JP2005320266A

JP2005320266A - Method for promoting plant growth, plant growth promoter and farm material

Info

Publication number: JP2005320266A
Application number: JP2004137970A
Authority: JP
Inventors: Tatsuhiko Kin; 辰彦金; Hiroshi Oonami; 弘大南
Original assignee: TA STEVIA CO Ltd; Houei Co Ltd
Current assignee: TA STEVIA CO Ltd; Houei Co Ltd
Priority date: 2004-05-07
Filing date: 2004-05-07
Publication date: 2005-11-17

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new method for promoting plant growth that is stably effective to plant growth, to obtain a new plant growth promoter that excels in quality uniformity and stability, and provide a farm material containing the plant growth promoter. <P>SOLUTION: Luteolin is given to a plant for promoting its growth. The plant growth promoter comprises luteolin. The farm material contains the plant growth promoter containing the luteolin and is provided in the form of powder, solid, grain or fluid. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

本発明は、農業、林業等において、植物の収量の向上のため、植物の生長を促進させる方法、植物生長促進剤及び農業資材に関する The present invention relates to a method for promoting plant growth, plant growth promoter and agricultural material for improving plant yield in agriculture, forestry and the like.

ステビアの葉及び茎の乾燥粉末を温水抽出し更にこれを発酵させたステビア抽出液を有効成分とする園芸用肥料が知られている（例えば特許文献１）。そしてこの園芸用肥料は、野菜や果実等の甘味の向上等に効果を有するとされる。
特許第２５７７１０５号公報 A horticultural fertilizer is known which contains a stevia extract obtained by warm water extraction of a dried stevia leaf and stem powder and further fermenting it (for example, Patent Document 1). This horticultural fertilizer is said to have an effect on improving sweetness such as vegetables and fruits.
Japanese Patent No. 2577105

公知の園芸用肥料は、ステビア抽出液の発酵に１８０〜３６０日の長期期間が必要である。更に、長期期間発酵させるため、種々の環境の変化により品質の均一性、安全性に欠けるという課題があり、使用に際しては水によって５００〜１５００倍に希釈するので、品質の均一性、安定性に欠け、植物の生長に対する有効性についても安全性に欠けるという課題がある。
本発明は，上記課題に鑑み、植物の生長に対して安定して有効性のある新規な植物の生長を促進する方法を提供すると共に、品質の均一性，安定性がある新規な植物生長促進剤及びこの植物生長促進剤を含む農業資材を提供するものである。 Known horticultural fertilizers require a long period of 180 to 360 days for fermentation of the stevia extract. Furthermore, since fermentation is performed for a long period of time, there is a problem of lack of quality uniformity and safety due to changes in various environments, and since it is diluted 500 to 1500 times with water during use, the quality uniformity and stability are improved. There is a problem of lack of safety in terms of lack and effectiveness on plant growth.
In view of the above problems, the present invention provides a method for promoting the growth of a novel plant that is stable and effective for the growth of the plant, and also promotes the growth of a novel plant having uniform quality and stability. An agricultural material containing the agent and the plant growth promoter is provided.

そのために、本発明の植物の生長を促進する方法は、下記［化学式１］のルテオリンを植物に与えることによる。
また、本発明の植物生長促進剤は、ルテオリンからなるものである。
さらに、本発明の農業資材は、ルテオリンからなる植物生長促進剤を含むことを特徴とするものである。
そして、本発明の農業資材は、粉末、固体、粒体、液体の材型であることを特徴とするものである。 Therefore, the method for promoting the growth of the plant of the present invention is by providing the plant with luteolin of the following [Chemical Formula 1].
The plant growth promoter of the present invention is composed of luteolin.
Furthermore, the agricultural material of the present invention is characterized by containing a plant growth promoter composed of luteolin.
And the agricultural material of this invention is a powder, a solid, a granule, and a liquid material type, It is characterized by the above-mentioned.

発明者は、植物生長促進効果の客体がルテオリンと判明し、肥料等の資材に適宣種々の作物等に応じてルテオリンを０．０１×１０^−９〜５×１０^−９Ｍの濃度範囲にて使用することにより、植物の成長に対する有効性について品質の均一な安定した効果を得ることができることに基づき本発明に想達した。 The inventor found that the object of the plant growth promoting effect was luteolin, and the luteolin in the concentration range of 0.01 × 10 ⁻⁹ to 5 × 10 ⁻⁹ M according to various crops suitable for materials such as fertilizers. The present invention has been conceived based on the fact that it is possible to obtain a uniform and stable effect on the effectiveness of plant growth.

本発明の植物の生長を促進する方法は、ルテオリンを植物に与えることによるから、植物の生長が安定して有効である効果がある。
本発明の植物生長促進剤はルテオリンからなり、本発明農業資材は上記植物生長促進剤を含むものであるから、品質が均一であり、安定性があるものを提供できる効果がある。 Since the method for promoting the growth of the plant of the present invention is by giving luteolin to the plant, there is an effect that the growth of the plant is stable and effective.
Since the plant growth promoter of the present invention is composed of luteolin and the agricultural material of the present invention contains the plant growth promoter, there is an effect of providing a product having uniform quality and stability.

本発明の植物の生長を促進する方法、植物生長促進剤及び農業資材の詳細を以下に説明する。 Details of the method for promoting plant growth, the plant growth promoter and agricultural materials of the present invention will be described below.

（ルテオリンの調整）
ステビア抽出液の生長促進効果について検証を行った結果、キュウリ発芽種子ではシャーレ粗寒天培地１０ｍｌに対してステビア乾燥量２ｍｇに相当する抽出物の濃度で、地上部・地下部ともに４０％以上の著しい生長促進効果が見られた。そこで、ステビアの農業用資材としての有効性をより明確にするために、その原因物質の検索、同定作業を行った。 (Luteolin adjustment)
As a result of verifying the growth promoting effect of Stevia extract, cucumber germinated seeds with a concentration of extract equivalent to 2 mg of Stevia dry for 10 ml of Petri dish agar medium, both above ground and below ground are markedly 40% or more. Growth promotion effect was seen. Therefore, in order to clarify the effectiveness of Stevia as an agricultural material, we searched and identified the causative substances.

植物生長促進物質の検索
図１は、植物生長促進物質の分類、精製行程を示したものである。当該各行程における生長促進の活性は、キュウリ発芽種子を用いたバイオアッセイで確認をした。図１中のＦｒ．７−３の分画で強い生長促進活性が見られた（図２参照）。 Search for Plant Growth Promoting Substances FIG. 1 shows the classification and purification process of plant growth promoting substances. The growth promoting activity in each step was confirmed by a bioassay using cucumber germinated seeds. In FIG. A strong growth-promoting activity was observed in the 7-3 fraction (see FIG. 2).

Ｆｒ．７−３（これをＦｒ．８と改名）の分画精製操作を進め、物質の構造決定を試みた。Ｆｒ．８の主成分であるＦｒ．８−２を起点としてＦｒ．１４までの分離精製を行った。 Fr. 7-3 (this was renamed as Fr. 8) was subjected to fraction purification, and an attempt was made to determine the structure of the substance. Fr. Fr. 8-2 as the starting point. Up to 14 separation and purification were performed.

植物生長促進物質の同定
Ｆｒ．８−２を再クロマトグラフィー（０〜６０ｍｉｎ：７０％メタノールイソクラティック溶出）に供した。バイオアッセイにより成長活性を確認したところ、Ｆｒ．１５−３という活性フラクションを得た。 Identification of plant growth promoting substances Fr. 8-2 was subjected to re-chromatography (0-60 min: 70% methanol isocratic elution). When growth activity was confirmed by bioassay, Fr. An active fraction of 15-3 was obtained.

Ｆｒ．１５−３をＳｅｐｈａｄｅｘＬＨ−２０クロマトグラフィー（１５×１５０ｍｍ、１００％メタノール）に供し、流速１ｍｌ／分で１０分間溶出分画を行った。この操作により、Ｆｒ．１５−３をＦｒ．１６−１からＦｒ．１６−３の３分画に分画した。 Fr. 15-3 was subjected to Sephadex LH-20 chromatography (15 × 150 mm, 100% methanol), and elution fractionation was performed at a flow rate of 1 ml / min for 10 minutes. By this operation, Fr. 15-3 in Fr. 16-1 to Fr. It fractionated into 3 fractions of 16-3.

Ｆｒ．１６−１からＦｒ．１６−３の分画についてバイオアッセイを行った結果、Ｆｒ．１６−２において肉眼で観察できるレベルでの活性が認められた（図３）。そこで、Ｆｒ．１６−２について活性の有無を確実に証明するためにＦｒ．１６−２の濃度依存性について検討した。その結果を図４に示す。図４を見ると、Ｆｒ．１６−２は１０ｐｐｂの濃度において根部・地上部でそれぞれ１５％・２５％の生長促進率を示した。これらの値は突出した生長促進効果を示すというには低い値であるが，統計学的には有意と認められる。また濃度依存性についてみると、４０ｐｐｂから１０ｐｐｂへと濃度が減少するに従って生長促進率が増大し、明確な濃度依存性が示唆される。こうした結果はＦｒ．１６−２が１０ｐｐｂ以下の濃度でさらに大きな活性を発現することを示唆している。 Fr. 16-1 to Fr. As a result of conducting a bioassay on the fraction of 16-3, Fr. In 16-2, activity at a level observable with the naked eye was observed (FIG. 3). Therefore, Fr. In order to reliably prove the presence or absence of activity for 16-2, Fr. The concentration dependency of 16-2 was examined. The result is shown in FIG. As can be seen from FIG. 16-2 showed a growth promotion rate of 15% and 25% in the root part and the above-ground part at a concentration of 10 ppb, respectively. These values are low to show outstanding growth-promoting effects, but are statistically significant. Further, regarding the concentration dependency, the growth promotion rate increases as the concentration decreases from 40 ppb to 10 ppb, suggesting a clear concentration dependency. These results are shown in Fr. This suggests that 16-2 expresses greater activity at a concentration of 10 ppb or less.

Ｆｒ．１６−２についてマススペクトルの結果分子量は２８６であると判明し、また ^１３Ｃ−ＮＭＲを測定した結果芳香族化合物であることが判明した。更にＮＭＲの特殊測定によりＦｒ．１６−２よりルテオリンを同定した。フラボノイドやフェノール類では同程度の濃度レベルで成長促進あるいは抑制効果が報告されている（Ｍａｃｉａｓｅｔａｌ．１９９７．１９９９）。そこで、Ｆｒ.１６−２の活性を詳細に検証するために標品のルテオリンを用いて低濃度領域における活性濃度依存性を調べた。 Fr. As a result of mass spectrum for 16-2, the molecular weight was found to be 286, and as a result of measuring ¹³ C-NMR, it was found to be an aromatic compound. Furthermore, Fr. Luteolin was identified from 16-2. Flavonoids and phenols have been reported to have growth-promoting or inhibitory effects at similar concentrations (Macias et al. 1997. 1999). Therefore, in order to verify the activity of Fr.16-2 in detail, the activity concentration dependency in the low concentration region was examined using the standard luteolin.

Demonstration example 2

（ルテオリンを用いた植物生長促進効果の確認）
市販ルテオリンを用いて図５に示す濃度でキュウリ発芽種子によるバイオアッセイを行い、その結果を表１に示す。 (Confirmation of plant growth promotion effect using luteolin)
Bioassay using cucumber germinated seeds was performed using commercially available luteolin at the concentrations shown in FIG. 5, and the results are shown in Table 1.

方法は、市販ルテオリンを更にゲルロ過にかけ不純物を取り除き、これを７．２０ｍｇにＴｗｅｅｎ８０（界面活性剤）１０ｍｌを加えて溶解させた。この液を脱塩水で希釈して寒天培養試験用に設立したモル濃度（５．０×１０^−８Ｍ）の原液を作成し、［表１］に示すルテオリン濃度に脱塩水で希釈することにより試験液を作成した。尚、各試験液のＴｗｅｅｎ８０濃度は同一になるようにＴｗｅｅｎ８０を適宣加えて調製した。試験液をそれぞれ１ｍｌずつシャーレに分注し、１％寒天水４ｍｌを加えて混合し、寒天培地を作成した。この寒天培地に、キュウリ発芽種子（青長四葉胡瓜、（株）オオタ種苗、発芽長２〜３ミリ）５粒播種して、２５℃暗所で３日間培養した。各濃度の試験は、３連で行った。Ｃｏｎｔｒｏｌとしてルテオリン無添加でＴｗｅｅｎ８０のみが入った区を設定した。そして、幼菌の根部及び下胚軸部の長さを測定して生長促進率（％）を算出した。 In the method, commercially available luteolin was further subjected to gel filtration to remove impurities, and this was dissolved in 7.20 mg by adding 10 ml of Tween 80 (surfactant). This solution was diluted with demineralized water to prepare a stock solution having a molar concentration (5.0 × 10 ⁻⁸ M) established for agar culture test, and diluted with demineralized water to the luteolin concentrations shown in [Table 1]. A test solution was prepared. Each test solution was prepared by appropriately adding Tween 80 so that the Tween 80 concentrations were the same. Each 1 ml of the test solution was dispensed into a petri dish, and 4 ml of 1% agar water was added and mixed to prepare an agar medium. On this agar medium, 5 seeds of cucumber germinated seeds (blue long four-leaf pepper, Ota seedling, germination length 2 to 3 mm) were sown and cultured in the dark at 25 ° C. for 3 days. Each concentration test was performed in triplicate. As a control, a section in which only Tween 80 was contained without the addition of luteolin was set. Then, the growth promotion rate (%) was calculated by measuring the length of the root part and the hypocotyl part of the larvae.

得られた結果を表１および図５に示した。これらの結果から０．０１〜５×１０^−９Ｍの濃度範囲でルテオリンの生長促進効果が認められ、０．３５×１０^−９Ｍ付近で活性最大値を示すことが明らかになった。 The obtained results are shown in Table 1 and FIG. From these results, it was clarified that the growth promoting effect of luteolin was observed in the concentration range of 0.01 to 5 × 10 ⁻⁹ M, and the maximum activity was observed in the vicinity of 0.35 × 10 ⁻⁹ M.

ステビアの植物生育促進物質の分離精製手順を示す図Diagram showing the separation and purification procedure of Stevia plant growth promoting substances Ｆｒ．７−３（Ｆｒ．８）のキュウリ種子生育促効果を示す図Fr. The figure which shows the cucumber seed growth promotion effect of 7-3 (Fr.8) Ｆｒ．１６−２のキュウリ種子生育促効果を示す図Fr. The figure which shows the cucumber seed growth promotion effect of 16-2 Ｆｒ．１６−２のバイオアッセイを示す図Fr. Diagram showing bioassay 16-2 ルテオリンの各濃度によるキュウリの幼苗の生長促進率を示す図Figure showing the growth promotion rate of cucumber seedlings at various concentrations of luteolin

Claims

A method for promoting plant growth by providing luteolin to plants.

A plant growth promoter consisting of luteolin.

An agricultural material comprising the plant growth promoter according to claim 2.

The agricultural material according to claim 3, wherein the agricultural material is a powder, solid, granule, or liquid material.