KR102169215B1 - Rhizobium tropici Sunchang180605 strain with nitrogen fixation and crop growth promotion effect, a method for improving soil using the same, and a method for promoting growth of leguminous crop - Google Patents

Abstract

The present invention relates to Rhizobium tropici Sunchang180605 strain with a nitrogen fixation power and an effect of promoting crop growth, a method for improving soil using the same, and a method for promoting growth of leguminous crops and, more particularly, to Rhizobium tropici Sunchang180605 strain with a nitrogen fixation power and an effect of promoting crop growth, a method for improving soil using the same, and a method for promoting growth of leguminous crops, in which leguminous crops can be cultivated in humid soils such as rice paddies and leguminous crops can be repeatedly cultivated by improving the early growth and activity of rhizome bacteria in legume crops, fixing nitrogen in the soil, and reducing salts.

Description

Rhizobium tropici Sunchang180605 strain with nitrogen fixation and crop growth promotion effect, a method for improving soil using it and promoting the growth of legume crops {Rhizobium tropici Sunchang180605 strain with nitrogen fixation and crop growth promotion effect, a method for improving soil using the same, and a method for promoting growth of leguminous crop}

The present invention relates to a Rhizobium tropici Sunchang180605 strain having a nitrogen-fixing power and a crop growth promoting effect, a soil improvement method using the same, and a method for promoting the growth of legume crops, and in particular, the early growth of rhizomes in legume crops. Rhizobium Trophy Seed, which improves activity, fixes nitrogen in the soil and reduces salts, enables cultivation of legume crops in humid soils such as paddy fields, and has nitrogen-fixing power and crop growth promotion effects that enable continuous cropping of legume crops. ( Rhizobium tropici ) Sunchang180605 strain, a soil improvement method using the same, and a method for promoting the growth of legume crops.

Existing conventional agriculture exacerbates rheumatoid activity and soil conditions for soybeans. The reason for this is that first, the use of soil fungicides can kill and reduce the existing rhizomes. Second, as the cultivation of rice paddies for soybeans has increased recently, humid soil conditions worsen the activity of rhizomes. Third, the simplification of the rhizome ecosystem is triggered by the soybean series.

It is known that several species of rhizomes coexist in various legume crops. Soybeans are Bradyrhizobium japonicum , Bradyrhizobium japonicum , B. elkanii , peanuts and eastern Rhizobium hainanese , kidney beans are Rhizobium leguminosarum ( R. leguminosarum ), R. elti , R. tropici , etc. Others include Brady Rhizobium , R. yuanmingense , and Azorhizobium caulinodans , which coexist with alfalfa, clover, and Sesbania .

In nature, these rhizomes do not coexist with one or two species rather than a single species.

On the other hand, for legumes, practical microbial preparations having excellent growth promoting effect and yield increasing effect have been proposed in Patent Documents 0001 to 0002, and the like.

Patent Document 0001 relates to a microbial preparation for legumes, and more specifically, to a microbial preparation for promoting the growth and increasing the yield of legumes such as soybeans, and a method of increasing the growth promoting yield, wherein the ratio of the number of cells is 0.1 to 10 It is characterized by containing azospiram brazilens NI-10 strain and rhizobacteria such as bacteria of the genus Rhizobium, bacteria of the genus Brazizobium or bacteria of the genus Azorizobium.

Patent Document 0002 relates to a microbial complex preparation for promoting the growth of soybean and green manure crops in a high salt soil environment and a method for manufacturing the same, and a microbial complex preparation obtained by mixing a strain of Rhizobium sp. and a strain of Pseudomonassp. Is by confirming the effect of increasing the germination rate and fresh weight of Hairy Vicia villosa Roth in promoting the growth of soybean and green manure crops in high salt environment soil, the Rhizobium sp. strain and Pseudomonas sp. By mixing strains, it can be usefully used as a microbial complex for promoting the growth of legumes and green manure crops and a method for producing the same.

JP3574686B2 (2004.07.09) JP10-1814073B1 (2017.12.26)

The present invention improves the early growth and activity of rhizomes in legume crops, and allows for cultivation of legumes in humid soils such as rice paddies through nitrogen fixation and salt reduction in the soil, and nitrogen fixation that allows continuous cropping of legumes Its purpose is to provide a Rhizobium tropici Sunchang180605 strain having an effect of promoting fruit and crop growth, a soil improvement method using the same, and a method of promoting the growth of legume crops.

The present invention for achieving the above object provides a ryzobium trophy seed ( Rhizobium tropici ) Sunchang180605 strain (KACC 92267P) having nitrogen-fixing power and crop growth promoting effect.

In particular, the crop is characterized in that the legume crop (leguminous crop).

In addition, the present invention provides a soil improvement method, characterized in that improving the soil by fixing nitrogen and reducing salts in the soil using the Rhizobium tropici Sunchang180605 strain (KACC 92267P).

In addition, the present invention provides a method for promoting the growth of legume crops, characterized in that promoting the growth of legume crops using Rhizobium tropici Sunchang180605 strain (KACC 92267P).

The present invention provides a method for coating a legume crop seed, characterized in that after soaking the seeds of a legume crop in a culture broth obtained by culturing Rhizobium tropici Sunchang180605 strain (KACC 92267P) and coating with a coating agent.

In addition, the present invention provides a coating seed of a legume crop, characterized in that produced by the coating method of the legume crop seed.

In addition, the present invention provides a method for promoting growth of legume crops, characterized in that the culture medium obtained by culturing the ryzobium trophy seed Sunchang180605 strain (KACC 92267P) is fertilized on the legume crop.

In particular, by diluting the culture medium of the Rhizobium trophy seed Sunchang180605 strain to 10 ^-6 cfu·ml ^-1 by diluting the R. It can be used by drenching inoculation on the branch of legume crops or by applying a topsoil layer.

The timing of use can be used in the whole process of mature plants from germinated seeds and young seedlings regardless of the growth stage of the legume crop. However, in the case of Rhizobium, the early inoculation is known to have a greater effect. Therefore, the effect of seed coating or early fertilization of R. tropici Sunchang180605 strain ( R. tropici Sunchang180605) can increase the effect of early inoculation.

The new Rhizobium tropici Sunchang180605 strain of the present invention improves the early growth and activity of rhizomes in legume crops, and improves the rhizosphere environment and root nodules deteriorated by conventional agriculture through nitrogen fixation and salt reduction in the soil. By greatly improving, it is possible to cultivate legume crops in humid soil, and there is an effect that it is possible to grow legume crops.

1 is a photograph showing 6 types of rhizomes selected from soil in the Sunchang-gun area.
FIG. 2 is a photograph of Daewon beans grown for 3 weeks by performing an in vitro experiment with four types of rhizomes having excellent room temperature activity.
Figure 3 is a photograph of Daewon beans grown for 3 weeks by performing an extracorporeal experiment of 40 new rhizomes treated with EMS.
FIG. 4 is a photograph of the roots of Daewon Beans after treatment with a new Rhizobium trophy seed, Sunchang180605 (R. tropici Sunchang180605), and then performing an in vitro fresh test for 3 weeks.
5 and 6 are gene sequence analysis results and relationship analysis results of the new Rhizobium trophy seed Sunchang180605 (R. tropici Sunchang180605).
7 is a photograph of soybean seeds coated with new Rhizobium trophy seed Sunchang180605 ( R. tropici Sunchang180605) using seedling and slaked lime powder.

Hereinafter, the Rhizobium tropici Sunchang180605 strain having nitrogen-fixing power and crop growth promoting effect of the present invention, a soil improvement method using the same, and a method for promoting the growth of legume crops will be described in detail as follows. The scope of the present invention is not limited to the following examples.

[Example 1] Collection of symbiotic rhizomes of soybean and isolation of useful rhizomes

84 soils in which soybeans and wild legumes grow naturally in the Sunchang-gun area were collected, and a number of native and native rhizomes that cause root nodules were collected. Among them, 6 types of rhizomes known to be effective in the occurrence of soybean root nodules were selected as shown in FIG. 1.

[Example 2] Ex vivo experiments of collected rhizome bacteria

Among the collected rhizomes, four species with excellent room temperature activity were reselected, and 1,000 strains of Daewon bean were disclosed, and an in vivo experiment was performed for 3 weeks, and a photograph of the grown Daewon bean is shown in FIG. 2.

R. tropici collected among reselected Rhizobacteria with excellent room temperature activity The strain (6-0) was excellent throughout the growth as shown in Table 1 below. The collected R. tropici strain 6-0 was superior in terms of plant height, fresh weight, chlorophyll, leaf tree, dry weight, and rhizomes compared to other strains as well as control.

4 kinds of rheumatoid fungi excellent in room temperature activity Plant length
(cm) Live weight
(g) chlorophyll Leaf tree Out of the building
(g) Rhizome Control (no treatment) 28.8 30.7 33.6 21.2 10.5 2.3 R.tropici 2-0 30.5 30.9 32.4 24.3 10.1 2.1 R.tropici 3-0 31.2 30.5 34.9 22.1 10.2 2.2 R.tropici 4-0 36.3 32.4 36.8 26.7 10.9 2.4 R.tropici 6-0 38.7 33.3 35.7 26.7 10.8 2.5

[Example 3] Development of a new Rhizobium trophy seed ( R. tropici )

R. tropici 6-0 ( R. tropici 6-0), which has superior overall growth such as plant height and live weight compared to the control, was selected, treated with 1 ppm EMS (Ethyl Methane Sulfonate) for 15 minutes, washed with water, and cultured in two layers for 4 hours to develop mutants. I did. Among these, R. tropici , which has excellent in-flight activity 40 strains were selected, and 8,200 strains of Daewon Kong were disclosed, and an in vitro bioassay was performed for 3 weeks as shown in FIG. 3.

It said emptying the rayijo collected in Doi trophy (R. tropici) to EMS (Ethyl Methane Sulfonate) the rayijo away trophy seed Sunchang180605 strain newly developed through the process (R. tropici Sunchang180605) is very long, as shown in Table 2 below, fresh weight, chlorophyll , Leaf tree, dry weight, and rhizome incidences were excellent.

In particular, the newly developed Rhizobium Trophy Seed Sunchang180605 strain has a height of 2.32cm (10.69% increase compared to the control) compared to the control, but fresh weight 1.00g (15.38% increase compared to the control), chlorophyll 1.78 (6.19% increase compared to the control), the main leaf Excellent with 0.53 sheets of number (6.62% increase compared to the control), 0.14g of dry matter (8.80% increase compared to the control), 5.6 days shortened the date of rooting bacteria (28.86% promoted compared to the control), and 0.67 (33.50% increased compared to the control) I did. In particular, the newly developed Rhizobium trophy seed Sunchang180605 strain, compared to the control, has an excellent growth promotion effect while maintaining very healthy growth without overgrowing because the plant height increases, but the live weight, dry weight, and the number of leaves increase.

In vitro test results of rheumatoid fungi with superior growth compared to control Plant length
(cm) Live weight
(g) chlorophyll Leaf tree Out of the building
(g) Rhizome
Birth (days) Rhizome
Incidence Control 21.70 6.50 28.73 8.00 1.59 19.4 2.00 Shin Gyunju* 24.02 7.50 30.51 8.53 1.73 13.8 2.67

* Rhizobium trophy seed Sunchang180605 strain ( R. tropici Sunchang180605)

Through the EMS treatment, early growth can be promoted through seed coating and fertilization using the new Rhizobium trophy seed Sunchang180605 strain ( R. tropici Sunchang180605).

As a result of confirming the roots of Daewon beans, which were tested for 3 weeks after treatment with the new Rhizobium trophy seed Sunchang180605 strain, the root lumps grown as shown in FIG. 4 were examined with an average of 5 mm or more with a size that can be identified with the naked eye.

In addition, the gene sequence of the new Rhizobium trophy seed Sunchang180605 strain ( R. tropici Sunchang180605) was analyzed, and the sequence analysis result is shown in FIG. 5. Based on this genetic information, a comparative analysis of the base sequence of the existing R. tropici and ITS16s rRNA gene registered in the National Center for Biotechnology Information and the phylogenetic analysis of FIG. tree), it was confirmed that it is a different rhizome bacteria compared to R. tropici , which has been revealed so far.

Next, the soil improvement effect of the new Rhizobium trophy seed Sunchang180605 strain ( R. tropici Sunchang180605) was verified. To this end, two types of rheumatoid control ( R. tropici KACC 10996) and treatment ( R. tropici Sunchang 180605 ) were disclosed, and soil changes on the 45th day after treatment with 10 ^-6 cfu·ml ^{-1 per} 1,000 weeks of soybeans (nitrogen fixation). , Salt reduction) was analyzed, and the results are shown in Table 3.

division
Soil improvement effect Nitrogen fixation Salt reduction Control ( R. tropici KACC 10996) 1.00 1.00 Shin, Kyunju ( R. tropici Sunchang180605) 2.53 2.81

As shown in Table 3, the new Rhizobium trophy seed Sunchang180605 strain ( R. tropici Sunchang180605) is compared with the standard R. tropici KACC 10996 owned by the Korean Agricultural Genetic Resource Center, which has a nitrogen fixation effect of 2.53 and a salt reduction effect of 2.81. As a result, effective nitrogen in the soil and salt reduction effect causing nutrient imbalance was high.

[Example 4] Seed Coating Using New Rhizobium Trophy Seed Sunchang180605 ( R. tropici Sunchang180605)

Through EMS treatment, the new Rhizobium trophy seed Sunchang180605 ( R. tropici Sunchang180605) was used for seed coating to induce growth promotion early.

Although existing technology seed coating absorbed moisture to the soybean from the soaking seeds, In Example 4, a new water Rhizobium seed rayijo away trophy Sunchang180605 (R. tropici Sunchang180605) in place of the seed used in the soaking liquid culture ^10- A suspension of ⁶ cfu·ml ^-1 was used to inoculate the rhizomes simultaneously with seed soaking for 6 hours. Next, the slaked lime powder and soybean seeds soaked for 6 hours were mixed in a weight ratio of 1:9, and seed coating was performed. The seed-coated soybean seeds were shaded for 24 hours to obtain coated seeds as shown in FIG. 7, and this was used for seed sowing.

[Example 5] Fertilization using a new Rhizobium trophy seed Sunchang180605 ( R. tropici Sunchang180605)

Through EMS treatment, a new R. tropici Sunchang180605 strain was fertilized to promote early growth of legume crops such as soybeans.

Existing technology does not fertilize a separate rhizome for soybeans, and there is no method of agricultural use for this.

The newly developed Rhizobium trophy seed Sunchang180605 strain ( R. tropici Sunchang180605) can be freely used according to the convenience of farmers, according to the convenience of farmers, through soil surface irrigation or foliar fertilization.

In particular, the newly developed Sunchang New Rhizobacterium strain was tested and selected through room temperature culture unlike other Rhizobacteria isolated under special conditions. Compared with the microorganisms used in other eco-friendly agricultural materials, the artificial high temperature conditions used in pure separation conditions are significantly different from the natural agricultural environmental conditions, which is very unfavorable in survival and competition in actual agricultural fields. It is prepared for the case of extinction in a short time.

The new Rhizobium trophy seed Sunchang180605 strain ( R. tropici Sunchang180605) has high activity in the competition with the microorganisms in real natural state. Therefore, new rayijo away trophy room temperature strain seed Sunchang180605 activity of Pseudomonas species (R. tropici Sunchang180605) (Pseudomonas sp .), Was black and the competitiveness of Bacillus (Bacillus sp.), The results are shown in Table 4.

Comparison of activity and tolerance Room temperature activity Pseudomonas sp. Bacillus sp. Control
( R. tropici KACC 10996) 1.03 0.57 0.88 Shin Gyunju
( R. tropici Sunchang180605) 3.24 1.74 1.96

In general, commonly present in Pseudomonas species (Pseudomonas sp.) And Bacillus (Bacillus sp.) And competition in the natural state, compared to the standard soil Rhizobium (R. tropici KACC 10996) between God and Rhizobium (R. tropici Sunchang180605) As a result, as shown in Table 4, the newly developed R. tropici Sunchang180605 strain ( R. tropici Sunchang180605) showed strong resistance.

In addition, the R. tropici Sunchang 180605 strain ( R. tropici Sunchang 180605 ) showed higher activity than the standard Rhizobacterium ( R. tropici KACC 10996) as shown in Table 4 in the growth activity of bacteria left at room temperature at 25°C for 15 days. It was confirmed that the fertilization effect can last for a long time because it has high resistance to soil drenching and foliar fertilization.

Claims (7)

Rhizobium tropici Sunchang180605 strain (KACC 92267P) having nitrogen-fixing power and crop growth promoting effect.
The method of claim 1,
The crop is a leguminous crop, characterized in that Rhizobium Trophy ( Rhizobium tropici ) Sunchang180605 strain (KACC 92267P), characterized in that.
Rhizobium tropici ( Rhizobium tropici ) using the Sunchang180605 strain (KACC 92267P) to improve the soil by fixing nitrogen and reducing salts in the soil, characterized in that the soil improvement method.
Rhizobium tropici seed ( Rhizobium tropici ) Sunchang180605 strain (KACC 92267P) using a legume crop growth promoting method, characterized in that to promote the growth of the legume crop.
Rhizobium tropici ( Rhizobium tropici ) Sunchang180605 strain (KACC 92267P) in the culture medium of the cultured legumes seeds after soaking the seeds of the legume crop, characterized in that coating with a coating method.

Coating seeds of a legume crop, characterized in that produced by the method of coating the seeds of the legume crop of claim 5.
Rhizobium tropici seed ( Rhizobium tropici ) Sunchang180605 strain (KACC 92267P) is a method for promoting the growth of legume crops, characterized in that fertilizing the culture medium of the cultured legume crop.

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