Example 1
The procedure was the same as Example 2 except that the light intensity was 25 μmol / m ² / sec.

  As shown in Table 1, compared with the rooting rate of Comparative Example 1, the rooting rates of Examples 1 and 2 were high, and the rooting rate of Example 2 was remarkable. This result shows that the rooting rate of cuttings of plants that accumulate secondary metabolites can be improved by the production method of the present invention.

Example 3
The procedure was the same as in Example 2 except that cinnamon (total alkaloid content: 0.7% or more) was used as a medicinal plant.

Example 4
Example 3 was repeated except that 50 mg / l of GSSG was added to the medium.

Comparative Example 2
The procedure was the same as Comparative Example 1, except that cinnamon (total alkaloid content 0.7% or more) was used as a medicinal plant.

Comparative Example 3
Comparative Example 2 was performed except that 50 mg / l of GSSG was added to the medium.
Table 2 shows the rooting rate per each test section after 6 weeks.

  As shown in Table 2, the rooting rates of Examples 3 and 4 were higher than those of Comparative Examples 2 and 3. This result shows that the rooting rate of cuttings of plants that accumulate secondary metabolites can be increased by the production method of the present invention.

Claims (5)

Including a step of treating from 2 weeks to 3 months under conditions of light intensity of 120 to 270 μmol / m ² / sec and humidity of 60% Rh or more, and rooting from cuttings that are branches including buds ,
A secondary metabolite containing at least one or more selected from alkaloids and derivatives thereof, or a secondary metabolite containing at least one selected from terpenoids, flavonoids, terpene glycosides and derivatives thereof How to produce medicinal plant cuttings that accumulate .   The production method according to claim 1, wherein the carbon dioxide concentration in the step of rooting from the cutting is 500 to 1500 ppm.   The production method according to claim 1 or 2, wherein 0.01 to 10 mg / L of auxin is added to the medium. The production method according to any one of claims 1 to 3 , wherein the medicinal plant is a licorice plant or a genus plant. Performing the step of rooting the cuttings in a culture vessel, the method of production according to any one of claims 1-4.