Dietzia and application thereof

Abstract

The invention discloses Dietzia sp., the Dietzia sp. Is WX3, the preservation number of the Dietzia sp. Is CGMCC No.32949, the Dietzia sp. Is preserved in China General Microbiological Culture Collection Center (CGMCC) on December 9, 2024, and the preservation address of the Dietzia sp. Is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China. The bacillus licheniformis has the capabilities of dissolving phosphorus, producing indoleacetic acid and producing siderophores, and has a remarkable effect in promoting the growth of rice root length, plant height, root dry weight and overground part dry matter weight as a growth-promoting bacterium.

Description

Technical Field

[0001] The invention relates to a strain of Dietzia and application thereof, belonging to the technical field of production of agricultural biological products. Background Art

[0002] Rhizosphere growth-promoting bacteria are beneficial bacteria that live freely in the rhizosphere soil, can promote plant growth and its absorption and utilization of mineral nutrients, and can inhibit harmful organisms. Rhizosphere growth-promoting bacteria can directly or indirectly promote plant growth through one or more mechanisms such as nitrogen fixation, phosphorus and potassium solubilization, and production of plant hormones. At present, the use of rhizosphere growth-promoting bacteria to promote plant growth and increase crop yields has attracted more and more attention and achieved good results.

[0003] Dietzia sp. has been confirmed to be a rhizosphere growth-promoting bacterium. So far, the research on Dietzia sp. has mainly focused on oil recovery and bioremediation of oil pollution, while there are few studies on promoting crop growth (DOI: 10.1038 / srep34768). Summary of the invention

[0004] Purpose of the invention: The purpose of the present invention is to provide a strain of Dietzia sp. and its application.

[0005] Technical solution: The present invention provides a strain of Dietzia sp., wherein the Dietzia sp. is WX3, the preservation number is CGMCC No.32949, and it was deposited in the General Microbiology Center of China Culture Collection Administration (CGMCC) on December 9, 2024, and the preservation address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China.

[0006] Furthermore, the 16S rRNA sequence of the Dietzia sp. is shown in SEQ ID NO.1.

[0007] The present invention also provides the use of the above-mentioned Dietzia sp. in preparing products for dissolving phosphate.

[0008] The present invention also provides the use of the above-mentioned Dietzia sp. in preparing products producing indoleacetic acid.

[0009] The present invention also provides the use of the above-mentioned Dietzia sp. in preparing iron-producing products.

[0010] The invention also provides the use of the above-mentioned Dietzia sp. in preparing products for promoting the growth of plants resistant to salt and alkali.

[0011] Furthermore, the promoting of salt-alkali tolerant growth of plants includes promoting the growth of plant root length, plant height, root dry weight and aboveground dry matter weight.

[0012] The invention also provides a method for promoting the salt-alkali tolerance growth of plants, which comprises the following steps: irrigating the roots of plants with a bacterial suspension containing the above-mentioned Dietzia sp.

[0013] Furthermore, the concentration of Dietzia sp. is 10 8 CFU / g soil.

[0014] Beneficial effects: Compared with the prior art, the present invention has the following outstanding and significant advantages: The present invention discloses a strain of Dietzia sp. WX3, which has the ability to solubilize phosphate, produce indoleacetic acid and produce siderophores, and as a growth-promoting bacterium, has a significant effect in promoting the growth of rice root length, plant height, root dry weight and aboveground dry matter weight. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The colonies of strain WX3 grown on Luria-Bertani solid medium.

[0016] Figure 2 Gram stain for strain WX3.

[0017] Figure 3 This is the phylogenetic tree of strain WX3.

[0018] Figure 4 The growth of strain WX3 on Montgena inorganic phosphate solid medium.

[0019] Figure 5 The growth of strain WX3 on CAS solid medium.

[0020] Figure 6 Shows the growth conditions of inoculated and non-inoculated rice.

[0021] Figure 7 This is a bar graph comparing the root lengths of rice with and without inoculation.

[0022] Figure 8 This is a bar graph comparing the plant heights of rice with and without inoculation.

[0023] Fig. 9 The bar graph shows the comparison of dry weight of rice roots with and without inoculation.

[0024] Fig.10 This is a bar graph comparing the dry matter weight of the aboveground parts of rice with and without inoculation. DETAILED DESCRIPTION

[0025] The technical solution of the present invention is further described below in conjunction with the accompanying drawings.

[0026] (I) Isolation and purification of strains

[0027] The bacterial isolates isolated from the rhizosphere soil of coastal saline-alkali rice were cultured on Luria-Bertani medium by the plate gradient dilution method and purified by the plate streak method until a pure culture was obtained, which was later called WX3.

[0028] (II) Identification of strains

[0029] 1. Morphological observation

[0030] The characteristics of WX3 colonies were observed, and the results were as follows Figure 1 As shown. The colonies of WX3 on Luria-Bertani solid medium are round, orange-red, opaque, with smooth edges and a moist and convex surface. Gram staining results show that WX3 is purple after Gram staining ( Figure 2 ), indicating that WX3 is a Gram-positive bacterium.

[0031] 2. Molecular identification

[0032] The 16S rRNA gene sequence of WX3 was PCR-generated using bacterial universal primers 27F and 1492R. PCR reaction system: 27F (20 μmol) 1 μL, 1492R (20 μmol) 1 μL, dNTP 4 μL, 10×Buffer 5 μL, Taq DNA polymerase 0.25 μL, DNA 2 μL, ddH2O to 50 μL. PCR reaction procedure: 95°C pre-denaturation for 3 min, 95°C denaturation for 30 s, 55°C annealing for 30 s, 72°C extension for 40 s, 35 cycles, and 72°C extension for 10 min. After purification, the PCR product was sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing. The sequencing results were submitted to NCBI and compared with the existing 16S rDNA sequences in the database, and the 16S rDNA gene sequences with high affinity were selected from Genebank to construct a phylogenetic tree. The results are shown in the figure. Figure 3 As shown, the sequence of WX3 belongs to the same minimal branch as Dietzias p.A14101. Therefore, WX3 was identified as Dietzia sp.

[0033]

[0034] (III) Identification of growth-promoting traits of WX3

[0035] 1. Identification of WX3's phosphorus dissolving ability

[0036] WX3 was inoculated onto the Montkina inorganic phosphate solid medium and incubated at 28°C for 48 hours. After the incubation, observe whether a transparent circle can be formed around the WX3 colony. If a transparent circle is formed, it is determined that the strain has the ability to dissolve phosphate. Figure 4 As shown, there is a transparent zone around the WX3 colony on the Montkina organophosphate medium, indicating that WX3 has the ability to solubilize phosphate.

[0037] 2. Identification of WX3's ability to produce indoleacetic acid

[0038] WX3 was inoculated into a nutrient broth liquid culture medium containing 100 mg / L L-tryptophan, and cultured at 30°C and 150 r / min for 48 h. The supernatant was collected by centrifugation at 4000 r / min for 10 min, and an equal volume of Salkowski colorimetric solution was added and mixed. The OD was measured after the reaction in the dark for 30 min. 530 , and according to the standard curve (y=0.0361x-0.0196, R 2 =0.9993) to quantify the ability of the strain to produce indoleacetic acid, and the results were repeated twice. The results are shown in Table 1. Each ml of WX3 culture solution contained 30.29 μg of indoleacetic acid.

[0039] Table 1 Indoleacetic acid production ability of strain WX3

[0040] strain <![CDATA[OD 530 ]]> Indoleacetic acid content (mg / L) WX3 1.07±0.10 30.29±2.80

[0041] 3. Identification of WX3's ability to produce siderophores

[0042] WX3 was spotted onto the CAS solid culture medium and inverted for 48 hours at 28°C. After the culture was completed, an orange-yellow halo was observed around the WX3 colony. If an orange-yellow halo was produced, it was identified as having the ability to produce iron carriers. Figure 5 As shown, there is an orange-yellow halo around the WX3 colony on the CAS test solid culture medium, indicating that WX3 has the ability to produce siderophores.

[0043] The above experimental results prove that Dietzia sp. WX3 strain has the ability to solubilize phosphate, produce indoleacetic acid and produce siderophores, and is a rhizosphere growth-promoting bacterium.

[0044] (IV) Effects of WX3 on rice seedlings

[0045] The seedlings of Huaidao No. 5 were transplanted into coastal saline soil for 7 days and then treated with a final concentration of 10 8CFU / g soil standard: WX3 bacterial suspension was evenly poured around the rice root system. At the same time, sterile water irrigation was set as a control. Rice was cultured under natural conditions. Watering was observed every day to maintain a 2-3 cm water layer. After 30 days of growth, the rice was photographed and the rice plant height, root length, root dry matter weight and aboveground dry matter weight were measured.

[0046] Depend on Figure 6 It can be seen that the growth of rice inoculated with WX3 is better than that of rice in the control treatment. Figures 7-10 The average root length, plant height, root dry weight and aboveground dry matter weight of rice inoculated with WX3 were higher than those of rice in the control treatment, indicating that inoculation with WX3 promoted the growth of rice in coastal saline soil.

Claims

1. A strain of Dietzia sp., characterized in that: The Dietzia sp. is WX3, with a deposit number of CGMCC No.32949. It was deposited in the General Microbiology Center of the China Culture Collection Administration (CGMCC) on December 9, 2024, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China.

2. The Dietzia sp. according to claim 1, characterized in that The 16S rRNA sequence of the Dietzias p. is shown in SEQ ID NO.

1.

3. Use of the Dietzia sp. according to any one of claims 1 to 2 in the preparation of phosphate-dissolving products.

4. Use of the Dietzia sp. according to any one of claims 1 to 2 in the preparation of products producing indoleacetic acid.

5. Use of the Dietzia sp. according to any one of claims 1 to 2 in the preparation of iron-producing products.

6. Use of the Dietzia sp. according to any one of claims 1 to 2 in the preparation of a product for promoting the growth of plants tolerant to salt and alkali.

7. The use according to claim 8, characterized in that: The method of promoting the salt-alkali tolerant growth of plants includes promoting the growth of plant root length, plant height, root dry weight and aboveground dry matter weight.

8. A method for promoting the growth of plants tolerant to salt and alkali, characterized in that: The method comprises the following steps: irrigating the roots of plants with a bacterial suspension containing the Dietzia sp. according to any one of claims 1 to 2.

9. The method for promoting the salt-alkali tolerance growth of plants according to claim 8, characterized in that: The concentration of Dietzia sp. is 10 8 CFU / g soil.

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