KR101144987B1 - Nematocide Compound containing amino acids extracted by using chicken feather-degrading bacterium Chryseobacterium sp. FBF-7 - Google Patents

Abstract

The present invention relates to a culture of a strain of Chrysobacteria FBF-7 and a composition for controlling root-knot nematodes containing the same, and more particularly, to a culture of a strain of Chrysophyta bacteria FBF-7 having a keratin- The present invention relates to a composition for controlling root-knot nematis, which is contained as an active ingredient. The composition is an excellent biological preparation capable of replacing a conventional chemical pesticide preparation. It can reduce the damage of pesticide residues in the soil and the undesirable bactericidal action of soil- In addition, the use of indigenous bacterium in Korea will make soil settlement possible and contribute to the practical use of the plantation area.

Description

[0001] The present invention relates to a culture of a strain of Chrysosobacterium FBF-7 and a composition for controlling root-knot nematis containing the same. [0002] Chryseobacterium sp. FBF-7}

The present invention relates to a culture of a strain of Chrysobacteria FBF-7 and a composition for controlling root-knot nematodes containing the same, and more particularly, to a culture of a strain of Chrysophyta bacteria FBF-7 having a keratin- The present invention relates to a composition for controlling root-knot nematodes containing an effective ingredient.

Plant-parasitic nematode and root-knot nematode are known as typical nematodes that damage the domestic horticultural crops and fruit and vegetable plantation sites. In particular, crop damage caused by root-knot nematodes Is serious.

The root-knot nematode is a pest which is the most difficult to control in the production of fruit and vegetables such as melon, cucumber, watermelon, melon, Currently, there are methods for control of root-knot nematodes such as cultivation control, physical control and chemical control. There are various kinds of cultivation control such as fresh water, fresh water, dry soil, deep ground, soil, rice cultivation and resistant cultivars. Heat treatment (steam, dry heat, hot dip method) and solar thermal disinfection are used for physical control. , And phoschia jat (nematoda) are sprayed to control nematodes. In the case of cultivated control, it is difficult to apply or costly depending on local conditions and cultivated crops. Resistant cultivars are limited to specific crops and are not practical. Chemical agents are highly effective in controlling pollutants, but most of them are toxic when they are left in the soil due to organic phosphorus and carbamates. They are harmful due to indiscriminate bactericidal action of beneficial microorganisms in the soil, which is beneficial to the growth of crops as well as environmental pollution. .

As an alternative to chemical control agents, studies on biological control using root-knot nematode microorganisms, biochemical agents and plant extracts have been actively conducted. Most of the nematode microorganisms reported so far are nematode fungicidal molds, which are extremely ineffective for practical use in farms and are difficult to commercialize for commercial use. In the case of natural bacteria, the microorganisms that have been proven to have biological control effect against root-knot nematode are Pasteuria penetrans and Bacillus thuringiensis , Bacillus penetrans , etc. However, there are many problems to be solved such as difficulty of cultivation and practical use of the cultivation land.

It is known that the biochemical agent studied as a biological agent for controlling nematodes in Korea is most effective in controlling nematodes by extracts from domestic native plants and medicinal plant extracts. However, since it is difficult to supply and receive raw materials, have.

Research on nematode control using biochemical agents such as various functional substances in addition to plant extracts has been actively conducted overseas. Especially, it has been confirmed that methionine amino acid and amino acid-containing substances such as "methionine soil blending effect of root-knot nematode reduction effect" have excellent nematocide effect, and the study is proceeding to practical use. However, in Korea, functional biochemical agents such as amino acid and amino acid- There is no study on nematode control, and it is required to develop the technology.

Accordingly, the inventors of the present invention have confirmed the surprising control effect against root-knot nematodes from a culture of a novel strain having a keratin-based protease-producing ability while carrying out continuous research on biological products for nematode control, and completed the present invention.

It is an object of the present invention to provide a composition for controlling root-knot nematodes containing a culture of a novel strain having a keratin-based protease-producing ability as an active ingredient.

In order to accomplish the above object, the present invention provides a composition for controlling root-knot nematodes comprising a culture of a novel strain having a keratin-based protease-producing ability as an active ingredient.

More particularly, the present invention provides a composition for controlling sweet potato root-knot nematodes from a culture of a novel strain having a keratin-based protease-producing ability, which is identified after separation from a pulmonary feather sample collected from a poultry farm.

The root-knot nematode controlling composition of the present invention is characterized in that it contains a culture of Chryseobacterium sp. FBF-7 (KACC 91463P) having a keratin-based protease-producing ability.

The composition for controlling root-knot nematodes of the present invention is an amino acid composition obtained by decomposition from feathers, wherein the amino acid is selected from the group consisting of valine, glutamic acid, aspartic acid, glycine, and at least one member selected from the group consisting of prorine, serine, leucine and essential amino acids.

The composition for controlling root-knot nematodes according to the present invention is a composition for controlling root-knot nematodes of Meloidogyne incognita . < / RTI >

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for controlling root-knot nematodes, which comprises a culture of a strain of Chryseobacterium sp. FBF-7 (KACC 91463P) having a keratin-based protease-producing ability.

The above-mentioned strain Chryseobacterium sp. FBF-7 (KACC 91463P) is a novel strain identified after separation from a feather sample collected from a poultry farm, and more particularly, a strain isolated from feathers and compost 125 strains were tested for their ability to produce keratinocyte proteases. Eight strains with excellent proteolytic enzyme-producing ability were isolated by primary cleavage at 48 hours after culture, Were isolated from pure keratin - derived proteins. Please refer to Fig.

16S rRNA sequencing analysis was carried out to identify the strain having excellent keratin protease activity, and it was confirmed to be a strain belonging to the genus Chryseobacterium described in SEQ ID NO: 1. See FIG.

Therefore, the present inventors named the newly isolated strain as Chryseobacterium sp. FBF-7 and deposited it on the Korean Agency for Cultural Heritage on Apr. 1, 2009 and assigned the accession number KACC 91463P received.

In the present invention, the protease produced from the strain Chryseobacterium sp. FBF-7 (KACC 91463P) has a strong keratinolytic activity.

More specifically the Cri Seo tumefaciens (Chryseobacterium sp.) FBF-7 (KACC 91463P) results confirmed the resolution of the waste feathers due to strain, the major amino acid produced is glutamic acid is the highest with 8.17 nmol / g (17%) (11%), valine 4.16 nmol / g (9%), proline 4.02 nmol / g (8%), leucine 3.84 (8%), arginine 5.8 nmol / nmol / g (8%) and 11 other amino acids were confirmed. A total of 17 amino acids were produced from the lung feathers by the novel strain Chryseobacterium sp. FBF-7 (KACC 91463P) Respectively.

From the above results, it was found that Chryseobacterium The strain FBF-7 (KACC 91463P) produces the keratin-based protease, thus completely decomposing the feathers of poultry including chicken feathers and downy feathers, thereby treating feathers from the poultry industry conveniently and at low cost There are advantages to be able to.

In addition, the above-mentioned Chryseobacterium The strain FBF-7 (KACC 91463P) (KACC 91463P) has the advantage that it can be used as a useful resource for industrial and agricultural uses by producing 17 amino acids from keratin protein byproducts such as lung feathers and claws.

In the present invention, the culture of the strain Chryseobacterium sp. FBF-7 (KACC 91463P) has an effect of controlling root-knot nematode. More specifically, the culture of poultry, (KACC 91463P) culture of the present invention cultured in a culture medium containing an organic nitrogen source selected from the group of feathers and a carbon source has an effect of controlling root-knot nematodes.

The carbon source may be at least one selected from the group consisting of galactose, lactose and sobitol. The culture may be cultivated at a culture temperature of 13 to 28 DEG C, a culture time of 65 to 80 hours, a pH 6.5 to pH 8.5. Please refer to Fig.

More specifically, the culture of the strain of Chryseobacterium sp. FBF-7 (KACC 91463P) is used for culturing the strain. After culturing the strain, 0.1 to 3.0% 0.05 to 0.2% by weight of organic nitrogen source and 0.025 to 0.1% by weight of carbon source, culturing under conditions of a culture temperature of 13 to 28 캜, a culture time of 65 to 80 hours, a pH of 6.5 to pH 8.5 .

In the present invention, the culture is an amino acid composition obtained by decomposing from feathers, wherein the amino acid is selected from the group consisting of valine, glutamic acid, aspartic acid, glycine, proline, serine, leucine, and essential amino acids.

In addition, in the present invention, the culture of the present invention can be used as a sweet potato root nodule ( Meloidogyne incognita ).

More specifically, the mortality and egg inhibition rates of larvae (J2) of sweet potato root-knot nematodes were investigated using an amino acid composition degraded by Chrysobacterium sp. FBF-7 from 2.0% % Of the amino acid composition of the present invention, the mortality rate of the larvae of 50% or more within 48 hours and the inhibition rate of egg incubation of 68% within 24 days can be confirmed. Thus, the amino acid composition (KAA) It was confirmed that this is a biological preparation. 8 and 9.

The inventive Chryseobacterium sp.) By ensuring that the amino acid and amino acid-containing material of the FBF-7 (KACC 91463P) culture is available as a functional biochemical agent for root-knot nematodes, it will contribute greatly to the control of nematodes at the plantation site.

The composition for controlling root-knot nematitis of the present invention is an excellent biologic agent that can replace the existing chemical pesticide preparation. It can reduce the damage of pesticide residues in the soil and the irremediable disinfection action of soil microorganisms, This will contribute to the practical use of the plantation site by making soil fixation possible.

1 is Cri Seo tumefaciens (Chryseobacterium in accordance with the present invention sp.) FBF-7 (KACC 91463P)
Figure 2 is a graph showing the activity of Chryseobacterium < RTI ID = 0.0 > sp.) FBF-7 (KACC 91463P) strain,
FIG. 3 is a graph showing the activity of Chryseobacterium sp.) FBF-7 (KACC 91463P) strain,
Figure 4 is a graphical representation of the results obtained with Chryseobacterium < RTI ID = 0.0 > sp.) FBF-7 (KACC 91463P), which is a major component of the pulmonary feathers,
FIG. 5 is a graph showing the results of the measurement of the activity of Chryseobacterium sp.) FBF-7 (KACC 91463P)
FIG. 6 is a graph showing the effect of the present invention on Chryseobacterium sp.) FBF-7 (KACC 91463P)
FIG. 7 is a graph showing the effect of the present invention on Chryseobacterium sp.) FBF-7 (KACC 91463P) strain,
Figure 8 is a graph showing the activity of Chryseobacterium < RTI ID = 0.0 > sp.) FBF-7 (KACC 91463P) strains,
FIG. 9 is a graph showing the activity of Chryseobacterium sp.) FBF-7 (KACC 91463P) strain cultured in the culture medium.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, it should be understood that the present invention is not limited by the following examples, and that various modifications or changes may be made therein without departing from the spirit and scope of the present invention.

Hereinafter, the technical and scientific terms used herein will be understood by those skilled in the art without departing from the scope of the present invention. Descriptions of known functions and configurations that may be unnecessarily blurred are omitted.

[Example 1] Isolation of a strain having keratinolytic activity

In order to isolate keratin - based proteolytic microorganisms, compost was collected from poultry farms located in Boryeong, Chungnam using pulmonary feathers and manure. The feathers were collected in the poultry house for a period of time and the feathers were collected. The compost was collected from 10 collection points in the poultry house and 100 g was collected and transported to the laboratory in a polyethylene vinyl bag. 10 g of each sample was weighed into 90 ml of sterilized water and sufficiently dispersed at 15,000 rpm for 2 minutes in a homogenizer (Ace M-10, Nihonseiki, Japan), and then placed in a test tube containing 9 ml of sterilized water to 10 ^-7 Were sequentially diluted. From the sequentially diluted final dilution water, 100 μl of the sample was added to a nutrient agar medium (composition: 10 g of beef extract, 10 g of peptone, 5 g of sodium chloride, 5 g of agar, 15 g and distilled water (1000 ml, pH 7.5), and then plated in a flat plate. All the specimens were cultured at 37 ℃ for 3 days. The colonies formed were counted and 125 colonies isolated from the colonies using a dissecting microscope (Zoom 2000, Leica) were separated into nutrient broth.

[ Example  2] Protease of the strain having keratinolytic activity Generation  Research

In order to investigate the ability of 125 strains isolated in Example 1 to isolate proteolytic enzymes, SMA (skim milk agar, Difco) medium, which is a selective medium for proteolytic strains, was used. (1% of skim milk, 0.03% of K ₂ HPO _4, 0.03% of KH ₂ PO ₄ , 0.01% of Na ₂ CO ₃ and 1.5% of agar) supplemented with 1% of skim milk and inorganic salt was prepared as a substrate, Were inoculated with 125 isolates from feather and compost samples and cultured at 37 ℃ for 48 hours. As a result, 48 strains showing proteolytic activity were isolated. As shown in FIG. 1, 48 isolates were isolated from 8 strains which showed a large transparent ring of 1 cm or more per 48 hours of culture and had excellent keratin protease production ability.

[ Example  3] of a strain having keratinolytic activity Lung feather  Resolution Measurement

In order to evaluate the lung feather resolution of 8 pure strains isolated in Example 2, a feather medium was prepared by adding 0.05% NH ₄ Cl, 0.05% NaCl, 0.03% K ₂ HPO _4, 0.03% KH ₂ PO ₄ , The microorganism was inoculated with each of the above protein degradation microorganisms in an amount of 0.01% MgCl ₂ 6H ₂ O, 0.01% yeast extract, 1% whole chicken feather powder, pH 7.5, The fledgling culture in which the strain was inoculated while shaking was cultured for 24 hours, 48 hours, and 72 hours.

The culture broth was centrifuged at 8,000 rpm for 20 minutes, and the remaining feathers were filtered with a filter paper (whattman filter paper No. 2) and washed four times with 10 ml of sterilized distilled water. The feathers filtered on the filter paper were dried in a dry oven at 60 ° C for 24 hours and the dry weight of the feathers was measured to calculate the feather degradation rate.

As a result, the FBF-7 strain having the highest feather decomposition rate was finally selected. As shown in Fig. 4, the strain FBF-7, which is a new strain, exhibited 35% of feathers added at 24 hours, 65% at 48 hours, and 67% at 72 hours.

[Example 4] Sequence analysis of 16S rRNA of FBF-7 strain

In order to confirm the phylogenetic position of the FBF-7 strain finally selected in Example 3, 16S rRNA was amplified by PCR to determine the nucleotide sequence as follows. The primers used to amplify 16S rRNA were 27F (SEQ ID NO: 2: 5'-AGAGTTTGATCCTGGCTCAG-3 ') and 1492R (SEQ ID NO: 3: 5'-AAGGAGGTGATCCAGCCGC-3').

PCR was performed according to the following conditions. After the reaction at 94 ° C for 5 minutes, the reaction was repeated 35 times at 94 ° C denaturation for 30 seconds, annealing at 55 ° C for 30 seconds and extension at 72 ° C for 35 minutes, final extension at 72 ° C for 7 minutes, Respectively. Perkin Elmer (GeneAmp PCR system 9700, Applied Biosystems) was used as a thermal cycler for DNA amplification. The PCR amplification products were electrophoresed in a 1% agarose gel, 0.5 × TAE buffer (0.045 M Tris-borate, 0.001 M EDTA) at 100 V and 25 mA for 30 min in an electrophoresis (Mupid-21, Gel documentation system, (Bio-Rad), stained with EtBr (ethidium bromide) for 15 minutes, and then amplified under UV, and purified with Qiagen PCR Purification Kit (Qiagen Inc).

16S rRNA was sequenced using ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (Applied Biosystems), and sequenced by ABI PRISM 310 Genetic Analyzer (Applied Biosystems). The homology of the 16S rRNA sequence obtained through the above experiment was investigated using the BLAST program of DDBJ / NCBI / RDP / GeneBank database. The alignment of each nucleotide sequence was aligned using the Clustal X algorithm and the phylogenetic location was determined based on the neuronal binding method.

As a result, the nucleotide sequence (1305 bp) of the 16S rRNA PCR amplification product was determined to confirm the phylogenetic position of the keratin-derived protein degrading microorganism FBF-7. As shown in FIG. 3, the strain FBF-7 was found to be a heterologous strain of Chrysobacterium sp. , Which is a flexible strain group of Chrysobacterium, confirmed Seo tumefaciens sense beurini (Chryseobacterium gambrini) homology, Cri Seo tumefaciens star Signature (Chryseobacterium stagni) and represented a 99.08% the FBF-7 strain of 99.15% is Cri Seo tumefaciens (Chryseobacterium) in the novel strain Respectively.

Then, the isolated strain was treated with Chryseobacterium sp.) FBF-7, deposited with the Korean Agricultural Microbiology Resource Center on April 1, 2009, and granted KACC 91463P.

[ Example  5] Cryptobacterium  genus FBF Production of keratin protease in strain -7

(1) causes of organic nitrogen Feather Badge  of mine Viability  Measure

Keratin family protein decomposition microorganism Chryseobacterium sp.) strain FBF-7 was inoculated on a feather medium prepared at a concentration of 1%, and then incubated at 37 ° C for 180 minutes with shaking for 3 days. Were added to a test tube and sequentially diluted to 10 ^-6 to 10 ^-8 . Then, 100 μl of the final dilution was dispensed into a nutrient agar, uniformly plated by a flat plate method, and cultured at 37 ° C. for 3 days And the colonies formed on the nutrient broth agar plate were counted to obtain Chryseobacterium sp.) strain FBF-7.

As a result, as shown in Figure 5, Chryseobacterium sp.) FBF-7 strain showed the maximum proliferative capacity to rapidly multiply to 1.7 × 10 ⁸ CFU / ㎖ within 48 hours using feathers as nutrients, and the growth rate decreased after 48 hours.

(2) From the lung feathers Cryptobacterium  genus FBF Quantitative qualitative analysis of amino acids produced by -7 strain

Keratin family protein decomposition microorganism Chryseobacterium The production of amino acids from the lung feathers by the strain FBF-7 belonging to the genus Chryseobacterium sp.) strain FBF-7 was inoculated and cultured at 37 ° C and 180 rpm for 24 hours, 48 hours, and 72 hours.

The cultured medium was centrifuged at 8,000 rpm for 20 minutes, and the supernatant, which was decomposed from the feathers and was thus made into an amino acid, was extracted. Quantitative analysis of the extracted amino acids was performed using a Waters UPLC ^TM amino acid analyzer. The results are shown in Table 1 below.

As shown in Table 1, the total content of keratin-based amino acids produced from the lung feathers by the strain of the keratin-family protein degrading microorganism Chryseobacterium sp., FBF-7 was measured. As a result, 7.03 nmol / g at 48 hours, 13.93 nmol / g at 48 hours and 48.25 nmol / g at 72 hours, respectively.

The major amino acids produced by the FBF-7 strain from lung feathers after 72 hours of incubation were the highest with 8.17 nmol / g (17%) of glutamic acid, 5.73 nmol / g (12%) with arginine, 5.28 (11%), valine 4.16 nmol / g (9%), proline 4.02 nmol / g (8%), leucine 3.84 nmol / g (8%) and other 11 amino acids , And 17 kinds of amino acids were produced from the lung feathers by the FBF-7 strain of Chrysobacterium sp.

[Example 6] Investigation of optimal culture condition for mass production of enzyme of strain Chrysobacterium FBF-7

Based on the results of Example 5 above, optimal culture conditions for the mass production of the enzyme of Chryseobacterium sp. FBF-7 were investigated.

(1) Investigation of optimum culture composition

(0.05% NH ₄ Cl, 0.05% NaCl, 0.03% K ₂ HPO ₄ , 0.03% KH ₂ PO ₄ , 0.01% MgCl ₂ 6H ₂ O, 0.01%, pH 7.5 ± 0.2 ) Showed a high growth ability of 2.3 x 10 ⁸ cells / ml within 48 hours of culturing at 20 ° C under the condition of growing at 15 to 50 ° C.

In order to find the optimal carbon source and nitrogen source for mass production of keratin protease produced by Chryseobacterium sp. Strain FBF-7, various carbon sources and nitrogen sources were used to evaluate the effect thereof.

First, in order to investigate the optimal carbon source necessary for the production of enzymes, 12 kinds of carbon sources such as glucose, fructose, rhamnose, saccharose, 0.05% each of maltose, lactose, glycerol, galactose, sorbitol, mannitol, starch and mannose was added to the medium .

After inoculation of the bacterium under the above conditions, the cells were incubated at 20 ° C and 150 rpm for 3 days, and the enzyme activity was examined by extracting the coenzyme.

As a result, as shown in FIG. 5, it was confirmed that the medium supplemented with galactose, lactose and sorbitol as the optimum carbon source for producing keratin protease from the strain increased the yield of the enzyme more than the control without the carbon source And the optimum carbon source was galactose.

In order to investigate the optimum nitrogen source, 14 kinds of nitrogen sources such as beef extract, casein, gelatin, peptone, skim (skim) were added to an inorganic salt medium supplemented with 2% milk), tryptone (tryptone), yeast extract (yeast extract), urea _{(urea), (NH 4)} HPO 4, NH 4 Cl, NH 4 NO 3, KNO 3, NaNO 3, and 0.1% of KNO _2, respectively . The medium was inoculated on the culture medium and cultured at 20 DEG C and 150 rpm for 3 days. Then, coenzyme was extracted to investigate the enzyme activity.

As a result, as shown in FIG. 6, in the case of the nitrogen source, the enzyme production was higher in the organic nitrogen source including the small-scale extract, casein, gelatin, peptone, skim milk, tryptone, yeast extract and urea than the control without the nitrogen source Especially, the enzyme production was increased about twice as much as that of the control in medium containing peptone and small - scale extract of organic nitrogen source.

From the above results, it was confirmed that the enzyme production was greatly increased by the addition of the nitrogen source to the keratin proteolytic enzyme production of the strain Chryseobacterium sp. FBF-7, which is essential for the production of the enzyme of the strain This is also the result of confirming that the composition is a medium.

(2) Investigation of optimum culture temperature

In order to investigate the optimum culture temperature for the mass production of the enzyme of Chryseobacterium sp. FBF-7, the strain was inoculated with an inorganic salt medium supplemented with 2% , 15 ° C, 20 ° C, 25 ° C, 30 ° C, 35 ° C, 40 ° C and 50 ° C for three days at 150 rpm and then the activity of the coenzyme extracted from the culture was measured.

As a result, the enzyme activity was highest at 16.8 unit / ml / min at 20 ° C, and 15.9 unit / ml / min at 25 ° C and 14.9 unit / ml / min at 15 ° C. This is also the result of confirming that the strain Chryseobacterium sp. FBF-7 produces a psychrotrophic keratinolytic protease.

(3) Optimal culture pH survey

In order to investigate the effect of pH on the production of keratin protease of Chryseobacterium sp. FBF-7 strain, the pH of the culture medium was adjusted to a range of 3.0 to 10.0, and then cultured. The activity of the extracted coenzyme was measured.

As a result, it was confirmed that the activity was the highest at 17.2 to 17.4 unit / ml / min at pH 7.0 to 8.0. This is the result of confirming that the neutral and weak alkaline culture conditions of Chrysobacterium sp. FBF-7 in the pH range of 7.0 to 8.0 are the optimum conditions.

(4) Chrysosobacterium ( Chryseobacterium  sp.) Fecal degradation rate of FBF-7 strain

As a result of measuring the degradation rate of the wheat after culturing the strain of Chrysobacterium sp. FBF-7 using the optimal culture conditions, it was confirmed that the degradation rate was about 75% within 72 hours of incubation.

[ Example  7] Investigation of amino acid composition decomposed from female moth

Amino acid extraction by biological degradation using Chrysobacterium sp. FBF-7 was carried out by inoculating the strain with an inorganic salt medium supplemented with 2.0% , 3 days), and the culture broth was centrifuged at 8,000 rpm for 20 minutes at 4 ° C to recover the offspring degradation product.

The quantitative analysis of the recovered degradation product was performed using an L-8900 amino acid analyzer (Hitachi, JAPAN), and the results are shown in Table 2 below.

As shown in Table 2, the total amino acid content was extracted at 1661.6 umol / ml, and among the 17 amino acids, valine (11%), glutamic acid (11%), aspartic acid (11%), glycine (10%), prorine (10%), serine (8%) and leucine ) (7%) were extracted as major amino acids.

It is also known that the eight essential amino acids of the extracted amino acids, namely histidine, threonine, lysine, methionine, valine, isoleucine, leucine, And phenylalanine contained 619.3 umol / ml, which contained about 37% of the total amino acid content.

[ Example  8] Sweet potato root-knot nematode ( Meloidogyne incognita ) Control effect

(1) Larval mortality of amino acid composition on sweet potato root nematode

(J2) of Sweet Potato Root-knot nematode was investigated using an amino acid composition degraded by Chrysobacterium sp. FBF-7 from 2.0% flax seed powder. The mortality was determined by 24-well plate indoor test method Respectively.

As a test group, the concentration of amino acid composition (KAA) obtained by decomposition from chicken feces was treated to 0.5%, 1% and 2%, and as a control group, a phos- phate-saponified emulsion Fosthiazate EC 30%; 0.025%) was used as a control, and distilled water sterilized as a control was treated.

As a result, as shown in FIG. 7, the test for the effect of the amino acid composition on the larval mortality of the sweet potato root nematode showed that the test group treated with the amino acid composition had a mortality rate of 41% at the concentration of 0.5% % Mortality and 67% mortality rate at 2% concentration and 81% mortality rate at 2% concentration.

In addition, the mortality rate of the phos- phate-azide emulsion of the control group was compared with that of the amino acid. As a result, the amino acid composition obtained by decomposing from the hair follicle was more than 50% And it was confirmed that there is an excellent effect in controlling.

(2) Inhibitory effect of amino acid composition on egg hatching on sweet potato root nematode

The inhibitory effect on the egg hatching of sweet potato root-knot nematodes was investigated by using the amino acid composition decomposed by 2.0% Chrysobacterium sp. FBF-7 from wheat flour, and the effect was investigated by measuring egg hatching rate . The egg hatching rate was determined by a 24-well plate assay.

As a test group, the concentration of the amino acid composition (KAA) obtained by decomposition from chicken feces was treated at 1% and 2%, and as a control group, Fosthiazate EC 30, which is used as a pesticide for controlling root nematode, %; 0.025%). And treated with distilled water sterilized as a comparative control.

As a result, it was confirmed that the egg hatching rate in sterilized distilled water as the control group was 8% at 3 days, 37% at 6 days, 59% at 18 days, and 90% at 24 days. When the concentration of the amino acid composition obtained from the decomposition of the chicken egg meal was 1%, it was 24% at 6 days, 55% at 12 days, 57% at 18 days, and 24% Day, the inhibition rate was 61%. In addition, when the amino acid composition was treated at a concentration of 2%, the inhibition of egg hatching was confirmed to be 34% at 6 days, 64% at 12 days, 68% at 18 days, and 68% at 24 days. On the contrary, when the control group, Phoschiaiet tanning was administered, the inhibition rate of egg hatching was 80% at day 6, 88% at day 12, 89% at day 18 and 92% at day 24.

The above results show that the amino acid composition (KAA) of the present invention is a superior biologic agent that can replace the existing chemical pesticide preparation, thereby reducing the damage of the pesticide residues in the soil and the undesirable bactericidal action of the soil- This is also the result of confirming the effect.

Agricultural Biotechnology Research Institute KACC91463 20090401

Attach an electronic file to a sequence list

Claims (7)

Chryseobacterium having a keratin-type protease-producing ability sp.) FBF-7 (KACC 91463P) culture. The method according to claim 1,
Wherein the culture is cultivated from a culture medium containing an organic nitrogen source and a carbon source selected from the group consisting of poultry feathers consisting of hair follicles, duck hair, and chicken hair. 3. The method of claim 2,
Wherein the carbon source is at least one selected from the group consisting of galactose, lactose and sobitol. 3. The method of claim 2,
Wherein the culture is cultivated at a culture temperature of 13 to 28 캜, a culture time of 65 to 80 hours, and a pH of 6.5 to pH 8.5. The method according to claim 1,
Wherein the culture is an amino acid composition obtained by decomposition from feathers. 6. The method of claim 5,
Wherein the amino acid is at least one selected from the group consisting of valine, glutamic acid, aspartic acid, glycine, prorine, serine, leucine and essential amino acids. Wherein the root-knot nematode inhibitor is a compound of formula (I). 7. The compound according to any one of claims 1 to 6,
The nematode was identified as Meloidogyne incognita . < / RTI >

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