JP7020424B2 - An isoprenoid production promoter using squalene as a precursor, and a method for producing an isoprenoid-rich plant using the same. - Google Patents

Description

This application claims priority based on Japanese Patent Application No. 2016-21705, which was filed on November 10, 2016, the entire disclosure of which is incorporated herein by reference. The present invention relates to a squalene-based isoprenoid production promoter used for plants. Hereinafter, in the present specification, "isoprenoid having squalene as a precursor" is abbreviated as IPSQ. The present invention also relates to a method for increasing the IPSQ content in a plant, in other words, a method for producing a plant having a high IPSQ content. Furthermore, the present invention relates to new uses of jasmonic acid compounds.

The present invention also relates to a medium suitable for tissue culture of plants belonging to the genus Licorice of the family Leguminosae among plants containing a high amount of IPSQ.

Glycyrrhizin, glycyrrhetinic acid, ginsenoside, soybean saponin and the like are known as crude drug components having more pharmacological actions than before. These are usually produced from medicinal plants such as the genus Licorice of the family Leguminosae and the genus Panax japonicus of the family Araliaceae. However, under normal cultivation methods and conditions thereof, there is a problem that it takes a long cultivation period to collect a plant containing a predetermined amount of crude drug components. For example, in the case of licorice plants, it is very difficult to cultivate glycyrrhizin so that it contains 2.0% by mass or more as specified by the Japanese Pharmacopoeia in the underground part, and even to make the content close to 2.0% 4 It takes up to 5 years to grow. As a method to solve this, an attempt was made to increase the amount of glycyrrhizin contained in the roots by applying squeezing restraint to the roots of licorice plants in advance or in the process of growth by a squeezing restraint mechanism and continuing to apply stress to the roots. (See, for example, Patent Document 1). As described above, conventionally, there has been a demand for a method for stably cultivating and collecting a plant containing a predetermined amount of crude drug components more efficiently.

In addition, for the purpose of stable production and supply of crude drug components, tissue culture of plants has been studied instead of the conventional cultivation of plants. For example, for the purpose of stable production of glycyrrhizin, an adventitious root culture method of a plant of the genus Licorice (Patent Document 2) and a tissue culture method using axillary buds (see Patent Document 3, Non-Patent Document 1, etc.) are available. Proposed.

Japanese Unexamined Patent Publication No. 2012-170344 Japanese Unexamined Patent Publication No. 1-291725 Japanese Unexamined Patent Publication No. 4-111824

Plant Tissue Culture Letters, 12 (2), 145-149 (1995)

In view of the above background, the present invention is used to promote the production of IPSQ in the above-mentioned squalene-based plant containing isoprenoid (IPSQ) and to increase the content thereof. It is an object of the present invention to provide a production promoter. Another object of the present invention is to provide a method for increasing the content of a plant containing IPSQ, in other words, a method for producing a plant containing a high IPSQ. Another object of the present invention is to provide the above-mentioned new uses for jasmonic acid compounds.

Further, the present invention provides a medium suitable for tissue culture of a plant belonging to the genus Licorice of the family Leguminosae among plants containing a high amount of IPSQ, particularly a medium capable of promoting rooting and improving the rooting rate. With the goal.

As a result of diligent studies to solve the above object, the present inventors have made a general formula (1) described later in the growth process of a plant containing isoprenoid (IPSQ) using squalene as a precursor, wherein at least a part of the plant is described later. It was found that the treatment with the jasmonic acid compound shown in (1) significantly increases the IPSQ content as compared with the untreated plant of the same species. That is, the jasmonic acid compound has an action of promoting the production of the IPSQ on the IPSQ-containing plant, and can be effectively used as an IPSQ production promoting agent. Therefore, by treating all or a part of the plant body of the plant that produces and contains IPSQ with a jasmonic acid compound in the growth process, a plant containing a large amount of IPSQ or a part thereof can be produced and obtained.

The present invention has been completed based on such findings, and has the following embodiments.
Hereinafter, in the present specification, the following abbreviations may be used for description.
Isoprenoid: IP
Squalene: SQ
Squalene-precursor isoprenoids: IPSQ
Triterpenes: TT
Triterpene glycosides: TTG
Steroid: ST
Steroid glycosides: STG
Glycyrrhizin: GL
Glycyrrhetinic acid: GA
3-Monoglucronyl glycyrrhetinic acid: 3MGA
Jasmonic acid compound: JA compound

（式中、Ｒ^１は１～２０の炭素を有する炭化水素基、Ｒ^２は水素原子または１～２０の炭素を有する炭化水素基を意味する）。
（Ｉ－２）さらに界面活性剤を含有する、（Ｉ－１）に記載する植物用ＩＰＳＱ生成促進剤。
（Ｉ－３）前記ＪＡ化合物が、ジャスモン酸、プロヒドロジャスモン、及びジャスモン酸メチルからなる群より選択される少なくとも１種である、（Ｉ－１）または（Ｉ－２）に記載する植物用ＩＰＳＱ生成促進剤。
（Ｉ－４）ＩＰＳＱが、トリテルペン（ＴＴ）、トリテルペン配糖体（ＴＴＧ）、及びステロイド配糖体（ＳＴＧ）からなる群より選択される少なくとも１種である、（Ｉ－１）～（Ｉ－３）のいずれかに記載する植物用ＩＰＳＱ生成促進剤。
（Ｉ－５）前記ＩＰＳＱがＴＴであり、当該ＴＴが、グリチルレチン酸、リキリチン酸、グラブリン酸、α－アミリン、トルメンティック酸、コロソリン酸、ウルソール酸、アリソール、ヘルボリン酸、フシジン酸、ラノステロール、エブリコ酸、ポリポレン酸、ツムロシン酸、パキマ酸、シクロアルテノール、シミシフゲノール、シミゲノール、ダマレンジオール、プロトパナキサジオール、プロトパナキサトリオール、ユーホール、ユーホルボール、エレモール酸、エレモン酸、ルペオール、ベツリン、ベツリン酸、リモニン、エボドール、ルタエビン、オバクノン、クァッシノイド、ブルセアンチン、マスリン酸、２α，１９α－ジヒドロキシ－３－オキソウルス－１２－エン－２８－オイック酸、β－アミリン、オレアノール酸、フペンジック酸、ヘラゲニン、プラティコディゲニン、ポリガラシン酸、プレセネゲニン、サイコゲニン、チャサポゲノール、及びこれらの塩からなる群より選択される少なくとも１種である、（Ｉ－４）に記載する植物用ＩＰＳＱ生成促進剤。
（Ｉ－６）前記植物が、マメ科カンゾウ属、バラ科マルメロ属、ナシ属、ナナカマド属、サンザシ属、シャリントウ属、ニガキ科ニガキ属、ミカン科キハダ属、ウコギ科オタネニンジン属、オモダカ科サジオモダカ属、マチン科マチン属、トウダイグサ科トウダイグサ属、イネ科イネ属、オオムギ属、チガヤ属、アブラナ科アブラナ属、キンポウゲ科サラシナショウマ属、フタバガキ科サラノキ属、ＨＯＰＥＡ属、マメ科ルピナス属、カバノキ科カバノキ属、クロウメモドキ科ナツメ属、カンラン科カンラン属、シソ科ウツボグサ属、ツツジ科アルクトスタフィロス属、オオバコ科オオバコ属、モチノキ科モチノキ属、リンドウ科リンドウ属、フトモモ科フトモモ属、キキョウ科キキョウ属、ヒメハギ科ヒメハギ属、セリ科ミシマサイコ属、ツバキ科ツバキ属、ミカン科キハダ属、またはゴシュユ属からなる群より選択される少なくとも１つに属するＴＴ含有植物である、（Ｉ－５）に記載する植物用ＩＰＳＱ生成促進剤。
（Ｉ－７）前記ＩＰＳＱがＴＴＧであり、当該ＴＴＧが、グリチルリチン（ＧＬ）、３－モノグルクロニルグリチルレチン酸（３ＭＧＡ）、バッカロシドＢ、アベナシンＡ－１、大豆サポニン、ギンセノシド、ウラルサポニン、チクセツサポニン、エレウテロシドＡ、アケボシド、プラティコジン、セネギン、オンジサポニン、サイコサポニン、テアサポニン、アスターサポニン、シビリコシド、スミラックスサポニン、エスチン、アラロシド、アストラガロシド、ギムネマ酸、ギベノサイド及びこれらの塩からなる群より選択される少なくとも１種である、（Ｉ－４）に記載する植物用ＩＰＳＱ生成促進剤。
（Ｉ－８）前記植物が、マメ科カンゾウ属、マメ科マメ属、マメ科アカシア属、マメ科ゲンゲ属、ウコギ科トチバニンジン属、ウコギ科ウコギ属、ウコギ科タラノキ属、アケビ科アケビ属植物、キク科シオン属、キキョウ科キキョウ属、ヒメハギ科ヒメハギ属、ユリ科アマドコロ属、ユリ科シオデ属、ツバキ科ツバキ属、セリ科ミシマサイコ属、ナデシコ科ドウカンソウ属、ナデシコ科サボンソウ属、トチノキ科トチノキ属、イネ科カラスムギ属、キョウチクトウ科ギムネマ属、及びウリ科アマチャズル属からなる群より選択される少なくとも１つに属するＴＴＧ含有植物である、（Ｉ－７）に記載する植物用ＩＰＳＱ生成促進剤。
（Ｉ－９）前記ＩＰＳＱがＳＴＧであり、当該ＳＴＧが、プレグナン配糖体、エクジステロン、イノコステロン、ストロファンチン、スミラックスサポニン、ヘレブリン、ジオスシン、オフィオポゴニン、スミラゲニン配糖体、ジギトニン、ギトニン、サルササポニン、ジギトキシン、ラナトシド、ギトキシン、ギタロキシン、コンバラトキシン、シラレン及びチモサポニンからなる群より選択される少なくとも１種である、（Ｉ－４）に記載する植物用ＩＰＳＱ生成促進剤。
（Ｉ－１０）前記植物が、ガガイモ科キジョラン属、ヒユ科イノコズチ属、キョウチクトウ科ストロファンツス属、キンポウゲ科オウレン属、ヤマノイモノ科ヤマノイモ属、ヒガンバナ科リュウゼツラン属、ユリ科シオデ属、ハナスゲ属、ジャノヒゲ属、ショウジョウバカマ属、ハラン属、ゴマノハグサ科ジギタリス属、ナス科ウィザニア属からなる群より選択される少なくとも１つに属するＳＴＧ含有植物である、（Ｉ－９）に記載する植物用ＩＰＳＱ生成促進剤。
（Ｉ－１１）前記式（１）で示されるＪＡ化合物またはそれを含む組成物の、植物用ＩＰＳＱ生成促進剤の製造のための使用。
（Ｉ－１２）前記組成物がさらに界面活性剤を含有するものである、（Ｉ－１１）に記載する使用。
（Ｉ－１３）ＩＰＳＱ含有植物に対して、当該植物体におけるＩＰＳＱの生成を促進するために使用される、前記式（１）で示されるＪＡ化合物またはそれを含む組成物。
（Ｉ－１４）前記組成物がさらに界面活性剤を含有するものである、（Ｉ－１３）に記載するＪＡ化合物またはそれを含む組成物。 (I) IPSQ production promoter (I-1) A plant IPSQ production promoter containing a jasmonic acid compound (JA compound) represented by the following formula (1):

(In the formula, R ¹ means a hydrocarbon group having 1 to 20 carbons, and R ² means a hydrogen atom or a hydrocarbon group having 1 to 20 carbons).
(I-2) The IPSQ production promoter for plants according to (I-1), which further contains a surfactant.
(I-3) The plant use according to (I-1) or (I-2), wherein the JA compound is at least one selected from the group consisting of jasmonic acid, prohydrojasmonate, and methyl jasmonate. IPSQ production promoter.
(I-4) IPSQ is at least one selected from the group consisting of triterpenes (TT), triterpene glycosides (TTGs), and steroid glycosides (STGs), (I-1) to (I-1). The IPSQ production promoter for plants according to any one of -3).
(I-5) The IPSQ is TT, and the TT is glycyrrhetinic acid, liquiric acid, gravurinic acid, α-amyrin, tolmentic acid, corosolic acid, ursoric acid, arisole, herboric acid, fusidic acid, lanosterol, Ebricoic acid, polyporenic acid, tumrosic acid, pachymaic acid, cycloartenol, amyrin, simigenor, damalendiol, protopanaxadiol, protopanaxatriol, euhole, euphorbol, elemolic acid, elemonic acid, lupeol, bethrin, bethrinic acid , Limonin, Evodol, Lutaebin, Obakunon, Quassinoid, Bruceanthin, Masphosphate, 2α, 19α-Dihydroxy-3-oxourus-12-en-28-Oic acid, β-Amyrin, Oleanolic acid, Fupendic acid, Heragenin, Platicodigenin The IPSQ production promoter for plants according to (I-4), which is at least one selected from the group consisting of polygalassic acid, presenegenin, psychogenin, chasapogenol, and salts thereof.
(I-6) The plants are the genus Kanzo, the genus Marumero, the genus Nanakamado, the genus Sanzashi, the genus Sharinto, the genus Nigaki, the genus Kihada, the genus Otaneninjin, and the genus Sagiomodaka. , Matin family Matin genus, Todaigusa family Todaigusa genus, Rice family Rice genus, Omugi genus, Chigaya genus, Abrana family Abrana genus, Kinpouge family Sarashinashoma genus, Futabagaki family Saranoki genus, HOPEA genus , Crow Meadow family Natsume genus, Kanran family Kanran genus, Shiso family Utsubogusa genus, Tsutsuji family Arctostaphyros genus, Oobaco family Oobako genus, Mochinoki family Mochinoki genus, Lindou family Lindou genus, Futomomo family Futomomo genus, Kikyo family IPSQ for plants according to (I-5), which is a TT-containing plant belonging to at least one selected from the group consisting of the genus Himehagi, the genus Mishimapsycho of the family Seri, the genus Tsubaki of the family Tsubaki, the genus Kihada of the family Mikan, or the genus Goshuyu. Production accelerator.
(I-7) The IPSQ is TTG, and the TTG is glycyrrhizin (GL), 3-monoglucuronyl glycyrrhetinic acid (3MGA), baccalocide B, avenacin A-1, soybean saponin, ginsenoside, uralsaponin, chiku. From the group consisting of setusaponin, eleutheroside A, akeboside, platicodin, senegin, ondisaponin, psychosaponin, theasaponin, astersaponin, cibilicoside, smilax saponin, estin, aralocid, astragaloside, gymnemic acid, givenoside and salts thereof. The IPSQ production promoter for plants according to (I-4), which is at least one selected.
(I-8) The plants include the genus Araliaceae, the genus Dianthus, the genus Acacia, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, and the genus Araliaceae. Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae The IPSQ production promoter for plants according to (I-7), which is a TTG-containing plant belonging to at least one selected from the group consisting of the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, and the genus Araliaceae.
(I-9) The IPSQ is STG, and the STG is pregnane glycoside, ecdysterone, inocosterone, strophanthin, smilax saponin, hellebrin, dioscin, offiopogonin, sumiragenin glycoside, digitonin, gitonin, sarsasapogenin. The IPSQ production promoter for plants according to (I-4), which is at least one selected from the group consisting of digitoxin, lanatoside, gitoxin, gitaroxin, convallatoxin, silalene and thymosapogenin.
(I-10) The plants are aspidistra, Achyranthes, Apocynaceae, Coptis genus, Ranunculaceae, Coptis genus, Higanbana, Ryuzetsuran, Yuri, Shiode, Anemarrhena, and Ophiopogon. The IPSQ production promoter for plants according to (I-9), which is an STG-containing plant belonging to at least one selected from the group consisting of the genus Apocynaceae, the genus Aspidistra, the genus Ophiopogon, and the genus Achyranthes. ..
(I-11) Use of the JA compound represented by the above formula (1) or a composition containing the same for producing an IPSQ production promoter for plants.
(I-12) The use according to (I-11), wherein the composition further contains a surfactant.
(I-13) A JA compound represented by the above formula (1) or a composition containing the same, which is used for promoting the production of IPSQ in a plant containing IPSQ.
(I-14) The JA compound according to (I-13) or a composition containing the same, wherein the composition further contains a surfactant.

（式中、Ｒ^１は１～２０の炭素を有する炭化水素基、Ｒ^２は水素原子または１～２０の炭素を有する炭化水素基を意味する）、
（ｂ）前記工程（ａ）で処理された植物を生育させる工程。
（II－２）前記ＪＡ化合物含有組成物がさらに界面活性剤を含有するものである、（II－１）に記載する作製方法。
（II－３）前記ＪＡ化合物が、ジャスモン酸、プロヒドロジャスモン、及びジャスモン酸メチルからなる群より選択される少なくとも１種である、（II－１）または（II－２）に記載する作製方法。
（II－４）ＩＰＳＱが、トリテルペン（ＴＴ）、トリテルペン配糖体（ＴＴＧ）、及びステロイド配糖体（ＳＴＧ）からなる群より選択される少なくとも１種である、（II－１）～（II－３）のいずれかに記載する作製方法。
（II－５）前記ＩＰＳＱがＴＴであり、当該ＴＴが、グリチルレチン酸、リキリチン酸、グラブリン酸、α－アミリン、トルメンティック酸、コロソリン酸、ウルソール酸、アリソール、ヘルボリン酸、フシジン酸、ラノステロール、エブリコ酸、ポリポレン酸、ツムロシン酸、パキマ酸、シクロアルテノール、シミシフゲノール、シミゲノール、ダマレンジオール、プロトパナキサジオール、プロトパナキサトリオール、ユーホール、ユーホルボール、エレモール酸、エレモン酸、ルペオール、ベツリン、ベツリン酸、リモニン、エボドール、ルタエビン、オバクノン、クァッシノイド、ブルセアンチン、マスリン酸、２α，１９α－ジヒドロキシ－３－オキソウルス－１２－エン－２８－オイック酸、β－アミリン、オレアノール酸、フペンジック酸、ヘラゲニン、プラティコディゲニン、ポリガラシン酸、プレセネゲニン、サイコゲニン、チャサポゲノール、及びこれらの塩からなる群より選択される少なくとも１種である、（II－４）に記載する作製方法。
（II－６）前記植物が、マメ科カンゾウ属、ルピナス属、バラ科マルメロ属、ナシ属、ナナカマド属、サンザシ属、シャリントウ属、ニガキ科ニガキ属、ミカン科キハダ属、ウコギ科オタネニンジン属、オモダカ科サジオモダカ属、マチン科マチン属、トウダイグサ科トウダイグサ属、イネ科イネ属、オオムギ属、チガヤ属、アブラナ科アブラナ属、キンポウゲ科サラシナショウマ属、フタバガキ科サラノキ属、ＨＯＰＥＡ属、カバノキ科カバノキ属、クロウメモドキ科ナツメ属、カンラン科カンラン属、シソ科ウツボグサ属、ツツジ科アルクトスタフィロス属、オオバコ科オオバコ属、モチノキ科モチノキ属、リンドウ科リンドウ属、フトモモ科フトモモ属、キキョウ科キキョウ属、ヒメハギ科ヒメハギ属、セリ科ミシマサイコ属、ツバキ科ツバキ属、ミカン科キハダ属、またはゴシュユ属からなる群より選択される少なくとも１つに属するＴＴ含有植物である、（II－５）に記載する作製方法。
（II－７）前記ＩＰＳＱがＴＴＧであり、当該ＴＴＧが、グリチルリチン（ＧＬ）、３－モノグルクロニルグリチルレチン酸（３ＭＧＡ）、バッカロシドＢ、アベナシンＡ－１、大豆サポニン、ギンセノシド、ウラルサポニン、チクセツサポニン、エレウテロシドＡ、アケボシド、プラティコジン、セネギン、オンジサポニン、サイコサポニン、テアサポニン、アスターサポニン、シビリコシド、スミラックスサポニン、エスチン、アラロシド、アストラガロシド、ギムネマ酸、ギベノサイド、及びそれらの塩からなる群より選択される少なくとも１種である、（II－４）に記載する作製方法。
（II－８）前記植物が、マメ科カンゾウ属、マメ科マメ属、マメ科アカシア属、マメ科ゲンゲ属、ウコギ科トチバニンジン属、ウコギ科ウコギ属、ウコギ科タラノキ属、アケビ科アケビ属植物、キク科シオン属、キキョウ科キキョウ属、ヒメハギ科ヒメハギ属、ユリ科アマドコロ属、ユリ科シオデ属、ツバキ科ツバキ属、セリ科ミシマサイコ属、ナデシコ科ドウカンソウ属、ナデシコ科サボンソウ属、トチノキ科トチノキ属、イネ科カラスムギ属、キョウチクトウ科ギムネマ属、及びウリ科アマチャズル属からなる群より選択される少なくとも１つに属するＴＴＧ含有植物である、（II－７）に記載する作製方法。
（II－９）前記ＩＰＳＱがＳＴＧであり、当該ＳＴＧが、プレグナン配糖体、エクジステロン、イノコステロン、ストロファンチン、スミラックスサポニン、ヘレブリン、ジオスシン、オフィオポゴニン、スミラゲニン配糖体、ジギトニン、ギトニン、サルササポニン、ジギトキシン、ラナトシド、ギトキシン、ギタロキシン、コンバラトキシン、シラレン、及びチモサポニンからなる群より選択される少なくとも１種である、（II－４）に記載する作製方法。
（II－１０）前記植物が、ガガイモ科キジョラン属、ヒユ科イノコズチ属、キョウチクトウ科ストロファンツス属、キンポウゲ科オウレン属、ヤマノイモノ科ヤマノイモ属、ヒガンバナ科リュウゼツラン属、ユリ科シオデ属、ハナスゲ属、ジャノヒゲ属、ショウジョウバカマ属、ハラン属、ゴマノハグサ科ジギタリス属、ナス科ウィザニア属からなる群より選択される少なくとも１つに属するＳＴＧ含有植物である、（II－９）に記載する作製方法。
（II－１１）前記工程（ａ）が、ＩＰＳＱ含有植物の植物体全体またはその一部を、ＪＡ化合物を０．００１～５質量％含む水溶液で処理する工程である、（II－１）～（II－１０）のいずれかに記載する作製方法。
（II－１２）前記工程（ａ）が、ＩＰＳＱ含有植物の地上部位を処理する工程である、（II－１）～（II－１１）のいずれかに記載する作製方法。
（II－１３）前記ＩＰＳＱ含有植物が、ＴＴＧを含有するマメ科カンゾウ属植物であって、第十七改正日本薬局方に準拠した方法で測定される地下部１００質量％中のＧＬ含量が乾燥物換算で２．０質量％以上のＩＰＳＱ含有植物を作製する方法である、（II－１）～（II－１２）のいずれかに記載する作製方法。
（II－１４）前記ＩＰＳＱ含有植物が、ＴＴＧを含有するウコギ科トチバニンジン属植物であって、第十七改正日本薬局方に準拠した方法で測定される主根１００質量％中のギンセノシドＲｂ１及びＲｇ１含量が、乾燥物換算でそれぞれ０．２質量％以上及び０．１質量％以上のＩＰＳＱ含有植物を作製する方法である、（II－１）～（II－１２）のいずれかに記載する作製方法。 (II) Method for producing IPSQ-rich plant and its processed product (II-1) Method for producing IPSQ-containing plant or its processed product having the following steps (a) and (b):
(A) A step of treating an entire plant or a part thereof of an IPSQ-containing plant with a JA compound represented by the following formula (1) or a composition containing the same.

(In the formula, R ¹ means a hydrocarbon group having 1 to 20 carbons, R ² means a hydrogen atom or a hydrocarbon group having 1 to 20 carbons),
(B) A step of growing the plant treated in the step (a).
(II-2) The production method according to (II-1), wherein the JA compound-containing composition further contains a surfactant.
(II-3) The production method according to (II-1) or (II-2), wherein the JA compound is at least one selected from the group consisting of jasmonic acid, prohydrojasmonate, and methyl jasmonate. ..
(II-4) IPSQ is at least one selected from the group consisting of triterpene (TT), triterpene glycoside (TTG), and steroid glycoside (STG), (II-1) to (II). The production method according to any one of -3).
(II-5) The IPSQ is TT, and the TT is glycyrrhetinic acid, liquiric acid, gravurinic acid, α-amyrin, tolmentic acid, corosolic acid, ursoric acid, arisole, herboric acid, fusidic acid, lanosterol, Everycic acid, polyporenic acid, tumrosic acid, pachymaic acid, cycloartenol, amyrin, simigenor, damalendiol, protopanaxadiol, protopanaxatriol, euhole, euphorbol, elemolic acid, elemonic acid, lupeol, bethrin, bethrinic acid , Limonin, Evodol, Lutaebin, Obakunon, Quassinoid, Bruceanthin, Masphosphate, 2α, 19α-Dihydroxy-3-oxourus-12-en-28-Oic acid, β-Amyrin, Oleanolic acid, Fupendic acid, Heragenin, Platicodigenin , Polygalassic acid, presenegenin, psychogenin, chasapogenol, and at least one selected from the group consisting of salts thereof, according to the production method according to (II-4).
(II-6) The plants include the genus Kanzo, the genus Lupinus, the genus Marumero, the genus Pear, the genus Nanakamado, the genus Sanzashi, the genus Sharinto, the genus Nigaki, the genus Kihada, the genus Otaneninjin, and the genus Omodaka. Family Sagiomodaka, Matin family Matin genus, Todaigusa family Todaigusa genus, Rice family Rice genus, Omugi genus, Chigaya genus, Abrana family Abrana genus, Kinpouge family Sarashinashoma genus, Futabagaki family Saranoki genus, HOPEA genus, Kabanoki family Family Natsume, Kanran, Kanran, Shiso, Arctostaphyros, Oobaco, Oobako, Mochinoki, Lindou, Futomomo, Kikyo, Himehagi The production method according to (II-5), wherein the TT-containing plant belongs to at least one selected from the group consisting of the genus Mishima Psycho of the family Seri, the genus Tsubaki of the family Tsubaki, the genus Kihada of the family Mikan, or the genus Goshuyu.
(II-7) The IPSQ is TTG, and the TTG is glycyrrhizin (GL), 3-monoglucuronyl glycyrrhetinic acid (3MGA), baccalocide B, avenacin A-1, soybean saponin, ginsenoside, uralsaponin, chiku. A group consisting of setusaponin, eleutheroside A, akeboside, platicodin, senegin, ondisaponin, psychosaponin, theasaponin, astersaponin, cibilicoside, smilax saponin, estin, aralocid, astragaloside, gymnemic acid, givenoside, and salts thereof. The production method according to (II-4), which is at least one selected from the above.
(II-8) The plants include the genus Araliaceae, the genus Spikenard, the genus Acacia, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, and the genus Araliaceae. Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae The production method according to (II-7), which is a TTG-containing plant belonging to at least one selected from the group consisting of the genus Araliaceae, the genus Spikenard, the genus Araliaceae, and the genus Araliaceae.
(II-9) The IPSQ is STG, and the STG is pregnane glycoside, ecdysterone, inocosterone, strophanthin, smilax saponin, hellebrin, dioscin, offiopogonin, sumiragenin glycoside, digitonin, gitonin, sarsasapogenin. , Digitoxin, lanatoside, gitoxin, gitaroxin, convallatoxin, silalene, and thymosapogenin, which is at least one selected from the group, according to (II-4).
(II-10) The plants are aspidistra, Achyranthes, Apocynaceae, Coptis genus, Ranunculaceae, Coptis genus, Higanbana, Ryuzetsuran, Yuri, Shiode, Anemarrhena, and Ophiopogon. The production method according to (II-9), which is an STG-containing plant belonging to at least one selected from the group consisting of the genus Apocynaceae, the genus Aspidistra, the genus Ophiopogon, the genus Ophiopogon, and the genus Achyranthes.
(II-11) The step (a) is a step of treating the whole or a part of the IPSQ-containing plant with an aqueous solution containing 0.001 to 5% by mass of the JA compound, (II-1) to The production method according to any one of (II-10).
(II-12) The production method according to any one of (II-1) to (II-11), wherein the step (a) is a step of treating the above-ground part of the IPSQ-containing plant.
(II-13) The IPSQ-containing plant is a leguminous licorice plant containing TTG, and the GL content in 100% by mass of the underground portion measured by a method in accordance with the 17th revised Japanese Pharmacopoeia is dry. The production method according to any one of (II-1) to (II-12), which is a method for producing a plant containing IPSQ in an amount of 2.0% by mass or more in terms of material.
(II-14) The IPSQ-containing plant is a TTG-containing plant belonging to the genus Panax japonicus in the family Araliaceae, and contains ginsenoside Rb1 and Rg1 in 100% by mass of the main root measured by a method in accordance with the 17th revised Japanese Pharmacopoeia. Is a method for producing IPSQ-containing plants of 0.2% by mass or more and 0.1% by mass or more in terms of dried matter, respectively, according to any one of (II-1) to (II-12). ..

（式中、Ｒ^１は１～２０の炭素を有する炭化水素基、Ｒ^２は水素原子または１～２０の炭素を有する炭化水素基を意味する）。
（III－２）上記組成物がさらに界面活性剤を含有するものである、（III－１）に記載する方法。
（III－３）前記ＪＡ化合物が、ジャスモン酸、プロヒドロジャスモン、及びジャスモン酸メチルからなる群より選択される少なくとも１種である、（III－１）または（III－２）に記載する方法。
（III－４）前記ＩＰＳＱが、トリテルペン（ＴＴ）、トリテルペン配糖体（ＴＴＧ）、及びステロイド配糖体（ＳＴＧ）からなる群より選択される少なくとも１種である、（III－１）～（III－３）のいずれかに記載する方法。
（III－５）前記ＩＰＳＱがＴＴであり、当該ＴＴが、グリチルレチン酸、リキリチン酸、グラブリン酸、α－アミリン、トルメンティック酸、コロソリン酸、ウルソール酸、アリソール、ヘルボリン酸、フシジン酸、ラノステロール、エブリコ酸、ポリポレン酸、ツムロシン酸、パキマ酸、シクロアルテノール、シミシフゲノール、シミゲノール、ダマレンジオール、プロトパナキサジオール、プロトパナキサトリオール、ユーホール、ユーホルボール、エレモール酸、エレモン酸、ルペオール、ベツリン、ベツリン酸、リモニン、エボドール、ルタエビン、オバクノン、クァッシノイド、ブルセアンチン、マスリン酸、２α，１９α－ジヒドロキシ－３－オキソウルス－１２－エン－２８－オイック酸、β－アミリン、オレアノール酸、フペンジック酸、ヘラゲニン、プラティコディゲニン、ポリガラシン酸、プレセネゲニン、サイコゲニン、チャサポゲノール、及びこれらの塩からなる群より選択される少なくとも１種である、（III－４）に記載する方法。
（III－６）前記植物が、マメ科カンゾウ属、ルピナス属、バラ科マルメロ属、ナシ属、ナナカマド属、サンザシ属、シャリントウ属、ニガキ科ニガキ属、ミカン科キハダ属、ウコギ科オタネニンジン属、オモダカ科サジオモダカ属、マチン科マチン属、トウダイグサ科トウダイグサ属、イネ科イネ属、オオムギ属、チガヤ属、アブラナ科アブラナ属、キンポウゲ科サラシナショウマ属、フタバガキ科サラノキ属、ＨＯＰＥＡ属、カバノキ科カバノキ属、クロウメモドキ科ナツメ属、カンラン科カンラン属、シソ科ウツボグサ属、ツツジ科アルクトスタフィロス属、オオバコ科オオバコ属、モチノキ科モチノキ属、リンドウ科リンドウ属、フトモモ科フトモモ属、キキョウ科キキョウ属、ヒメハギ科ヒメハギ属、セリ科ミシマサイコ属、ツバキ科ツバキ属、ミカン科キハダ属、またはゴシュユ属からなる群より選択される少なくとも１種のＴＴ含有植物である、（III－５）に記載する方法。
（III－７）前記ＩＰＳＱがＴＴＧであり、当該ＴＴＧが、グリチルリチン（ＧＬ）、３－モノグルクロニルグリチルレチン酸（３ＭＧＡ）、バッカロシドＢ、アベナシンＡ－１、大豆サポニン、ギンセノシド、ウラルサポニン、チクセツサポニン、エレウテロシドＡ、アケボシド、プラティコジン、セネギン、オンジサポニン、サイコサポニン、テアサポニン、アスターサポニン、シビリコシド、スミラックスサポニン、エスチン、アラロシド、アストラガロシド、ギムネマ酸、ギベノサイド、及びこれらの塩からなる群より選択される少なくとも１種である、（III－４）に記載する方法。
（III－８）前記植物が、マメ科カンゾウ属、マメ科マメ属、マメ科アカシア属、マメ科ゲンゲ属、ウコギ科トチバニンジン属、ウコギ科ウコギ属、ウコギ科タラノキ属、アケビ科アケビ属、キク科シオン属、キキョウ科キキョウ属、ヒメハギ科ヒメハギ属、ユリ科アマドコロ属、ユリ科シオデ属、ツバキ科ツバキ属、セリ科ミシマサイコ属、ナデシコ科ドウカンソウ属、ナデシコ科サボンソウ属、トチノキ科トチノキ属、イネ科カラスムギ属、キョウチクトウ科ギムネマ属、及びウリ科アマチャズル属からなる群より選択される少なくとも１種のＴＴＧ含有植物である、（III－７）に記載する方法。
（III－９）前記ＩＰＳＱがＳＴＧであり、当該ＳＴＧが、プレグナン配糖体、エクジステロン、イノコステロン、ストロファンチン、スミラックスサポニン、ヘレブリン、ジオスシン、オフィオポゴニン、スミラゲニン配糖体、ジギトニン、ギトニン、サルササポニン、ジギトキシン、ラナトシド、ギトキシン、ギタロキシン、コンバラトキシン、シラレン、及びチモサポニンからなる群より選択される少なくとも１種である、（III－４）に記載する作製方法。
（III－１０）前記植物が、ガガイモ科キジョラン属、ヒユ科イノコズチ属、キョウチクトウ科ストロファンツス属、キンポウゲ科オウレン属、ヤマノイモノ科ヤマノイモ属、ヒガンバナ科リュウゼツラン属、ユリ科シオデ属、ハナスゲ属、ジャノヒゲ属、ショウジョウバカマ属、ハラン属、ゴマノハグサ科ジギタリス属、ナス科ウィザニア属からなる群より選択される少なくとも１種のＳＴＧ含有植物である、（III－９）に記載する方法。
（III－１１）処理工程が、ＩＰＳＱ含有植物の植物体全体またはその一部を、ＪＡ化合物を０．００１～５質量％含む水溶液で処理する工程である、（III－１）～（III－１０）のいずれかに記載する方法。
（III－１２）処理工程が、ＩＰＳＱ含有植物の地上部位を処理する工程である、（III－１）～（III－１１）のいずれかに記載する方法。 (III) Use of JA compound or composition containing it (III-1) The present invention comprises a step of treating an IPSQ-containing plant using the JA compound represented by the following formula (1) or a composition containing it. Method for promoting the production of IPSQ in plants

(In the formula, R ¹ means a hydrocarbon group having 1 to 20 carbons, and R ² means a hydrogen atom or a hydrocarbon group having 1 to 20 carbons).
(III-2) The method according to (III-1), wherein the composition further contains a surfactant.
(III-3) The method according to (III-1) or (III-2), wherein the JA compound is at least one selected from the group consisting of jasmonic acid, prohydrojasmonate, and methyl jasmonate.
(III-4) The IPSQ is at least one selected from the group consisting of triterpenes (TT), triterpene glycosides (TTGs), and steroid glycosides (STGs), (III-1) to (III-4). The method described in any of III-3).
(III-5) The IPSQ is TT, and the TT is glycyrrhetinic acid, liquiric acid, gravurinic acid, α-amyrin, tolmentic acid, corosolic acid, ursoric acid, arisole, herboric acid, fusidic acid, lanosterol, Everycic acid, polyporenic acid, tumrosic acid, pachymaic acid, cycloartenol, amyrin, simigenor, damalendiol, protopanaxadiol, protopanaxatriol, euhole, euphorbol, elemolic acid, elemonic acid, lupeol, bethrin, bethrinic acid , Limonin, Evodol, Lutaebin, Obakunon, Quassinoid, Bruceanthin, Masphosphate, 2α, 19α-Dihydroxy-3-oxourus-12-en-28-Oic acid, β-Amyrin, Oleanolic acid, Fupendic acid, Heragenin, Platicodigenin , Polygalassic acid, presenegenin, psychogenin, chasapogenol, and at least one selected from the group consisting of salts thereof, according to (III-4).
(III-6) The plants include the genus Kanzo, the genus Lupinus, the genus Marumero, the genus Pear, the genus Nanakamado, the genus Sanzashi, the genus Sharinto, the genus Nigaki, the genus Kihada, the genus Otaneninjin, and the genus Omodaka. Family Sagiomodaka, Matin family Matin genus, Todaigusa family Todaigusa genus, Rice family Rice genus, Omugi genus, Chigaya genus, Abrana family Abrana genus, Kinpouge family Sarashinashoma genus, Futabagaki family Saranoki genus, HOPEA genus, Kabanoki family Family Natsume, Kanran, Kanran, Shiso, Arctostaphyros, Oobaco, Oobako, Mochinoki, Lindou, Futomomo, Kikyo, Himehagi , The method according to (III-5), wherein the plant is at least one TT-containing plant selected from the group consisting of the genus Mishima Psycho, the genus Tsubaki, the genus Kihada of the family Mikan, or the genus Goshuyu.
(III-7) The IPSQ is TTG, and the TTG is glycyrrhizin (GL), 3-monoglucuronyl glycyrrhetinic acid (3MGA), baccalocide B, avenacin A-1, soybean saponin, ginsenoside, uralsaponin, chiku. A group consisting of setusaponin, eleutheroside A, akeboside, platicodin, senegin, ondisaponin, psychosaponin, theasaponin, astersaponin, cibilicoside, smilax saponin, estin, aralocid, astragaloside, gymnemic acid, givenoside, and salts thereof. The method according to (III-4), which is at least one selected from the above.
(III-8) The plants are the genus Kanzo, the genus Araliaceae, the genus Acacia, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, and the genus Kiku. Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae The method according to (III-7), which is at least one TTG-containing plant selected from the group consisting of the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, and the genus Araliaceae.
(III-9) The IPSQ is STG, and the STG is pregnane glycoside, ecdysterone, inocosterone, strophanthin, smilax saponin, hellebrin, dioscin, offiopogonin, sumiragenin glycoside, digitonin, gitonin, sarsasapogenin. , Digitoxin, lanatoside, gitoxin, gitaroxin, convallatoxin, silalene, and thymosapogenin, which is at least one selected from the group, according to (III-4).
(III-10) The plants are: Gagaimo family Kijoran genus, Hiyu family Inokozuchi genus, Kyochikuto family Strophanthus genus, Kinpouge family Auren genus, Yamanoimono family Yamanoimo genus, Higanbana family Ryuzetsuran genus, Yuri family Shiode genus, Anemarrhena genus , The method according to (III-9), wherein the plant is at least one STG-containing plant selected from the group consisting of the genus Anemarrhena, the genus Anemarrhena, the genus Anemarrhena, the genus Anemarrhena, and the genus Anemarrhena.
(III-11) The treatment step is a step of treating the whole or a part of the IPSQ-containing plant with an aqueous solution containing 0.001 to 5% by mass of the JA compound, (III-1) to (III-). The method according to any one of 10).
(III-12) The method according to any one of (III-1) to (III-11), wherein the treatment step is a step of treating the above-ground part of the IPSQ-containing plant.

(IV) Medium for tissue culture of Licorice genus plant, method for promoting rooting using the same, and method for molding and production of Licorice genus plant, the present inventors, medium for tissue culture of Licorice genus plant. As a result, the rooting rate can be significantly increased by using WPM (Woody Plant medium) medium, which is conventionally used for tissue culture of wood (tree) but not for tissue culture of herbaceous plants. I found. Specifically, as shown in the experimental examples described later, the rooting rate is significantly increased by using the WPM medium as compared with the case of using the MS medium conventionally used for tissue culture of herbs. It has been found that it is possible to improve the rooting rate to 100% or nearly 100% without using the plant hormone auxin. The present invention has been completed based on such findings, and has the following embodiments.
(IV-1) WPM medium used for tissue culture of plants of the genus Licorice of the family Leguminosae. The present invention can be rephrased as the use of a WPM medium for tissue culturing a legume Licorice plant, or the use of a WPM medium for producing a tissue culture medium for a legume Licorice plant.
(IV-2) The medium according to (IV-1), wherein the WPM medium further contains amino acids.
(IV-3) The amino acid is at least one selected from the group consisting of proteinogenic amino acids, preferably glycine, glutamine and asparagine, more preferably glycine and glutamine, (IV-1) or (IV-2). The medium according to.
(IV-4) The medium according to any one of (IV-1) to (IV-3), wherein the WPM medium further contains sugar.
(IV-5) At least one selected from the group in which the sugar is glucose, sucrose, mannose, maltose, ribose, arabinose, galactose, meliviose, trehalose, cellobiose, lactose, raffinose, sorbitol and glycerol, and fructose, preferably. The medium according to any one of (IV-1) to (IV-4), wherein is sucrose or glucose.
(IV-6) The medium according to any one of (IV-1) to (IV-5), wherein the WPM medium further contains vitamins.
(IV-7) The vitamins are PABA (p-aminobenzoic acid), vitamin K (biotin), vitamin B5 (D-calcium pantothenate), folic acid, I-inositol, nicotinic acid, niacinamide (nicotinic acid amide). ), Vitamin B6 (Pyrdoxine HCl) (and Pyrodoxal HCl), Vitamin B2 (riboflavin), Vitamin B1 (thiamine) and Vitamin B12 (cyanocobalamine), Vitamin C (L-ascorbic acid), choline chloride, At least one selected from the group consisting of choline dihydrogenate citrate, orthoic acid, putresin, timine, vitamin A (retinol), and vitamin D (cholecalciferol), (IV-1) to (IV-6). ). The medium according to any one of.
(IV-8) The medium according to any one of (IV-1) to (IV-7), wherein the WPM medium further contains a solidifying agent.
(IV-9) At least selected from the group consisting of gellan gum (gellan gum, fighter gel, gellan gum), native gellan gum, agar, gelatin, pectin, carrageenan, starch, guar gum, xanthan gum, carboxymethyl cellulose, and tamarind gum. The medium according to any one of (IV-1) to (IV-8), which is a kind.
(IV-10) The medium according to any one of (IV-1) to (IV-9), which is substantially free of plant hormones, preferably auxin.
(IV-11) Described in any one of (IV-1) to (IV-10), wherein the licorice genus plant is a plant selected from Glycyrrhiza uralensis Fisch. And G. glabra L. Medium.
(IV-12) The medium according to any one of (IV-1) to (IV-11), which is a medium used for improving the rooting rate in a tissue culture of a plant belonging to the genus Licorice of the family Leguminosae.
(IV-13) A method for producing molding of a plant according to any one of (IV-1) to (IV-11), which comprises a step of culturing a tissue of a licorice plant using the medium according to any one of (IV-1) to (IV-11).
(IV-14) The production method according to (IV-13), wherein the tissue is a neighboring tissue containing a growth point of a plant of the genus Licorice.
(IV-15) A method for promoting rooting of a licorice plant, which comprises a step of culturing a tissue of a licorice plant using the medium according to any one of (IV-1) to (IV-11).
(IV-16) The method for promoting rooting according to (IV-15), wherein the tissue is a nearby tissue containing a growth point of a plant of the genus Licorice.

According to the present invention, it is possible to provide an IPSQ production promoter that can be suitably used for a plant containing isoprenoid (IPSQ) having squalene as a precursor. In particular, when the plant is a plant belonging to the genus Licorice of the family Mame, which is one of the medicinal plants, the whole plant or a part thereof is treated with the IPSQ production promoter to grow the plant, and the plant is grown in the underground part thereof. Licorice plants with high IPSQ content by increasing the production of triterpene glycosides (TTG) such as glycyrrhetin (GL) or 3-monoglucuronyl glycyrrhetinic acid (3MGA) or triterpene (TT) such as glycyrrhetinic acid and their You can get the underground part. The underground part is useful as a raw material for crude drugs (licorice) and as a raw material for sweeteners (glycyrrhizin). When the plant is a plant belonging to the genus Ginseng of the family Araliaceae, the production amount of ginsenoside in the underground portion thereof is increased by treating all or a part of the plant with the IPSQ production promoter to grow the plant. , A plant of the genus Ginseng of the family Araliaceae with a high content of ginsenoside and its underground part can be obtained. The underground part is useful as a raw material for crude drugs (medicinal carrots). As described above, according to the present invention, IPSQ-containing plants such as legumes of the genus Licorice and plants of the genus Ginseng of the family Araliaceae are treated with the IPSQ production promoter in whole or in part thereof to obtain a plant. By growing, it becomes possible to efficiently produce a plant containing a high content of IPSQ and its site.

Further, according to the WPM medium for tissue culture of legumes of the genus Licorice (medium of the present invention) of the present invention, the rooting rate of tissue-cultured Licorice plants, which had a low rooting rate in the past, can be significantly improved. .. As a result, it becomes possible to efficiently produce a plant of the genus Licorice (molding) of the family Leguminosae, which is useful as a raw material for producing glycyrrhizin. Furthermore, since the medium of the present invention does not need to contain auxin, which is a plant hormone, there is no risk of plant hormones accumulating in plants of the genus Licorice of the family Leguminosae according to the medium of the present invention, and production of glycyrrhizin that is safe for humans and the environment. It is possible to produce and provide a plant of the genus Licorice of the family Leguminosae as a raw material.

(I) IPSQ Production Accelerator The IPSQ production accelerator according to the embodiment of the present invention contains a jasmonic acid compound (JA compound) represented by the following general formula (1) as an active ingredient. Here, the active ingredient means an ingredient that exerts the effect of the invention. In the present invention, it means a component that exerts an IPSQ production promoting action on an IPSQ-containing plant and contributes to the effect of the present invention.

［式中、Ｒ^１は炭素数１～２０の炭化水素基を表し、Ｒ^２は水素原子又は炭素数１～２０の炭化水素基を表す。］

[In the formula, R ¹ represents a hydrocarbon group having 1 to 20 carbon atoms, and R ² represents a hydrogen atom or a hydrocarbon group having 1 to 20 carbon atoms. ]

The hydrocarbon group in R ¹ may be an aliphatic hydrocarbon group or an aromatic hydrocarbon group. It is preferably an aliphatic hydrocarbon group.

Here, the aliphatic hydrocarbon group may be a saturated aliphatic hydrocarbon group or an unsaturated aliphatic hydrocarbon group. Further, the aliphatic hydrocarbon group may be a linear or branched aliphatic hydrocarbon group, or may be an aliphatic hydrocarbon group having a ring in its structure. Further, the aromatic hydrocarbon group is a group obtained by removing one hydrogen atom from an aromatic hydrocarbon group such as benzene and naphthalene, and examples thereof include a phenyl group and a naphthyl group.

Specific examples of the saturated aliphatic hydrocarbon group in R ¹ include linear, branched or cyclic alkyl groups. It is preferably a linear or branched alkyl group, and more preferably a linear alkyl group. The linear or branched alkyl group preferably has 1 to 20 carbon atoms, more preferably 2 to 10 carbon atoms, and further preferably 3 to 7 carbon atoms. The cyclic alkyl group preferably has 3 to 20 carbon atoms, more preferably 3 to 12 carbon atoms.

Examples of such alkyl groups include, but are not limited to, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, t-butyl group, n-pentyl group, 2 -Methylbutyl group, 1-methylbutyl group, neopentyl group, 1,2-dimethylpropyl group, 1,1-dimethylpropyl group, cyclopentyl group, n-hexyl group, 4-methylpentyl group, 3-methylpentyl group, 2 -Methylpentyl group, 1-methylpentyl group, 3,3-dimethylbutyl group, 2,3-dimethylbutyl group, 1,3-dimethylbutyl group, 2,2-dimethylbutyl group, 1,2-dimethylbutyl group , 1,1-dimethylbutyl group, 3-ethylbutyl group, 2-ethylbutyl group, 1-ethylbutyl group, 1,2,2-trimethylbutyl group, 1,1,2-trimethylbutyl group, 1-ethyl-2- Methylpropyl group, cyclohexyl group, n-heptyl group, 2-methylhexyl group, 3-methylhexyl group, 4-methylhexyl group, 5-methylhexyl group, 2,4-dimethylpentyl group, n-octyl Group, 2-ethylhexyl group, 2,5-dimethylhexyl group, 2,5,5-trimethylpentyl group, 2,4-dimethylhexyl group, 2,2,4-trimethylpentyl group, n-nonyl group, 3, 5,5-trimethylhexyl group, n-decyl group, 4-ethyloctyl group, 4-ethyl-4,5-dimethylhexyl group, undecyl group, dodecyl group, tridecyl group, tetradecyl group, pentadecyl group, hexadecyl group, heptadecyl group. Examples thereof include a group, an octadecyl group, a nonadecyl group, and an icosyl group.

The unsaturated aliphatic hydrocarbon group in ^R1 is preferably a linear or branched unsaturated aliphatic hydrocarbon group. More preferably, it is a linear unsaturated aliphatic hydrocarbon group. The number of carbon atoms of the unsaturated aliphatic hydrocarbon group is not limited, but is preferably 2 to 20, more preferably 2 to 10, and even more preferably 3 to 7. As the unsaturated aliphatic hydrocarbon group, specifically, a group in which one or more single bonds between carbon atoms in the above-mentioned alkyl group are replaced with an unsaturated bond of a double bond or a triple bond is exemplified. be able to. The number of unsaturated bonds in the unsaturated aliphatic hydrocarbon group may be one or two or more. When the number of unsaturated bonds is two or more, these unsaturated bonds may be only double bonds or only triple bonds, and further, double bonds and triple bonds are mixed. May be good. The position of the unsaturated bond in the unsaturated aliphatic hydrocarbon group is not particularly limited.

Examples of the unsaturated aliphatic hydrocarbon group in R ¹ include, but are not limited to, an alkenyl group and an alkynyl group. More preferably, it is an alkenyl group. Examples of the alkenyl group include, but are not limited to, an ethenyl group, a propenyl group, a butenyl group, a pentenyl group, a hexenyl group, a heptenyl group and the like.

The hydrocarbon group in R ² is not particularly limited, and the hydrocarbon group exemplified in R ¹ can be similarly mentioned. It is preferably an aliphatic hydrocarbon group. Here, the aliphatic hydrocarbon group may be a saturated aliphatic hydrocarbon group or an unsaturated aliphatic hydrocarbon group. It is preferably a saturated aliphatic hydrocarbon group. The saturated aliphatic hydrocarbon group may be a linear, branched or cyclic saturated aliphatic hydrocarbon group, but is preferably a linear or branched saturated aliphatic hydrocarbon group. , More preferably a linear saturated aliphatic hydrocarbon group.

Specific examples of the saturated aliphatic hydrocarbon group in ^R2 include an alkyl group having 1 to 20 carbon atoms. An alkyl group having 1 to 10 carbon atoms is preferable, and an alkyl group having 2 to 5 carbon atoms is more preferable. As a specific example thereof, the above-mentioned alkyl group of R ¹ can be mentioned in the same manner.

Among the compounds represented by the general formula (1), a compound in which R ¹ is a linear unsaturated hydrocarbon group, preferably an alkenyl group, and R ² is a hydrogen atom (hereinafter, this compound group is referred to as "this compound group". Also referred to as "compound (1-1)"); a compound in which R ¹ is a linear alkyl group and R ² is a linear alkyl group (hereinafter, this compound group is referred to as "compound (1-2)". (Also also referred to as); and a compound in which R ¹ is a linear unsaturated hydrocarbon group, preferably an alkenyl group, and R ² is a linear alkyl group (hereinafter, this compound group is referred to as "compound (1-3)". ) ”) Is included.

As an example of the compound contained in the compound (1-1), jasmonic acid represented by the following formula (1-1) can be mentioned.

Further, as an example of the compound contained in the compound (1-2), prohydrojasmon represented by the following formula (1-2) can be mentioned.

Further, as an example of the compound contained in the compound (1-3), methyl jasmonate represented by the following formula (1-3) can be mentioned.

These are JA compounds suitable as an active ingredient of the IPSQ production promoter according to the embodiment of the present invention. That is, it is preferable that the IPSQ production promoter of the present invention contains at least one JA compound selected from the group consisting of the above-mentioned jasmonic acid, prohydrojasmonate, and methyl jasmonate as an active ingredient. More preferably, prohydrojasmon and methyl jasmonate, and particularly preferably prohydrojasmon.

The JA compound targeted by the present invention includes various isomers. For example, for prohydrojasmon, the (1R, 2R) body represented by the following formula (1-2-1), the (1S, 2S) body represented by the following formula (1-2-2), and the following formula (1-2S) are used. There are various isomers such as the (1R, 2S) isomer represented by 2-3) and the (1S, 2R) isomer represented by the following formula (1-2-4). All of these are included as prohydrojasmon in the JA compound targeted by the present invention.

Such an isomer may be blended alone in the IPSQ production promoter of the present embodiment, or may be blended in the form of a mixture of two or more kinds without being isolated.

The JA compound may form a salt. The salts of JA compounds include alkali metal salts such as sodium salt, potassium salt and lithium salt; alkaline earth metal salts such as calcium salt and magnesium salt; metal salts such as zinc salt, iron salt and manganese salt; Ammonium salts; examples thereof include nitrates such as nitrates, sulfates, acetates, carbonates, hydrogencarbonates, and phosphates.

The JA compound used in the present invention may be isolated from nature or artificially synthesized regardless of its origin. Commercially available products can be conveniently used. For example, as prohydrojasmon, a commercially available jasmomate (registered trademark) liquid agent (Meiji Seika Pharma Co., Ltd.) can be used. The jasmomate solution is a preparation containing both trans-form and cis-form prohydrojasmon represented by the above formulas (1-2-1) to (1-2-4) (“propyl (1RS, 2RS)). -(3-oxo-2-pentylcyclopentyl) acetate "includes 10 ± 2%).

The ratio of the JA compound contained in the IPSQ production promoting agent is not particularly limited as long as the prepared IPSQ production promoting agent exerts the IPSQ production promoting effect, and is usually appropriately in the range of 0.1 to 99.9% by mass. You can choose. For example, it can be appropriately selected from the range of 1 to 50% by mass, preferably from the range of 3 to 20% by mass.

In addition to the JA compound described above, the IPSQ production accelerator includes a carrier, an emulsifier, a dispersant, a spreading agent, a thickener, a disintegrant, a binder, a pH adjuster, etc. Any component of the above can be appropriately contained in a proportion not exceeding 100% by mass in total. These may be used individually by 1 type and may be used in combination of 2 or more type.

Examples of the liquid carrier include a solvent for dissolving or dispersing the JA compound, for example, water; alcohols such as ethanol, 1-propanol, and butanol; polyhydric alcohols such as ethylene glycol and propylene glycol; xylene and the like. Hydrocarbons can be mentioned. Examples of the solid carrier include kaolin, attapulsite, bentonite, calcium carbonate, talc, zeolite and the like. The ratio of the carrier in the IPSQ production promoting agent is not particularly limited as long as the prepared IPSQ production promoting agent exerts the IPSQ production promoting effect, and is usually selected from the range of 0.1 to 99.9% by mass. can. For example, it can be selected from the range of 1 to 99% by mass, preferably from the range of 3 to 80% by mass, and more preferably from the range of 10 to 60% by mass.

As the emulsifier, dispersant and spreading agent, a surfactant is usually used. Therefore, a surfactant can be added to the IPSQ production accelerator as another optional component. Examples of the surfactant include nonionic surfactants (for example, polyoxyethylene alkyl phenyl ether, polyoxyethylene alkyl ether, polyoxyalkylene fatty acid ester, polyoxyethylene hextan fatty acid ester, polyalkylene glycol alkyl ether, and sorbitan fatty acid). Esters, polyoxyethylene resin acid esters, polyoxyethylene fatty acid esters, etc.), anionic surfactants (eg, higher alcohol sodium sulfate, polynaphthylmethane sulfonate sodium, dioctyl sulfosuccinate sodium, alkylbenzene sulfonate sodium, lignin sulfonate calcium Etc.), Cationic surfactants (eg dialkyldimethylammonium polynaphthylmethanesulfonate, stearyltrimethylammonium chloride, etc.), Silicon-based surfactants (polyoxyalkyleneoxypropylheptamethyltrisiloxane, polyoxyethylenemethylpolysiloxane), And amphoteric surfactants. It is preferably a nonionic surfactant, more preferably a polyoxyethylene alkyl phenyl ether (including, for example, polyoxyethylene octylphenyl ether, polyoxyethylene nonylphenyl ether, etc.), a polyoxyethylene alkyl ether (for example, for example). Polyoxyethylene dodecyl ether and the like are included), polyoxyalkylene fatty acid ester (for example, polyoxyethylene fatty acid ester and the like are included) and the like can be mentioned. The nonionic surfactant can also be used in combination with an anionic surfactant or a cationic surfactant. The combination of the nonionic surfactant and the anionic surfactant includes a combination of a polyoxyethylene alkyl phenyl ether such as polyoxyethylene nonylphenyl ether and calcium lignin sulfonate, a polyoxyethylene alkyl phenyl ether and a dioctyl sulfosuccinate. The combination with sodium acid can be exemplified.

The ratio of the surfactant to be blended in the IPSQ production promoter is not particularly limited as long as the prepared IPSQ production promoter exerts the IPSQ production promoting effect, and is usually in the range of 0.1 to 99.9% by mass. You can choose from. For example, it can be selected from the range of 1 to 99% by mass, preferably from the range of 3 to 80% by mass, and more preferably from the range of 10 to 50% by mass.

As the surfactant, preferably, a surfactant used as a component of an agricultural spreading agent can be used. The spreading agent is an additive containing a surfactant as a main component, and is usually used for the purpose of improving the adhesion, spreading or adhesion of the chemical solution to the surface of the plant. Surfactants used in spreading agents can be classified into three types, nonionic surfactants, anionic surfactants, and cationic surfactants, based on their physicochemical structure and properties. Although not limited, the nonionic surfactant has a high effect of lowering the surface tension, and can improve the adhesion to a plant that is difficult to get wet and enhance the action effect of the chemical solution. Anionic surfactants mainly have the effect of improving the suspension of chemicals, and are usually used in combination with nonionic surfactants. The cationic surfactant can enhance the action and effect of the chemical solution by increasing the adsorptivity or permeability. It is preferably a nonionic surfactant or a mixture of a nonionic surfactant and an anionic surfactant.

The spreading agent is a general exhibition having a function of improving the spreadability of the chemical solution by mainly lowering the surface tension to improve the adhesion to plants that are difficult to get wet, or improving the mixability with wettable powders and emulsions. Adhesive; A functional spreading agent that has the function of enhancing the permeation and transferability of chemical components into plants in addition to the functions of the above general spreading agent; and the adhesive spreading having the function of enhancing rain resistance and extending the residual effect. It can be classified as a dressing agent.

Examples of the main component of the general spreading agent include a nonionic surfactant, a mixture of a nonionic surfactant and an anionic surfactant, or a silicon-based surfactant. For example, general spreading agents containing nonionic surfactants as the main component are not limited, but are Agra (Agro-Kanesho Co., Ltd.), Admix (Nissan Chemical Industry Co., Ltd.), and Xarino 10 (Nihon Nohyaku Co., Ltd.) Althorpe. 30 (Sumitomo Chemical Gardening Co., Ltd.), Myrineau (Nihon Nohyaku Co., Ltd.), High Ten Power (Hokuko Chemical Industry Co., Ltd.) and the like can be mentioned. Diecoat (Nippon Soda Co., Ltd.), Dine (Sumitomo Chemical Garden Products Co., Ltd.), Cindyne (Sumitomo Chemical Garden Products Co., Ltd.), although not limited, general spreading agents mainly composed of a mixture of a nonionic surfactant and an anionic surfactant. Sumitomo Chemical Garden Products Co., Ltd.), Sprayer (RIKEN Green Co., Ltd.), Gramin S (Mitsui Chemical Agro Co., Ltd.), Kumiten (Kumiai Chemical Co., Ltd.) and the like can be mentioned. Examples of the general spreading agent containing a silicon-based surfactant as a main component include, but are not limited to, breakthrough (Sankei Chemical Co., Ltd.) and Makupika (Toyo Green Co., Ltd.). Dyne is preferred.

The main components of the functional spreading agent are nonionic surfactants, mixtures of nonionic surfactants and anionic surfactants, anionic surfactants, and nonionic surfactants and cationics. A mixture with a surfactant can be mentioned. For example, as a functional spreading agent containing a nonionic surfactant as a main component, Mix Power (Singenta Japan Co., Ltd.), Approach BI (Maruwa Biochemical Co., Ltd.), Supply (OAT Agrio Co., Ltd.), Squash (Maruwa Co., Ltd.) Biochemical Co., Ltd.), Surfactant WK (Maruwa Biochemical Co., Ltd.), Platen 80 (Sankei Chemical Co., Ltd.), Santokuten 80 (Sumitomo Chemical Gardening Co., Ltd.) and the like. Examples of the functional spreading agent containing a mixture of a nonionic surfactant and an anionic surfactant as a main component include Wide Coat (Nippon Kayaku Co., Ltd.) and the like. Examples of the functional spreading agent containing a mixture of a nonionic surfactant and a cationic surfactant as a main component include needs (Kumiai Chemical Industry Co., Ltd.), Bravo (Agro-Kanesho Co., Ltd.) and the like. .. Examples of the functional spreading agent containing anionic surfactant as a main component include Submerge (Syngenta Co., Ltd.) and the like.

Paraffin and polyoxyethylene resin acid ester can be mentioned as the main components of the adhesive spreading agent. Examples of the adhesive spreading agent containing paraffin as a main component include Avion E (Abion Corporation), Petan V (Agro-Kanesho Co., Ltd.), and Stickel. Examples of the adhesive spreading agent containing a polyoxyethylene resin acid ester as a main component include K.K. K sticker (Agro-Kanesho Co., Ltd.) and the like can be mentioned.

The shape (dosage form) of the IPSQ production promoter is not limited, but depending on the application method (usage) to plants, powders, DL powders, FDs (above, powders); granules; powders, fine granules. , Fine granules (above, powder granules); powder; tablets; wettable powder, flowable agent, granule hydrate, dry flowable (above, hydrate); water solvent, granule water solvent (above, water solvent) Emulsion, EW agent (above, emulsion); liquid agent, ME solvent (above, liquid agent); oil agent, surf agent (above, oil agent); aerosol; paste agent; fumigant agent; fumigant agent; gas agent; microcapsule agent; It can be prepared in various shapes such as a packing agent. Examples of the properties at the time of use include solids, liquids and gases, but it is preferable to use them in a liquid state. Therefore, the shape (dosage form) of the IPSQ production accelerator is preferably a liquid shape such as a liquid agent, a flowable agent, an emulsion, and an oil agent.

Examples of the timing and method of using the IPSQ production promoter for plants include spraying, spraying, applying, and soaking the whole plant or a part of the IPSQ-containing plant during the seed or growing period. It is preferably applied to IPSQ-containing plants during the growing season. It is necessary to appropriately adjust the ratio of JA compound in the IPSQ production promoter when used in plants, depending on the type of JA compound used, the type of target plant, the mode of use, the method of use, the time of use, etc. Although it cannot be unconditionally specified, when it is used in a liquid state, the JA compound is usually adjusted in the range of 0.001 to 5% by mass. It is preferably 0.002 to 1% by mass, more preferably 0.003 to 0.1% by mass, and even more preferably 0.004 to 0.01% by mass. The usage and dosage of the IPSQ production promoter will be described in detail in the column (II) below as a method of using the JA compound-containing composition relating to the method for producing an IPSQ-containing plant.

A plant to which an IPSQ production promoter is applied is a plant containing IPSQ in at least a part of the plant body. Specifically, IPSQ is generated and contained (stored / accumulated) in at least a part of the plant body in the growth process of the plant. That is, it can be said that the plant is an IPSQ-producing plant, but in the present invention, the plant is referred to as an IPSQ-containing plant.

Here, squalene-precursor isoprenoids (IPSQ) include triterpenoids and steroids (ST). Triterpenoids include triterpene glycosides (TTGs) and triterpenes (TTs), and steroids (STs) include steroid glycosides (STGs) (also referred to as steroid saponins).

Here, examples of the triterpene glycoside (TTG) include glycyrrhizin (GL), a salt thereof (dipotassium glycyrrhizinate, etc.), baccaroside B, avenacin A-1, soybean saponin, and ginsenoside. Uralsaponin, Chixetsusaponin, Eleutheroside A, Akeboside, Platicodin, Senegin, Ondisaponin, Psychosaponin, Theasaponin, Astersaponin, Cibilicoside, Smiraxsaponin, Estin, Araloside, Astragaloside, Gymnemaic acid, Gibenoside. Here, ginsenoside includes ginsenoside Rb group such as ginsenoside Rb1 and ginsenoside Rb2; ginsenoside Rg group such as ginsenoside Rg1 and ginsenoside Rg2; ginsenoside Re; ginsenoside Ra; ginsenoside Rc; ginsenoside Rc; ginsenoside Rd. Although not limited, ginsenosides are preferably ginsenosides Rb group and ginsenoside Rg group, and more preferably ginsenosides Rb1 and Rg1. Further, the soybean saponin includes, for example, soybean saponin Aa, soybean saponin βg and the like. Ural saponin includes Ural saponin B, Ural saponin MY and the like. Tixetsusaponin includes Tixetsusaponin Ib, Tixetsusaponin IV (aralocid A), Tixetsusaponin IVa, Tixetsusaponin V and the like. Akeboshid includes Akeboshid St _b , Akeboshid St _c , Akeboshid St _d , Akeboshid St _f , Akeboshid St _h , Akeboshid St _e , Akeboshid St _j and the like. Platycodin includes Platycodin A, Platycodin C, Platycodin D, Platycodin D2 and the like. Senegin is also called ondisaponin, and senegin includes senegin II, senegin III (ondisaponin B), senegin IV (ondisaponin A) and the like. Psychosaponin includes psychosaponin E, psychosaponin F, psychosaponin G and the like. Theasaponins include A1, A2, A3, A4, A5, B2, C1, E1 and the like. Cibilicoside includes cibilicoside A, B and the like. Smilax saponins include Smilax saponins AC and the like. Astragaloside includes astragaloside I, II, III, IV and the like.

Among the IPSQ-containing plants targeted by the present invention, the TTG-containing plant produces and contains at least one of the above-mentioned TTGs, and may contain any two or more of them. These TTG-containing plants are not limited, but for example, the genus Araliaceae, the genus Araliaceae, the genus Acacia, the genus Araliaceae, the genus Araliaceae, the genus Araliaceae, Araliaceae, Araliaceae, Akebi, Kiku, Zion, Araliaceae, Himehagi, Araliaceae, Amadokoro, Araliaceae, Shiode, Tsubaki, Seri, Legume, Plants belonging to the genus Sabonsou of the family Araliaceae, the genus Tochinoki of the family Araliaceae, the genus Karasumugi of the family Rice, the genus Gymnema of the family Araliaceae, the genus Araliaceae or the genus Fudansou of the family Araliaceae, particularly medicinal plants can be mentioned.

Here, the plants belonging to the genus Glycyrrhiza of the family Glycyrrhiza include Glychyrrhiza uralensis, Glychyrrhiza glabra, and the like. The TTG contained in the underground part of the licorice plant includes the above-mentioned glycyrrhizin (GL), dipotassium glycyrrhizinate, 3-monoglucuronyl glycyrrhetinic acid (3MGA), vaccaroside B, and avenacin A-1 (AVENACIN). a-1), soybean saponin; uralsaponin can be mentioned. Glycyrrhizin (GL), dipotassium glycyrrhizinate, 3-monoglucuronyl glycyrrhetinic acid (3MGA) are preferable, and GL is more preferable. The underground part of the licorice plant also contains glycyrrhetinic acid, gravurinic acid, and liquiritin acid among the triterpenes (TT) described later. Preferably, glycyrrhetinic acid is included.

In addition, plants belonging to the genus Panax of the Araliaceae family include Panax ginseng CAMey (also known as Korean ginseng or Korean ginseng) and American ginseng (Panax quinquefolius L.) (also known as Western ginseng and flower flag ginseng). ), Panax notoginseng Berk. (Also known as ginseng), Panax japonicus (also known as ginseng), etc. are included. Examples of the TTG contained in the roots of plants of the genus Panax japonicus in the family Araliaceae include ginsenoside, which is a triterpene saponin. The types of ginsenosides are as described above. It is preferably a ginsenoside Rb group such as ginsenoside Rb1 and ginsenoside Rb2; and a ginsenoside Rg group such as ginsenoside Rg1 and ginsenoside Rg2, and more preferably ginsenoside Rb1 and ginsenoside Rg1. In terms of containing a large amount of these ginsenosides Rb1 and Rg1, ginseng, American ginseng, and Panax notoginseng are preferable, and ginseng is more preferable. Panax japonicus further contains chixetsu saponin as TTG. The types of chixetsu saponins are as described above.

Examples of triterpenes (TT) include ursan-type triterpenes (eg, α-amylin, tormentic acid, corosolic acid, ursoric acid), protostan-type triterpenes (eg, arisole, herboric acid, fushidic acid), and lanostane-type triterpenoids (eg, lanosterol, ebricco). Acids, polyporenic acid, tumrosic acid, pachymaic acid, cycloartenol, simisifgenol, simigenor), damaran triterpenes (eg damarenediol, protopanaxadiol, protopanaxatriol), euphan-tilkaran triterpenes (eg euhole) , Euphorbol, elemolic acid, elemonic acid), lupine-type triterpenes (lupeol, bethrin, bethuric acid), limonine-type triterpenes (eg limonin, evodol, rutaevin, obakunon), quassin-type triterpenes (eg quassinoids, bruseantin), oleanan-type triterpenes (eg For example, masphosphate), 2α, 19α-dihydroxy-3-oxourus-12-en-28-oic acid, β-amylin, oleanolic acid, fupensic acid, heragenin, platicodigenin, polygalassic acid, presenegenin, psychogenin, chasapogenol, liquiritin. Examples include acids, gravurinic acid, glycyrrhetinic acid, and salts thereof. Here, Arisol includes Arisol A, B, C and their monoacetates and the like. Polypolenic acid includes polypolene acids A, C and the like. Damarene diol includes Damarene diol, Damarene diol I, II and the like. Psychogenins include psychogenins E, F, G and the like. Chasapogenol includes teasapogenols A, B, C, D, E and the like. The quassinoids include quassin (nigakilactone D), neoquassin, nigakilactones A to N (excluding D), picrasma A, C, F, nigaki hemiacetal A, B, C and the like. Glycyrrhetinic acid is preferable.

Among the IPSQ-containing plants targeted by the present invention, the TT-containing plant produces and contains at least one of the above-mentioned TTs, and may contain any two or more of them.
These TT-containing plants include, but are not limited to, for example, the genus Kanzo, the genus Marumero, the genus Pear, the genus Nanakamado, the genus Sanzashi, the genus Sharinto, the genus Nigaki, the genus Kihada, and the genus Otaneninjin. , Omodaka family Sagiomodaka genus, Matin family Matin genus, Todaigusa family Todaigusa genus, Rice family rice genus, Omugi genus, Chigaya genus, Abrana family Abrana genus, Kinpouge family Sarashinashoma genus, Futabagaki family Saranoki genus , Kanzo, Kabanoki, Kabanoki, Kuromedoki, Natsume, Kanran, Kanran, Shiso, Arctostaphyros, Oobaco, Oobako, Mochinoki, Lindou, Futomomo , The genus Kikyo of the family Kikyo, the genus Himehagi of the family Himehagi, the genus Mishimapsycho of the family Seri, the genus Tsubaki of the family Tsubaki, the genus Kihada of the family Mikan, or the genus Goshuyu.

Steroid glycosides (STGs) include pregnane glycosides, ecdysterone, inocosterone, strophanthin, smilax saponin, hellebrin, dioscin, offiopogonin, smiragenin glycosides, digitonin, gitonin, sarsasapogenin, digitoxin, lanatoside, and gitoxins. , Gitaloxin, convallatoxin, silalene and thymosaponin. Pregnane glycosides include conzlangoglycosides A, A1, A0, B0, C, C0, C1, D1, conzlangogenins A, C and the like. Strophantin includes G-strophantin (ouabain), K-strophantin and the like. Smilax saponins include Smilax saponins A, B, C and the like. Offiopogonins include offiopogonins A to D, B'to D'and the like. Lanatoside includes lanatoside A, B, C and the like. Timosaponin includes Timosaponin A-III and the like.

Among the IPSQ-containing plants targeted by the present invention, the STG-containing plant produces and contains at least one of the above-mentioned STGs, and may contain any two or more of them. These STG-containing plants include, but are not limited to, for example, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena, Anemarrhena. Plants belonging to the genus Anemarrhena, the genus Janohige, the genus Anemarrhena, the genus Haran, the genus Digitalis of the family Anemarrhena, or the genus Wizania of the family Anemarrhena can be mentioned.

According to the IPSQ production promoter of the present invention, the content of IPSQ at at least one site in the plant body of the above-mentioned IPSQ-containing plant can be increased by the action of the JA compound. The sites containing IPSQ (preferably accumulation sites) include leaves, stems, flowers, fruits, seeds (hereinafter collectively referred to as "shoot type"), and roots (taproot, lateral root), depending on the type of IPSQ-containing plant. ) (Hereinafter, collectively referred to as "taproot"). For example, the GL-containing site (accumulation site) in a plant belonging to the genus Licorice of the family Leguminosae is a root system (preferably taproot) and stron. Stron is a stem (shoot system), but since it is a rhizome that crawls sideways from the root to the ground near the ground surface, the roots (taproot, lateral roots) and stron are collectively referred to as "underground parts" here. The ginsenoside-containing site (accumulation site) in the Araliaceae Panax japonicus plant is a root system, preferably a taproot.

Although the mechanism of action of the IPSQ production promoter of the present embodiment to increase the IPSQ content in IPSQ-containing plants is unconfirmed and not limited, the JA compound acts on the metabolic pathway (production pathway) for producing IPSQ in these plants. It is presumed that they are doing it. In addition, in the plants of the genus Panax japonicus of the family Leguminosae and the plants of the genus Panax japonicus in the family Araliaceae, it has been confirmed that the weight of the underground part containing IPSQ increases due to the action of the JA compound. ) Is promoted, and as a result, it can be inferred that the IPSQ content is also increased. However, regardless of such a mechanism of action, the use of the IPSQ production promoter of the present invention can significantly increase the IPSQ content in the IPSQ-containing plant as compared with the case where the IPSQ production promoter is not used. It is possible to obtain a plant body or a part thereof of a plant containing more IPSQ. That is, by using the IPSQ production promoter of the present embodiment, a plant body of a plant containing a large amount of IPSQ efficiently in a short cultivation period or a plant thereof, as compared with a conventional cultivation method that does not use the IPSQ production promoter. You can get the part.

（式中、Ｒ^１は１～２０の炭素を有する炭化水素基、Ｒ^２は水素原子または１～２０の炭素を有する炭化水素基を意味する）、
（ｂ）前記工程（ａ）で処理された植物を生育する工程。 (II) Method for Producing IPSQ-Containing Plant and Processed Product The method for producing IPSQ-containing plant or processed product thereof, which is one embodiment of the present invention, can be carried out by a method having the following steps:
(A) A step of treating all or a part of an IPSQ-containing plant with a JA compound represented by the following formula (1) or a composition containing the same.

(In the formula, R ¹ means a hydrocarbon group having 1 to 20 carbons, R ² means a hydrogen atom or a hydrocarbon group having 1 to 20 carbons),
(B) A step of growing the plant treated in the step (a).

Hereinafter, the steps (a) and (b) will be described.
(1) Processing step (a):
Step (a) is a step of treating the target plant with a JA compound or a composition containing the JA compound.

The JA compound used in this step (a) is as described in (I) above, and the description thereof is incorporated herein by reference. It is preferably at least one selected from the group consisting of jasmonic acid, prohydrojasmonate, and methyl jasmonate, and these include various isomers. Although not limited, it is preferably prohydrojasmon and methyl jasmonate, more preferably prohydrojasmon and methyl jasmonate, and even more preferably prohydrojasmon.

Examples of the composition containing the JA compound (JA compound-containing composition) include the IPSQ production promoter described in (I) above, and the description thereof is incorporated herein by reference. That is, in this step (a), the above-mentioned IPSQ production promoter can be used as the JA compound-containing composition.

The JA compound-containing composition preferably contains a surfactant in addition to the JA compound. The surfactant can be added to the above composition as an emulsifier, a dispersant or a spreading agent. By adding a surfactant to the JA compound-containing composition, it is possible to enhance the uniform dispersibility and solubility of the JA compound, enhance the adhesion of the JA compound to the plant body, and enhance the permeability. can. The types of surfactants used and their blending ratios are as described in (I), and the description can be incorporated herein by reference.

The shape of the JA compound-containing composition used in this step (a) can be any of solid, liquid, and gas, but is preferably liquid. The liquid may be any liquid and includes any of a solution, a dispersion and a milky lotion, but from an environmental point of view, an aqueous solution, an aqueous dispersion, an O / W or a W / O / W type milky lotion is preferable. The JA compound-containing composition having the said shape is prepared by appropriately dissolving, dispersing or diluting the IPSQ production accelerator of various shapes (forms) described in (I) with water or the like to adjust the concentration to be suitable for the treatment. It can also be prepared with.

Here, the concentration suitable for the treatment, that is, the concentration of the JA compound in the composition containing the JA compound may be usually in the range of 0.001 to 5% by mass. It is preferably 0.002 to 1% by mass, more preferably 0.003 to 0.1% by mass, and even more preferably 0.004 to 0.01% by mass. By treating a plant of an IPSQ-containing plant with a composition containing a JA compound in these concentration ranges, the IPSQ content in the plant can be effectively increased.

Although the amount of the JA compound-containing composition having the above concentration per treatment is not limited, for example, 10 to 1000 L can be mentioned per planting area 10a (1000 m ² ) of the IPSQ-containing plant. In this range, for example, it may be 50 to 700 L / 10a, 100 to 400 L / 10 a, or 100 to 200 L / 10 a.

The plant to be treated in this step (a) is the IPSQ-containing plant described in (I) above. As an IPSQ-containing plant, IPSQ, particularly TTG-containing genus Araliaceae, genus Araliaceae, genus Araliaceae, genus Araliaceae, genus Araliaceae, genus Araliaceae, genus Araliaceae, genus Araliaceae Genus, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae, Araliaceae It is a plant belonging to the genus, the genus Spikenard, the genus Araliaceae, the genus Spikenard, the genus Araliaceae, the genus Araliaceae, or the genus Araliaceae. It is more preferably a medicinal plant, and more preferably a plant belonging to the genus Licorice such as Ural licorice and licorice, and a plant belonging to the genus Araliaceae such as Panax ginseng.

As a method for treating these IPSQ-containing plants with a JA compound or a composition containing a JA compound, the JA compound comes into contact with the whole plant or at least a part thereof, and as a result, the effect of the present invention (promotion of IPSQ production, The method is not particularly limited as long as it is a method for achieving IPSQ content increase). For example, soil spraying, soil injection, soil mixing, compounding in hydroponic cultivation liquid, compounding in cultivation medium, immersion treatment of plant underground part (above, treatment mainly for plant underground part), leaf stem spraying, foliage application, plant Examples thereof include immersion treatment, gas treatment, smoke treatment (above, treatment mainly for the above-ground part of the plant) of the above-ground part. A method of contacting the JA compound with the above-ground part of the plant (leaf stem spraying, foliage coating, above-ground part immersion treatment, gas treatment, and smoke treatment) is preferable, and more preferably, leaf stem spraying is performed because of the convenience of treatment. Is. Leaf stem spraying can be performed using conventional equipment and methods such as sprayers or sprayers.

The treatment conditions (environmental conditions at the time of treatment) are not limited as long as the conditions exert the effect of the present invention, but for example, when the JA compound is a compound that easily vaporizes, the effect of the JA compound is obtained more effectively. From the viewpoint, it is preferable to perform the treatment in the closed space. Specifically, it is a method of treating an IPSQ-containing plant with a JA compound or a JA compound-containing composition in a closed space and growing (cultivating, culturing) the plant. Here, examples of the closed space include a test tube, a petri dish, a cultivation container, a cultivation room, a greenhouse, a plant factory, and the like. Further, when the IPSQ-containing plant is gas-treated or smoke-treated using the JA compound or the JA compound-containing composition, it is preferably performed in the closed space.

The treatment time with the JA compound or the composition containing the JA compound can be appropriately set according to the type of the plant containing IPSQ. The time is not limited, but includes the time before sowing, at the same time as sowing, after sowing, before planting, at the same time as planting, after planting, and the like. It is preferably during the growing period of the IPSQ-containing plant. Specifically, it is in the period from germination to harvest (excluding the period required for the growth step (b) described later), preferably the period from germination to harvest after 3 months (however). , Excluding the period required for the growth step (b) described later).

The growth period of the above-mentioned IPSQ-containing plant varies depending on the type, growth environment, growth state, etc. of the IPSQ-containing plant and cannot be unconditionally determined. The period can be mentioned. Therefore, it is desirable to treat it within this growing period. It is preferably treated within a growth period of 1 to 4 years, more preferably 1 to 3 years, still more preferably 1 to 2 years, and particularly preferably 1 to 1 year and 6 months after germination. The number of processes during the period may be one or more, and can be appropriately selected. Although not limited, it is preferable to do it at least once a year.

For example, the growth period of a plant belonging to the genus Panax japonicus in the family Araliaceae can be one to six years after germination. Therefore, it is desirable that the above plants be treated within this growing period. It is preferably treated within a growth period of 1 to 5 years, more preferably 1 to 4 years, still more preferably 1 to 3 years after germination. The number of processes during the period may be one or more, and can be appropriately selected. Although not limited, it is preferable to do it at least once a year.

(2) Growth step (b):
The step (b) is a step of growing the plant treated in the above-mentioned step (a).
The step can be carried out by cultivating the IPSQ-containing plant in a viable method, and even if the conditions and the like are appropriately set and adjusted according to the conventional method according to the type, growth state and growing time of the IPSQ-containing plant. good. As such a cultivation method, any of the conventional methods such as open field cultivation, house cultivation, greenhouse cultivation, tunnel cultivation, hydroponics and the like may be used, and can be appropriately selected.

It is preferable that such a growth step is carried out for a period until a desired amount of IPSQ is produced and accumulated in the IPSQ-containing site of the IPSQ-containing plant. The growth period in the step (b) varies depending on the type and growth state of the IPSQ-containing plant and the treatment method in the step (a), but is 14 days to 6 years from the treatment in the treatment step (a). It can be appropriately selected from the range. It is preferably 30 days to 4 years, and more preferably 30 days to 2 years.

For example, in the case of plants of the genus Licorice of the family Leguminosae, until the taproot and taproot, preferably the GL content (total amount) contained in the taproot, reaches the standard value of 2.0% by mass or more specified by the 17th revised Japanese Pharmacopoeia in terms of dry matter. It is preferable to cultivate and grow during the period of. Further, in the case of a plant belonging to the genus Panax japonicus of the family Araliaceae, the taproot ginsenoside Rb1 and Rg1 contents may be cultivated and grown for a period of 0.2% by mass or more and 0.1% by mass or more, respectively, in terms of dry matter. preferable. The GL content and the ginsenoside Rb1 and Rg1 contents can be measured and determined in accordance with the quantitative method described in the 17th revised Japanese Pharmacopoeia.

Thus, the IPSQ-containing plant that has undergone the growth step (b) is harvested. Next, a site containing IPSQ (site containing IPSQ) can be collected from the harvested IPSQ-containing plant. The IPSQ-containing site is underground, preferably taproot and stron for legumes of the genus Licorice, and root system, preferably taproot for plants of the genus Panax japonicus of the family Araliaceae. These IPSQ-containing sites can be further subjected to various processing treatments such as drying, shredding, crushing, crushing, squeezing, extraction, heating, and sterilization, if necessary, and prepared as processed products of IPSQ-containing plants. Can be obtained.

Hereinafter, the production method of the present invention will be described in more detail by taking as examples of plants of the genus Licorice of the family Leguminosae and plants of the genus Panax japonicus of the family Araliaceae as IPSQ-containing plants. However, for matters not described here, the above descriptions (1) and (2) are used as they are.

[Method of producing licorice plant of the family Leguminosae or its processed product]
The method for producing a licorice plant according to an embodiment is a step of treating the whole plant or a part thereof with a JA compound or a composition containing a JA compound at least once during the growing period of the licorice genus plant (step (a). )), It can be carried out according to the conventional cultivation method. The cultivation method is not particularly limited as long as the licorice plant can grow. Examples of the cultivation method include open field cultivation, house cultivation, greenhouse cultivation, row cover cultivation, hydroponics and the like. Further, the licorice genus plant may be one grown from seeds, or may be grown using cultured seedlings or cuttings. Usually, the period from cultivating a licorice plant to harvesting the underground part containing GL can be set within 1 to 5 years from germination of seeds or planting of cuttings or the like.

Licorice plants are perennials, and it is known that in cold regions, the above-ground parts die in winter and sprout again in spring. Therefore, it is preferable that the treatment period with the JA compound or the composition thereof (the implementation period of step (a)) is the growth period of the above-ground part from the viewpoint of increasing the GL content in the underground part. The growing season of the above-ground part is a period in which the above-ground part is not dead or dormant. Therefore, it is preferable to treat at least once within the period from the start of growth of the above-ground part to the harvest (excluding the period required for the growing step (b)) after the above-ground part of each year has died or dormant. The treatment in this case may be treatment to the underground part or treatment to the above-ground part as shown in the experimental example, but due to the simplicity of the treatment, spraying (including spraying) to the above-ground part or It is an application.

The method for producing a Licorice plant includes, after the above-mentioned steps (a) and (b), a step of harvesting the Licorice plant and further collecting the underground part (taproot, taproot), preferably the taproot. May be good. The time when the licorice plant is harvested is preferably the time when a desired amount of GL is produced and accumulated in the underground part. As described above, it is preferable that the GL content in the underground portion reaches the standard value of 2.0% by mass or more specified in the 17th revised Japanese Pharmacopoeia. To this extent, although not limited, the harvest time of licorice plants is within 1 to 5 years after germination of the plant and within 14 days to 6 years after the treatment of step (a). Is preferable. It is preferably within 30 days to 2 years, more preferably within 30 days to 1 year after the treatment of step (a). The younger the germination of the plant and the shorter the cultivation period from planting to harvesting, the lower the cultivation cost, and the advantage is that the risk of weather damage and pest damage is reduced.

The underground part (taproot, stron) of the licorice plant thus harvested and collected is peeled as necessary, and dried as it is or in a peeled state, and is used as a crude drug (GL) or the like. Used as a raw material for pharmaceuticals or as a raw material for sweeteners. It is preferably used as a raw material for pharmaceutical products such as crude drugs.

In the licorice genus plant obtained by the above-mentioned production method, the total amount of GL contained in the underground part is 2.0% by mass or more in terms of dried matter by the measuring method in accordance with the 17th revised Japanese Pharmacopoeia. It is preferable to have. More preferably, the total amount contained in the taproot and stron is 2.0% by mass or more, and particularly preferably, the total amount contained in the taproot is 2.0% by mass or more.

Conventionally, when trying to obtain a Licorice plant containing nearly 2.0% by mass of GL in the underground part, it took about 4 to 5 years to cultivate the plant, and after all, the GL was dried on the taproot. No plant of the genus Licorice containing 2.0% by mass or more by mass was obtained.

On the other hand, according to the production method according to the present embodiment, the GL content in the underground part of the licorice genus plant is increased by treating the licorice genus plant with the JA compound or the JA compound-containing composition in the growing step. Can be made to. Therefore, as shown in Examples described later, even in a relatively short cultivation period of 1 year and 6 months, a licorice genus plant containing 2.0% by mass or more of GL in terms of dry matter can be obtained in the underground part. Can be done. Here, the increase means the underground part (taproot and stron) of one plant of the licorice genus plant prepared by treating with the JA compound as compared with the licorice genus plant prepared without the treatment with the JA compound. It means that the GL content (absolute amount) contained in the taproot) is high. In this case, the method and conditions for producing the licorice plant shall be unified except for the treatment with the JA compound. The plant in this case is called a GL high-content plant (IPSQ high-content plant). As a GL high content plant, as described above, it is a licorice genus plant containing 2.0% by mass or more of GL in the underground portion in terms of dry matter.

According to the production method according to the present embodiment, not only the licorice genus plant (GL high content licorice genus plant) containing GL in a high content of 2.0% by mass or more in the dry mass in the underground part but also the GL is high content. You can get the product of the underground part including. Further, by processing them, it is possible to obtain a plant having a high IPSQ content or a processed product in the basement thereof. Therefore, the production method according to the present embodiment can be suitably used as a method for producing the underground portion containing a high content of IPSQ and its processed product.

Further, according to the production method according to the embodiment, the weight of the taproot of the licorice genus plant can be increased by the step of treating the licorice genus plant with the JA compound. Here, the increase means the weight of the taproot per strain of the licorice genus plant treated with the JA compound (wet weight) as compared with the licorice genus plant prepared under the same conditions under the same conditions without being treated with the JA compound. ) Means that there are many.

From the above, according to the production method according to the embodiment, the GL content contained in the underground part of the licorice genus plant can be increased. It can also increase the weight of taproots of licorice plants. Therefore, the yield of GL obtained comprehensively can be significantly increased, and the roots of licorice plants containing GL abundantly can be obtained from a practical point of view in a shorter cultivation period than before. .. Further, as shown in Experimental Example 2 described later, according to the production method according to the embodiment of the present invention, a licorice genus plant (Ural licorice) is treated with prohydrojasmon as a JA compound to treat the licorice genus plant. The content of glycyrrhetinic acid (GA) and 3-monoglucuronyl glycyrrhetinic acid (3MGA) can be increased as well as the content of GL contained in the underground portion of licorice.

[Method of producing Panax japonicus plant of Araliaceae or its processed product]
One embodiment of the method for producing a Panax japonicus plant is a step of treating all or a part of the plant body with a JA compound or a composition containing a JA compound at least once during the growing period of the Panax japonicus plant (step (a). )), It can be carried out according to the conventional cultivation method. The cultivation method is not particularly limited as long as it is a method in which Panax japonicus can grow. Examples of the cultivation method include open field cultivation, house cultivation, greenhouse cultivation, row cover cultivation, hydroponics and the like. Further, the Panax japonicus plant may be one grown from seeds, or may be grown using cultured seedlings or cuttings. Usually, the period from the cultivation of Panax japonicus plants to the harvesting of the underground part containing ginsenoside can be mentioned within 1 to 6 years from the germination of seeds or the planting of cuttings and the like.

Panax japonicus is a perennial plant, and it is known that in cold regions, the above-ground part withers in winter and buds reappear in spring. Therefore, it is preferable that the treatment period with the JA compound or the composition thereof (the implementation period of step (a)) is the growth period of the above-ground part from the viewpoint of increasing the ginsenoside content in the underground part. The growing season of the above-ground part is a period in which the above-ground part is not dead or dormant. Therefore, it is preferable to treat at least once within the period from the start of growth of the above-ground part to the harvest (excluding the period required for the growing step (b)) after the above-ground part of each year has died or dormant. The treatment in this case may be treatment to the underground part or treatment to the above-ground part as shown in the experimental example, but due to the simplicity of the treatment, spraying (including spraying) to the above-ground part or It is an application.

The method for producing a Panax japonicus plant is a step of harvesting the Panax japonicus plant after the above-mentioned steps (a) and (b), and further collecting the underground part (root or rhizome excluding fine roots), preferably taproot. You may have. It is preferable that the plants of the genus Panax japonicus are harvested after the above-ground parts have died. To this extent, although not limited, the harvest time of Panax japonicus is within 1 to 6 years after germination of the plant and within 30 days to 5 years after the treatment of step (a). Is preferable. It is preferably within 60 days to 3 years, more preferably within 60 days to 1 year after the treatment of step (a). The younger the germination of the plant and the shorter the cultivation period from planting to harvesting, the lower the cultivation cost, and the advantage is that the risk of weather damage and pest damage is reduced.

The underground part of the Panax japonicus plant thus harvested (roots or rhizomes excluding fine roots) is peeled as necessary, and is dried as it is or in a peeled state. It is used as a raw material for pharmaceuticals such as carrot, ginseng, and Panax japonicus, or as a raw material for processed foods. It is preferably used as a raw material for pharmaceutical products such as crude drugs.

Among the plants of the genus Panax japonicus obtained by the above-mentioned production method, the total amount of ginsenoside Rb1 and ginsenoside Rg1 contained in the underground part of Panax japonicus (root excluding fine roots), lightly boiled or steamed product is the tenth. (7) It is preferable that the measurement method conforms to the revised Japanese Pharmacopoeia and is 0.10% by mass or more and 0.20% by mass or more, respectively, in terms of dried product. More preferably, the total amount contained in the taproot is 0.10% by mass or more and 0.20% by mass or more, respectively, and particularly preferably, the total amount contained in the taproot is 0.10% by mass or more and 0.20% by mass or more. ..

According to the production method according to the present embodiment, by treating a Panax japonicus plant with a JA compound in its growth step, an IPSQ high-content plant containing ginsenoside or chixetsu saponin in the underground part of the Panax japonicus plant is to be increased. Can be done. Here, the increase means the rhizome of one plant of the genus Panax japonicus prepared by treating with the JA compound (excluding fine roots) as compared with the plant of the genus Panax japonicus prepared without treatment with the JA compound. It means that the IPSQ content (absolute amount) contained in the root or rhizome is high. In this case, the method and conditions for producing Panax japonicus plants shall be the same except for the treatment with JA compound. As described above, the IPSQ content Panax ginseng preferably contains ginsenoside Rg1 in an underground portion (roots excluding fine roots) in an amount of 0.01% by mass or more in terms of dry matter and ginsenoside Rb1 in an amount of 0.02% by mass or more in terms of dry matter. Panax ginseng.

Further, according to the production method according to the embodiment, the weight of the taproot of the Panax japonicus plant can be increased by the step of treating the Panax japonicus plant with the JA compound. Here, the increase means the weight of the taproot per strain of the Panax japonicus plant treated with the JA compound (wet weight) as compared with the Panax japonicus plant prepared under the same conditions under the same conditions without being treated with the JA compound. ) Means that there are many.

From the above, according to the production method according to the embodiment, the IPSQ content contained in the underground part of the Panax japonicus plant can be increased. It can also increase the weight of the taproots of Panax japonicus plants. Therefore, the yield of IPSQ obtained comprehensively can be remarkably increased, and the roots of Panax japonicus plants containing IPSQ abundantly can be obtained from a practical point of view.

The invention described above also includes the following embodiments:
Item 1: A step of applying at least one compound selected from the group consisting of jasmonic acid and a derivative of jasmonic acid to a plant of the genus Licorice is characterized by increasing the TTG content contained in the root of the plant of the genus Licorice. Cultivation method of licorice genus plant.
Item 2: The cultivation method according to Item 1, wherein the site where the compound is brought into contact with the licorice genus plant is the above-ground part of the licorice genus plant.
Item 3: From the jasmonic acid and the jasmonic acid derivative contained in the agent, which comprises a step of applying an agent containing at least one compound selected from the group consisting of jasmonic acid and a derivative of jasmonic acid to a plant belonging to the genus Kanzo. Item 3. The cultivation method according to Item 1 or 2, wherein the concentration of at least one compound selected from the group is 0.001 to 5% by mass.
Item 4: A step of applying an agent containing at least one compound selected from the group consisting of jasmonic acid and a derivative of jasmonic acid to a plant of the genus Licorice, which is characterized by further containing a surfactant. The method for cultivating a plant of the genus Licorice according to any one of Items 1 to 3.
Item 5: The cultivation method according to any one of Items 1 to 4, wherein the triterpene glycoside is glycyrrhizin.
Item 6: The cultivation method according to any one of Items 1 to 5, wherein the compound is at least one selected from the group consisting of jasmonic acid, prohydrojasmonate, and methyl jasmonate.
Item 7: The cultivation method according to any one of Items 1 to 6, wherein the licorice plant is Glychyrrhiza uralensi or Glychyrrhiza glabra.
Item 8: Any one of 1 to 7, wherein the TTG content contained in the root of the licorice plant, measured by a method according to the Japanese Pharmacopoeia, is 2% by mass or more in dry mass. The cultivation method described in the section.
Item 9: An agent for plants of the genus Licorice, which comprises at least one compound selected from the group consisting of jasmonic acid and a derivative of jasmonic acid.
Item 10: The agent for plants of the genus Licorice according to Item 9, which is a liquid.
Item 11: The agent for plants of the genus Licorice according to Item 9 or 10, further comprising a surfactant.
Item 12: For plants of the genus Licorice according to any one of Items 9 to 11, wherein the compound is at least one selected from the group consisting of jasmonic acid, prohydrojasmon, and methyl jasmonate. Agent.
Item 13: Contains at least one compound selected from the group consisting of jasmonic acid and derivatives of jasmonic acid, and is characterized by increasing the GL content contained in the roots of Licorice plants by applying it to Licorice plants. An agent for plants of the genus Licorice.
Item 14: The agent for plants of the genus Licorice according to Item 13, which is a liquid.
Item 15: The agent for a licorice plant according to Item 13 or 14, further comprising a surfactant.
Item 16: The plant of the genus Licorice according to any one of Items 13 to 15, wherein the derivative of jasmonic acid is at least one selected from the group consisting of prohydrojasmon and methyl jasmonate. Agent.

(III) A medium for tissue culture of a plant belonging to the genus Licorice of the family Licorice, a method for promoting rooting using the same, and a method for producing molding of a plant of the genus Licorice of the family Licorice. The present invention relates to a method for promoting rooting using the same, and a method for producing molding of a plant belonging to the genus Licorice of the family Licorice. The target legumes of the genus Licorice are as described in (I) above as IPSQ-containing plants, and include Glycyrrhiza uralensis Fisch. And G. glabra L.

[Medium for tissue culture of legumes]
As a medium for tissue culture of plants of the genus Licorice of the family Leguminosae (hereinafter, simply referred to as "medium of the present invention"), a WPM (Woody Plant medium) medium containing the components shown in Table 1 below can be used as a basic medium. .. The WPM medium has been conventionally used as a basic medium for culturing organs and callus of woody plants (trees) whose target is different from that of the present invention, and its composition and preparation method can follow it.

For tissue culture of legumes of the genus Licorice, a WPM medium composed of the above components can be used as it is, but at least one selected from the group consisting of sugars, amino acids, and vitamins is further blended therein. WPM medium can also be used.

Here, the sugar is not limited as long as it is used as a carbon source, but glucose, sucrose, mannose, maltose, ribose, arabinose, galactose, melibiose, trehalose, cellobiose, lactose, raffinose, sorbitol and glycerol, and fructose. Can be mentioned. Of these sugars, one type alone or two or more types may be arbitrarily combined and added to WPM for use. It is preferably sucrose or glucose, and more preferably sucrose. It is also possible to combine sucrose with other sugars and add them to the WPM medium. The proportion of sugar to be blended in the WPM medium is not limited as long as it does not interfere with the effects of the present invention, but can generally be in the range of 0.01 to 10% by mass per 100% by mass of the final medium. It is preferably 0.1 to 5% by mass, and more preferably 0.5 to 3% by mass.

Amino acids include, but are not limited, such as glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, lysine, δ-hydroxylysine, arginine, cysteine, cystine, phenylalanine, tyrosine, tryptophan. , Histidine, proline, and at least one proteinogenic amino acid selected from the group consisting of 4-hydroxyproline. Preferred examples thereof include monoaminodicarboxylic acid amides such as asparagine and glutamine, and glycine. Glutamine is more preferable as the monoaminodicarboxylic acid amide. These amino acids may be used alone or in any combination of two or more kinds and added to WPM. The ratio (total amount) of amino acids to be blended in the WPM medium is not limited as long as it does not interfere with the effects of the present invention, but is usually more than 0 and 20 mg / L or less, preferably 15 mg / L or less per 100% by mass of the final medium. It is more preferably 10 mg / L or less.

Vitamins include vitamins and enzyme cofactors. Vitamin and enzyme cofactors are not limited as long as they are used for tissue culture, for example, PABA (p-aminobenzoic acid), vitamin K (biotin), vitamin B5 (D-calcium pantothenate), folic acid, I-. Inositol, nicotinic acid, niacinamide (nicotinic acid amide), vitamin B6 (pyrdoxine HCl) (and pyrodoxal HCl), vitamin B2 (riboflavin), vitamin B1 (thiamine) and vitamin B12 (cyanocobalamine), vitamins Examples thereof include C (L-ascorbic acid), choline chloride, choline dihydrogenic citrate, orthoic acid, putresin, timine, vitamin A (retinol), and vitamin D (cholecalciferol). The ratio (total amount) of vitamins to be blended in the WPM medium is not particularly limited as long as it does not interfere with the effects of the present invention, and can be appropriately adjusted.

The medium of the present invention may be liquid, semi-solid, or solid. In the semi-solid or solid form, especially when used in static culture (eg, in vitro culture), the medium of the present invention may contain a solidifying agent in addition to the WPM medium. In this case, the medium of the present invention may further contain at least one selected from the group consisting of the above-mentioned sugars, vitamins and amine acids. Here, as the solidifying agent, at least one selected from the group consisting of gellan gum (gellan gum, fighter gel, gellan gum), native gellan gum, agar, gelatin, pectin, carrageenan, starch, guar gum, xanthan gum, carboxymethyl cellulose, and tamarind gum is mentioned. be able to. These solidifying agents may be used alone by adding and blending them to the WPM medium, or may be used by arbitrarily combining two or more kinds and adding and blending them to WPM. Gellan gum and agar are preferred. The ratio of the solidifying agent to be blended in the WPM medium can be appropriately set according to the type of the solidifying agent, the hardness of the medium to be prepared, and the like. Although not limited, a range of more than 0 and 1% by mass or less per 100% by mass of the final medium can be mentioned. It is preferably 0.5% by mass or less, and more preferably 0.4% by mass or less. When these solidifying agents are used, it is preferable to combine them with monovalent cations such as lithium, sodium or potassium, or divalent cations such as calcium, magnesium or barium, depending on the type of solidifying agent.

The medium of the present invention may contain a plant hormone, but the amount thereof is preferably a small amount in consideration of the influence on the human body and the environment. Here, examples of the plant hormone include auxin, gibberellin, brassinosteroid, ethylene, cytokinin and the like. It is preferably auxin. Examples of auxins include indole-3-acetic acid (IAA), indole-3-butyric acid, naphthalene acetic acid (NAA), naphthoxyacetic acid, phenylacetic acid, 2,4-dichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxyacetic acid. Can be mentioned. These may be used alone by adding and blending to the WPM medium, or two or more kinds may be arbitrarily combined and used by adding and blending to WPM. The ratio (total amount) of plant hormones, especially auxin, to be blended in the WPM medium is 0 to less than 0.005 mg / L, preferably 0 to less than 0.002 mg / L, more preferably 0 to 0, per 100% by mass of the final medium. .001 mg / L or less. As shown in the experimental examples described later, according to the above-mentioned medium of the present invention, the rooting rate of a plant belonging to the genus Licorice of the family Leguminosae can be significantly improved without adding a plant hormone. Therefore, the medium of the present invention more preferably does not contain plant hormones in terms of both rooting efficiency and effects on the human body and the environment.

[Method of promoting rooting of plants of the genus Licorice of the family Leguminosae / Method of producing molding]
By culturing a tissue piece containing one or more axillary buds or shoot apex of a leguminous licorice genus plant using the above-mentioned medium of the present invention, rooting of a legume licorice genus plant is promoted as shown in an experimental example described later. And the rooting rate can be improved. As a result, it becomes possible to more efficiently produce the molding of plants of the genus Licorice of the family Leguminosae.

Here, for the culture of the tissue piece, a method used in the conventional tissue culture of a plant of the genus Licorice of the family Leguminosae can be used except that the medium of the present invention described above is used as the medium. The culturing method may be static culturing or shaking culturing, and can be appropriately selected depending on the tissue piece used for culturing.

Examples of the tissue pieces used for culturing include nearby tissues including growth points of sterilized plants obtained by organ differentiation from callus or aseptic seeding. The tissues containing the growth points include, for example, shoot apical tissue, axillary bud tissue, stem including axillary bud, root head, embryo, stron-like tissue and culture containing growth point (polyblast, shoot primordium, redifferentiated). Callus, etc.) can be mentioned. Tissue pieces containing stem apex or axillary buds, preferably isolated from sterilized licorice plants, are used for primary culture. The tissue piece containing the stem apex or axillary bud can be collected from the stem of a plant of the genus Licorice by an appropriate means such as cutting, and is prepared so as to leave the stem and the leaf or a part of the leaf. When placing the tissue piece in a medium, the stem portion of the tissue piece is placed so that the shoot apex and axillary buds of the tissue piece are oriented upward from the medium, and the tissue culture medium (for example, MS medium) is usually used. ). By culturing the tissue pieces, one shoot can be obtained from one shoot apex and one axillary bud contained therein. Since the shoot grows while forming a plurality of axillary buds, a large number of tissue pieces containing one shoot apex or one axillary bud can be collected from one shoot.

By using these as materials and culturing using the medium of the present invention in the same manner as described above, rooting can be promoted and rooting can be performed at a high rate. By repeating these series of culture operations, the rooted shoots of the licorice genus can be grown in a series in a short period of time.

The seedlings of Licorice plants obtained by the culture method using the medium of the present invention are taken out from the medium, and then the medium attached to the roots is removed with running water or the like so that the roots are not damaged by a support such as vermiculite or rock wool. After planting and growing under appropriate acclimation conditions for a certain period of time, it can be grown as a healthy plant (molding) by transplanting it to soil and cultivating it normally according to the conventional method.

As described above, in the present specification, the term "contains" is used to include the terms "consisting of" and "consisting of".

The present invention will be described in more detail below with reference to Experimental Examples and Examples, but the present invention is not limited to these Experimental Examples and Examples. Unless otherwise specified, the steps, treatments, or operations in the following experimental examples are carried out under room temperature and atmospheric pressure conditions. Room temperature means a temperature in the range of 10 to 40 ° C.

Experimental Example 1
Promotion of IPSQ production with respect to GL content using a licorice genus plant (Glychyrrhiza uralensis) containing glycyrrhizin (GL), which is a triterpene glycoside (TTG), as an isoprenoid (IPSQ) using squalane as a precursor. The effect of agent (TTG production promoter) treatment was investigated. A jasmomate solution (Meiji Seika Pharma Co., Ltd.) was used as an IPSQ production promoter. The composition of the jasmomate solution is shown in Table 2.

(1) Experimental method The seeds of Glycyrrhiza uralensis were sown in a medium in a test tube in a sterile state to obtain tissue culture seedlings that had been cloned and proliferated. Specifically, the tissue culture seedlings were prepared as follows.
1. 1. The seeds of Glycyrrhiza uralensis are soaked in 10-fold diluted antiformin for 5 minutes to sterilize the seed surface, and then rinsed 3 times with sterile water.
2. 2. The surface-sterilized seeds are allowed to stand (24 ± 1 ° C., 16 hours, 3000 to 6000 lux, dark period 8 hours) on a medium (MS medium or WPM (Woody Plant Medium) medium) in vitro to germinate.
3. 3. After germination (after rooting), the axillary buds that have been cultured and grown for about 2 to 4 weeks are aseptically cut out and inserted into a new medium.
4. After cutting and germination, repeat steps 3 and 4 to increase the number of clones.
After the insertion of No. 4, cultured seedlings with sufficient root development were used for the test.

The two clones (clone 1 and 2) of the tissue culture seedlings obtained by the above clone proliferation were divided into a jasmomate-treated group (test group: Example 1) and a jasmomate non-treated group (test group: Comparative Example 1), respectively, and described below. An experiment was conducted. In the following experiment, the medium was 20 mL each of WPM medium containing 1% sucrose (w / v), 0.1 mg / L L-glutamic acid, and 0.2% gellan gum (w / v). The one prepared by pouring was used.

On the medium of the tissue culture seedlings (clone 1 and 2) of the jasmomate-treated group, 500 μL of a diluted solution of the jasmomate solution (hereinafter, simply referred to as “jasmomate solution”) was added dropwise so as not to cover the seedlings. The jasmomate solution was prepared by diluting the jasmomate solution with water 1000 times in terms of volume and sterilizing and filtering so that the prohydrojasmon concentration was 50 ppm. The specific gravity of prohydrojasmon to 1 mass of water is 0.97.

This tissue culture seedling was cultured and grown under the conditions of 24 ± 1 ° C. and a day length of 16 hours (dark period 8 hours) of 3000 to 6000 lux for 1 week from under the jasmomate droplets. It is considered that at least a part of the component (prohydrojasmon) contained in the jasmomate solution dropped on the medium during the culture growth process volatilized and came into contact with the above-ground part of the seedling. One week after the dropping, the underground part (taproot) of the grown tissue culture seedling was collected, and the total weight (raw weight) of the underground part and the GL content contained in the underground part were measured. On the other hand, the tissue culture seedlings (clones 1 and 2) of the jasmomate untreated group were similarly cultured and grown for one week without treatment, and the GL content contained in the underground part was measured.

The GL content in the underground part was measured by the following method in accordance with the 17th revised Japanese Pharmacopoeia.
[Measurement method of GL content]
The mass of the dried underground part is precisely weighed and powdered using CT193 Cyclotech (manufactured by Foss Japan). Weigh accurately about 1 g of the powder, add 30 ml of dilute ethanol, shake for 15 minutes, centrifuge, and separate the supernatant. Add 15 ml of dilute ethanol to the residue and operate in the same manner. Combine all the extracts and add dilute ethanol to make exactly 50 ml, and use this as the sample solution. Separately, weigh accurately about 20 mg of the standard glycyrrhizin product (separately, the water content of 10 mg is measured by the potentiometric titration method) and dissolve it in dilute ethanol to make exactly 50 ml. Weigh accurately 10 ml of this solution and add dilute ethanol to make 50 ml, which is used as the standard solution. Accurately take 20 μl each of the sample solution and the standard solution, subject them to HPLC under the following conditions, and calculate the glycyrrhizin content (mg) contained in the sample solution from the obtained peak area of glycyrrhizin based on the following formula.

<Calculation formula>
Amount of glycyrrhizin (GL) (mg)
= Amount of GL standard product converted to dehydrated product (mg) x (Ar / Ar) x (1/5)
Ar: GL peak area of sample solution As: GL peak area of standard solution

<HPLC conditions>
HPLC device: Alliance HPLC 2695 (manufactured by Waters)
Column: 5 μm octadesul silylated silica gel (inner diameter 4.6 mm, length 15 cm)
Column temperature: Around 40 ° C. Mobile phase: 3.85 g of ammonium acetate is dissolved in 720 ml of water, and 5 ml of acetic acid (100) and 280 ml of acetonitrile are added.
Flow rate: 1 to 1.5 ml / min (so that the GL retention time is about 15 minutes)
Detector: Ultraviolet absorptiometer (measurement wavelength: 254 nm)

(2) Experimental results Table 3 shows the measurement results. The table shows the GL concentration (ppm) calculated from the amount of GL (mg) contained in the total weight (wet weight mg) of the underground part (raw) measured by the above method as the GL content.

As shown in Table 3, in clone 1 and clone 2 of the jasmomate treatment group (Example 1), the weight (mg) and GL content (absolute amount [mg] and unit of the underground part) of the underground part were about 1 week after the dropping of the jasmomate solution. The amount contained per weight [ppm]) was increased as compared with those in the jasmomate untreated group (Comparative Example 1). From this, by treating a licorice plant containing GL, which is a TTG, in the underground part with a JA compound in the growth process, the production of GL is promoted, the amount of storage in the underground part is increased, and the GL is stored. It was confirmed that the weight of the underground part containing licorice also increased.

Experimental Example 2
Based on the results of Experimental Example 1, a cultivation test in soil was carried out, and in the same manner, the effect of the IPSQ production promoter treatment on the GL content of Glycyrrhiza uralensis was investigated. We also investigated the effect of IPSQ production promoter treatment on the contents of glycyrrhetinic acid (GA) and 3-monoglucuronyl glycyrrhetinic acid (3MGA) contained in the underground part of Glycyrrhiza uralensis.

(1) Experimental method Three months after sowing the seeds of Glychyrrhiza uralensis, seedlings are planted in a field (owned by Oji Holdings Co., Ltd. in Kitagakuden, Kuriyama-cho, Yubari-gun, Hokkaido), and about 1 The seedlings (planted seedlings) grown in the year were used for the test.

The planted seedlings (growth period: January 1 year) were divided into a jasmomate-treated group (Example 2) and a jasmomate-untreated group (comparative example 2). In the jasmomate-treated group, a diluted solution of the jasmomate solution (diluted 500 times with water) was sprayed on the above-ground part at an amount of 30 ml / seedling so as to adhere uniformly to the surface of the leaves. The number of sprays was one. It was cultivated and grown in a natural environment for about one month. On the other hand, the untreated Jasmomate group was similarly cultivated and grown in the natural environment without being treated. Then, the underground part was randomly dug up from each of the jasmomate-treated group and the non-treated group, and the taproots of 5 individuals in each group were collected. The dry weight of the taproots of 5 individuals in each group (total amount of 5 individuals) was measured, and the GL content (total amount of 5 individuals) contained in these taproots was reported to the 17th revised Japanese Pharmacopoeia in the same manner as in Experimental Example 1. Measured in a compliant manner. In addition, glycyrrhetinic acid (GA) and 3-monoglucuronyl glycyrrhetinic acid (3MGA) contained in the taproot were also analyzed using the LC-MS / MS method, and a calibration curve prepared with each standard having a known concentration was obtained. Quantified using.

[Quantitative determination of GA and 3MGA]
Equipment: LC-MS / MS (3200QTRAP: manufactured by ABSciex) Fixed phase: ODS column (Cadenza CD-C18: manufactured by Intact) (inner diameter 2 mm, length 25 cm)
Mobile phase: Mixture of water and acetonitrile Flow rate: 0.3 ml / min
Detection: Multiple Reaction Monitoring.

In addition, the same experiments were performed on the planted seedlings (growth period: February 1 year), and each was randomly selected from each jasmomate-treated group (Experimental Example 3) and non-treated group (Comparative Example 3). Five taproots were collected and their dry weight and GL content, glycyrrhetinic acid (GA) content, and 3-monoglucuronyl glycyrrhetinic acid (3MGA) content contained in the taproot were measured (all as the total amount of the five individuals). Calculation). The GL content was measured according to the method described in Experimental Example 1, and the GA content and 3MGA content were measured according to the above method.

(2) Experimental results Table 4 shows the measurement results. The table shows the GL concentration (%) calculated from the amount of GL (mg) contained in the total weight (dry weight) (mg) of the taproot measured by the above method as the GL content. The GA content and the 3MGA content are shown as the GA content and the 3MGA content (μg) contained in the total weight (dry weight) (g) of the taproot measured by the above method.

As shown in Table 4, all of the jasmomate-treated groups (Experimental Examples 2 and 3) were treated with jasmomate in all of the GL amount, GA amount, and 3MGA amount contained in the taproot about one month after the application of the jasmomate solution. It was clearly increased as compared with the untreated jasmomate group (Comparative Examples 2 and 3). In addition, the weight of the taproot was increased in the jasmomate-treated group as compared with the non-treated group. From this, as in Experimental Example 1, by treating the licorice plant with the JA compound, the production of GL, GA and 3MGA in the underground is promoted and the content thereof is increased by soil cultivation. It was confirmed that the weight of the underground part also increased.

Experimental Example 3
Using methyl jasmonate as an IPSQ production promoter (TTG production promoter), the same test as in Experimental Example 1 was carried out to investigate the effect on the glycyrrhizin (GL) content.

(1) Experimental method Similar to Experimental Example 1, seeds of Glychyrrhiza uralensis were sown in a medium in vitro in a sterile state to obtain tissue culture seedlings that had been cloned and proliferated.

The three clones (clone 1-3) of the tissue culture seedlings obtained by clone proliferation were divided into a methyl jasmonate-treated group (test group: Example 4) and a methyl jasmonate-untreated group (test group: Comparative Example 4), respectively. , The following experiment was performed.

500 μL of a methyl jasmonate solution was added dropwise to the medium of the tissue culture seedlings (clone 1 to 3) of the methyl jasmonate-treated group so as not to cover the seedlings. Methyl jasmonate solution is a spreading agent per 1 L of methyl jasmonate aqueous solution prepared by diluting a methyl jasmonate product (manufactured by Wako Pure Chemical Industries, Ltd.) with water so that the concentration of methyl jasmonate becomes 0.05% by mass. Was added at a ratio of 0.3 ml and prepared. Dyne (containing polyoxyethylene nonylphenyl ether and calcium lignin sulfonate: Sumitomo Chemical Garden Products Co., Ltd.) was used as the spreading agent.

This tissue culture seedling was added to the methyl jasmonate solution dropped onto the medium during the culture growth process of 3000 to 6000 lux at 24 ± 1 ° C. for 16 hours (dark period 8 hours) for 1 week after the addition of the methyl jasmonate solution. It is considered that at least a part of the contained component (methyl jasmonate) volatilized and came into contact with the above-ground part of the seedling. One week after the dropping, the roots (taproot and lateral roots) of the grown tissue culture seedlings were collected, and the total weight (wet weight mg) of the roots (raw) and the GL content (mg) contained in the roots were measured with Experimental Example 1. Similarly, the measurement was performed in accordance with the 17th revised Japanese Pharmacopoeia.

(2) Experimental results The measurement results are shown in Table 5. The table shows the GL concentration (ppm) calculated from the amount of GL (mg) contained in the total weight (wet weight mg) of the underground part (raw) as the GL content.

As shown in Table 5, in any of the clones treated with methyl jasmonate (Example 4), the GL content at the root was about 1 week after the addition of the methyl jasmonate solution, and the GL content at the root was not treated with methyl jasmonate (Comparative Example 4). Was increasing compared to. In particular, it was confirmed that the raw weight of the root of clone 2 treated with methyl jasmonate was significantly increased as compared with the group not treated with methyl jasmonate. From this, it was confirmed that the same effect as in Experimental Example 1 can be obtained even when methyl jasmonate is used as the JA compound.

Experimental Example 4
Using licorice (Glychyrrhiza glabra) as a plant of the genus Licorice, the same test as in Experimental Example 1 was carried out to investigate the effect of IPSQ production promoter (TTG production promoter) treatment on glycyrrhizin (GL) content.

(1) Experimental method Tissue culture seedlings cloned and proliferated were obtained in the same manner as in Experimental Example 1 except that licorice (Glychyrrhiza glabra) was used as a plant of the genus Licorice. The three clones (clone 1 to 3) of the tissue culture seedlings obtained by clone proliferation were divided into a jasmomate-treated group (test group: Example 5) and a jasmomate non-treated group (test group: Comparative Example 5), respectively, and Experimental Example 1 The same experiment was performed. One week after the jasmomate droplets were dropped, the GL content (mg) contained in the roots (taproot and lateral roots) of the cultured seedlings was measured by a method according to the Japanese Pharmacopoeia. In addition, the weight (wet weight mg) of the root (raw) of licorice was measured.

(2) Experimental results Table 6 shows the measurement results. The table shows the GL concentration (ppm) calculated from the amount of GL (mg) contained in the total weight (wet weight mg) of the root (raw) as the GL content.

As shown in Table 6, even when licorice is used as a plant of the genus Licorice, the jasmomate-treated group (Example 5) has the raw weight of the root and GL about one week after the jasmomate droplets, as in Experimental Example 1. The contents were all increased as compared with those in the jasmomate untreated group (Comparative Example 5). From this, by treating the licorice genus plant containing GL in the underground part with the JA compound, the production of GL is promoted, the storage amount in the underground part is increased, and the weight of the underground part containing GL is also increased. It was confirmed that it increased.

Experimental Example 5
Using Panax ginseng, which is a plant belonging to the genus Panax japonicus in the family Araliaceae, as a plant containing TTG as IPSQ, the effect of IPSQ production promoter (TTG production promoter) treatment on the weight of the underground part (main root) was investigated. Panax ginseng is a medicinal plant containing ginsenosides in its taproot. A jasmomate solution (Meiji Seika Pharma Co., Ltd.) was used as the IPSQ production accelerator (TTG production accelerator).

(1) Experimental method The shaking culture callus of Panax ginseng was weighed by removing the water on the surface with a filter paper, and then planted on a solid medium in a petri dish at a ratio of 5 per petri dish. Here, the shake-cultured callus is a callus induced by using a root tissue piece of Panax ginseng, which is cultured in an MS medium (pH 5.6) containing sucrose in a dark place at 24 ° C. to maintain the passage. It was used. As the solid medium, MS medium (pH 5.6) containing sucrose and gellite (w / v) (Wako Pure Chemical Industries, Ltd.) was used.

Prepare three of the above-mentioned petri dishes in which shaking culture callus was planted (for jasmomate treatment and for non-jasmomate treatment), and add 500 μl of a 500-fold diluted solution of jasmomate solution to the callus in the petri dish for jasmomate treatment. 500 μl of sterile water per individual was dropped from above onto the callus in the petri dish for jasmomate treatment, and each was irradiated with 1600 lux light for 16 hours a day under 24 ° C. and statically cultured for 4 weeks. ..
The weight of the callus was measured every week from the start of the culture.

(2) Experimental results Table 7 shows changes in callus weight over time. The numerical values in parentheses in Table 7 indicate the increased amount (mg) obtained by subtracting the callus weight at the start of culturing from the callus weight after culturing.

As shown in Table 7, it was confirmed that the weight of the callus increased significantly with time by the jasmomate treatment as compared with the case without the jasmomate treatment. Specifically, the weight of callus treated with jasmomate at 4 weeks of culture was 124% higher in absolute weight ratio of callus and 160% higher in weight increase ratio than in the case without jasmomate treatment. From this, it was confirmed that the jasmonic acid compound has a growth promoting effect on the taproot tissue of a plant belonging to the genus Panax japonicus in the family Araliaceae.

Experimental Example 6
Based on the results of Experimental Example 5, ginseng 3rd grade individuals were cultivated in soil, and the effect of IPSQ production promoter (TTG production promoter) treatment on ginsenoside content was investigated.

(1) Experimental method (1-1) Preparation of ginseng third-year-old individual Panax ginseng third-year individual was cultivated and prepared as follows.
(A) In 2014, seeds of Panax ginseng were sown in the soil of the land of Hokkaido.
(B) In 2015, the grown seedlings were harvested as first-year autumn seedlings and transplanted to the soil of the same land to continue cultivation.
(C) In 2017, acquired a third-year individual of Panax ginseng.

Cultivation was carried out in a natural environment under light-shielding conditions.

(1-2) Experimental method The planting test plot of Panax ginseng 3rd grade individuals was divided into 2 plots (jasmomate treated plot and jasmomate non-treated plot) (unit area of 1 plot: 0.9 m ² ) and planted in the jasmomate treated plot. 200 ml of jasmomate solution (jasmomate solution diluted 500-fold with water) was sprayed on the above-ground part of Panax ginseng so as to evenly adhere to the surface of the leaves (implemented in July 2017) (Example 7). The number of sprays was one. After that, it was cultivated and grown in a natural environment under shading conditions for about 3 months. On the other hand, Panax ginseng planted in the Jasmomate untreated plot was cultivated and grown in a natural environment under the same light-shielding conditions without being treated (Comparative Example 7).

Then, in September 2017, the underground part of all ginseng plants planted in the jasmomate treated area and the untreated area was dug up and the taproot was collected. The wet weight (total amount g) of the taproot of Panax ginseng in each test group was measured, and the yield per unit area of soil (wet weight g / m ² ) was calculated. In addition, the amount of ginsenoside (ginsenoside Rg1 and ginsenoside Rb1) contained in the taproot of Panax ginseng collected from each test group was measured by a method according to the 17th revised Japanese Pharmacopoeia.

(2) Experimental results The measurement results are shown in Tables 8 and 9.

表中、括弧内の数値は、ジャスモメート非処理区の結果を100とした場合におけるジャスモメート処理区の含有量（比率）を示す。

In the table, the numerical values in parentheses indicate the content (ratio) of the jasmomate-treated group when the result of the jasmomate-untreated group is 100.

As shown in Table 9, in the ginseng treated group (Experimental Example 7), the amount of ginsenosides (Rb1 and Rg1) contained in the taproot about 1 month after the spraying of jasmomate was not treated with jasmomate. It was significantly increased (about 1.4 times) as compared with Panax ginseng (Comparative Example 7). Further, as shown in Table 8, the weight of the taproot was increased in the jasmomate-treated group as compared with the non-treated group (about 1.4 times). From this, as in Experimental Example 6, by treating the Araliaceae Panax japonicus plant with the JA compound, the production of ginsenoside in the root is promoted and its content is increased, and the root portion is also cultivated in soil. It was confirmed that the weight of the ginseng also increased.

Experimental Example 7 Promotion of rooting of Licorice plants of the family Licorice Axillary buds (including shoot apex) prepared from seeds of Licorice plants of the family Licorice (Ural licorice, licorice) were cultivated using various media to determine the rooting rate. Compared.
Specifically, according to the description of 1 to 3 of "(1) Experimental method" of Experimental Example 1, the axillary buds (specimen) of a leguminous plant belonging to the genus Licorice, which was cultured for 2 to 4 weeks after germination from seeds and then aseptically cut out. ) Was inserted into the medium shown in Table 11 below and cultured. Among the basal media shown in Table 11, the MS medium (Murashige-Skoog medium) is a basal medium commonly used for tissue culture of herbaceous plants. The medium 1 using this as the basic medium is M. This medium is used in the paper by Kohjyouma et al. (Non-Patent Document 1), and was used as a comparative medium (comparative example) for the present invention. On the other hand, the WPM medium used in the media 2 to 5 is a medium used for tissue culture of woody plants, and is not usually used for tissue culture of herbaceous plants. The composition is shown in Table 10. As the basic medium, those to which vitamins were added were used. Specifically, the MS medium (Wako Pure Chemical Industries, Ltd.) used was a basic MS medium supplemented with Murashige and Skoog vitamin powder (Sigma-Aldrich), and the WPM medium was McCown containing vitamins and the like. Woody Plant medium including vitamins (Duchefa) was used (see Table 11).

The culture was carried out for 1 month under the conditions of 24 ± 1 ° C., 16 hours day length 3000 to 6000 lux, and dark period 8 hours. One month after culturing, the presence or absence of rooting was visually confirmed for each sample, and the rooting rate was calculated by the following formula.
[Number 1]
Rooting rate (%) = (number of rooted samples / total number of samples) x 100

The results are also shown in Table 12.

As shown in Table 12, by culturing using WPM medium as the basal medium, rooting of licorice genus plants in tissue culture is promoted and rooting is promoted regardless of the presence or absence of auxin, which is a kind of plant hormone. It was confirmed that the rate was high. In particular, the effect was significantly observed by adding a nitrogen source such as glutamine or sugar such as sucrose to the WPM medium. Further, by adding a solidifying agent in addition to the nitrogen source and sugar to the WPM medium to prepare a solid medium, rooting can be promoted and the rooting rate can be significantly improved without adding auxin. It was confirmed that it could be done. From this, the rooting rate in tissue culture of Licorice plants, which had a low rooting rate by the conventional method, can be improved by using WPM medium as the basal medium without using auxin. In addition, since this method does not require the use of plant hormones such as auxin for culturing, according to the culturing method using the medium of the present invention, there is no accumulation of plant hormones in the plant body, and safe crude drugs and sweeteners are used. It becomes possible to produce a plant of the genus Licorice as a raw material.

Claims (16)

An IPSQ production promoter for plants containing prohydrojasmon represented by the following formula (1) .

(In the formula, R ¹ means an alkyl group having 5 carbons and R ² means an alkyl group having 3 carbons):
IPSQ is at least one selected from the group consisting of triterpenes and triterpene glycosides .
An IPSQ production promoter, wherein the plant is a plant of the genus Licorice of the family Araliaceae containing a triterpene and a triterpene glycoside, or a plant of the genus Panax japonicus of the family Araliaceae containing a triterpene glycoside. The IPSQ production promoter for plants according to claim 1, further comprising a surfactant. The IPSQ is a triterpene, and the triterpene is glycyrrhetinic acid, liquiric acid, grabric acid, α-amyrin, tormentic acid, corosolic acid, ursoric acid, arisole, herboric acid, fucidic acid, lanosterol, ebriconic acid, polyporenic acid. , Tumrosic acid, pachymaic acid, cycloartenol, amyrinol, amyrinol, damalendiol, protopanaxadiol, protopanaxatriol, euhole, euphorbol, elemolic acid, eremonic acid, lupeol, bethrin, bethrinic acid, limonine, evodol, Lutaebin, obakunone, quassinoid, bruceanthin, masphosphate, 2α, 19α-dihydroxy-3-oxourus-12-en-28-euic acid, β-amyrin, oleanolic acid, fupensic acid, heragenin, platycodigenin, polygalasic acid, presenegenin The IPSQ production promoter for plants according to claim 1 or 2, which is at least one selected from the group consisting of amyrin, chasapogenol, and salts thereof. The IPSQ production promoter for plants according to claim 3 , wherein the plant is a triterpene-containing plant belonging to the genus Licorice of the family Leguminosae. The IPSQ is a triterpene glycoside, and the triterpene glycosides are glycyrrhizin, 3-monoglucuronyl glycyrrhetinic acid, baccalocid B, avenacin A-1, soybean saponin, ginsenoside, uralsaponin, tixetsusaponin, and eleutheroside A. , Akevoside, Platicodin, Senegin, Ondisaponin, Psychosaponin, Theasaponin, Astersaponin, Cibilicoside, Smiraxsaponin, Estin, Araloside, Astragaloside, Gymnemic acid, Gibenoside and at least one selected from the group consisting of salts thereof. The IPSQ production promoter for plants according to claim 1 or 2 , which is a species. The IPSQ production promoter for plants according to claim 5 , wherein the plant is a triterpene glycoside-containing plant belonging to the genus Licorice of the family Leguminosae or the genus Panax japonicus of the family Araliaceae. The use for producing an IPSQ production promoter for plants, which is a prohydrojasmonic acid represented by the following formula (1) or a composition containing the same.

(In the formula, R ¹ means an alkyl group having 5 carbons and R ² means an alkyl group having 3 carbons):
IPSQ is at least one selected from the group consisting of triterpenes and triterpene glycosides .
Use, wherein the plant is a plant of the genus Licorice of the family Mame containing a triterpene and a triterpene glycoside, or a plant of the genus Ginseng of the family Araliaceae containing a triterpene glycoside . The use according to claim 7 , wherein the composition further contains a surfactant. A composition containing prohydrojasmon represented by the following formula (1), which is used for promoting the production of IPSQ in an IPSQ-containing plant .

(In the formula, R ¹ means an alkyl group having 5 carbons and R ² means an alkyl group having 3 carbons) :
IPSQ is at least one selected from the group consisting of triterpenes and triterpene glycosides.
The composition, wherein the IPSQ-containing plant is a licorice genus plant containing triterpenes and triterpene glycosides or a Panax japonicus plant containing triterpene glycosides . The composition containing prohydrojasmon according to claim 9 , wherein the composition further contains a surfactant. A method for producing an IPSQ-containing plant or a processed product thereof, which comprises the following steps (a) and (b).
(A) A step of treating an entire plant or a part thereof of an IPSQ-containing plant with prohydrojasmon represented by the following formula (1) or a composition containing the same.

(In the formula, R ¹ means an alkyl group having 5 carbons and R ² means an alkyl group having 3 carbons),
(B) Step of growing the plant treated in the step (a):
The above-mentioned production method, wherein the IPSQ-containing plant is a plant of the genus Licorice of the family Mame containing a triterpene and a triterpene glycoside or a plant of the genus Ginseng of the family Araliaceae containing a triterpene glycoside . The production method according to claim 11 , wherein the step (a) is a step of treating the whole plant or a part of the IPSQ -containing plant with an aqueous solution containing 0.001 to 5% by mass of prohydrojasmon. The production method according to claim 11 or 12 , wherein the step (a) is a step of treating a ground portion of an IPSQ-containing plant. The IPSQ-containing plant is a plant of the genus Licorice of the family Leguminosae containing a triterpene glycoside, and the GL content in 100% by mass of the underground portion measured by a method in accordance with the 17th revised Japanese Pharmacopoeia is converted into a dry matter. The production method according to any one of claims 11 to 13 , which is a method for producing a plant containing 2.0% by mass or more of IPSQ. The IPSQ-containing plant is a plant of the genus Panax japonicus containing a triterpene glycoside, and the ginsenoside Rb1 and Rg1 contents in 100% by mass of the main root measured by a method in accordance with the 17th revised Japanese Pharmacy Law are The production method according to any one of claims 11 to 13 , which is a method for producing IPSQ-containing plants of 0.2% by mass or more and 0.1% by mass or more, respectively, in terms of dried matter. Using the prohydrojasmon represented by the following formula (1) or a composition containing the same, a plant belonging to the genus Panax japonicus or Araliaceae containing at least one IPSQ selected from the group consisting of triterpenes and triterpene glycosides. A method for promoting the production of IPSQ in a plant having a step of treating a plant belonging to the genus Panax japonicus.

(In the formula, R ¹ means an alkyl group having 5 carbons, and R ² means an alkyl group having 3 carbons).