KR20120119375A - Dioscoreacea extract for preventing storage disease of fruits and manufacturing method thereof - Google Patents

Abstract

PURPOSE: A Dioscorea nipponica Makino extract for preventing fruit storage and a manufacturing method thereof are provided to enhance antibiosis activity for Penicillium digitatum. CONSTITUTION: A manufacturing method of Dioscorea nipponica Makino extract comprises the following steps: preparing dried Dioscorea nipponica Makino; shake culturing the prepared Dioscorea nipponica Makino after mixing the same with a solvent; filtering the cultured extract into a gauze for oriental medicine; removing solvent from the filtrate by using a rotary vacuum evaporator; and freeze-drying the extracts in which the solvent is removed by using a vacuum evaporator. The second step mixes 1 kg of Dioscorea nipponica Makino with 500 liters of the solvent. The solvent is methanol or butanol. The second step is shaking and cultivating at 20-30 deg. Celsius for 48 hours.

Description

Extract of cheonsan for fruit storage bottle control and its manufacturing method {Dioscoreacea extract for preventing storage disease of fruits and manufacturing method

The present invention relates to, and more particularly, to extract and a method of manufacturing for the fruits storage disease Penny chamber ryumsok (Penicillium) antimicrobial activity with excellent medicinal plants for the pathogens Tianshan relates to a for Tianshan extract and a method of manufacturing the same.

In fruits, diseases caused by Penicillium spp. Are not caused during the growing period of the crop, but are mainly caused by wounds during harvesting, packaging, storage and transportation. 10-30%, and some underdeveloped countries make up more than 30%. In addition, if the disease occurs during the storage of fruits, such as pears, apples, citrus fruits, and grapes, which have high value added, the merchandise will be greatly degraded, resulting in economic losses.

Among citrus stockpiles , the loss caused by Penicillium digitatum ( P. digitatum ) , which we commonly call a mixture of blue mold, and penicillium Italicum ( P. italicum) , which causes blue mold disease, is lost during the entire storage. It accounts for more than 90% of the losses incurred. Among the bottle inflict the greatest damage during the storage of citrus green mold disease is a P. Green fungal disease caused by digitatum causes millions of spores to form at the infected site and scatters, causing them to act as inoculum, and the optimal temperature for disease development can be grown at 6 ~ 37 ℃ within 25 ℃.

In contrast, P. italicum is P. It can grow at lower temperatures than the digitatum and even at 0 ° C. Among these two pathogens, P. The digitatum, Penicillium app. , which causes fruit storage disease, has the strongest pathogenicity among pathogens, and the amount of spores is also very high. Therefore, green mold disease control is very important.

The most effective control agents for the green and blue mold diseases, which are the main citrus storage rots, are organic biological pesticides such as sodium bicarbonate, Imazaryl (IMZ: imazalil), TBZ (thiabendazole), and pyrimethaneyl ( PYR: pyrimethanil), flu-dioxo-carbonyl. ((FLUD: fludioxonil) and sodium o-phenyl phenate are danje and mixtures has been widely used in gotta (sodium o -phenylphenate) but imazalil, drug for thiabendazole and sodium o- phenylphenate With the development of resistant P. digitatum bacteria, new control agents have been required.

In Korea, it is reported that the drug for the control of citrus reservoirs is not activated, and water spraying 1,000 fold of thiophanate-methyl, an organic synthetic pesticide 10 days before harvesting, is effective for the control. However, this drug is also resistant to the development of resistant bacteria is very far from the situation. Therefore, there is a need for a new control method that can replace organic synthetic pesticides and solve problems such as the emergence of resistant bacteria.

 The use of chemical synthetic pesticides in the control of plant diseases around the world is on the decline due to the Rio Convention, and the use of low-toxic and eco-friendly pesticides to prevent the corruption of storage fruits is required due to the need for harmless and eco-friendly disease control methods. Development of control measures is becoming important.

As a low-toxic and eco-friendly penicillium control method, plant extract control and microorganism control are attempted. In addition, P. Natural compounds such as amino butyric acid (β -aminobutyric acid) - for regulating plant growth in order to improve the resistance to the disease of fruit for digitatum Jass monsan (jasmonic acid), amino acid, beta (amino acid) and its derivatives Research on physical bottle control methods such as use and washing with UV and hot water is being conducted.

 In order to minimize the use of organic synthetic pesticides and to develop excellent natural pesticides that are harmless to humans and highly active against pests, the main components of pyrethrin, nicotine and rotenone, which are the main components of Pesticides, Tobacco and Deris, Organophosphates or carbamate systems have been developed as a related compound pesticide with lower toxicity than organic synthetic pesticides. In addition, in line with this trend, many disease control substances are searched for from various plant species, and strong antimicrobial properties of some substances have been identified.

However, the development of biological pesticides derived from natural products is very few compared to the vast resources of natural products. In addition, Penicillium bacteria are very difficult to develop and use microbial pesticides because they are antimicrobial due to their secreted antibiotics. In this sense, the control method of the natural product-derived biopesticides can solve the problems of microbial preparations, and the development cost is cheaper than chemical pesticides. In addition, comprehensive pest management methods are essential in the future, and biological control methods must be used together, and therefore, development of eco-friendly natural products-derived pesticides is required.

In accordance with the necessity of the above, the present invention provides a natural pesticide derived from natural products that can control environmentally friendly storage bottles of fruits.

Another object of the present invention is to provide a manufacturing method that can replace organic synthetic pesticides and can be mass-produced in order to solve the problem of the emergence of resistant bacteria.

The method for preparing cheonsan extract according to the present invention for solving the above problems, (1) preparing a dried cheonsanyong, (2) mixing the prepared cheonsanyong with a solvent, followed by shaking culture, (3) Filtering the cultured extract to a medicinal gauze, (4) removing the solvent from the filtered material using a rotary pressure reducer, and (5) lyophilizing the extract from which the solvent is removed using a pressure reducer. It is characterized by including.

In a preferred embodiment of the present invention is mixed with 1 kg for cheonsan at a rate of 500 L of solvent, the solvent is a culture of shaking at least for 48 hours at 20 ~ 30 ℃ using methanol or butanol, the main component is protodioscin (protodioscin) And a composition for controlling fruit storage bottles.

The extract for cheonsan according to the present invention is a green fungal bacterium Penicillium Excellent antimicrobial activity against digitatum has the effect of preventing the storage bottle from fruit.

In addition, the extract for producing cheonsan has the effect of mass-producing the extract for cheonsan in a simple process.

1 is a photograph showing the mycelia and spore-like identification pathogens found from infected citrus fruits.
2 Penicilliun The antifungal activity of medicinal plant extracts against digitatum .

Hereinafter, the present invention will be described in detail with reference to the drawings and examples, but the present invention is not limited to the drawings and examples.

1. Identification of Pathogens

For the study of the control of storage diseases caused by the genus Penicillium , Penicillium microorganisms were identified and identified. To separate pathogens from diseased fruit, isolate spores and mycelium from pathogens on citrus fruit surface aseptically, inoculate in PDA medium, grow 24 hours incubator in incubator at 24 ℃, isolate potato and agar Transfer to medium (PDA: Potato Dextrose Agar medium) and incubated for 3 days in a 24 ℃ incubator. For the identification of the cultures, the form and color of spores growing on the medium were confirmed, and spores and mycelia were observed by microscopic examination at 400 times. Confirmation, Penicillium from infected citrus fruit Pure separation of digitatum and Penicillium italicum could be identified.

Figure 1 is a photograph showing the mycelia and spore-like identification pathogens found from infected citrus, A is infected citrus, B is Penicilliun of PDA medium digitatum , C is Penicillium in PDA medium Italicum is shown and D to E are microscopic images of mycelium and spores.

In addition to the two pathogens isolated above, five penicillium genera, as shown in Table 1, were used in the experiment from the Korea Agricultural Culture Collection Center (KACC) to secure strains of the genus Penicillium which causes storage diseases. Presale Penicillium genus is a major Penicillium genus pathogen that occurs during storage of mushrooms, garlic and vegetables as well as high value added fruits such as citrus fruits, pears, grapes and apples.

2. Selection of antimicrobial active medicinal crops

Penicillium , a storage disease in 32 medicinal plants, including Tianshan, which is expected to contain useful antimicrobial substances In order to test the antifungal activity against digitatum , the inhibition of mycelial growth was examined.

Each of the 32 plants was crushed and dried in a grinder, and then 100 g of 100% methanol was added to 5 l and incubated for 48 hours at 24 ° C., and the extract was filtered three times using gauze and filter paper. The filtered material was completely removed by using a rotary pressure reducer and lyophilized.

The lyophilized plant extracts were adjusted to 100-fold concentration and then placed on a paper plate and placed on a PDA medium coated with Penicillium digitatum . As a result of the assay, mycelial growth was suppressed in the extract of No. 18 in Table 2 and FIG. It could be confirmed. Table 2 shows the Penicillium Antifungal activity of medicinal plants at the concentration of 5 mg / ml against digitatum .

(-: Promote growth, +: less than 25%, ++: 25 ~ 50%, +++: 50 ~ 75%)

3. Cheonsan Dragon  Mass Production of Extracts

Cheonsan used in the present invention is a medicinal plant of the horse as its scientific name is Dioscorea nipponica Makino.

As we saw above, Cheonsan Yong Penicillium The antibacterial activity against digitatum was excellent. The drug used in the experiment was purchased from a Chinese herbal medicine store, and the experiment was disclosed, but the appearance and form of the ingredients were different each time they were purchased. Currently, the majority of Chinese herbal medicines are distributed in China, and sometimes domestically, are very expensive. The results of the antimicrobial activity test in Cheonsan were domestic Cheonsan and it was possible to obtain a stable and active substance, but the results were not constant when using the Chinese Cheonsan. Commercially available Cheonsan Dragon was distributed under the generic names of Beijing Cheonsan and Chinese Cheonsan. As a result of examining the characteristics of native Cheonsan Dragon and cultivated farms, it was confirmed that the possibility of mass-cultivation of Cheonsan Dragon in Korea. The artificial cultivation method for cheonsan use requires more specific research results, but it seems to be very similar to the ginseng cultivation method.

Such cheonsan extract can be mass-produced in the following examples.

1) Into a 500 liter reactor, 1 kg of dried Cheonsan and 100 liters of methanol or butanol (500 liters) were put and incubated for 48 hours at 24 ° C.

2) The cultured extract is filtered through medicinal gauze five times.

3) The filtered material is removed from the solvent using a 20 L rotary depressurizer, and freeze-dried in a 200 L large depressurizer.

4. Analysis of Active Ingredients of Extracts

As a result of analyzing the active ingredient of the extract for cheonsan, it was confirmed that the active ingredient of the substance having antimicrobial activity was protodioscin.

[ Experimental Example  One]

In order to examine the antibacterial activity ability of the ethereal extract (protodioscin) for were manufactured with each of 50ppm, 100ppm, 200ppm, 300ppm, 400ppm, the concentration of 500ppm to melt the protodioscin obtained from extracts of JIAHERB obtained in Example 1 in the TDW, P . The inhibition of mycelial growth was measured in PDA medium inoculated with digitatum and P. italicum .

P. Inhibition of mycelial growth of protodioscin against digitatum showed that inhibition of mycelial growth from 50 ppm of protodioscin was inhibited by 21.3% at 50 ppm, compared to 4.2% of 50% inhibition of protodioscin from JIAHERB. Inhibitory effect was shown. However, it was confirmed that the average 6.3% activity is lowered at the concentration of 100ppm or more, which is considered to be due to the difference in extract production process with a professional extract production company.

Inhibition of mycelial growth of protodioscin against P. italicum showed no inhibitory effect on mycelial growth at 50 ppm. In 100ppm, 200ppm, 300ppm, 400ppm, 500ppm of protodioscin extracted from Cheonsan, 34.6%, 45.8%, 56.4%, 62.5%, 62.8% showed mycelial growth inhibition, and 100ppm, 200ppm, 300ppm of protodioscin purchased from JIAHERB. , 400ppm, 500ppm showed 35.5%, 47.7%, 60.1%, 61.2%, 63.9% inhibition of mycelial growth. Protodioscin inhibited the mycelial growth of P. italicum .

The antimicrobial activity of protodioscin on the medium showed that P. digitatum It was confirmed that the average mycelial suppression effect of P. italicum was reduced by 7.25%. Based on these results, the target for field evaluation and formulation is P. The digitatum was deemed appropriate, and it was evaluated that more than 200ppm of protodioscin should be used for field experiments and commercialization, or the search and study of supplements to enhance the effect were necessary.

Claims (6)

(1) preparing the dried cheonsanyong
(2) mixing the prepared cheonsanyong with a solvent and then shaking culture
(3) filtering the cultured extract to a medicinal gauze
(4) removing the solvent from the filtered material using a rotary pressure reducer
(5) a method for producing an extract for cheonsan, comprising lyophilizing the extract from which the solvent has been removed using a vacuum concentrator. The method of claim 1,
Step (2) is a method for producing an extract for cheonsan, characterized in that the mixture of the solvent 500ℓ in 1kg for cheonsan The method of claim 2,
The solvent is a method for producing an extract for cheonsan, characterized in that methanol or butanol The method of claim 1,
Step (2) is a method for producing an extract for cheonsan, characterized in that the culture in shaking for at least 48 hours at 20 ~ 30 ℃ A composition for extracting cheonsan prepared by the method according to any one of claims 1 to 4, wherein the component is protodioscin. The composition according to claim 5, wherein the composition is for controlling fruit storage bottles.

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