KR101135133B1 - Method for Decreasing Biomass of Plant or Reprssing Plant Flowering and Compositions Thereof - Google Patents

Abstract

The present invention relates to methods and compositions for reducing the size of plants or inhibiting flowering. Specifically, the present invention discloses methods and compositions for reducing the size of plants or inhibiting flowering using 2- (4-diethylamino) -2-hydroxybenzoyl) benzonic acid.

2- (4-diethylamino) -2-hydroxybenzoyl) benzonic acid, plant, size, flowering

Description

Method for Decreasing Biomass of Plant or Reprssing Plant Flowering and Compositions Thereof}

The present invention relates to methods and compositions for reducing the size of plants or inhibiting flowering.

Plant growth is an overall phenomenon that occurs during the course of life, from plant germination to death.

From the beginning of germination, the plant is divided into two parts of shoot apical meristem and root apical meristem through continuous cell division to form parts of the entire plant such as root, leaf and flower. Its size makes it grow genetically up to its intended portion.

The growth of these plants is genetically planned as a series of consecutive biochemical and physiological phenomena, which are very sophisticated and active at the level of cells, tissues and organs.

Plant growth control is very important socio-economically, because it is directly and indirectly related to the increase in farmers' harvest.

This is why biotech workers are looking for genes, hormones and other chemicals to control plant growth.

On the other hand, the technique of suppressing the flowering of plants is important in terms of being able to prevent the gene transfer of genetically modified crops.

Genetically modified plants have the risk of disrupting and destroying the ecosystem by changing the traits of allogeneic and related species by pollen scattering. In fact, herbicide-tolerant canola ( Brassica napus ) has been reported to cause gene transfer by pollen scattering at 3 km (Rieger et al., 2002), and herbicide-resistant Bentgrass at 21 km. Gene transfer has been reported (Watrud et al., 2004). As a result, most countries, including Korea, have a risk assessment system in place when they permit the manufacture, sale, and import of genetically modified plants.In some cases, they are enclosed spaces to prevent exposure to the natural environment. I'm growing in. Therefore, inhibition of flowering in genetically modified plants has an important meaning in that they are a trait that can fundamentally block the risk of disturbing or destroying the ecosystem by pollen scattering.

On the other hand, there are three major methods of inducing mutations in living organisms: the first is the treatment of chemicals in living organisms, the other is the irradiation of X-rays or gamma rays in living organisms, and the third A method of introducing a vector that activates or inactivates a gene into a living organism.

The present inventors have completed the present invention by producing a mutant plant with reduced size and suppressed flowering using the first method.

It is an object of the present invention to provide a method for reducing the size of plants or inhibiting flowering.

Another object of the present invention is to provide a composition capable of reducing the size of a plant or inhibiting flowering of the plant.

Other objects of the present invention will be presented below.

In one aspect, the invention relates to a method for inhibiting flowering of a plant.

In another aspect, the invention relates to a method for reducing the size of a plant.

When the inventor (s) receives a variety of compounds from the Korea Compound Bank and treats each of the compounds with plants, the flowering is suppressed (i.e., infertile) and reduced in size compared to wild-type plants in certain compounds. It was confirmed that the plant was obtained (see FIG. 1).

As a result of confirming the structure of the compound through NMR analysis, the compound has an IUPAC name 2- (4-diethylamino) -2-hydroxybenzoyl) benzonic acid [2-((4-diethylamino) -2 -hydroxybenzoyl) benzoic acid] is a compound represented by the following <Formula 1>.

<Formula 1>

The present inventors maintain the basic skeleton of the compound of <Formula 1>, by using a variety of methods known in the art to substitute and modify the ethyl group and to process the modified compound to the plant treated the modified compound It was also confirmed that flowering was suppressed compared to wild-type plants and the size of the plants was smaller.

Including the compound of <Formula 1>, if the modified compound is expressed collectively, it may be represented by the compound of the formula (2).

<Formula 2>

In the above, R ₁ and R ₂ are each independently H, a straight or pulverized alkyl group, a straight or pulverized alkenyl group, a straight or pulverized alkynyl group, a cyclo alkyl group, a cyclo alkenyl group, an aryl group, an alkaryl group or an aralkyl group.

The present invention is provided based on the above experimental results.

Therefore, the method of inhibiting the flowering of the plant of the present invention or the method of reducing the size of the plant is characterized in that it comprises the step of contacting the compound of the formula (2) or a salt thereof to the plant.

As used herein, the term "plant" includes all plants that can produce useful results to humans, including cultivated plants, whose flowering is inhibited or the size of the plants is reduced. The most direct examples include genetically modified crops (especially nutrient-producing genetically modified crops), various weed plants that inhibit the growth of crops, or bonsai plants for ornamental purposes, and plants that humans may or may not eat. It will be included in the range of plants for reasons of simplicity of food, convenience of transportation and the like. Specifically, the plant includes weed plants such as arable land, bonsai plants such as roses, pine, pine, bamboo, rice, wheat, barley, corn, soybeans, potatoes, wheat, red beans, oats, sorghum, cabbage, radish, red pepper, strawberries , Tomato, watermelon, cucumber, cabbage, melon, pumpkin, green onion, onion, carrot, ginseng, sesame, sugar cane, beet, perilla, peanut, rapeseed, apple tree, pear tree, jujube, peach, bile, grape, Edible plants such as tangerines, persimmons, plums, apricots and bananas. Preferably it means a plant belonging to the cruciferaceae in the same way as the Arabidopsis used in the examples below. The plants belonging to the Cruciferaceae include cabbage, rapeseed and Chinese cabbage (Brassica campestris L. ssp.pekinensis or Brassica rapa L. ssp.pekinensis), cabbage (Brassica oleracea L.), broccoli (broccoli), cauliflower, kale ), Gat, turnip or Brassica rapa, rapeseed, radish and the like.

In addition, the "plant" is understood as a meaning including not only mature plants but also plant cells, plant tissues, plant seeds, and the like that can develop into mature plants.

Therefore, in the method of the present invention, contacting the compound of the above-mentioned <Formula 2> as an active substance to a plant means not only contacting the compound to a mature plant, but also plant cells, plant tissues, and plant seeds capable of developing into a mature plant. It is meant to include contact with the back.

In addition, in the present specification, the “size of a plant” may be easily understood as a biomass, and is understood to include the weight and / or size of organs such as leaves, stems, and roots of plants. Therefore, "reducing the size of a plant" can be understood to reduce the weight and / or size of organs such as leaves, stems, roots, etc. of a plant.

In addition, in the present specification, "inhibition of flowering" means that flowers (or buds) are not formed, and can be understood as meaning of traits in which seeds are not produced by not forming flowers. Therefore, plants whose flowering is suppressed will have traits that cannot be sexually reproduced.

In addition, in the present specification, the meaning of "contact" is meant to include not only direct contact by spraying the compound of <Formula 2> to the plant, but also indirect contact by spraying on the soil and the like on which the plant is growing. Therefore, in order to reduce the size of the plant or to suppress flowering, it is to be understood that the act of spraying the compound of <Formula 2> together with the fertilizer or by spraying it in the fertilizer is included in the scope of the present invention.

In the method of the present invention, the compound of <Formula 2> which is an active substance is preferably R ₁ and R ₂ are each independently H, C ₁ -C ₉ linear or pulverized alkyl group, C ₂ -in <Formula 2> C _{9 is a} straight chain or pulverized alkenyl group, C ₂ -C ₉ linear or pulverized alkynyl compound, more preferably R ₁ and are each independently H, C ₁ -C ₉ linear or pulverized alkyl group in <Formula 2> Compound, and more preferably R ₁ and R ₂ are each independently H, C ₁ -C ₄ straight chain or a branched alkyl group, and more preferably R ₁ and R ₂ are each independently H, C ₁ and -C ₄ straight chain alkyl group, and more preferably R ₁ and R ₂ may be each independently is identified as H, methyl or ethyl a compound. Most preferably, the compound of <Formula 1> wherein R ₁ and R ₂ are both ethyl groups.

On the other hand, in order to suppress the flowering of the plant or to reduce the size of the plant, how much amount of the compound of the formula <2> should be sprayed on the plant to determine the type of plant and the plant to suppress the flowering. The growth will depend on the environment of the soil, the purpose of inhibiting flowering of the plant (eg, the purpose of regulating the timing of harvesting).

As long as it is based on techniques known in the art, it is within the range of its usual ability for a person skilled in the art to determine in what amount the compound of formula 2 should be sprayed on a particular plant.

Typically, the spread amount of the compound of Formula 2 will range from 0.1 kg to 100 kg per 1000 m ^{2 of} land regardless of the type of plant.

On the other hand, in the contact of the compound of the formula (2) as an active substance with the plant, the compound of the formula (2) may be in contact with the plant, but may be further contacted with a suitable carrier or the compound of the formula (2) It is preferable to mix and contact. This is because the effect of inhibiting flowering or reducing the size of the plant can be further enhanced by facilitating the absorption of the compound of the above formula by the plant or the contact of the plant with the compound.

The suitable carrier will be embodied in the description of the composition for inhibiting flowering or reducing the size of the plant of the present invention as described below.

In another aspect, the present invention relates to a plant with reduced flowering or a plant with reduced size obtained by a method of inhibiting flowering of a plant as described above, or a method of reducing the size of a plant.

In another aspect, the present invention relates to a composition for inhibiting flowering or a composition for reducing the size of a plant, including the compound of Formula 2 or a salt thereof.

The composition of the present invention may have the effect of inhibiting flowering or reducing the size of the plant by contacting the plant.

In the above, the meanings of "plant", "flowering of the plant" and "contact" are effective as they are.

In addition, the above-mentioned also is effective as it is about the preferable aspect of the compound of <Formula 2> which is an active substance. Therefore, the compound of the most preferable aspect of the compound of <Formula 2> included in the composition of the present invention may be referred to as the compound of <Formula 1>.

On the other hand, the composition of the present invention may further comprise a suitable carrier in addition to the compound of <Formula 2> to further increase the effect of suppressing the flowering of the plant or reducing the size of the plant.

The carrier may be a natural or synthetic organic or inorganic material that can facilitate the absorption of the compound of Formula 2 by the plant or the contact of the plant with the compound. The carrier may be a solid (e.g. clay, natural or synthetic silicate, silica, resin, wax or solid fertilizer) and may be a liquid (e.g. water, alcohol, ketone, petroleum fraction, aromatic or paraffinic hydrocarbon, Chlorinated hydrocarbons, or liquefied gases).

The compositions of the present invention may also contain any components such as protective colloids, adhesives, thickeners, thixotropic agents, penetrants, stabilizers, cryoprotectants, waterproofing agents, corrosion inhibitors, desiccants, UV-stabilizers, pigments, dyes, antisettling agents and the like. May contain one or more.

On the other hand, the composition form of the present invention may be in the form of a solid, dispersible powder, wettable powder, dispersible granules or soluble granules.

In the case of a solid or dispersible powder in the form of the composition, the content of the compound of <Formula 2> may be up to 100%.

Compositions in the form of wettable powders (or spray powders) and dispersible granules generally comprise from about 20% to 95% of the compound of Formula 2, from about 0.1% to about 5% of wetting agent, from about 3% to about 10% dispersant and, if desired, other additives such as stabilizers, pigments, dyes, penetrants, adhesives or antisettling agents.

The composition in the form of the wettable powder or the dispersible granules may be used as a liquid composition at the end of use.

The composition of the invention may also take the form of a liquid from the outset, in particular in the form of a water soluble concentrate, a suspension concentrate or a paste.

The water soluble concentrate will typically comprise from about 10% to about 80% of the compound of formula 2 above.

Compositions of the present invention in liquid form that can be used directly can include from about 0.01% to about 20% of the compound of Formula 2 above.

Suspension concentrates which can be used by spraying are preferably not high in viscosity or forming precipitates during the storage period, generally from about 10% to about 75% of the compound of formula 2, about 0.5% to about 15 0.1% to about 10% of additives such as surfactants, thixotropic agents, pigments, dyes, corrosion inhibitors, stabilizers, penetrants, adhesives, anti-settling agents, and the like.

As described above, the present invention can provide a method and composition for reducing the size of a plant or inhibiting flowering.

Hereinafter, the present invention will be described with reference to Examples. However, the scope of the present invention is not limited to these examples.

Example 1 Collection of Compounds for Variant Selection

For the purpose of screening compounds that may affect various phenotypes of plants, various compounds were provided from Korea Compound Bank and used for screening.

<Example 2> Selection of Arabidopsis size reduction and blooming suppression mutants from (Arabidopsis thaliana)

In order to search for compounds affecting the phenotype of plants using Arabidopsis, 1 / 2B ₅ medium diluted with B ₅ medium (Duchefa, USA) was first added to each well of a 24 well plate. Dispense and solidify ml. In this medium, three seeds of Arabidopsis seeds were seeded, and the plants were grown for one week in a temperature-controlled culture room at 23 ° C., and then each compound provided to the plants on a clean bench (concentration was about 1 mM) (DMSO) Dissolved in an organic solvent) was dispensed at about 3 μl so that the concentration of the compound in the well containing 1.5 mL of medium was about 2 μM.

After treatment with the compound, every two days, all the phenotypes of the plant that can be observed with the naked eye, such as the growth pattern of the plant root, the shape of the leaves, the timing of flowering or not, the length of the internode, the degree of aging, and the control plants (DMSO) was compared with the phenotype.

Control plants refer to plants grown in medium treated with DMSO only. The reason for observing the plants grown in the DMSO-treated medium as a control is to confirm whether the change in the phenotype is a result of processing the compound provided by the Korea Compound Bank or the result of DMSO, the dissolved organic solvent. To do that.

As a result, as shown in Figs. 1 to 3, the size of leaves, intercalates, roots, etc. of the compounds treated as shown in Figs. 1 to 3 are smaller than those of the control plants, and no plants are formed. I could confirm it.

Figure 1 is a picture of growing the Arabidopsis for about 1 week by treating the compound, Figure 2 and Figure 3 is a picture of the leaves and plants as a whole when treated with the compound and growing Arabidopsis for 3 weeks, respectively.

In order to confirm that the above results were reproduced, the same experiment was repeated with respect to the compound causing the change in the phenotype, and as a result, it was confirmed that the change in the phenotype is reproduced.

Example 3 Compound Structure Analysis

As a result of analyzing the structure of the compound through NMR analysis after the experiment as described above, the compound of the structure shown in the above formula (2-((4-diethylamino) -2-hydroxybenzoyl) benzoic acid, CAS No .: 5809-23-4).

1 is a photograph when the Arabidopsis grows for about 1 week by treating the compound.

2 and 3 are photographs of the whole leaves and plants when the compound was treated and the Arabidopsis larvae were grown for three weeks.

Claims (24)

A method of inhibiting flowering of a plant, comprising contacting the plant with a compound of Formula 2 or a salt thereof <Formula 2>

In the above, R ₁ and R ₂ are each independently an ethyl group. The method of claim 1, The plant is a method of inhibiting the flowering of the plant, characterized in that the plant belonging to the cruciferaceae. delete delete delete Plants with suppressed flowering obtained by the method of claim 1. A method for reducing the size of a plant comprising contacting the plant with a compound of Formula 2 or a salt thereof <Formula 2>

In the above, R ₁ and R ₂ are each independently an ethyl group. The method of claim 7, wherein The plant is a method of reducing the size of the plant, characterized in that the plant belonging to the cruciferaceae. delete delete delete Plants of reduced size obtained according to the method of claims 7 or 8. A composition for inhibiting flowering of a plant comprising the compound of Formula <2> or a salt thereof as an active ingredient. <Formula 2>

In the above, R ₁ and R ₂ are each independently an ethyl group. The method of claim 13, The plant is a composition for inhibiting flowering of a plant, characterized in that the plant belongs to the cruciferaceae. delete delete delete The method of claim 13, The composition comprises at least one selected from the group consisting of protective colloids, adhesives, thickeners, thixotropic agents, penetrants, stabilizers, cryoprotectants, waterproofing agents, corrosion inhibitors, desiccants, UV-stabilizers, pigments, dyes and antisettling agents. Composition for inhibiting flowering of a plant, characterized in that. A composition for reducing the size of a plant comprising the compound of Formula <2> or a salt thereof as an active ingredient. <Formula 2>

In the above, R ₁ and R ₂ are each independently an ethyl group. The method of claim 19, The plant is a composition for reducing the size of a plant, characterized in that the plant belonging to the cruciferaceae. delete delete delete The method of claim 19, The composition comprises at least one selected from the group consisting of protective colloids, adhesives, thickeners, thixotropic agents, penetrants, stabilizers, cryoprotectants, waterproofing agents, corrosion inhibitors, desiccants, UV-stabilizers, pigments, dyes and antisettling agents. Composition for reducing the size of a plant, characterized in that.

Images