NOVEL STRAIN AND PLANT GROWTH PROMOTER COMPRISING THE SAME

Abstract

To provide a strain having a plant growth promoting action and a plant growth promoting agent comprising the strain.SOLUTION: The present invention was completed by finding that the strain Pantoea isolated from Quinoa has a plant growth promoting effect.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a novel strain and a plant growth promoter containing said strain. [Background technology]

[0002] Quinoa (Chenopodium quinoa) is an allopolyploid plant native to the Andes that belongs to the genus Chenopodium and the subfamily Chenopodiaceae of the family Amaranthaceae. Quinoa seeds are rich in protein, essential amino acids, and minerals, and are expected to become a substitute for rice (Non-Patent Document 1, Non-Patent Document 2). Furthermore, quinoa's excellent resistance to environmental stresses, such as salt tolerance and cold tolerance, makes it possible to cultivate it in harsh environments (Non-Patent Document 3), and the Food and Agriculture Organization of the United Nations (FAO) has evaluated it as an important means of resolving the world's food crisis (Non-Patent Document 4). Quinoa, which was an important food source in the Andes, has now spread around the world, and is being researched and cultivated commercially. Furthermore, the quinoa genome has been decoded (Non-Patent Document 5).

[0003] In order to respond to the food crisis, it is necessary to promote the growth of quinoa and increase the yield. Chemical fertilizers are used to promote plant growth. Chemical fertilizers reduce beneficial soil bacteria, which can have a negative impact on plant growth and on human health. For this reason, plant growth promoters that use microorganisms are being developed. As a plant growth promoter utilizing a microorganism, a Lactobacillus strain isolated from salted squid has been reported to have a plant growth promoting effect (Patent Document 1). However, no strains have been reported that have growth-promoting effects on quinoa. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Re-tabled publication 2016 / 021204 [Non-patent literature]

[0005]

Linked Book1

Outdoor Tool2

Outdoor Tools3

Outdoor Tools 4

Direct Environment 5

[0006] An object of the present invention is to provide a bacterial strain having a plant growth-promoting effect and a plant growth-promoting agent containing said bacterial strain. [Means for solving the problem]

[0007] As a result of intensive research aimed at solving the above problems, the present inventors discovered that a Pantoea genus strain isolated from quinoa has a plant growth-promoting effect, and thus completed the present invention.

[0008] That is, the present invention is as follows. 1. A Pantoea agglomerans strain having the following properties: (1) Gram-negative (2) The shape is rod-shaped. (3) Glucose is the carbon source (4) Bacillus (5)Aerobic (6) Having the 16S rRNA sequence set forth in SEQ ID NO:1 (7) Can grow on trypticase soy medium (8) Can grow at temperatures between 26 and 35°C 2. The strain having accession number NITE P-03213. 3. A strain according to the above item 1 or 2, which has a plant growth-promoting effect. 4. The strain described in the above 2 or 3, wherein the plant is quinoa. 5. A plant growth promoter comprising one or more of the strains described in 1 to 4 above. 6. A plant inoculated with one or more of the strains of 1 to 4 above. Effect of the Invention

[0009] The present invention can provide a bacterial strain having a plant growth-promoting effect and a plant growth-promoting agent containing said bacterial strain. [Brief description of the drawings]

[0010] [Figure 1] Phylogenetic analysis of strains. Bootstrap values ​​from 1000 replicates are shown for each branch. Bars indicate 0.002 substitutions per site. [Diagram 2] Measurements of fresh and dry weights of control plants and strain-inoculated plants (N=40). [Diagram 3] Representative results of control and inoculated plants. 40 plants were tested. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] (The present invention) The present invention relates to a bacterial strain having a plant growth-promoting effect and a plant growth-promoting agent containing said bacterial strain.

[0012] (Strain) The strain of the present invention has a plant growth-promoting effect and has the following characteristics: (1) Gram-negative (2) The shape is rod-shaped. (3) Glucose is the carbon source (4) Bacillus (5)Aerobic (6) Having the 16S rRNA sequence set forth in SEQ ID NO:1 (7) Can grow on trypticase soy medium (8) Can grow at temperatures between 26 and 35°C Specifically, examples of such microorganisms include those belonging to the genus Pantoea agglomerans. The strains of the present invention include wild-derived strains, strains into which known mutations have been introduced, and genetically modified strains.

[0013] A genetically modified bacterium of a microorganism belonging to Pantoea agglomerans has a 16SrRNA that has an identity of 80% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more to the 16SrRNA sequence set forth in SEQ ID NO:1.

[0014] The strain of the present invention includes the bacterium designated "Panotea agglomerans J1" in the following examples (deposited under the accession number NITE P-03213 on April 28, 2020 at the Patent Microorganisms Depositary Center of the National Institute of Technology and Evaluation (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture)).

[0015] As is clear from the results of the Examples described below, when a plant is inoculated with the strain of the present invention, the strain has the effect of increasing the weight of the plant (particularly the growth of the plant's stems and leaves (fresh weight and dry weight)) by about 1.1 times, 1.2 times, 1.3 times, or 1.3 times or more, compared to when the strain is not inoculated.

[0016] (Plant growth promoter) The plant growth-promoting agent of the present invention contains the bacterial strain having plant growth-promoting activity of the present invention. The strain of the present invention may be a killed cell, but live cells are preferably used because they exert an excellent plant growth promoting effect.

[0017] The form of the plant growth promoter is not particularly limited, and may be any form that a typical microbial agent can take, such as a bacterial suspension, liquid, granule, powder, hydrated agent, pack, granular hydrate, microcapsule, emulsion, etc., and can be prepared into any dosage form and used according to the purpose. For example, the strain of the present invention can be suspended in water or the like and provided as a bacterial suspension or liquid. Alternatively, the strain of the present invention can be adsorbed onto a pharmaceutically acceptable carrier and provided as a wettable powder, dust, or granule. In this case, the carrier can be white carbon, diatomaceous earth, clay, talc, perlite, rice husk, bone powder, etc. In addition, surfactants, dispersants, adjuvants, and the like can be used as pharma- ceutically acceptable additives.

[0018] The concentration of the strain of the present invention contained in the plant growth promoter of the present invention is not particularly limited. In the case of a liquid formulation, the concentration is, for example, 1.0×10 3 ~10 12 Cells / mL, 1.0 x 10 5 ~10 9 The bacteria may be in a culture solution itself. For solid preparations, for example, 1.0x 10 3 ~10 12 Cells / g, 1.0 x 10 5 ~10 9 It may also be cells / g. For example, in the case of a formulation that is diluted before use when treating seeds, plants, or soil, the concentration of the strain of the present invention in the formulation before dilution is 1.0 x 10 8 ~10 10 It may also be cells / g. In addition, in the case of a dispersion of the strain (for example, a dispersion obtained by diluting the above-mentioned formulation), the concentration of the bacteria when applied to seeds, plants, or soil is 1.0 x 10 5 ~10 9 It may be cells / mL.

[0019] The method for using the strain of the present invention or the plant growth promoter of the present invention is to treat at least one selected from the group consisting of seeds, plants, and soil with the strain of the present invention. The plant may be at any stage, preferably after germination. More preferably, the plant is a growing plant having stems, leaves, and roots, and more preferably, a young plant. For example, quinoa seedlings 1 to 7 days after germination are transferred to soil containing vermiculite and inoculated with the strain of the present invention.

[0020] The method of inoculating the strain of the present invention into a plant is not particularly limited, and includes, for example, immersing the roots of the plant in the plant growth promoter of the present invention (e.g., a culture solution of the strain, a bacterial suspension, or a powder of the strain), spraying the plant growth promoter of the present invention onto the plant (particularly the leaves, stems, etc.), spraying the plant growth promoter of the present invention onto the soil for growing the plant, etc.

[0021] Plants to which the plant growth promoter of the present invention can be applied include various cereals or pseudocereals, such as wheat, oats, corn, rye, barley, spelt, millet, sorghum, kamut, triticale, buckwheat, quinoa (Chenopodium quinoa), and / or Amaranthaceae plants, preferably Amaranthaceae plants, more preferably quinoa. EXAMPLES

[0022] The present invention will be described in detail below with reference to specific examples, but the present invention is not limited to these examples. EXAMPLES

[0023] (Strain isolation and phylogenetic analysis) Seven species of Pantoea agglomerans were isolated from seeds of the Kd strain of quinoa, which has been cultivated experimentally in Japan for more than 30 years. Based on the base sequences of the 16SrRNA gene of previously reported Pantoea agglomerans strains (named NR or KX) obtained from the US National Center for Biotechnology Information (NCBI) and the Panotea agglomerans strain isolated in this example (named M8), sequence alignment (multiple alignment) was performed using ClustalW software, and a molecular phylogenetic tree was created using the Neighbor-Joining method with MEGA7 software (Figure 1). This molecular phylogenetic tree confirmed that the seven Panotea agglomerans strains J1-J7 (J strains) isolated in this example form a clade different from previously reported Pantoea agglomerans strains and are new strains. EXAMPLES

[0024] (Plant growth promoting effect of Pantoea agglomerans J1 strain) Among the seven strains of Panotea agglomerans confirmed to be novel strains in Example 1 above, the plant growth promoting effect of the Pantoea agglomerans J1 strain was confirmed. Quinoa (Chenopodium quinoa, accession: Kd) cotyledons grown on GM agar medium for 4 days were transferred to a mixture of vermiculite and professional (Professional soil for cuttings and buds No. 2, Daio Kasei Co., Ltd.) in a ratio of 9:1, and then Pantoea agglomerans J1 strain (1.0 x 10 7 ~10 8 The plants were inoculated with 1.0 mL of vermiculite / mL and grown in trays containing vermiculite:professional (9:1). After 4 weeks of cultivation, the inoculated plants were photographed next to control plants (fed with 1.0 mL of MilliQ water instead of the bacterial suspension), and the fresh and dry weights of the control plants and the inoculated plants were measured.

[0025] (Confirmation of plant growth promotion effect) It was confirmed that four weeks after transplanting into pots, the plants inoculated with Panotea agglomerans J1 had significantly increased fresh and dry weights compared to the control plants (Figure 2). Inoculum concentration 1.0 x10 8 Plants inoculated with Panotea agglomerans J1 at 1.5 cells / mL showed an average increase in fresh weight and dry weight of 1.3 times that of control plants, confirming promotion of plant growth (especially in stems and leaves) (Figure 3). From the above, it was confirmed that Panotea agglomerans J1 has a plant growth-promoting effect. [Industrial Applicability]

[0026] The present invention can provide a novel Pantoea agglomerans fungus having a plant growth-promoting effect.

Claims

1. Pantoea agglomerans J1 (accession number: NITE P-03213) Add to cultivation soil, Direct spray or immersion in a suspended liquid. Either by How to grow quinoa.

2. Pantoea agglomerans J1 (accession number: NITE P-03213) Add to cultivation soil, Direct spray or immersion in a suspended liquid. Either by Pantoea agglomerans J1 (accession number: NITE P-03213) is allowed to exist in quinoa. How to grow quinoa.

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