Streptomyces omiyaensis gc01 having the alleviation of salt damage in soybean plants, and uses thereof

Abstract

The present invention relates to a strain of Streptomyces omiyaensis GC01 (accession number: KACC 92474P) having a salt damage reduction effect on soybeans and its use, and provides a strain of Streptomyces omiyaensis GC01 (accession number: KACC 92474P) having a salt damage reduction effect on soybeans.

Description

Technology Field

[0001] The present invention relates to Streptomyces omiansis ( Streptomyces omiyaensis It is about ) strains. Background Technology

[0002] Since the 1960s, the area of ​​reclaimed farmland has reached 135,100 ha, but the reclaimed land has poor soil quality and high salinity, making it very unsuitable for crop growth. Therefore, in order to secure stable crop yields and cultivate a variety of crops, it is urgent to improve the soil and develop cultivation techniques suitable for it, as well as salt-tolerant varieties.

[0003] Since reclaimed land has a very high salt content, the utilization of reclaimed land can be enhanced by cultivating rice, which is more salt-tolerant than other crops. However, due to the rapid increase in rice stockpiles caused by decreased consumption, a system is needed to secure stable farm income by cultivating field crops or fodder crops instead of rice.

[0004] Eco-friendly agriculture refers to a farming method that minimizes pollution of the biological environment, including water, air, and soil, by using minimal amounts of chemical pesticides, fertilizers, and antibiotics during the production of agricultural, forestry, and livestock products. It is garnering attention as a method that produces agricultural products safe for human health while maintaining agricultural productivity and ecosystems.

[0005] For a long time, agriculture has used chemicals such as pesticides and fertilizers to increase crop cultivation efficiency; however, this has caused toxicity problems due to pesticide residues, and the emergence of pests and diseases resistant to chemical components has led to the persistent issue of having to periodically change the compounds used.

[0006] Accordingly, eco-friendly agriculture utilizing beneficial microorganisms is emerging as a means to solve these problems. Beneficial microorganisms used in agriculture include photosynthetic bacteria, Bacillus subtilis, lactic acid bacteria, yeast, and Chlorella, which are utilized as 'biological promoters' and 'biopesticides,' respectively.

[0007] The demand for agricultural microorganisms is gradually increasing as a means to enable stable agriculture while being free from the harmful effects of chemical pesticides. Agricultural microorganisms are produced in various forms, such as beneficial microbial activated solutions, organic fermented compost, and organic acid fermented solutions, and are being utilized in agricultural fields. Prior art literature

[65535] Korean Published Patent Application No. 10-2023-0085601 Korean Published Patent Application No. 10-2023-0091483 The problem to be solved

[0008] The objective of the present invention is to provide Streptomyces omiansis having a salt damage reduction effect ( Streptomyces omiyaensis ) It provides the GC01 strain (accession number: KACC 92474P). means of solving the problem

[0009] The present invention relates to a Streptomyces omiyansis GC01 strain, wherein Streptomyces omiyansis has a salt damage reduction effect on soybeans ( Streptomyces omiyaensis ) Provides the GC01 (accession number: KACC 92474P) strain.

[0010] In one embodiment of the present invention, the strain may be characterized by having 16S rRNA of the nucleotide sequence represented by SEQ ID NO. 1.

[0011] The present invention relates to a composition for reducing salt damage to soybeans, comprising a strain according to one embodiment of the present invention, a culture solution of said strain, or all of these as an active ingredient.

[0012] In one embodiment of the present invention, the composition can be extracted with butanol.

[0013] In one embodiment of the present invention, the composition for reducing salt damage to soybeans may be characterized by reducing at least one of the lipid peroxide content, hydrogen peroxide content, initial fluorescence value, reduction amount of normalized vegetation index, and reduction amount of photochemical reflectance index of soybeans.

[0014] In one embodiment of the present invention, the composition for reducing salt damage to soybeans may be characterized by enhancing at least one of the maximum fluorescence value and the maximum fluorescence / initial fluorescence ratio of the soybeans.

[0015] The present invention relates to a method for reducing salt damage in soybeans, wherein Streptomyces omiansis ( Streptomyces omiyaensis The method comprises the step of treating a soybean, a soybean surrounding area, or both of the group consisting of the GC01 (accession number: KACC 92474P) strain, a culture medium of said strain, a composition containing said strain, and a composition containing a culture medium of said strain.

[0016] In one embodiment of the present invention, the method for reducing salt damage to soybeans may be characterized by reducing at least one of the lipid peroxide content, hydrogen peroxide content, initial fluorescence value, reduction amount of normalized vegetation index, and reduction amount of photochemical reflectance index of the soybeans.

[0017] In one embodiment of the present invention, the method for reducing salt damage to soybeans may be characterized by enhancing at least one of the maximum fluorescence value and the maximum fluorescence / initial fluorescence ratio of the soybeans.

[0018] In one embodiment of the present invention, the processing step may treat any one of the strain, the culture solution, and the composition with the soybean by a seed immersion method.

[0019] The present invention relates to a Streptomyces omiyansis GC01 strain, wherein Streptomyces omiyansis reduces at least one of lipid peroxidation content, hydrogen peroxide content, initial fluorescence value, reduction in normalized vegetation indicators, and reduction in photochemical reflectance index of soybeans grown under salt conditions ( Streptomyces omiyaensis ) Provides the GC01 (accession number: KACC 92474P) strain.

[0020] The present invention relates to a Streptomyces omiyansis GC01 strain, wherein Streptomyces omiyansis enhances at least one of the maximum fluorescence value and the maximum fluorescence / initial fluorescence ratio of soybeans grown under salt conditions ( Streptomyces omiyaensis ) Provides the GC01 (accession number: KACC 92474P) strain. Effects of the invention

[0021] Streptomyces omiansis according to one embodiment of the present invention ( Streptomyces omiyaensis The GC01 strain can reduce salt damage to soybeans. Brief explanation of the drawing

[0022] Figure 1 is a graph showing the plant lipid peroxidation content with and without the presence of the GC01 strain according to one embodiment of the present invention. Figure 2 is a graph showing the plant hydrogen peroxide content with and without the GC01 strain according to one embodiment of the present invention. Figure 3 is a graph showing the normal vegetation index of plants based on the presence or absence of the GC01 strain according to one embodiment of the present invention. Figure 4 is a graph showing the photochemical reflectance index based on the presence or absence of the GC01 strain according to one embodiment of the present invention. Figure 5 is a graph showing the initial fluorescence value based on the presence or absence of the GC01 strain according to one embodiment of the present invention. Figure 6 is a graph showing the maximum fluorescence / initial fluorescence ratio according to the presence or absence of the GC01 strain according to one embodiment of the present invention. FIG. 7 is a phylogenetic tree of the GC-1 strain prepared according to one embodiment of the present invention. Specific details for implementing the invention

[0023] Hereinafter, with reference to the attached drawings, the Streptomyces omiyansis strain GC01, which has a salt damage reduction effect on soybean plants according to the present invention, and its uses will be described in detail.

[0024] Prior to this, the terms used in this specification and claims shall not be interpreted as being limited to their dictionary meanings; rather, based on the principle that the inventor may appropriately define the concepts of the terms to best describe their invention, they shall be interpreted in a meaning and concept consistent with the technical spirit of the invention.

[0025] The exemplary embodiments described in the detailed description, drawings, and claims are not intended to be limiting, and other embodiments may be used, and other modifications are possible without departing from the spirit or scope of the technology disclosed herein.

[0026] Therefore, it should be understood that the embodiments described in this specification and the configurations illustrated in the drawings are merely preferred embodiments of the present invention and do not represent all of the technical ideas of the present invention, and that various equivalents and modifications that can replace them may exist at the time of filing this application.

[0027] Those skilled in the art will readily understand that the components of this disclosure, namely those generally described herein and those depicted in the drawings, can be arranged, configured, combined, and designed in various different configurations, all of which are obviously devised and form part of this disclosure.

[0028] The present invention relates to Streptomyces omiansis ( Streptomyces omiyaensis The present invention is characterized by providing a composition for reducing salt damage to soybeans comprising the strain GC01 (accession number: KACC 92474P), the strain, or a culture solution thereof, and a method for reducing salt damage to soybeans using the strain or the composition thereof.

[0029] The present invention relates to Streptomyces omiansis having a salt damage reduction effect on soybeans ( Streptomyces omiyaensis ) Provides the GC01 (accession number: KACC 92474P) strain.

[0030] The term “salt damage reduction” is used herein in its ordinary sense to refer to the reduction of the phenomenon in which the germination rate or growth of plants is lowered due to salts. Since soybeans are used as the plant in this invention, soybeans are disclosed as an example of the plant below.

[0031] Therefore, 'reduction of salt damage to soybeans' generally refers to a decrease in phenomena such as reduced germination rate, reduced yield, reduced fresh weight, reduced weight of specific parts, reduced number of specific parts, or growth inhibition caused by salt concentration when soybeans are exposed to salt concentrations higher than the physiologically permissible concentration for soybeans (in their environment, such as soil or water); this implies an improvement in the soybeans' ability to withstand or tolerate salt concentrations.

[0032] In one embodiment of the present invention, the degree of such salt damage reduction can be measured using various salt stress indices including yield, biomass, plant height, number of leaves, number of roots, weight of roots, weight of root nodules, number of root nodules, lipid peroxidation content, hydrogen peroxide content, initial fluorescence value, reduction in normalized vegetation indicators, reduction in photochemical reflectance index, maximum fluorescence value, and maximum fluorescence / initial fluorescence ratio, and the extent to which plant individuals suffer damage from salt can be objectively evaluated according to the increase or decrease of each indicator.

[0033] For example, the salt stress index may apply at least one of the following: yield, weight of root nodules, number of root nodules, lipid peroxidation content, hydrogen peroxide content, initial fluorescence value, reduction in normalized vegetation index, reduction in photochemical reflectance index, maximum fluorescence value, and maximum fluorescence / initial fluorescence ratio. In one embodiment of the present invention, the salt stress index may apply at least two of the following: yield, weight of root nodules, number of root nodules, lipid peroxidation content, hydrogen peroxide content, initial fluorescence value, reduction in normalized vegetation index, reduction in photochemical reflectance index, maximum fluorescence value, and maximum fluorescence / initial fluorescence ratio.

[0034] Accordingly, Streptomyces omiansis having a salt damage reduction effect on soybeans provided in the present invention ( Streptomyces omiyaensis The GC01 strain (accession number: KACC 92474P) can reduce at least one of the lipid peroxidation content, hydrogen peroxide content, initial fluorescence value, reduction in normalized vegetation index, and reduction in photochemical reflectance index of soybeans grown under salt conditions, and can increase at least one of the maximum fluorescence value and the maximum fluorescence / initial fluorescence ratio of soybeans grown under salt conditions. The extent to which the GC01 strain of the present invention has an effect on each salt stress index will be specified through examples described later, and through the disclosed examples, it will become more evident through the changes in each salt stress index of soybeans disclosed later that the GC01 strain of the present invention has a significant effect in reducing salt damage to soybeans.

[0035] In addition, in the following examples, Streptomyces omiansis of the present invention ( Streptomyces omiyaensis As a result of 16S rRNA sequencing analysis for the microbial identification and classification of strain GC01 (Accession No.: KACC 92474P), it was found to have the sequence of SEQ ID NO. 1, and through the results of phylogenetic analysis of the sequence of the said strain, Streptomyces omyansis ( Streptomyces omiyaensis By confirming that it is most closely related to the ) strain, the GC01 strain of the present invention is Streptomyces omiyansis ( Streptomyces omiyaensis It was identified as the GC01 strain.

[0036] In addition, the identified Streptomyces omyansis ( Streptomyces omiyaensis In order to identify the GC01 strain, the strain was deposited with the National Institute of Agricultural Sciences (Agricultural Genetic Resources Center, KACC) on November 17, 2022, and was assigned accession number KACC 92474P.

[0037] The present invention relates to Streptomyces omiansis ( Streptomyces omiyaensisA composition for reducing salt damage in soybeans is provided, comprising the GC01 strain (accession number: KACC 92474P), a culture solution of the strain, or both of these as active ingredients.

[0038] The term “culture solution” refers to a product obtained by inoculating a strain into a culture medium and culturing it for a certain period of time. It includes, but is not limited to, the culture solution itself obtained by culturing the strain according to the present invention in a suitable liquid medium; a filtrate (filtrate or supernatant after centrifugation) obtained by filtering or centrifuging the culture solution to remove the strain; a cell lysate obtained by ultrasonically treating the culture solution or treating the culture solution with a lysozyme; and a concentrate obtained by concentrating the culture solution, filtrate, lysate, etc.

[0039] In addition, the above-mentioned probiotic or salt damage reduction composition may contain a solid culture medium, and a culture such as a dried product or extract of the culture medium, but is not limited thereto, and may further include a suitable excipient or carrier.

[0040] The composition of the present invention may be prepared in the form of a liquid fertilizer, or may be used in the form of a powder by adding a filler, or may be formulated into granules. However, the formulation is not particularly limited.

[0041] In addition, the composition of the present invention is Streptomyces omiansis ( Streptomyces omiyaensis It can be prepared by extracting the GC01 strain (accession number: KACC 92474P), the culture medium of the strain, or both of these with an organic solvent.

[0042] In one embodiment of the present invention, the composition of the present invention is Streptomyces omiansis ( Streptomyces omiyaensis It can be prepared by extracting the GC01 strain (accession number: KACC 92474P), the culture medium of the strain, or both of these with butanol.

[0043] A composition for reducing salt damage to soybeans according to one embodiment of the present invention can reduce salt damage to soybeans. For example, the composition for reducing salt damage to soybeans according to the present invention can reduce salt damage occurring in at least one of soybean seeds and soybean seedlings.

[0044] As previously disclosed, the composition for reducing salt damage to soybeans according to one embodiment of the present invention can be confirmed to have a salt damage reduction effect on soybeans through at least one of the lipid peroxide content, hydrogen peroxide content, initial fluorescence value, reduction amount of normalized vegetation index, reduction amount of photochemical reflectance index, maximum fluorescence value, and maximum fluorescence / initial fluorescence ratio of soybeans, and can be confirmed to have a salt damage reduction effect on soybeans through the increase or decrease of each disclosed salt stress index.

[0045] More specifically, a composition for reducing salt damage in soybeans according to one embodiment of the present invention can reduce salt damage by reducing at least one of the lipid peroxide content, hydrogen peroxide content, initial fluorescence value, reduction amount of normalized vegetation index, and reduction amount of photochemical reflectance index of soybeans.

[0046] In addition, a composition for reducing salt damage to soybeans according to one embodiment of the present invention can reduce salt damage by enhancing at least one of the maximum fluorescence value and the maximum fluorescence / initial fluorescence ratio of soybeans.

[0047] A composition for reducing salt damage to soybeans according to one embodiment of the present invention may additionally include any other component used in the field to reduce salt damage to soybeans or increase or decrease the salt stress index, in addition to the strain of the present invention described above, the culture medium of said strain, or a mixture thereof. The other optional component may be a chemical substance or another strain of bacteria.

[0048] In addition to the previously disclosed composition for reducing salt damage to soybeans, if at least one of the following effects can be utilized: a salt damage reduction effect for soybeans, a lipid peroxidation content reduction effect, a hydrogen peroxide content reduction effect, an initial fluorescence value reduction effect, a normalized vegetation index reduction effect, a photochemical reflectance index reduction effect, a maximum fluorescence enhancement effect, and a maximum fluorescence / initial fluorescence ratio enhancement effect, Streptomyces omyansis ( Streptomyces omiyaensis The use of a composition comprising the GC01 strain (accession number: KACC 92474P), a culture medium of the said strain, or both as an active ingredient is not limited.

[0049] For example, Streptomyces omiansis ( Streptomyces omiyaensis A composition comprising the GC01 strain (accession number: KACC 92474P), a culture medium of the strain, or all of these as active ingredients may be named as a composition for alleviating salt damage (of soybeans), a composition for removing salt damage, a salt damage alleviator, a salt damage reduction agent, a salt damage removal agent, a composition for reducing lipid peroxidation, a lipid peroxidation reduction agent, a composition for reducing hydrogen peroxide, a hydrogen peroxide reduction agent, a composition for reducing salt stress, a composition for reducing salt stress, a composition for alleviating salt stress, a salt stress reduction agent, a salt stress alleviator, a composition for enhancing the salt stress index, a salt stress index enhancer, etc.

[0050] The present invention relates to Streptomyces omiansis ( Streptomyces omiyaensis A method for reducing salt damage to soybeans is provided, comprising the step of treating one or more selected from the group consisting of the GC01 strain (accession number: KACC 92474P), a culture medium of said strain, a composition containing said strain, and a composition containing a culture medium of said strain on soybeans, a soybean surrounding area, or both.

[0051] The parts of the soybean to which the salt damage reduction method of the present invention is applied are not limited. Accordingly, it includes not only the whole soybean but also parts of the soybean, such as seeds, sprouts, stems, roots, leaves, fruits, and all of these, as well as parts thereof.

[0052] The present invention relates to Streptomyces omiansis ( Streptomyces omiyaensis The GC01 strain (accession number: KACC 92474P), a culture medium of the strain, and a composition containing the strain can be provided to soybean seeds, soybean seedlings, and soybeans by immersion, drenching, spraying, etc.

[0053] For example, soybean seeds may be immersed in the culture solution or the composition. Additionally, for example, the culture solution of the strain or the composition of the strain may be applied as a drench treatment to soybean seedlings.

[0054] The surrounding area disclosed in the present invention refers to a range in which any one of the strain of the present invention, the culture medium of said strain, and a composition comprising said strain and / or said culture medium can exert an effect, and includes both above ground and underground.

[0055] For example, it may be an area within about 2m, about 1m, about 70cm, about 50cm, about 25cm, about 10cm, or about 5cm around a bean, part of a bean, or fruit.

[0056] The method for reducing salt damage to soybeans according to the present invention is Streptomyces omiansis of the present invention ( Streptomyces omiyaensis ) GC01 (accession number: KACC 92474P) strain, a culture medium of said strain, and any one of said strain and / or said culture medium is applied continuously at any point in the life cycle of a soybean, at one or more stages in the life cycle of a soybean, at regular points in the life cycle of a soybean, and / or throughout the entire life cycle of a soybean.

[0057] Therefore, Streptomyces omiansis ( Streptomyces omiyaensis The application time of any one of the GC01 (accession number: KACC 92474P) strain, the culture medium, and the composition may be changed as needed.

[0058] For example, Streptomyces omiansis ( Streptomyces omiyaensis Any one of the GC01 (accession number: KACC 92474P) strain, a culture medium of said strain, and a composition comprising said strain and / or said culture medium may be applied during the growth of soybeans, before and / or during flowering, before and / or during seed development.

[0059] Also, for example, Streptomyces omiansis ( Streptomyces omiyaensis Any one of the GC01 strain (accession number: KACC 92474P), the culture medium of the strain, and the composition comprising the strain and / or the culture medium may be applied multiple times according to a set interval.

[0060] Also, for example, Streptomyces omiansis ( Streptomyces omiyaensis Any one of the GC01 (accession number: KACC 92474P) strain, a culture medium of said strain, and a composition comprising said strain and / or said culture medium may be applied before, during, and / or immediately after soybeans are transplanted from one location to another, for example, from a greenhouse or hotbed to a field.

[0061] In one embodiment of the present invention, the culture solution or the composition may be applied before sowing soybean seeds.

[0062] A method for reducing salt damage to soybeans according to one embodiment of the present invention may be characterized by reducing at least one of the lipid peroxide content, hydrogen peroxide content, initial fluorescence value, reduction amount of normalized vegetation index, and reduction amount of photochemical reflectance index of soybeans, or increasing at least one of the maximum fluorescence value and the maximum fluorescence / initial fluorescence ratio of soybeans.

[0063] The present invention will be explained in more detail below through examples. However, these examples are intended only to explain the invention in more detail and do not limit the scope of the invention.

[0065] Example 1: Effect of the present invention's GC01 strain on alleviating salt damage in soybean plants

[0066] In order to confirm the salt damage mitigation effect of the GC01 strain in the present invention, the degree of salt damage in soybean plants was measured according to the presence or absence of the GC01 strain.

[0067] Example 1-1: Changes in lipid peroxide content and hydrogen peroxide content according to salt conditions

[0068] First, to extract the culture filtrate of the actinomycete GC01 strain using a solvent, the culture was cultured in TSB (tryptic soy broth, Difco, USA) medium for 3 days (28 ℃, 150 rpm), centrifuged at 6,000 rpm for 20 minutes, and the supernatant was filtered using a 0.45 µm filter. The filtered culture was extracted with an organic solvent (n-butanol) and concentrated under reduced pressure, and each extract was diluted to a concentration of 10 ppm. Soybeans were immersed in the butanol extract (10 ppm) of actinomycete GC01 for 1 hour, then sown in pots (14 cm in diameter) filled with potting soil, and the soybeans were grown in a glass greenhouse for 5 to 6 leaves. After sowing, salt stress (or salt stress) was induced by drenching with 40 mL of 200 mM NaCl solution for 3 days, and 1 week after the final salt solution drenching, leaf samples were taken to evaluate lipid peroxidation and hydrogen peroxide accumulation in order to assess plant cell destruction caused by salt damage in soybean plants.

[0069] Figure 1 is a graph showing the plant lipid peroxidation content with and without the presence of the GC01 strain according to one embodiment of the present invention.

[0070] Referring to FIG. 1, the degree of reduction in lipid peroxidation due to the presence or absence of the GC01 strain according to one embodiment of the present invention can be confirmed. Considering that lipid peroxidation increases due to the death of plant leaves, it is desirable to consider that salt damage is reduced when the degree of lipid peroxidation decreases.

[0071] More specifically, Comparative Example 1-1 (Control), which did not contain the GC01 strain of the present invention, was measured to have a lipid peroxidation (MDA; Malondialdehyde) value of 4.07±0.18 due to salt damage, whereas Experimental Example 1-1, which was treated with the GC01 strain of the present invention, was measured to have a lipid peroxidation value of 2.98±0.13 due to salt damage. In other words, the lipid peroxidation value increased by salt damage differed significantly depending on the presence or absence of the GC01 strain, and it was confirmed that when the GC01 strain was included, the damage caused by salt damage (lipid peroxidation value) decreased by approximately 26.8%.

[0072] Figure 2 is a graph showing the plant hydrogen peroxide content with and without the GC01 strain according to one embodiment of the present invention.

[0073] Referring to FIG. 2, the degree of reduction in hydrogen peroxide based on the presence or absence of the GC01 strain according to one embodiment of the present invention can be confirmed. Considering that hydrogen peroxide is a cause of cell death, it is desirable to consider that a decrease in the hydrogen peroxide level reduces salt damage.

[0074] More specifically, Comparative Example 1-2 (Control), which did not contain the GC01 strain of the present invention, showed a hydrogen peroxide value of approximately 0.93±0.08 due to salt damage, whereas the experimental example treated with the GC01 strain of the present invention showed a hydrogen peroxide value of 0.59±0.07 due to salt damage. In other words, the hydrogen peroxide value increased due to salt damage varied depending on the presence or absence of the GC01 strain, and it was confirmed that when the GC01 strain was included, the salt damage (hydrogen peroxide value) decreased by approximately 36.6%.

[0075] Example 1-2: Changes in vegetation index values, photochemical reflectance index values, initial fluorescence values, and maximum fluorescence / initial fluorescence ratio according to salt conditions

[0076] Soybeans treated with the actinomycete GC01 strain were sown in 1 / 2000 (ar) Wagner pots and cultivated until the flowering stage, which is the reproductive growth stage. Afterward, 500 mL of 200 mM NaCl was applied three times at 2-day intervals. Subsequently, on the 28th day, a non-destructive optical sensor (PolyPen, Czech) and a chlorophyll fluorescence meter (fluropen, Czech) were used to evaluate the plant stress index caused by salt damage. The stress index was defined as the Normalized Difference Vegetation Index (NDVI), Photochemical Reflectance Index (PRI) value, initial fluorescence (Fo; initial fluorescence in dark adapted tissue) value, and the ratio of maximum fluorescence (Fm; maximum fluorescence in dark adapted tissue) to initial fluorescence (Fm / Fo).

[0077] Figure 3 is a graph showing the normal vegetation index of plants based on the presence or absence of the GC01 strain according to one embodiment of the present invention.

[0078] Referring to FIG. 3, the difference in plant normalized vegetation indicators (NDVI) due to the presence or absence of the GC01 strain according to one embodiment of the present invention can be observed.

[0079] More specifically, Comparative Examples 1-3 (Control), which did not contain the GC01 strain of the present invention, were found to have a normalized vegetation index value of approximately 6.8 in the absence of salt (NaCl 0 mM) and a normalized vegetation index value of approximately 6.0 in the presence of salt (NaCl 200 mM). That is, it was confirmed that the normalized vegetation index value, which is generally reduced by salt, is approximately 12%.

[0080] Experimental Examples 1-3, which included the GC01 strain of the present invention, were found to have a normalized vegetation index value of approximately 6.8 in the absence of salt (NaCl 0 mM) and a normalized vegetation index value of approximately 6.3 in the presence of salt (NaCl 200 mM). That is, it was confirmed that the normalized vegetation index value decreased by approximately 7.4% due to salt, and it was confirmed that the amount of decrease in the normalized vegetation index value was somewhat reduced due to the GC01 strain.

[0081] Figure 4 is a graph showing the photochemical reflectance index based on the presence or absence of the GC01 strain according to one embodiment of the present invention.

[0082] Referring to FIG. 4, the difference in plant photochemical reflectance index due to the presence or absence of the GC01 strain according to one embodiment of the present invention can be observed. The photochemical reflectance index is related to the xanthophyll cycle of carotenoid pigments responsible for the heat energy dissipation of plants, and the PRI value decreases when the total amount of xanthophyll decreases due to environmental stress. That is, considering that the photochemical reflectance index decreases when a crop is stressed, it is desirable to consider that salt damage is reduced when the amount of decrease in the photochemical reflectance index decreases or when the photochemical reflectance index increases.

[0083] More specifically, Comparative Examples 1-4 (Control), which did not contain the GC01 strain of the present invention, were found to have a photochemical reflectance index (PRI) value of approximately 0.019 in the absence of salt (NaCl 0 mM) and a photochemical reflectance index value of approximately 0.006 in the presence of salt (NaCl 200 mM). That is, it was confirmed that the photochemical reflectance index value, which is generally reduced by salt, is approximately 78%.

[0084] Experimental Examples 1-4, which included the GC01 strain of the present invention, were found to have a photochemical reflectance index value of approximately 0.020 in the absence of salt (NaCl 0 mM) and a photochemical reflectance index value of approximately 0.012 in the presence of salt (NaCl 200 mM). In other words, it was confirmed that the photochemical reflectance index value reduced by salt was approximately 40%, and the amount of reduction in the photochemical reflectance index value due to the GC01 strain was significantly reduced (approx. -78% -> approx. -40%). Accordingly, it is evident that when the GC01 strain is treated, the stress caused by salt can be significantly reduced compared to when the GC01 strain is not treated.

[0085] Figure 5 is a graph showing the initial fluorescence value based on the presence or absence of the GC01 strain according to one embodiment of the present invention.

[0086] Referring to FIG. 5, the difference in the initial fluorescence value of a plant due to the presence or absence of the GC01 strain according to one embodiment of the present invention can be observed. Considering that the initial fluorescence value increases when the crop is under stress, it is desirable to consider that salt damage is reduced when the initial fluorescence value decreases or the degree of increase in initial fluorescence due to salt is reduced.

[0087] More specifically, Comparative Examples 1-5 (Control), which did not contain the GC01 strain of the present invention, were found to have an initial fluorescence (F0) value of approximately 6,000 in the absence of salt (NaCl 0 mM) and an initial fluorescence (Fo) value of approximately 7,977±710 in the presence of salt (NaCl 200 mM). That is, it was confirmed that the initial fluorescence value, which generally increases due to salt, is approximately 33%.

[0088] Experimental Examples 1-5, which included the GC01 strain of the present invention, were found to have an initial fluorescence (Fo) value of approximately 6,500 in the absence of salt (NaCl 0 mM) and an initial fluorescence (Fo) value of approximately 5,937±230 in the presence of salt (NaCl 200 mM). That is, although the initial fluorescence value should have increased due to the salt, the initial fluorescence value was significantly reduced due to the GC01 strain, resulting in a decrease of approximately 8.3%. Considering that the initial fluorescence value increased by approximately 33% due to the salt previously, it is evident that the GC01 strain had a significant effect on significantly reducing salt stress, as it reduced the initial fluorescence value by approximately 41.3% compared to the condition without the GC01 strain.

[0089] Figure 6 is a graph showing the maximum fluorescence / initial fluorescence ratio according to the presence or absence of the GC01 strain according to one embodiment of the present invention.

[0090] Referring to FIG. 6, the difference in the maximum fluorescence / initial fluorescence ratio of a plant with or without the GC01 strain according to one embodiment of the present invention can be observed. Considering that, as previously disclosed, the initial fluorescence value is proportional to the stress of the crop, the maximum fluorescence value is inversely proportional to the stress of the crop, and the maximum fluorescence / initial fluorescence ratio is inversely proportional to the stress of the crop, it is desirable to consider that salt damage is reduced when the maximum fluorescence / initial fluorescence ratio increases or the degree of decrease in the maximum fluorescence / initial fluorescence ratio due to salt is reduced.

[0091] More specifically, Comparative Examples 1-6 (Control), which did not contain the GC01 strain of the present invention, were found to have a maximum fluorescence / initial fluorescence ratio of approximately 4.9 in the absence of salt (NaCl 0 mM) and a maximum fluorescence / initial fluorescence ratio of approximately 3.8 ± 0.3 in the presence of salt (NaCl 200 mM). That is, it was confirmed that the maximum fluorescence / initial fluorescence ratio, which is generally reduced by salt, is approximately 23%.

[0092] Experimental Examples 1-6, which included the GC01 strain of the present invention, were found to have a maximum fluorescence / initial fluorescence ratio of approximately 4.4 in the absence of salt (NaCl 0 mM) and a maximum fluorescence / initial fluorescence ratio of approximately 4.7 ± 0.2 in the presence of salt (NaCl 200 mM). That is, although the maximum fluorescence / initial fluorescence ratio should have decreased due to salt, the GC01 strain increased the maximum fluorescence / initial fluorescence ratio by approximately 6.8%. Considering that the maximum fluorescence / initial fluorescence ratio decreased by approximately 23% due to salt as previously mentioned, it is evident that the GC01 strain had a significant effect on significantly reducing salt stress, as it increased the maximum fluorescence / initial fluorescence ratio by approximately 29.8% compared to the condition without the GC01 strain.

[0093] Examples 1-3: Changes in soybean yield and root nodules according to salt conditions

[0094] Soybeans treated with the actinomycete GC01 strain were sown in 1 / 2000 (ar) Wagner pots and cultivated until the flowering stage, which is the reproductive growth stage. Subsequently, 500 mL of 200 mM NaCl was applied three times at 2-day intervals. Afterward, the soybean yield, root nodule weight, and number of root nodules were measured, and the results are shown in Table 1 below. The soybean yield was measured at the time of harvesting the soybean fruits from the cultivated plants.

[0095] Treatment Weight (g) / plant Node weight (g) / plant Number of nodes / plants Comparative Examples 1-7 22.5 ± 0.7 1.9 ± 0.3 153.7 ± 15.2 Experimental Example 1-7 24.2 ± 0.5 * 3.9 ± 0.6 * 223.5 ± 27.4 *

[0096] Referring to Table 1 above, differences in soybean yield, root nodule weight, and number of root nodules can be observed depending on the presence or absence of GC01 in salt-treated soybean plants.

[0097] Regarding the soybean yield, it was confirmed that Comparative Examples 1-7, in which salt-treated soybean plants were not treated with the GC01 strain, had a yield of 22.5 g ± 0.7 g per plant, while Experimental Examples 1-7, in which salt-treated soybean plants were treated with the GC01 strain, had a yield of 24.2 g ± 0.5 g per plant. In other words, under salt conditions, the soybean yield of the plants increased by approximately 7.6% as they were treated with the GC01 strain.

[0098] When examining the weight of root nodules, it was confirmed that Comparative Examples 1-7, in which salt-treated soybean plants were not treated with the GC01 strain, had a root nodule weight of 1.9 g ± 0.3 g per plant, while Experimental Examples 1-7, in which salt-treated soybean plants were treated with the GC01 strain, had a root nodule weight of 3.9 g ± 0.6 g per plant. In other words, under salt conditions, the root nodule weight of soybean plants increased significantly (approximately 105.3%) upon treatment with the GC01 strain.

[0099] When examining the number of root nodules, it was confirmed that Comparative Examples 1-7, in which salt-treated soybean plants were not treated with the GC01 strain, had 153.7 ± 15.2 root nodules per plant, and Experimental Examples 1-7, in which salt-treated soybean plants were treated with the GC01 strain, had 223.5 ± 27.4 root nodules per plant. In other words, under salt conditions, the number of root nodules in soybean plants increased significantly (approximately 45.4%) as a result of treatment with the GC01 strain.

[0101] Example 2: Identification of GC01 strain

[0102] In the present invention, genetic analysis was performed on the GC01 strain, which was previously confirmed to have a salt damage reduction effect, to identify the GC01 strain of the present invention.

[0103] First, after obtaining the 1,494 bp 16S rRNA sequence of the GC01 strain, the said 16S rRNA sequence was analyzed and searched on EZBioCloud. As a result of the search, the GC01 strain is Streptomyces zamiseticus ( Streptomyces zaomyceticus ) NBRC T Strain and 99.5%, Streptomyces exfoliatus ( Streptomyces exfoliatus ) NRRL B-2924 T Strain and 99.86%, Streptomyces narbornensis ( Streptomyces narbonensis ) NBRC 12801 T Strain and 99.86%, to Streptomyces venezuela ( Streptomyces venezuelae ) ATCC 10712 T Strain and 99.79%, Streptomyces gardeneri ( Streptomyces gardneri ) NBRC 12865 T Strain and 99.79%, Streptomyces omiansis ( Streptomyces omiyaensis ) NBRC 13449 T 99.79% similarity with the strain was observed. Therefore, to confirm the identification, phylogenetic analysis was performed using the statistical method Neighbor-joining (applying the Maximum composite likelihood model).

[0104] FIG. 7 is a phylogenetic tree of the GC-1 strain prepared according to one embodiment of the present invention.

[0105] Referring to FIG. 7, the phylogenetic classification of the GC-1 strain according to one embodiment of the present invention can be confirmed. As a result of the classification, the GC01 strain of the present invention is Streptomyces omyansis ( Streptomyces omiyaensis ) 13449 T It was confirmed that they were grouped into the group closest to the strain.

[0106] Therefore, the GC01 strain of the present invention (Streptomyces omiansis) Streptomyces omiyaensis It was identified as the GC01 strain, and the identified Streptomyces omyansis ( Streptomyces omiyaensis The GC01 strain was deposited with the National Institute of Agricultural Sciences (Agricultural Genetic Resources Center; KACC) on November 17, 2022, and was assigned accession number KACC 92474P.

[0107] As described above, Streptomyces omiansis according to one embodiment of the present invention ( Streptomyces omiyaensis The GC01 strain (Accession No.: KACC 92474P) has the effect of reducing salt damage in soybeans and alleviating salt stress, thereby mitigating the reduction in soybean yield, root nodule weight, and root nodule number caused by salts. In particular, Streptomyces omyansis ( Streptomyces omiyaensis When the GC01 strain (accession number: KACC 92474P) is cultured and the culture filtrate is extracted with butanol and then applied to crop seeds, significant relief of salt stress and reduction of salt damage in soybeans may be observed.

[0108] When the strain of the present invention, the composition for reducing salt damage to soybeans of the present invention, and the method for reducing salt damage to soybeans of the present invention are applied, soybean cultivation can proceed smoothly even in areas with high salt content, such as reclaimed land. Accordingly, this can make a significant contribution to the commercialization of environmentally stress-resistant soybeans, enable a response to the reduction of cultivated land area, and improve soybean production efficiency.

[0109] In particular, in Korea, where the land area is small and mountainous terrain is developed, reclaimed farmland must be created to stably produce food, and this has great significance in terms of food security as it enables the production of soybeans even on such reclaimed farmland.

[0110] From the foregoing, those skilled in the art will understand that various changes and modifications are possible within the scope of the technical concept of the present invention. Accordingly, the technical scope of the present invention is not limited to the contents described in the detailed description of the specification, but must be determined by the claims.

[0111] Depository Name: National Institute of Agricultural Sciences, Rural Development Administration, Microbial Bank (KACC) Accession Number: KACC92474P Date of Deposit: 2022-11-17

Claims

Claim 1 Streptomyces omyansis, which has a salt damage reduction effect on soybeans and is deposited under accession number KACC 92474P ( Streptomyces omiyaensis ) GC01 strain. Claim 2 In claim 1, the strain is characterized by having 16S rRNA of the nucleotide sequence represented by SEQ ID NO. 1, and Streptomyces omyansis (deposited under accession number KACC 92474P) Streptomyces omiyaensis ) GC01 strain. Claim 3 A composition for reducing salt damage to soybeans, comprising a strain according to claim 1, a culture solution of said strain, or all of these as an active ingredient. Claim 4 In claim 3, the composition is a composition for reducing salt damage to soybeans, extracted with butanol. Claim 5 delete Claim 6 delete Claim 7 Streptomyces omyansis, deposited under deposit number KACC 92474P ( Streptomyces omiyaensis A method for reducing salt damage to soybeans, comprising the step of treating one or more selected from the group consisting of a GC01 strain, a culture medium of said strain, a composition containing said strain, and a composition containing a culture medium of said strain on a soybean, a region surrounding the soybean, or both. Claim 8 delete Claim 9 delete Claim 10 In claim 7, the processing step is a method for reducing salt damage to soybeans, wherein any one of the strain, the culture solution, and the composition is treated to the soybeans by a seed immersion method. Claim 11 Reducing the early fluorescence value of soybeans grown under salt conditions, Streptomyces omyansis, deposited under accession number KACC 92474P ( Streptomyces omiyaensis ) GC01 strain. Claim 12 delete Claim 13 In claim 11, the strain further comprises the effect of reducing the lipid peroxidation content of soybeans, and Streptomyces omyansis, deposited under accession number KACC 92474P ( Streptomyces omiyaensis ) GC01 strain. Claim 14 In claim 11, the strain further comprises the effect of reducing the hydrogen peroxide content of soybeans, and Streptomyces omyansis, deposited under accession number KACC 92474P ( Streptomyces omiyaensis ) GC01 strain. Claim 15 In claim 11, the strain further comprises the effect of reducing the reduction in the normalized vegetation indicator of soybeans, and Streptomyces omyansis, deposited under accession number KACC 92474P ( Streptomyces omiyaensis ) GC01 strain. Claim 16 In claim 11, the strain further comprises the effect of reducing the reduction in the photochemical reflectance index of soybeans, and Streptomyces omyansis, deposited under accession number KACC 92474P ( Streptomyces omiyaensis ) GC01 strain. Claim 17 In claim 11, the strain further comprises the effect of enhancing the maximum fluorescence / early fluorescence ratio of soybeans, and Streptomyces omyansis, deposited under accession number KACC 92474P ( Streptomyces omiyaensis ) GC01 strain.

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