Dittmariella sp. and application thereof

Abstract

The application discloses a Dietzia sp., which is WX3, has a preservation number of CGMCC No. 32949, and is preserved in the China General Microbiological Culture Collection Center (CGMCC) on December 9, 2024, and has an address of No. 1, Beichen West Road, Yard 3, Chaoyang District, Beijing. The Dietzia sp. has the abilities of dissolving phosphorus, producing indole acetic acid and producing iron carrier, and has a significant effect as a growth-promoting bacterium in promoting the growth of rice root length, plant height, root dry weight and aboveground dry matter weight.

Description

Technical Field

[0001] This invention relates to a strain of Dietrichia coli and its application, belonging to the field of agricultural biological product manufacturing technology. Background Technology

[0002] Rhizosphere growth-promoting bacteria are beneficial bacteria that live freely in the rhizosphere soil, promoting plant growth and the absorption and utilization of mineral nutrients while inhibiting harmful organisms. These bacteria can directly or indirectly promote plant growth through one or more mechanisms, such as nitrogen fixation, phosphorus solubilization, potassium solubilization, and the production of plant hormones. Currently, the use of rhizosphere growth-promoting bacteria to enhance plant growth and increase crop yields is receiving increasing attention and has yielded positive results.

[0003] Dietzia sp. has been confirmed as a rhizosphere growth-promoting bacterium. To date, research on Dietzia sp. has mainly focused on oil recovery and bioremediation of oil pollution, with relatively little research on its role in promoting crop growth (DOI: 10.1038 / srep34768). Summary of the Invention

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

[0005] Technical solution: This invention provides a strain of Dietzia sp., namely Dietziasp. WX3, with accession number CGMCC No.32949, which was deposited on December 9, 2024 at the China General Microbiological Culture Collection Center (CGMCC), located at No.3, Courtyard 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 application of the above-mentioned Dietzia sp. in the preparation of products for phosphate solubilization.

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

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

[0010] The present invention also provides the application of the above-mentioned Dietzia sp. in the preparation of products that promote the salt-tolerant growth of plants.

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

[0012] The present invention also provides a method for promoting salt-alkali tolerant growth of plants, the method comprising the following steps: drenching the roots of plants with a bacterial suspension containing the aforementioned Dietzia sp.

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

[0014] Beneficial effects: Compared with the prior art, the present invention has the following outstanding advantages: The present invention discloses a strain of Dietzia sp. WX3, which has the ability to solubilize phosphorus, produce indoleacetic acid and produce siderophores. As a growth-promoting bacterium, it has a significant effect on promoting the growth of rice in terms of root length, plant height, root dry weight and aboveground dry matter weight. Attached Figure Description

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

[0016] Figure 2 Gram staining for strain WX3.

[0017] Figure 3 Phylogenetic tree of strain WX3.

[0018] Figure 4 The growth of strain WX3 on Monkina inorganic phosphorus solid medium.

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

[0020] Figure 6 The growth status of rice with and without inoculation.

[0021] Figure 7 A bar chart comparing the root lengths of rice plants that were inoculated and those that were not.

[0022] Figure 8 A bar chart comparing the plant height of inoculated and uninoculated rice.

[0023] Figure 9 A bar chart comparing the root dry weight of rice plants that were inoculated and those that were not.

[0024] Figure 10 A bar chart comparing the aboveground dry weight of rice plants that were inoculated and those that were not. Detailed Implementation

[0025] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0026] (I) Isolation and purification of strains

[0027] Bacterial isolates from the rhizosphere soil of coastal saline-alkali rice were cultured on Luria-Bertani medium using a serial dilution method. The isolates were then purified using the streak plate method until a pure culture was obtained, which was subsequently designated WX3.

[0028] (II) Identification of strains

[0029] 1. Morphological observation

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

[0031] 2. Molecular identification

[0032] PCR was performed on the 16S rRNA gene sequence of WX3 using universal bacterial primers 27F and 1492R. The PCR reaction mixture consisted of: 1 μL 27F (20 μmol), 1 μL 1492R (20 μmol), 4 μL dNTPs, 5 μL 10×Buffer, 0.25 μL Taq DNA polymerase, 2 μL DNA, and ddH2O to a final volume of 50 μL. The PCR program was: 95℃ pre-denaturation for 3 min, 95℃ denaturation for 30 s, 55℃ annealing for 30 s, 72℃ extension for 40 s, for 35 cycles, followed by a final extension at 72℃ for 10 min. The purified PCR product was sent to Shanghai Sangon Biotech Co., Ltd. for sequencing. The sequencing results were submitted to NCBI for comparison with existing 16S rDNA sequences in the database, and a phylogenetic tree was constructed using highly related 16S rDNA gene sequences selected from Genebank. The results are shown below. Figure 3 As shown, the sequence of WX3 belongs to the same minimal branch as Dietziasp. A14101. Therefore, WX3 was identified as Dietzia sp.

[0033]

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

[0035] 1. Identification of phosphorus solubility of WX3

[0036] WX3 was inoculated onto Monkina inorganic phosphorus solid medium and incubated upside down at 28°C for 48 hours. After incubation, the presence of a clear zone around the WX3 colony was observed. If a clear zone was present, the strain was considered to possess phosphate-solubilizing function. Results are as follows: Figure 4 As shown, a clear zone exists around the WX3 colony on the Monkina organic phosphorus medium, indicating that WX3 has the ability to dissolve phosphorus.

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

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

[0039] Table 1. Indoleacetic acid production capacity of strain WX3

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

[0041] 3. Evaluation of WX3's ability to produce ferrocarriers

[0042] WX3 was spot-inoculated onto CAS detection solid medium and incubated upside down at 28℃ for 48 hours. After incubation, the presence of an orange-yellow halo around the WX3 colonies was observed. The presence of an orange-yellow halo indicated that the colony possessed siderogenic capacity. Results are as follows: Figure 5 As shown, the WX3 colonies on the CAS solid culture medium have an orange-yellow halo around them, indicating that WX3 has the ability to produce siderophores.

[0043] The above experimental results demonstrate that Dietzia sp. WX3 strain possesses the ability to solubilize phosphorus, produce indoleacetic acid, and produce siderophores, and belongs to the rhizosphere growth-promoting bacteria.

[0044] (iv) Effects of WX3 on rice seedling stage

[0045] The seedlings of Huai Dao No. 5 were transplanted into coastal saline soil and cultured for 7 days, then treated with a final concentration of 10. 8The standard CFU / g soil concentration was achieved by uniformly irrigating the WX3 bacterial suspension around the rice roots. A sterile water irrigation treatment was also set up as a control. The rice was cultivated under natural conditions. Watering was observed daily, maintaining a water layer of 2-3 cm. After 30 days of growth, the rice was photographed, and 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 rice inoculated with WX3 showed better growth than the control rice. See the detailed results below. Figures 7-10 The average root length, plant height, root dry weight, and aboveground dry weight of rice inoculated with WX3 were all higher than those of the control rice, indicating that inoculation with WX3 promoted the growth of rice in coastal saline soil.

Claims

1. A method for promoting rice growth in saline-alkali land, characterized in that, The method includes the following steps: using bacteria containing Dietzia Dietzia sp. The bacterial suspension was used for root irrigation of rice, and the bacteria mentioned are *Dizzyella*. Dietzia sp. The strain described is WX3, with accession number CGMCC No. 32949, deposited on December 9, 2024, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China; [The description of *Dizzyella* is missing from the original text.] Dietzia sp. The concentration is 10 8 CFU / g soil.

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