UA109939C2 - IMIDASOL DERIVATIVES - Google Patents

Abstract

A compound having the effect of regulating plant growth is selected from the group consisting of: (A) 3H-imidazo [4,5-d] [1,2,3] triazine-4,6 (5H, 7H) -dione; and (B) 3-methyl-3H-imidazo [4,5-d] [1,2,3] triazine-4,6 (5H, 7H) -dione.

Description

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The field of technology

ИО0О01) This invention relates to an imidazole derivative.

Technical level

І0002| For example, phytohormones are known as compounds that regulate plant growth.

Phytohormones are derived from plants themselves, but phytohormones can also be synthesized as compounds and used as agricultural chemicals and activators. As such plant growth regulating compounds, imidazole derivatives are described in patent literature 1 - 4. For example, in patent literature 4, 7H-imidazo|4,5-4|1,2,3)griazin-4(ZN)-one (another name: 2-azahypoxanthine, hereinafter sometimes referred to as "AGK") is described. and at the same time, it is also described that AHK exhibits the effect of stimulating growth or suppressing the growth of plants.

List of references

Patent literature

I0003)| Patent Literature 1: Japanese patent application Mo 63-104965 is set forth

Patent Literature 2: Japanese patent application Mo 63-68570 is set forth

Patent Literature 3: Japanese patent application Mo 4-210680 is set forth

Patent literature 4: Japanese patent Mo 4565018

Brief description of the invention

Technical problem

I0004| There is a need for a compound such as AHA that can regulate plant growth and can be used as an agricultural chemical activator for agriculture and horticulture. Thus, the task of the present invention is to provide a new compound that can regulate plant growth.

Problem solving

ИО005| During the study of AHA, the inventors discovered a metabolite of AHA, ZN-imidazoi|4,5- 9111,2,3Igriazine-4,6(5H, 7H)-dione (another name: 2-aza-8-oxo-hypoxanthine, which sometimes referred to as "AOG"), which has plant growth-regulating effects similar to AHA. In addition, the inventors synthesized 3-methyl-ZN-imidazo|4,5-4|(1,2,3)griazine-4,6(5H,7H)-dione (hereinafter sometimes referred to as "3-methyl-AOG" ) with AOG and found that 3-methyl-AOG also has an effect

On the regulation of plant growth. For this reason, the present invention provides a compound selected from the group consisting of the following compounds (A) and (B): (A) 3H-imidazo|4,5-4|01,2,3)griazine-4,6 (5H, 7H)-dione; and (B) 3-methyl-ZN-imidazo|4,5-4|(1,2,3Hgriazine-4,6(5H,7H)-dione.

II0006| The present invention also provides a plant growth regulator containing a compound selected from the group consisting of compounds (A) and (B) described above. The compound of the present invention can be effectively used as a plant growth regulator, since the compound can strongly stimulate plant growth.

II0007| The present invention also provides a method for producing AOG, which includes enabling xanthine oxidase to act on AGC to produce AOG. Applying the specified manufacturing method, it is possible to easily obtain AOG with a high yield.

ИЙО008| The present invention also provides a method for the production of AOG, comprising the steps of: extracting the plant organism in order to prepare the extract; and isolation of AOH from the extract. Applying the specified production method, AOG can be obtained without resorting to an enzymatic reaction.

Beneficial effect of the invention

I0009| According to the present invention, a compound is provided that can regulate plant growth.

Brief description of graphic materials 00101 Figure 1 is a chromatogram obtained as a result of chromatography of an extract of zoisia japonica grown in soil to which AHA was added.

Figure 2 is a chromatogram obtained as a result of chromatography of an extract of rice grown in a liquid nutrient medium to which AHA was added.

Figure C is a diagram showing the crystal structure of an AHA metabolite determined using X-ray diffraction analysis.

Figure 4 is a graph showing the effect of adding AHA or AOG to the medium on rice growth.

Figure 5 represents (a) tandem mass spectroscopy spectrum and (b) full mass spectrum of standard AOG.

Figure b represents (a) tandem mass spectroscopy spectrum and (b) full mass spectrum of 60 rice root extract.

Figure 7 represents (a) tandem mass spectroscopy spectrum and (b) full mass spectrum of tomato root extract.

Figure 8 is a graph showing the effect on rice growth of addition to the medium

AOG or 3-methyl-AOG.

Description of variants of implementation

I0011| The present invention provides (A) AOG, that is, ZN-imidazo|4,5-41(11,2,3)griazine-4,6(5H,7H)-dione. AOG is a compound represented by the following formula (1). mouth 12|. and

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I0015| Xanthine oxidase, which is used for the production of AOG from AHA, is not particularly limited, and at the same time, xanthine oxidase, which comes from, for example, buttermilk, can be used. In order to give xanthine oxidase the ability to act, for example, on AHA, AHA is dissolved in a reaction solvent, resulting in a reaction mixture, and at the same time, xanthine oxidase can be added to the reaction mixture. The amount of xanthine oxidase to be added is, but is not particularly limited to, preferably 20-200 Units, more preferably 50-200 Units, and more preferably 100-200 Units per 1 g of AGC. The reaction conditions are not particularly limited, and it is possible to allow the xanthine oxidase to act, for example, within its optimal reaction conditions. For example, the reaction temperature is preferably 20 - 35 "C, more preferably 25 - 32 "C, and also more preferably 28 - 30 "C. For example, the reaction time is preferably 6-50 hours, more preferably 24-50 hours, and and even more preferably 48-50 hours.The reaction solvent includes, but is not particularly limited to, water-based solvents such as water or a buffer solution, and the buffer solution may include, for example, phosphate-buffered saline solution, phosphate buffer solution as well as distilled water Additives such as

Reaction activators can be added to the reaction mixture. It is possible to obtain AOG according to the present invention from the reaction mixture by performing operations such as extraction, evaporation, purification, as well as drying the reaction mixture if necessary after the completion of the reaction.

ІІ0016| It is also possible to obtain AOG from the plant body. Plants are not particularly limited, provided they produce AOG, and may be spermatophytes (seed plants), pteridophytes (fern-like plants), or bryophytes (mosses). Spermatophytes can be gymnosperms or angiosperms, and angiosperms can be monocots or dicots.

I0017| Such plants include, in particular, plants of the Roaceae family, the 5oiIapaaceae family, the Tpeaaceae family, the Sotroziae family, the Cozaaceae family, and the I Chaseae family.

Among the above, due to the large amount of AOG produced, plants of the Roaceae family or the Zoiapaaceae family are preferred. Plants of the Roaceae family include the genus

RpuPoviaspuv, the family of Nogdeit, the family of Tgyisut, the family of Ogua, the family of Adgovii5, the family of 2ousia, the family of Zasspagyt, and also the family of 7ea. Among the mentioned, the genus Ogula is predominant, and rice (Ogula zayima) is more predominant. Plants of the BoiIapaceae family include the genus Husoregvisop, the genus ZoiIapit, the genus

Sarzisite, genus Misoiapa, genus Oaishga, genus RPuzaiy5, as well as genus Reishpia. Among the mentioned, kind

Husoreghissop is predominant and tomato (Zoiaphyte iusoregsisit) is more preferred. 0018) Plants to which AGC is applied can be used as plants for obtaining AOG. When AHAs are applied to plants, AOG is produced as a metabolite

AGK This AOG can be isolated from the plant body. If applied to plants

AGC, then more AOG is obtained from plants than when it is normally produced by plants. 0019) As a method of obtaining AOG from a plant organism, a method containing the steps of extracting the plant organism in order to prepare an extract can be used; and isolation of AOH from the extract. A method of producing an extract by extracting a plant organism includes, but is not particularly limited to, for example, a method in which a plant organ is collected from a plant organism containing AOG, the plant organ is ground into a powder, and the powdered organ is extracted using an organic solvent such as ethanol or acetone to obtain an extract. Method of selection

AOG from the extract is not particularly limited, but at the same time, for example, it is possible to isolate AOG according to the present invention by performing such operations as evaporation, purification, and drying of the obtained extract if necessary. (0020) A plant organ containing AOG may be, but is not limited to, the entire plant organism, or anything from roots, stems, leaves, flowers, reproductive organs, and seeds, and may additionally be are cell cultures. Among the mentioned, due to the content of a large amount of AOG, the roots are predominant.

I0021| The present invention also provides (B) 3-methyl-AOG, i.e., 3-methyl-3H-imidazoi|4,5-91,2,3|griazine-4,6(5H,7H)-dione. C3-methyl-AOG is a compound represented by the following formula (I). 0022 (9) ns M

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0023) 3-methyl-AOG can be synthesized by methylation of AOG, and 3-methyl-AOG can be synthesized, for example, according to the method described in

Examples.

I0024| By applying AOG or 3-methyl-AOG, it becomes possible to regulate plant growth. The growth of a plant in the present invention is not particularly limited, provided that it is a phenomenon that includes the normal differentiation or proliferation of plant cells, and also includes not only the growth and enlargement of the organs that make up plant organisms, but also the germination from a seed, the formation of a flower , as well as seed binding. AOG and 3-methyl-AOG can stimulate or inhibit plant growth, depending on the target plant species, the concentration of AOG or 3-methyl-AOG, and the fraction of plants that will be affected by AOG or

Z-methyl-AOH. Thus, it is possible to use a plant growth regulator containing AOG or 3-methyl-AOG as a growth promoter or growth inhibitor for various plant organs, depending on the target plants and objectives. (0025) The target plants are not particularly limited, provided that they are plants that undergo growth regulation by AOG or C3-methyl-AOG. The target plants may be spermatophytes, pteridophytes, or bryophytes, and the spermatophytes may be gymnosperms or angiosperms, (p angiosperms may be monocotyledonous or dicotyledonous. 0026) Such plants include, in particular, plants of the family Roaceae, family 5oiIapaceae, family Tpeaceae, family Sotroziae, families of Kozaseae, as well as families | asses

Among the mentioned, in view of the significant effect of growth regulation with the use of AOG, plants of the Roaceae family are preferred. Plants of the Roaceae family include the genus

RpuPoviaspuv, genus Nogdeit, genus Tgyisut, genus Ogua, genus Adgovii5, genus 2ouvzia, genus Zzasspaguit, and also genus 7ea. Among the mentioned, the genus Ogula or the genus 7ousia is predominant.

I0027| The target plant organ can be, but is not limited to,

anything from roots, stems, leaves, flowers, reproductive organs, and seeds, and may also be cell cultures. Among the above, given the significant effect of growth regulation using AOG, roots, stems, leaves, or seeds are preferred. (0028) The concentration and application method of AOG or 3-methyl-AOG to be applied to plants can be selected according to circumstances, depending on the species of target plants, their organs, purposes, and the like. For example, in the case where the target plant is rice (Ogua zei), the target organ is a stem, and the goal is to increase the length of the stem, it is preferable to grow the plant using a liquid nutrient medium in which

AOG is dissolved to obtain 5-2000 μM in a normal liquid nutrient medium. In this case, the concentration of AOG in the liquid nutrient medium is more preferably 100-1500 μM, and even more preferably 500-1000 μM. In the case where the target plant is Ogula zayima, the target organ is a root, and the goal is to increase the length of the root, it is preferable to grow the plant using a liquid nutrient medium in which AOG is dissolved to 50-2000 µM in normal liquid nutrient medium. In this case, the concentration of AOG in the liquid nutrient medium is more preferably 250-1500 μM, and even more preferably 500-1000

MKM. In the case where the target plant is Ogula zayima, the target organ is a seed, and the objective is to increase the size or number of seeds in order to increase seed yield, it is preferred to grow the plant using a liquid nutrient medium in which the AOG is dissolved to obtain 5-1000 μm in a normal liquid nutrient medium. Alternatively, if the goal is to increase the seed yield of Ogula zaeima, it is also preferable to grow Ogula zaeima in soil using a nutrient medium in which AOG is dissolved to 5-500 µm.

I0029| In the case of using 3-methyl-AOG, for example, when the target plant is

In general, the target organ is the root, and while the goal is to increase the length of the root, it is preferable to grow the plant using a liquid nutrient medium in which 3-methyl-AOH is dissolved to obtain 50-1000 µM in a normal liquid nutrient medium. In this case, the concentration of Z-methyl-AOG in the liquid nutrient medium

Zo is more preferably 200-500 µM. 0030) The plant growth regulator containing AOG or 3-methyl-AOG according to the present invention, in addition to AOG or 3-methyl-AOG, may contain bactericides, anti-fungal substances, insecticides, or compounds having the effect of regulating plant growth, other than AOG or 3-methyl-AOG. In addition, the regulator may contain known additives for the formulation of the preparation form. As such additives for the preparation form, it is possible to use, but in particular not limited to them, for example, auxiliary substances, emulsifying substances, as well as moisturizing substances. Types of species of plant growth regulators according to the present invention may include, but are not limited to, for example, emulsions, wetting powders, water-soluble powders, solutions, granules, powders, microcapsules, fumigants, fumigants, aerosols, liquid-liquid substances, pastes, tablets, coating substances, ultra-low volume spray substances, oily substances, as well as complex fertilizers, and at the same time, consumers can choose them according to the circumstances, depending on the species of the target plants, their organs, purposes, and the like. The plant growth regulator in the specified species can be produced using known methods.

Examples

I0O31| Hereinafter, the present invention is described more specifically with reference to the Examples, but this invention is not limited to the Examples.

И0ОоЗ2И (Example 1. Detection of the AGC metabolite of Japanese zoysia)

Zoysia japonica (Adgoviiz 5i0Iopitega) was grown for 30 days on a control plot of normal agar medium (100 ml) and on a plot of agar medium with AHA, to which 1 mM of AHA (100 ml) was added. The growth of zoisia japonica shoots in the plot with AHK was clearly activated, compared to the increase of zoysia japonica shoots in the control plot.

The shoots and roots of cultivated Zoysia japonica were harvested, extracted using ethanol, and the shoot extract and root extract were subjected to reversed-phase high-performance liquid chromatography (reverse-phase HPLC). The conditions of the study were as follows.

Column: YuRemeiovzia C30-OS-5 column (size 4.6 x 250 mm), flow rate: 0.5 ml/minute, mobile phase: gradient elution of 2 95 methanol in 0.05 95 trifluoroacetic acid (Liquid A) for 12 minutes; 2-100,956 methanol in liquid A for 120 minutes; 100 90 methanol bo for 20 minutes, detection: determination of absorbance at UV 254 nm. chromatogram,

obtained by chromatographic study, shown in Figure 1. From the eluate of the shoot extract from the AHA site (Figure 1 (b)), the AHA peak was not detected, but the metabolite peak

AGC was detected. Peaks of AHA and metabolite of AHA were detected from the eluate of the root extract from the area with AHA (Figure 1(c)). This is thought to be because AHA in the environment, other than the AHA metabolite in the root, is present in small amounts in the eluate of the root extract. No rush

Neither AHA nor the AHA metabolite peak was detected from eluates of shoot extracts and root extracts of the control plot (Figure 1a) and (c)).

I0OZZI (Example 2. Detection of rice AHA metabolite)

Rice (Ogula zaina) was grown on a control plot of normal liquid nutrient medium and on a plot of nutrient medium with AHAC to which AHAC was added to 0.2 mM in normal liquid nutrient medium for 14 days. The increase of rice shoots in the plot with AGC was clearly activated, compared to the increase of rice shoots in the control plot. The shoots and roots of the grown rice were collected, extracted and analyzed using reversed-phase HPLC as in Example 1. The chromatogram obtained using the chromatography is shown in Figure 2. In the AHAC plot, the AHAC peak was not detected even from the eluate of the shoot extract , nor from the root extract eluate, but the metabolite peak

AGC was detected (Figure 2(b6) and (d)). Neither the AHA peak nor the AHA metabolite peak was detected from the eluates of shoot extracts and root extracts of the control plot (Figure ha and (c)).

I0OZA4I (Example 3. Isolation of the AHA metabolite)

Rice (Ogula zayima) was grown on a control plot of normal liquid nutrient medium and on a plot of nutrient medium with AHAC, to which AHAC was added up to 1 mM, in normal liquid nutrient medium for 20 days. Shoots (360 g) of grown rice were collected and extracted using ethanol. The ethanol-soluble fraction was evaporated under reduced pressure and extracted using dichloromethane.

The dichloromethane-insoluble fraction was extracted using ethanol to obtain the ethanol-soluble fraction (9.8 g). The fraction soluble in ethanol was subjected to chromatography with silica gel (filler: 350 g of silica gel 60 M, column size: 4 x 60 cm) and successively

Zo was eluted using mixtures of dichloromethane: methanol - 9:1, 7:3, 5:5, with the preparation of eight fractions. Among the above, fraction C (288 mg) was successively purified using

HPLC (column: column Yuemeiovzii C30-OS1-15/30, size: 50 x 500 mm, flow rate: 25 ml/min, mobile phase: 595 methanol, detection: UV 310 nm) and HPLC (column: column

Remeiovzii C30-0O00-5, size: 20 x 250 mm, flow rate: 5 ml/minute, mobile phase: 10 95 methanol, detection: UV 310 nm), until the separation at the end of 10.5 mg of the AHA metabolite.

I0OZ5I (Example 4. Structure determination using X-ray structural analysis)

X-ray structural analysis was carried out on the isolated metabolite of AHA in the following way.

X-ray diffraction studies on single crystals were carried out using 5Rhipa-8 (structural analysis on a beam of VI O281 single crystals). The conditions of the study were as follows. wavelength: 0.8260(4) A, beam size: length 140 x width 159 µm, photon flux: 1.81 x 108 photons/sec, and photon flux density: 8.13 x 103 photons/sec/µm?. Figure C is a diagram showing the crystal structure of an AHA metabolite obtained by X-ray crystallography. As a result of X-ray crystallography, it was confirmed that the isolated metabolite of АГК was ЗН-imidazo|4,5-41(11,2,3)griazine-4,6(5Н, 7Н)-dione (another name: 2-aza-8 -oxo-hypoxanthine, "HOG"), having the structure of formula (1).

I0OZ6) (Example 5. Production of AOG)

AOG was obtained by enabling xanthine oxidase to act on AGC in the following manner. In 1 L of phosphate-buffered saline solution (10 mM, pH 7.4), 137 mg of AHA was additionally dissolved, 25 mg of xanthine oxidase (derived from buttercup, 0.28 Units/mg) was added, and the resulting mixture was left to settle at at a temperature of 30 "C. To the mixture, 25 mg of xanthine oxidase was added three times every 24 hours. After the last addition, the mixture was left to settle for the next 24 hours. That is, in general, 100 mg of xanthine oxidase was allowed to act on 137 mg of AHA for 96 hours. As a result, by using an HPLC assay (Yumeiovzia column C30-OS-5 (size 4.6 x 250 mm), flow rate: 0.5 ml/min, mobile phase: gradient elution of 295 methanol in 0.0595 trifluoroacetic acid (Liquid A) for 12 minutes; 2-100 956 methanol in liquid A for 120 minutes; 100 90 methanol for 120 minutes; detection: absorbance at UV 254 nm) it was confirmed that AGC was completely converted to AOG. The product was purified using flash

chromatography with ODS gel (octadecyl silica gel) (filler: 350 g of ODS gel, column size: 4 x 60 cm, mobile phase: water, water/methanol - 9:1) to isolate 120 mg of AOG (yield 78.4 95). Using HPLC retention time, absorption wavelength, and mass spectrometry, it was confirmed that the isolated substance was AOG.

I0OZ7I (Example 6. Effect of AOG on rice)

Sterilized rice (Mirroprage) seeds (Ogula zaini /. see Mirroprage) were allowed to germinate at a temperature of 28 °C for three days. Germinated seeds (four seeds per tube) were grown in a tube to which a control liquid nutrient medium or nutrient medium was added, until to which AGK or AOG was added to achieve different concentrations, and left at a temperature of 28 "C for a week. AOG was used, which was obtained in accordance with the method of Example 5. The control liquid nutrient medium contained 0.5 mM

MNAMOz, 0.3 mM MagnHPO, 0.15 mM Cu»5O», 0.2 mM MaosiI», 0.1 mM SaCi», 23 μM HE-ethylenediaminetetraacetic acid (Be-EDTC), 25 μM NzVoO»z, 4.5 µM Mn5O", 0.15 µM Si5Oox, 0.35 µM 2p5054, as well as 0.05 µM Mag2Mobh. As in the case of liquid nutrient media containing AHA or AOG, the control liquid media described above were supplemented with

AGC or AOG so that the final concentration was 50 μM, 200 μM, or 1000 μM. The liquid nutrient medium was replaced with a new liquid nutrient medium every other day. After growing, the length of shoots and roots was measured. The measured shoot and root enlargement is shown in Figure 4. It was confirmed that AOG, similarly to AGC, activates shoot and root enlargement in a concentration-dependent manner, and that AOG was a metabolite of AGC (in Figure 4, "Kon" stands for control. """ indicates the value of P "0.05 and "77" indicates the value of P "0.01. n-11.).

I0OZ8)I (Example 7. Isolation of AOG from rice)

Rice (Ogula zaina) was grown hydroponically using a normal liquid rice nutrient medium for two months, and the roots (52 g) were harvested. After the roots were powdered with a mixer, the powdered roots were extracted using ethanol and acetone, and then the extract was evaporated to dryness to obtain a root extract (340 mg). The specified extract was dissolved in a solvent

Zo (95 95 acetonitrile and 0.05 95 formic acid) to bring it to a concentration of 10 mg/ml, and the resulting solution was analyzed using a liquid chromatograph with a tandem mass spectrometer (LC-MS/MS). The AOG reference standard has a peak at a retention time of 2.5 minutes using LU, a molecular mass peak of 152.9 M-NO was detected from the full mass spectrum at the indicated retention time, and a fragment ion peak molecular weight of 97 was detected from the spectrum of tandem mass spectroscopy. Similar to the AOG reference standard, the rice root extract has a peak at a retention time of 2.5 minutes, using LC, a molecular mass peak of 152.0|M-NO| was detected from the full mass spectrum at the indicated retention time, and the fragment ion peak of molecular mass 97 was detected from

MS/MS spectrum. Since the same molecular weight peak as that of the AOG reference standard was detected from the rice extract, it was confirmed that the rice roots contained endogenous AOG.

The tandem mass spectrum (a) and full mass spectrum (b) of the AOG reference standard are shown in Figure 5. The tandem mass spectrum (a) and full mass spectrum (b) of the rice root extract are also shown in Figure 6. Amount The AOG contained in the rice root, which was calculated based on the calibration graph and the AOG peak area, was approximately 2.5 ng per 52 g of rice root. The conditions of LC-MS/MS research and the equipment used are as follows. (0039) "LC-MS/MS research conditions and equipment" (Part of LC)

Pump: I S-20A0 (ZNIMAYU2HO SOCROKATIOM company)

Column: RS NICHIS (size: 2.0 mm x 100 mm, company Zpizeido Co., tsa.)

Flow rate: 0.2 mL/minute Injection volume: 10 µL (MS/MS fraction)

Mass spectrometer: STO OMKVITKAR OIZSOMEKU (type with ion trap, detection mode of negatively charged ions, TNEKMO ZSIEMTIRIS company) 0040) (Example 8. Isolation of AOG from tomato)

Tomato (Zoiaphyte Iusoregvzisit) was grown hydroponically using normal liquid nutrient medium for tomatoes for two months, and the roots (17 g) were collected. After the roots were powdered with a mixer, the powdered roots were extracted using ethanol and acetone, then the extract was evaporated to dryness, and the dichloromethane-soluble part was freed to obtain the root extract (4.7 mg) . Said extract was dissolved in a solvent (95 95 acetonitrile, 0.05 95 formic acid) to bring it to a concentration of 10 mg/ml, and the resulting solution was analyzed using LC-MS/MS as in Example 7. The conditions of the study and the equipment used for the analysis were the same as in Example 7. As a result, the tomato root extract, similar to the AOG reference standard, has a peak at a retention time of 2.5 minutes using LC, a molecular weight peak, which is 152.9|M-NI- was detected from the full mass spectrum at the indicated retention time, and a fragment ion peak with a molecular weight of 97 was detected from the tandem mass spectroscopy spectrum. Thus, it was confirmed that tomato roots also contained endogenous AOG. Tandem mass spectroscopy spectrum (a) and total mass spectrum (b) of tomato root extract are shown in Figure 7. The amount of AOG contained in the tomato root, which was calculated from the calibration plot and the peak area of AOG, was approximately 0.1 ng, or less, for 17 g of tomato roots, which was less than in the case of rice.

IO0ATI (Example 9. Effect of AOG on growing rice in soil (growing in pots))

Rice (Mirroprage) (Ogula zaina I. see Mirroprage) was sown on April 29, 2011, and all grown seedlings were transplanted on June 7 into pots (1/5000 pots) filled with soil including fertilizer containing M ( 1440 mg), Pg2O5 (12 mg), KgO (760 mg) and Cao (806 mg"), and were grown in the soil in a vegetation house (28 "C) under any of the seven types of growing conditions described below (1) - (7), until September 24. Water was provided by providing two liters of tap water or tap water to which AOG was added weekly (see below).

Cultivation conditions (1) Tap water was provided during cultivation in pots. (2) Tap water to which AOG was added so that the final concentration was 50

MKM was provided for two weeks (from June 7 to June 20) during the stage

From landing Tap water was provided at a different time period. (3) Tap water to which AOG was added so that the final concentration was 50

MKM was provided for two weeks (from July 4 to July 17) during the tillering phase.

Tap water was provided at a different time period. (4) Tap water to which AOG was added so that the final concentration was 50

MKM was provided for two weeks (from July 25 to August 7) during the surface feeding stage for panicle formation. Tap water was provided at a different time period. (5) Tap water to which AOG was added so that the final concentration was 50

MKM was provided for two weeks (from August 15 to August 28) during the surface feeding stage for ripening. Tap water was provided at a different time period. (6) Tap water to which AOG was added so that the final concentration was 5 μM was provided during pot cultivation. (7) Tap water to which AOG was added so that the final concentration was 50

MKM, provided during cultivation in pots.

Paddy rice and plant organisms grown in soil were dried at 307 for 15 days, and paddy rice mass (both the weight of one plant and the weight of 100 paddy grains), panicle length, stem length, number of panicles, number of shoots, were determined. as well as the mass of the ground part. The results are shown in Table 1. Due to the fact that AOG was applied throughout the period, the number of panicles and the weight of the aerial part were higher than in the control plants. In addition, due to the application of AOG at the surface fertilization stage during the panicle formation phase or later, the mass of paddy rice per plant was higher than that of the control plants (In Table 1, numerical values indicate the mean values of their standard deviation. "Increased level" indicates increased level (95) in relation to control plants. """ indicates the value of Р « 0.05. The number of samples (pots) - 6.). 0042

ITable 11) 50 μm 50 μm AOG

SHE CHE I'm an troll. and

Stage The phase of bush feeding for planting, formation of panicles

Unpolished rice rice (g/plant) o Increased level (90)

Sho 1 |znyui ze» | Rice starch 2.19-0.06 2.11-0.07 2.12520.12 2.12520.07 (g/1100 grains)

Plant organism o Raised level(0)! (77771111 (Increased level (90)Ї /////717111111111Ї11111111111111Ї1111111111111111

Stage of surface 5 MEM AOG 50 μm AOG feeding for All time All time rice ripening (g/plant) rice (g/100 grains) o Increased level(90) | ////////7777711111111111111111010241 11111123 (Increased level(90) | //////777777777ii177111180111111111271

ИО04ЗИ (Example 10. Impact of AOG on growing rice in the soil (growing in the field))

They used rice (Mirropbage) (Oguga zayiima I. sm. Mirropbage) and grew it in the field in the following way. As growing conditions, five species from (1) - (5) were established.

Rice (Mirroprage) (Ogula zayima I. see Mirroprage) was sown in a seedling box on April 29, 2011, and provided with a total of 15 liters of tap water or tap water to which AOG was added for two weeks from May 24 , while seedlings were grown in a seedling box. On June 7, all grown seedlings were transplanted into the field.

Planting density was set according to the inter-row space, which was 30 cm, and the width of the inter-rows, which was 15 cm (three growing areas of 3 x 3.3 m per plot). On June 7, 50 liters of tap water were supplied to the site, or tap water to which was added

AOG, as the main fertilizer. Cultivation continued, and 50 liters of tap water, or tap water to which AOG was added, was applied to the plot on July 25 as a top dressing for panicle formation. Plant organisms were collected on October 12.

Cultivation conditions (1) Tap water was used as a liquid nutrient medium during the cultivation of seedlings, the main fertilizer, and surface fertilization for the formation of panicles. (2) Tap water to which AOG was added so that the final concentration was 0.5

MM, provided as a liquid nutrient medium during seedling cultivation.

Tap water was used as the main fertilizer, as well as as a top dressing for the formation of panicles. (3) Tap water to which AOG was added so that the final concentration was 1.0

MM, provided as a liquid nutrient medium during seedling cultivation.

Tap water was used as the main fertilizer and top dressing for the formation of panicles. (4) Tap water to which AOG was added so that the final concentration was 0.5

MM, provided as the main fertilizer. Tap water was used as a liquid nutrient medium during the cultivation of seedlings and surface feeding for the formation of panicles. (5) Tap water to which AOG was added so that the final concentration was 0.5

MM, was provided as surface nutrition for the formation of panicles. Tap water was used as a liquid nutrient medium during the cultivation of seedlings and as the main fertilizer.

Paddy rice was dried, and the total weight of paddy rice, husk weight, weight of unsieved paddy rice and sifted paddy rice, per 10 plant organisms, as well as the weight of 1000 grains were determined. The results are shown in Table 2. Due to the application of AOG, the total weight of paddy rice, the weight of husk, the weight of unsieved paddy rice, as well as the weight of sifted paddy rice were higher than those of the control plants (In Table 2, the numerical values indicate the mean values x standard deviation. "Increased level" indicates an increased level (90) relative to control plants. """ indicates a value of P 15 "0.05.). 00441

ITable 21 0.5 mm AOG 1.0 mm AOG

Per 10 plants Control During During the cultivation of seedlings paddy rice seedlings (g) paddy rice (g) o Raised level(90) | 77777777 1777/7859 | sh-B 102: pen (nty | eEshetn

Per 10 plants o Surface top dressing with a dormant fertilizer We formation of panicles of paddy rice (g) paddy rice (g) 0045)

Zo (Example 11. Production of Z-methyl-AOH)

According to the following method, 3-methyl-AOG was obtained from AOG.

In 5 ml of DMSO (anhydrous dimethylsulfoxide), 153 mg of AOG was dissolved at a temperature of 50 "С, 0.075 ml of iodomethane was added to this mixture, and the resulting mixture was subjected to a reaction for 4 hours. The fraction obtained using preparative thin-layer chromatography (TLC, mobile phase SNecSig: methanol - 9:1), additionally subjected to HPLC (column

C30-O0-5 reagents (size 20 x 250 mm, flow rate: 5 ml/minute, mobile phase: 10 95 methanol in 0.0 5 95 trifluoroacetic acid, detection: UV 310 nm) to obtain 10.2 mg (yield 6.11 95) Z-methyl-AOG.

Using the results of mass spectrometry using H-NMR and 3C-NMR, it was confirmed that the obtained substance is 3-methyl-AOG, and the details were as follows.

When the sample was examined using a mass spectrometer (mass spectrometer UM5-

T100 C) in the mode of determining positively charged ions, then t/2168|M'NI and t/2190|(M--Mai" were determined.

In addition, using H-NMR and 3D-NMR, the sample showed the following values. "H-NMR (500 MGu) 5 3.88 1300-NMR (125 MGu) b 37.9, 112.8, 142.1, 148.0, 152.7 0046) (Example 12. Effect of AOG and Z-methyl-AOG on rice)

Sterilized rice (Mirroprage) seeds (Ogula zain /. see Mirroprage) were allowed to germinate at a temperature of 28 "C for three days. The germinated seeds (four seeds per tube) were grown in a tube to which was added a control liquid nutrient medium, to which 0.2 mm AOG was added, or nutrient medium to which 0.2 mm 3-methyl-AOG was added, and left at a temperature of 28 "C for a week. The control liquid nutrient medium contained 0.5 mm MNAMO»z, 0.3 mm MagNRO», 0.15 mm 20 K»50O5, 0.2 mm

Massi»g, 0.1 mm Sassi», 23 μM ERe-ethylenediaminetetraacetic acid (He-EDTC), 25 μM NzVOz, 4.5

MKM Mpy5O», 0.15 µM Si5O», 0.35 µM 2p50»5, as well as 0.05 µM Mag2Mob». The liquid nutrient medium was replaced with a new liquid nutrient medium every other day. After growing, the length of shoots and roots was measured. The measured root enlargement is shown in Figure 8.

It was confirmed that 3-methyl-AOG, similarly to AOG, had increased activity for roots. (In Figure 8, """ indicates a P value of 0.05, and "77" indicates a P value of 0.01. p-16.) In contrast, 3-methyl-AOG has no effect on shoot growth .

Industrial suitability

I0047| Since AOG and Z-methyl-AOG according to the present invention have the effect of regulating growth, they can be effectively used as plant growth regulators. Such regulators

Plant growth can be widely used in agriculture and horticulture.

Claims (12)

FORMULA OF THE INVENTION 1. A compound selected from the group consisting of the following compounds (A) and (B): Dion; and (B) 3-methyl-ZN-imidazoi4,5-4|1,2,3|Hygriazin-4,6(5H,7H)-dione. 2. The compound according to claim 1, where the compound is (A) ZH-imidazo(|4,5-41(11,2,3)griazine-4,6(5H,7H)-dione. 3. Plant growth regulator containing the compound according to claim 1. 4. A method of regulating plant growth, which includes the action of a compound selected from the group consisting of the following compounds (A) and (B): (A) ZH-imidazo|4,5-4111,2,3)griazine-4 ,6(5H, 7H)-dione; and (B) 3-methyl-ZN-imidazo|4,5-4|1,2,3)|Itriazine-4,6(5H,7H)-dione, per plant. 5. The method according to claim 4, where the plant is a plant of the Roaceae family. 6. The method according to claim 4, where the plant is a plant of the genus Ogula or genus 20ousia. 45 7. The method of obtaining ZH-imidazo(4,5-41(11,2,3|Hygriazin-4,6(5H, 7H)-dione, which includes the stage: enabling xanthine oxidase to act on 7H-imidazo|4,5- 411,2,3)griazin-4(3H)-one to obtain ZH-imidazo|4,5-4111,2,3)griazin-4,6(5H,7H)-dione, or which includes stages: 50 extraction plant organism in order to prepare an extract; and isolation of ZH-imidazoi|4,5-41(/1,2,3)triazine-4,6(5H,7H)-dione from the extract. 8. The method according to claim 7, where the method includes the stage: enabling xanthine oxidase to act on 7H-imidazo|4,5-411,2,3)griazin-4(3H)-one to obtain 3H-imidazo|4,5-4111 ,2,3)griazine-4,6(5H,7H)-dione. 55 9. The method according to claim 7, where the method includes the steps of: extracting the plant organism in order to prepare the extract; and isolation of ZH-imidazoi|4,5-41(/1,2,3)triazine-4,6(5H,7H)-dione from the extract. 10. Use of a compound selected from the group consisting of the following compounds (A) and (B): (A) ЗН-imidazo|4,5-4|1,2,3)griazine-4,6(5Н, 7Н )-dione; and because (B) 3-methyl-ZH-imidazo|4,5-4111,2,3)Itriazine-4,6(5H,7H)-dione, to regulate plant growth. 11. Application according to claim 10, where the plant is a plant of the Roaceae family. 12. Application according to claim 10, where the plant is a plant of the genus Ogula or the genus 2ousia. I 7 AND RUN away from the fault of Gei - VONEKIVK TE KE Z U nn ХВ хх E ц је в ХЕ i: Ш і ho E o Tі i є Mня SEZ SUDNA: cheh more Ж I ЖЕ I ХО Х; Eh - "oh hehi he STILL EE in ZU OO OKO KK ooo puppy Kk ho how poh "k.. xx I h a oheki KeYA - x. Ж - и ги бух ЕЕ как I ТАЖ AMKU KE N Я e B ny shi shi nyny rak Azh aka o a zannia as ve zh de ah Uh Teslya zh BUUya xx SCREEN Ky "OK. Z 2 shock MKMOL NI - Y Y "Z shchi y xx M I, these "their "Fipppyipitsipy i upmoom ho n orkhen B EK mm g hm VUK Mu 5 SCHE in i; them. Zhe N xx NI i ON Y ELE oh ХЕ ше: ; ЗАКУТЕКИЯВ ТИКИХ Ш т х к и ЗЕМ И По ооее ро о о ее. So "M. te U m ko Be kh zh se ih mk I suk KI ZE 3 ih ko pit N heavy ov Boi i JHE zate M m tmykdvoooooih shi zh zhk xx KOHAN XX ZK B MOM o VOM i pe Po oo VN Ky MO VE OK On o 1.