KR100621917B1 - Composition for control of harmful algae and dukweed comprising polygonatum or its extract - Google Patents

Abstract

The present invention relates to a harmful algae and duckweed control composition containing the Polygonatum plants or extracts thereof as an active ingredient and a method for controlling the same, the poisonous plants or extracts of the present invention is harmless to beneficial algae or aquatic organisms In addition, since only the harmful algae and red algae and other harmful algae selectively exhibits an excellent control effect and exhibits an additive effect with the compounds having other control activities, the control composition containing the same is natural harmful and excellent algae control agent It can be usefully used as.

Description

COMPOSITION FOR CONTROL OF HARMFUL ALGAE AND DUKWEED COMPRISING POLYGONATUM OR ITS EXTRACT}             

1 is a result of comparing the inhibitory activity of the growth of the rice frog frogs of the extracts extracted by the extraction method A to E in Example 2,

FIG. 2 is a view of the extract of the roundworm extract extracted by the extraction method A of Example 2, Chlorella vulgaris ( Clorella vulgaris), which is a beneficial green algae, with microcystis aeruginosa and Anabaena affinis , which are harmful cyanobacteria in freshwater algae. vulgaris ) and the effects on the growth of the Scenedesmus spp. at low algae concentration (A) and high algae concentration (B),

Figure 3 shows the effect on the proliferation of Cochlodinium polykrikoides of red algae of the extract of the dongleule extracted by the extraction method A of Example 2,

Figure 4 shows the effect on the motility of Cochlodinium polykrikoides of the extract of the hulling extract extracted by the extraction method A of Example 2,

Figure 5 shows the interaction of the extract of the goulle and momilactone B (momilactone B) extracted by the extraction method A of Example 2 for the control of the rice frog.

The present invention relates to a harmful algae and duckweed control composition containing the plant as an active ingredient or a fungus and its control method.

Algae inhabit seawater or freshwater and affect ecosystems. Especially, algae, which adversely affects the environment and economic activities, are called harmful algae. It is divided into red tide phenomenon and green algae phenomenon.

Red Tide phenomenon nose keulrodi is generated from the enemy mass proliferation of algae is the cause, mainly sea of nyumsok (Cochlodinium) to give the fatal order in the aquaculture industry, including fish, shellfish, algal blooms are micro-hour seutiseu in (Microcystis), in Ana Vena ( Anabaena ) , Oscillatoria , Aphanizomenon or One species or several species of cyanobacteria, such as Nodularia , grow in large quantities, mainly in freshwater eutrophicated waters.

Mass propagation of these harmful algae causes several damages:

1) Generates odorants, lowers the value of drinking water and releases toxins that are harmful to humans and animals (Haider et al., Chemosphere , 52 ( 1 ), 1-21, 2003);

2) It causes the death of fish and aquatic organisms due to lack of dissolved oxygen, toxin generation and the accumulation of foreign substances in gills, and it causes the off-flavor in the fish meat even if the algae is not proliferated. Falling (Duke et al . , Weed Sci. , 50, 138-151, 2002);

3) economic loss of overchlorination due to filter paper closure and inhibition of flocculation precipitation during water treatment;

4) destroy ecosystems by killing native animals, moving habitats, changing populations, and losing food; And

5) It causes discomfort and impedes leisure and industrial activities by coloring of water and formation of scum.

In Korea, a large amount of algae occurs in the sea, lakes, ponds, rivers, and fish farms due to the increase of emission sources after climate change and industrialization, which is a big social and economic problem. In recent years, microcystin toxins of harmful algae have also been frequently detected in lakes and rivers (Park et al. The Korean J. of Phycol. , 11 ( 1 ), 149-154, 1996).

Therefore, researches to control such harmful algae are actively conducted, and the existing reported related technologies are largely divided into physical control, chemical control, and biological control. Physical control includes the installation of a filtration device or an air injector and the method of precipitating algae by spraying ocher. However, this method is non-selective, and when the flow of the algae is strong, the control effect is lowered, and when the amount of sunshine and water temperature increase, red tide occurs again. Biological control methods include methods using natural bacteria or viruses of harmful algae (Walker et al., Biological Control, 18, 71-78, 2000; Korean Patent Publication No. 2002-13923). In this case, the effect is low, slow release and lack of host specificity. Chemical control refers to a method of inhibiting or removing algae by treating an inorganic or organic compound in water, and such chemical control can be divided into three types according to the characteristics of the compound used.

First, copper sulfate, chlorine, alkaline earth metal peroxides and hydrogen peroxide are treated, and are fast-acting and have a pronounced treatment effect, but non-selective and inorganic matters have a problem of salt accumulation. Second is the method of treating herbicides such as simazine and diuron, which are also non-selective and toxic to humans, which is a safety issue for non-target organisms.

The third is a method of treating natural products or derivatives thereof, which is an emerging natural friendly technique. As natural products studied so far, potassium ions (potassium ion, Parker et al. , Appl. Envir. Microbiol. , 63, 2324-2329, Lysine and its derivatives (Hehmann et al., J. Applied Phycology , 14, 85-89, 2002), ferulic acid, trans- cinnamic acid, quinone compounds (Schrader et al . , Bull Environ. Contam.Toxicol . , 60, 651-658, 1998), basililamide (bacillamide, Jeong et al., Tetrahedron Letters , 44, 8005-8007, 2003), fischerellin A and B, Hagmann et al., Tetrahedron Letters , 37 ( 36 ), 6539-6542, 1996; Papke et al., Tetrahedron Letters , 38 ( 3 ), 379-382, 1997), oxygenated fatty acids and potassium ricinoleate, Duke et al., Weed Sci. , 50, 138-151, 2002), and plants include ripe barley straw and Lolium perenne (Duke et al . , Weed Sci. , 50, 138-151, 2002; Schrader, Agricultural Research , 51 ( 4 ), 21, 2003).

However, existing algae control techniques using natural products are low in efficiency, effective only in non-selective or extremely limited species of algae, and have difficulty in mass production. Therefore, there is a demand for technology development that can compensate for these shortcomings.

On the other hand, the perennial herb is a perennial herb belonging to the family Liliaceae, with about 10 species growing in Korea. Young shoots and rhizomes are edible, and they are also used as medicines for pneumonia, ophthalmosis, lung spear, cardiac, nourishment, tonic, diabetes, intestinal life, nominal and customs in ornamental or oriental medicine (Kim, Korean Resource Plant V , 171-). 174, 1996). It is also used to manufacture health drinks (Korean Patent Publication No. 2002-361092), tea, herbal kimchi (Korean Patent Publication No. 2003-12546), or Nunglongji containing Donggul (Korean Patent Publication No. 2002-64744). However, no use as an algicide has been reported.

Accordingly, the present inventors have diligently studied natural-friendly natural product control agents selectively acting only on harmful birds, and as a result, freshwater and seawater can be protected while protecting beneficial algae and aquatic organisms containing polygonatum plants or extracts thereof. Only harmful algae were found to be effective.

SUMMARY OF THE INVENTION An object of the present invention is to provide a safe and nature-friendly natural product-containing control composition and a method for controlling the same, which protects beneficial aquatic organisms and controls only harmful algae or duckweed.

In accordance with the above object, the present invention provides a harmful algae and duckweed control composition containing a polygonatum plants or extracts growing in Korea and a control method using the same.

In Polygonatum in plants can be used in the present invention jukdae (Polygonatum lasianthum var. Coreanum Nakai) , yellowfin Polygonatum (Polygonatum humile Fisher ex Maximowicz), dogwood Polygonatum (Polygonatum stenophyllum Maximowicz), Tung Polygonatum (Polygonatum inflatum Komarov), Polygonatum odoratum (Polygonatum odoratum var.pluriflorum Owhi), Polygonatum falcatum A. Gray, Polygonatum involucratum Maximowicz, Polygonatum koreanum , and Polygonatum robustum , etc. Do.

In addition, the Lepidoptera plant may be used as the plant tissue itself or as an extract obtained by extracting the tissue of the plant with water and / or an organic solvent.

When using the plant tissues themselves, the dry gourd plants are dried at room temperature or, if necessary, dried in a dryer of less than 100 ° C. and then powdered and stored. The dried powder can be used by directly spraying it where it is necessary to control.

When using the extract of the plant, it is possible to exemplify the plant of the plant that can be used for the extraction process, such as living tissue, shaded tissue or completely dried tissue. It is preferable to use the root diameter which shows the outstanding control activity.

Organic solvents that can be used for extraction include methanol, ethanol and propanol, and the extraction process of the present invention may include a process of extracting in various ways using water and / or an organic solvent. For example, in consideration of long-term storage and transportation, extract the tissues of the plant of the genus Hungle, and then concentrated under reduced pressure after filtration, or separated by adding water and butanol to the methanol extract (dry) An extraction method comprising the step of drying the fractions under reduced pressure is preferred.

In addition, a surfactant may be mixed with the extract of the hulling agent for stability of the effect. The surfactant can be ionic or nonionic. Examples of suitable surfactants include salts such as sodium or calcium of polyacrylic acid and lignin sulfonic acid; Condensation products with fatty acids, aliphatic amines, amides, ethylene oxide and / or propylene oxide containing at least 12 carbon atoms in the molecule; Fatty acid esters of glycerol, sorbitan, sucrose or pentaerythritol; Ethylene oxide and / or propylene oxide and their condensates; fatty alcohols or alkyl phenols such as p-octylphenol, p-octylcresol, ethylene oxide and / or propylene oxide and condensation products; Sulfates or sulfonates of these condensation products; Alkali or alkaline earth metal salts of sulfuric acid or sulfonic acid esters containing at least 10 carbon atoms in the molecule, preferably sodium salts such as sodium lauryl sulfate, secondary alkyl sodium sulfate, sodium salts of sulfonated castor oil and dodecylbenzene sulfonate Sodium alkylarylsulfonates such as; And polymers of ethylene oxide and copolymers of ethylene oxide and propylene oxide. The surfactant may be mixed in an amount of 0.0001 to 0.05% by weight, preferably 0.005 to 0.03% by weight based on the extract. In addition, the control composition of the present invention may further include a carrier and additives commonly used for formulation in the pesticide field.

Controlling composition of the present invention harmful birds and shows a preventive effect on duckweed, toxic algae include micro during seutiseu (Microcystis), Ana vena (Anabaena), come la thoria (Oscillatoria), Aphanizomenon Menon (Aphanizomenon), no dyulra Liao ( Nodularia ) and Cochlodinium , and the like. Therefore, the present invention provides a method for controlling harmful algae or duckweed, including treating the extract of the genus Lepidoptera to the area where the harmful algae and duckweed occurred or expected to occur.

When the dry powder of the plant of the present invention is treated in the control area, it can be treated in the range of 0.05 mg / ml to 1.25 mg / ml based on the plant tissue raw weight. In the case of the plant extract, from 0.003 g / L (3 ppm) to 0.012 g / L (12 ppm), preferably 0.005 g for the control of harmful algae based on the final concentration of the treatment area. / l (5 ppm) to 0.0.010 g / l (10 ppm), and 0.010 g / l (10 ppm) to 0.060 g / l (60 ppm), preferably 0.030 for duckweed control g / l (30 ppm) to 0.050 g / l (50 ppm).

The invention is illustrated in detail by the following examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example 1 Comparison of Control Effects According to Species or Tissues of the Plant Species

Of the Polygonatum genus (Polygonatum) plants, jukdae (Polygonatum lasianthum var. Coreanum Nakai) , yellowfin Polygonatum (Polygonatum humile Fisher ex Maximowicz), dogwood Polygonatum (Polygonatum stenophyllum Maximowicz), Tung Polygonatum (Polygonatum inflatum Komarov), Polygonatum (Polygonatum odoratum var. pluriflorum Owhi), Polygonatum falcatum A. Gray and Polygonatum involucratum Maximowicz were collected from the wild and divided into outposts, underground roots and ground (leaves and stems), and their extracts were extracted from Lemna paucicostata 381, The effect on proliferation was investigated.

300 g of donggles' fresh tissue was added to 0.5 L of methanol, and the active ingredient was extracted at room temperature for 1 day, and then dried under reduced pressure. The dried material obtained was obtained from the Frogweed rice growth medium (Hutner's medium; Growth and Differentiation in Plants, ed by WE Loomis, The Iowa State College Press, Ames, Iowa. P. 417, 1958). As shown in the figure, it was dissolved at a concentration of 2 to 250 ppm, and dispensed into a test container at 2 ml, 3 for each concentration. At this time, the growth medium without the dry matter as a control was dispensed in the same manner in a test container. In each test vessel, one individual was mothballed with four fronds, and it was cultured for 5 days under a photoperiod (50 μmolm ⁻² s ⁻¹ ) at 26 ° C. for 14 hours. Based on the results, the relative activity of each Rhododendron plant extract is shown in Table 1 below with the activity of 100 completely and 0 in the control group.

As a result, as shown in Table 1, all the extracts of the fungus plant used showed control activity, in particular, the root extracts of the fungus, tortilla and layered tortilla were up to a concentration ranging from about 4 to 30 ppm. Activity was shown.

Example 2 Comparison of Active Differences According to Extraction Methods

In order to determine a method of extracting a plant of the genus Oleum, which can be usefully used as a control composition, the following five extraction methods were compared with the inhibitory effect on photosynthesis of the rice frog.

1) Organic Solvent Extraction (Extracting Method A): 1.2 kg of a round root diameter was cut to a thickness of about 5 mm, and then immersed in 2 L of methanol and extracted at room temperature for 2 days. The obtained extract was filtered through gauze, concentrated under reduced pressure, and re-dissolved in water to 500 ml. When 500 ml of butanol was added to the aqueous solution of the extract and mixed, the mixture was left for 1 hour to perform layer separation. The separated water layer was taken to perform layer separation in the same manner as above, and the water layer was taken off to remove the solvent. The mixture was dried under reduced pressure sufficiently to obtain 60 g of the extract. It was diluted in the growth medium of the frogs to the concentration of 62.5, 31, 16, 8, 4 ppm.

2) Hot water extraction (extraction method B): 20 g of the round root diameter was cut to about 5 mm, put into an appropriate amount of water, and extracted at 100 ° C. for 2 hours. After extraction, the remaining rhizome sections were discarded and the extract was collected and water was added and filtered until the final volume was 20 ml. 6 ml of water was added to 4 ml of this extract filtrate to obtain an extract. This was diluted in the growth medium of some frogs so that the final concentration is 500, 250, 125, 62.5, 31 ppm.

3) Extraction at room temperature (extraction method C): 20 g of round root diameter was cut into 5 mm size, put in 20 ml of water, extracted at room temperature for 2 days, and filtered. 6 ml of water was added to 4 ml of this extract filtrate to obtain a round extract. This was diluted in the growth medium of some frogs so that the final concentration is 500, 250, 125, 62.5, 31 ppm.

4) Bio-extraction (extraction method D): 4 g of round gourd root was put in 6 ml of distilled water, ground and centrifuged (15,000 g, 10 minutes) to obtain a supernatant extract of about 1 ml. Diluted in the medium of the frog frog growth so that the final concentration is 250, 125, 62.5, 31, 16 ppm.

5) Dry powder extraction (extracting method E): 1.04 g of dried gourd root powder was mixed with 9 ml of distilled water, followed by centrifugation (15,000 g, 10 minutes) to obtain a round gourd extract as a supernatant. This was diluted to 250, 125, 62.5, 31, 16 ppm to the growth medium of the frog.

The test solution prepared in this way was dispensed three by 2 ml each, and each test container was wounded with one individual Frogweed plant having four fronds. After that, the chlorophyll content was investigated by incubating for 5 days at 26 ° C. for 14 hours in photoperiod (luminosity: 50 μmolm ^-2 s ^-1 ), and the method of extracting DMSO extracted with solvent (DMSO) was investigated. (DMSO extraction technique, Hiscox and Israelstam method (Can. J. Bot. 57: 1332-1334, 1979) were used.

As a result, as shown in Figure 1, the extraction method E to extract the dry powder immersed in water showed the best effect, the extraction method D to extract the biological sample directly showed the next excellent effect. Extraction method B or extraction method C showed a low extraction yield but less drug efficacy due to high temperature. On the other hand, extraction method A, which is extracted with methanol, shows relatively low efficacy, which is considered to be due to some loss of active ingredient during extraction.

However, extraction method A requires an organic solvent handling facility, but can be easily concentrated, which is advantageous for long-term storage and transportation. Extraction method E can be extracted with a simple facility. There are disadvantages. Therefore, for practical use, extraction methods A and E may be usefully used among the above methods, but it may be necessary to select an extract production method in consideration of their advantages and disadvantages.

Example 3: Control effect of fresh oyster extract on freshwater algae

In order to investigate the control effect of the extract of the donggule on freshwater algae, two kinds of harmful algae (microsistis aerugii), which were distributed by the Korea Institute of Bioscience and Biotechnology, using the donggule extract extracted by the extraction method A of Example 2 The control and algae effects of the labor and management ( Microcystis aeruginosa (UTEX 2388) and Anabaena affinis ) and two beneficial green algae ( Chlorella vulgaris and Scenedesmus spp.) Were investigated. .

Divided into the case of low algae concentration and high algae concentration of 4 to 8 times the concentration of low algae, it was confirmed the control (low algae concentration) and algae (high algae concentration) effect on the harmful algae of the extract. The initial concentration of each algae was adjusted by absorbance measurement using a spectrophotometer (Shimazu UV6504PC). In the case of Microcistis aeruginosa, the low concentrations of A680 nm to A800 nm were 0.086 and the high concentrations were 0.390. Low concentrations of A665.5 nm to A800 nm were 0.08 and high concentrations were 0.32. In the case of Chlorella Bulgari The low concentration of A687 nm to A800 nm was 0.0463 and the high concentration was 0.6503. The low concentration of A688 nm to A800 nm was 0.090 and the high concentration was 0.4413. The round goose extract extracted by the extraction method A of Example 2 was diluted in a growth medium (Allen's medium) at a concentration of 1 to 1000 ppm, and supplied to a test vessel of each algal species at low or high algal concentration conditions. . This was incubated at 25 ℃, photoperiod 14 hours (luminance: 60 μmol ^-2 s ^-1 ), 140 rpm conditions. In order to analyze the growth of algae 4 days after the extract treatment, the absorbance or pigment content was calculated as the relative growth inhibition (%) relative to the control group not treated with the extract.

As a result, as shown in Figure 2, the extract was shown to selectively control and killing algae only harmful algae without affecting the beneficial green algae. In addition, the low initial concentration of algae showed a good activity even in the case of low concentration extracts.

Example 4 Control Effect of Red Algae Extract on Red Tide

In order to confirm the control effect of the roundworm extract on the red algae which is harmful to the sea, Cochlodinium polykrikoides is known to cause the most problems in Korea by using the roundworm extract extracted by the extraction method A of Example 2. The control effect on) was confirmed.

Coclodinium polycricoides were obtained from the National Fisheries Research and Development Institute and cultured in F / 2 medium prepared with seawater (Guillard and Ryther, 1962) at 20 ° C., photoperiod for 24 hours, and brightness of 65 μmolm ⁻² s ⁻¹ . It was. 50 ml each of subcultured coclodinium polycricoides was added to the Erlenmeyer flask so as to be 3 × 10 ³ cells / ml, and the donggulae extract was filtered using a 0.20 μm filter, and the final concentration of the extract was 6.25 to 100 ppm. The flasks were administered to each flask, and the flask not administered with the extract was used as a control. This was incubated for 4 days at 20 ° C. and continuous light conditions (luminosity: 65 μmolm ⁻² s ⁻¹ ), and 5 μl each day was taken from the culture medium, and the number and motility were examined using an optical microscope (150 ×). Repeat 10 times per treatment.

As a result, as shown in FIG. 3, the degree of suppression was about 35% in 6.25 ppm of the extract, but from 12.5 ppm, the red algae growth was suppressed almost 100%. At concentrations above 25 ppm, it inhibited division as well as induced cell destruction, resulting in an excellent killing effect. In addition, the inhibitory effect on the motility of the red tide (Fig. 4), the extract concentration was 12.5 ppm lowered to 23.4% until the third day, and then recovered slightly after that. However, it was confirmed that at the concentration of 25 ppm or more of the extract, the activity was sharply decreased and almost 100% was inhibited on the second day after the treatment. Therefore, it can be seen that red tide is effectively controlled when the concentration of the extract is more than 10 ppm.

Example 5 Mixture Treatment Effect of Dongle Extract and Momilactone B

Momilactone B (Tetrahedron Letters 39: 3861-3864, 1973; Biosci. Biotech. Biochem. 63: 1318-1320, 1999), a natural substance that is known to have inhibitory effects on weeds such as blood or cob The interaction between the liver and the control of the frog was confirmed. A mixture of a round gourd extract (13, 16, 19, 22, and 25 ppm) and Momilactone B (0.19, 0.38, 0.75 ppm) extracted by the extraction method A of Example 2 was treated to Jok-frog rice as described in Example 2. , The interaction of the two compounds was confirmed. The evaluation of the interactions was calculated by the Colby method (Colby SR, Weeds , 15, 20-22, 1967).

As a result, as shown in Figure 5, the extract of the donggulle and Momolactone B showed an additive effect. Therefore, by treating the extract with the other compound having a similar effect, it can be seen that the effect of improving the efficacy, lowering the treatment concentration and reducing the production cost.

As discussed above, the fungus or its extract extracted according to the production method of the present invention is harmless to beneficial algae or aquatic organisms, and selectively shows only excellent harmful effects to harmful algae such as southern algae and red algae, and other control activities. Since it exhibits an additive effect with the compounds having the above, the control composition containing the same can be usefully used as a natural algae and excellent efficacy of algae control agents.

Claims (11)

A harmful algae and duckweed control composition containing Polygonatum plants or extracts thereof as an active ingredient. The method of claim 1, The Polygonatum spp., Jukdae (Polygonatum lasianthum var. Coreanum Nakai) , yellowfin Polygonatum (Polygonatum humile Fisher ex Maximowicz), dogwood Polygonatum (Polygonatum stenophyllum Maximowicz), Tung Polygonatum (Polygonatum inflatum Komarov), Polygonatum (Polygonatum odoratum var. Pluriflorum Owhi) , Polygonatum falcatum A. Gray, Polygonatum involucratum Maximowicz, Polygonatum koreanum , Polygonatum robustum , and mixtures thereof. The method of claim 1, Harmful birds are selected from the group consisting of algae of the genus Microcystis , Anabaena , Oscillatoria , Aphanizomenon , Nodularia and Cochlodinium The composition characterized in that. The method of claim 1, The plant of the genus Dungle plant is a dry powder powdered after drying or drying in a dryer of less than 100 ℃. The method of claim 1, The extract is a composition, characterized in that the extract of the tissue of the plant of the genus Oleum is extracted with a solvent selected from the group consisting of water, an organic solvent and a combination thereof. The method of claim 5, wherein And wherein the organic solvent is selected from the group consisting of methanol, ethanol, propanol and mixtures thereof. The method of claim 1, The extract is prepared by the method comprising extracting the tissues of the plant of the genus Hungle with methanol and then concentrated under reduced pressure after filtration, separating the concentrate by adding water and butanol to the concentrate and then obtaining a water fraction. A method of controlling harmful algae or duckweed, comprising treating the composition of claim 1 to an area where the harmful algae and duckweed occurred or are expected to occur. The method of claim 8, The control method according to claim 1, wherein the final concentration of the tissue of the plant of the genus Oleum in the region to be controlled is 0.05 mg / ml to 1.25 mg / ml based on the fresh tissue weight. The method of claim 8, The composition of claim 1 for the control of harmful algae, characterized in that the final concentration of the extract of the plant of the genus Leung in the control area to be treated from 0.003 g / l (3 ppm) to 0.012 g / l (12 ppm), characterized in that Control method. The method of claim 8, The composition of claim 1 for the control of duckweed, characterized in that the final concentration of the extract of the plant in the control area to be treated so that the final concentration of 0.010 g / l (10 ppm) to 0.060 g / l (60 ppm) Way.

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