Basfia succinat-10 and its application in solubilizing phosphorus and promoting growth

Abstract

The present application relates to the field of microbial technology, and particularly relates to a rhizosphere basing cupriavidus basellens - 10 and application thereof in dissolving phosphorus and promoting growth. The rhizosphere basing cupriavidus basellens - 10 provided by the present application has a preservation number of GDMCC No. 65501, and the rhizosphere basing cupriavidus basellens - 10 can increase the contents of chlorophyll a, chlorophyll b, carotenoids and total chlorophyll in leaves of cinnamomum chekiangense seedlings, increase the plant height, ground diameter, total fresh weight, root fresh weight, stem fresh weight, leaf fresh weight, total root length and thick root number of the cinnamomum chekiangense seedlings, and promote the accumulation of soluble sugar and effective phosphorus in roots and leaves.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbial technology, and particularly relates to a rhizosphere Cupriavidus basellae-10 and application thereof in phosphorus dissolution and growth promotion. BACKGROUND

[0002] Phellodendron chinense Schneid is an important perennial woody medicinal material, which is widely distributed in Hunan, Hubei and Sichuan, etc. in southern China. This plant can usually grow to 10-12 m high, with purple-brown young branches and dark brown-gray bark, and has white pitted lenticels on the surface without hair. The leaves of P. chinense are odd-pinnately compound, in opposite condition, with leaf axis, petiole and leaf back covered with fine pubescence. As a dioecious plant, P. chinense blooms yellowish single sex flower in May-June. The stem bark of P. chinense contains active ingredients such as alkaloids, flavonoids, sterols, etc., among which the content of alkaloids is the highest. The alkaloids in the stem bark of P. chinense mainly include berberine, phellodendrine, palmatine, etc., among which berberine has the highest content and has significant anti-tumor, anti-inflammatory and antibacterial, hypoglycemic and hypolipidemic effects, and is widely used in the treatment of cancer, enteritis, diabetes, arthritis, etc. The growth and medicinal ingredient content of P. chinense are affected by rhizosphere growth-promoting bacteria, nutrient elements, site conditions, etc., thereby affecting its yield and economic value.

[0003] Phosphorus (P) is one of the key nutrients required for plant growth, and plays an important role in photosynthetic phosphorylation, nucleic acid synthesis, protein phosphorylation modification, etc., thereby regulating plant growth, development, stress response and secondary metabolism. Plants mainly absorb phosphorus from soil, but effective phosphorus is easily combined with metal ions such as Fe 2+ , Ca 2+ , Al 3+ to form insoluble compounds, which cannot meet the needs of plant growth. The soil in Hunan Province is mainly red soil, which has high total phosphorus content but low effective phosphorus content (≤0.01%), limiting the growth of plants and the synthesis of important secondary metabolites. Therefore, the exogenous application of phosphorus fertilizer can effectively supplement the content of effective phosphorus in soil, but can easily cause environmental problems such as eutrophication. On the other hand, the addition of rhizosphere phosphorus-dissolving bacteria can effectively alleviate the limitation of soil phosphorus, and Pseudomona s moraviensis, Bacillus safensis, Falsibacillus pallidus, etc. have significant phosphorus-dissolving ability and growth-promoting function.

[0004] However, different types of plants enrich different types of rhizosphere growth-promoting bacteria, and there is no related report on the isolation and identification of rhizosphere growth-promoting bacteria of Chinese medicinal material P. chinense and the growth-promoting function thereof. Therefore, it is urgent to research a kind of bacteria which can improve the phosphorus-dissolving capacity of the rhizosphere soil of P. chinense and promote the growth of seedlings. SUMMARY

[0005] To solve the above technical problems, the application provides a rhizosphere Cupriavidus basilensis-10 and its application in phosphorus dissolution and growth promotion, which can increase the contents of chlorophyll a, chlorophyll b, carotenoids and total chlorophyll in the leaves of the young seedlings of C. fortunei, increase the plant height, ground diameter, total fresh weight, root fresh weight, stem fresh weight, leaf fresh weight, total root length and thick root number of the young seedlings of C. fortunei, and promote the accumulation of soluble sugar and available phosphorus in the roots and leaves.

[0006] To this end, the application provides the following technical solutions,

[0007] In the first aspect, the application provides, in optional embodiments, a rhizosphere Cupriavidus basilensis-10, the preservation number of which is GDMCC No: 65501.

[0008] In the application, the preservation number of the rhizosphere Cupriavidus basilensis-10 is GDMCC No: 65501, the preservation date is November 19, 2024, the preservation center is the Guangdong Microbial Culture Collection Center, the preservation address is the 5th floor of No. 59 Building, 100 Middle Martyrs Road, Guangzhou, the Institute of Microbiology of Guangdong Academy of Sciences, the preserved classification name is Cupriavidus basilensis CB-10, and the preserved survival state is survival.

[0009] Preferably, the nucleotide sequence of the rhizosphere Cupriavidus basilensis-10 is shown as SEQ ID NO. 1.

[0010] In the second aspect, the application provides, in optional embodiments, a rhizosphere Cupriavidus basilensis-10 inoculant prepared from the above-mentioned rhizosphere Cupriavidus basilensis-10.

[0011] In the third aspect, the application provides, in optional embodiments, a preparation method of the above-mentioned rhizosphere Cupriavidus basilensis-10 inoculant, comprising the following steps:

[0012] The rhizosphere Cupriavidus basilensis-10 is inoculated into a culture medium for culture, and when the OD 600 of the bacterial liquid is 0.7, the rhizosphere Cupriavidus basilensis-10 inoculant is obtained.

[0013] Preferably, the culture temperature is 25-30℃, the culture time is 8-12h, and the rotation speed is 150-250rpm; and / or, the culture medium is R2A liquid medium.

[0014] In the third application, the application provides, in optional embodiments, an application of the above-mentioned rhizosphere Cupriavidus basilensis-10 or the above-mentioned rhizosphere Cupriavidus basilensis-10 inoculant in promoting the growth of young seedlings of C. fortunei.

[0015] Preferably, the indexes for promoting the growth of the C. wilsoniana seedlings include the plant height, ground diameter, total fresh weight, leaf fresh weight, stem fresh weight, root fresh weight, total root length and thick root number of the C. wilsoniana seedlings; and the method for promoting the growth of the C. wilsoniana seedlings by the rhizosphere B. solisilvae-10 inoculum is as follows: 20-30 mL of the rhizosphere B. solisilvae-10 inoculum is used to inoculate each C. wilsoniana seedling growing to the four-leaf stage by the root irrigation method.

[0016] In a fourth aspect, the present application provides, in optional embodiments, an application of the above-mentioned rhizosphere B. solisilvae-10 or the above-mentioned rhizosphere B. solisilvae-10 inoculum in improving the pigment content of the C. wilsoniana seedling leaves.

[0017] Preferably, the pigments include chlorophyll a, chlorophyll b, carotenoids and total chlorophyll.

[0018] In a fifth aspect, the present application provides, in optional embodiments, an application of the above-mentioned rhizosphere B. solisilvae-10 or the above-mentioned rhizosphere B. solisilvae-10 inoculum in regulating the soluble substance content of the C. wilsoniana seedling leaves and roots.

[0019] Preferably, the soluble substances include soluble sugar and fructose. In the present application, the rhizosphere B. solisilvae-10 or the rhizosphere B. solisilvae-10 inoculum can improve the content of the soluble sugar in the C. wilsoniana seedling leaves and roots, and can also improve the content of the fructose in the C. wilsoniana seedling roots.

[0020] In a sixth aspect, the present application provides, in optional embodiments, an application of the above-mentioned rhizosphere B. solisilvae-10 or the above-mentioned rhizosphere B. solisilvae-10 inoculum in improving the effective phosphorus content of the C. wilsoniana seedling leaves.

[0021] The nucleotide sequence represented by the SEQ ID NO. 1 is as follows:

[0022]

[0023] Compared with the prior art, the present application has one of the following beneficial effects:

[0024] 1. The rhizosphere Cupriavidus basellens-10 provided by the present application can increase the contents of chlorophyll a, chlorophyll b, carotenoids and total chlorophyll in the leaves of the C. fortunei seedlings, increase the plant height, ground diameter, total fresh weight, root fresh weight, stem fresh weight, leaf fresh weight, total root length and thick root number of the C. fortunei seedlings, and promote the accumulation of soluble sugar and available phosphorus in the roots and leaves. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 It is a photo of the rhizosphere Cupriavidus basellens-10 separated and purified in Example 1 of the present application, wherein Figure 1 A is a front view of the rhizosphere Cupriavidus basellens-10, Figure 1 B is a back view of the rhizosphere Cupriavidus basellens-10.

[0026] Figure 2 It is a schematic diagram of the results of analyzing the homology of the rhizosphere Cupriavidus basellens-10 based on the adjacency method in Example 1 of the present application;

[0027] Figure 3 It is a schematic diagram of the results of analyzing the homology of the rhizosphere Cupriavidus basellens-10 based on the minimum evolution method in Example 1 of the present application;

[0028] Figure 4 It is a schematic diagram of the results of analyzing the homology of the rhizosphere Cupriavidus basellens-10 based on the maximum parsimony method in Example 1 of the present application;

[0029] Figure 5 It is a schematic diagram of the results of the phosphorus solubilizing effect of the rhizosphere Cupriavidus basellens-10 in Example 2 of the present application;

[0030] Figure 6 It is a schematic diagram of the results of the effect of the rhizosphere Cupriavidus basellens-10 on promoting the growth of the C. fortunei seedlings in Example 4 of the present application;

[0031] Figure 7 It is a schematic diagram of the results of the effect of the rhizosphere Cupriavidus basellens-10 on increasing the pigment content of the leaves of the C. fortunei seedlings in Example 4 of the present application;

[0032] Figure 8 It is a schematic diagram of the results of the effect of the rhizosphere Cupriavidus basellens-10 on regulating the soluble substance content of the leaves and roots of the C. fortunei seedlings in Example 4 of the present application;

[0033] Figure 9 It is a schematic diagram of the results of the effect of the rhizosphere Cupriavidus basellens-10 on increasing the available phosphorus content of the leaves of the C. fortunei seedlings in Example 4 of the present application;

[0034] Figure 10 Figure 1 shows the phenotype of the seedlings of P. davidiana var. bungeanae cultivated in the CK group and CB-10 group in Example 4 of the present application. DETAILED DESCRIPTION

[0035] In order to make the objectives, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with specific examples. It should be understood that the specific examples described herein are only used to explain the present application and do not limit the present application.

[0036] The experimental methods used in the following examples are conventional methods unless otherwise specified.

[0037] The materials, reagents and the like used in the following examples can be obtained from commercial channels unless otherwise specified.

[0038] The preparation method of the R2A solid culture medium is as follows: 18.12 g of R2A agar culture medium dry powder (purchased from Haibo Biological Company) is accurately weighed and dissolved in distilled water, and then the volume is made up to 1 L. Then the prepared medium is transferred to a sterilization pot and sterilized at 121°C for 15 min. After the medium is sterilized, 75% ethanol solution is taken out for sterilization and placed on an ultraclean workbench for cooling. When the temperature drops to 40-50°C, sterile amphotericin B is added (the addition amount is 0.8 mg / L), mixed uniformly, and then divided into culture dishes for standby.

[0039] The preparation method of the R2A liquid culture medium is as follows: 3.2 g of R2A liquid culture medium dry powder (purchased from Haibo Biological Company) is accurately weighed and dissolved in pure water, and then the volume is made up to 1 L with pure water. The prepared medium is transferred to a sterilization pot and sterilized at 121°C for 15 min. After sterilization, the medium is cooled to 40-50°C on an ultraclean workbench, then sterile amphotericin B is added (the addition amount is 0.8 mg / L), and then the mixture is shaken thoroughly and divided into 50 mL sterilized triangular flasks for standby.

[0040] The main components of the inorganic phosphorus bacterial culture medium are as follows: glucose 10 g / L, (NH4)2SO4 0.5 g / L, NaCl 0.3 g / L, KCl 0.3 g / L, MgSO4·7H2O 0.3 g / L, FeSO4·7H2O 0.03 g / L, MnSO4·4H2O 0.03 g / L, Ca3(PO4)2 5 g / L, distilled water 1 L, and pH is 7.0.

[0041] Example 1

[0042] The present embodiment provides a preparation method of rhizosphere Cupriavidus basellae-10, which comprises the following steps:

[0043] S1: 0.2 g of a fresh soil sample (rhizosphere soil taken from 1 mm of the root of a one-year-old Phellodendron amurense in the experimental field of the West Garden of Central South University of Forestry and Technology) was weighed into a conical flask containing 50 mL of sterilized water, the flask mouth was covered with a sealing film, and the sealing film was fixed with a rubber band. Then the conical flask containing the rhizosphere soil sample was placed in an ultrasonic cleaner, and was shaken at room temperature for 3 min at a shaking speed of 100 rpm / min to obtain a soil bacteria suspension.

[0044] S2: 1 mL of the soil bacteria suspension was centrifuged at a speed of 6000 rpm for 3 min, and the supernatant was collected. 50 μL of the supernatant was uniformly coated on the surface of R2A solid culture medium, the culture dish was sealed with a sealing film, and was placed in a constant temperature incubator (temperature: 28°C) for 24 h. Then single colonies on the R2A solid culture medium were picked, and were repeatedly purified and observed for growth morphology to obtain single rhizosphere bacteria. A total of 20 independent rhizosphere bacterial strains were isolated.

[0045] S3: The isolated independent rhizosphere bacterial colonies were inoculated into R2A liquid culture medium, and after 11 h of culture, 500 μL of the bacterial solution was mixed with 500 μL of a glycerol solution, was quickly frozen in liquid nitrogen, and then was transferred to a -80°C refrigerator for storage. The results are shown in Figure 1 .

[0046] S4: The rhizosphere bacterial genomic DNA was extracted using a bacterial genomic DNA extraction kit (Omega Bio-Tek (Beijing) Biotechnology Co., Ltd.), and the 16S rDNA sequence was amplified by PCR using primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-CTACGGTACCTT GTTACGA-3'). Then the amplified fragments were sequenced on an Illimina NovaSeq 6000 platform (Shenzhen Microskol Technology Group Co., Ltd.). The 16S rDNA sequence of the strain obtained by sequencing was subjected to BLAST comparison analysis with the NCBI nucleic acid database, and a phylogenetic tree was constructed based on the Neighbor-joining method, Minimum Evolution and Maximum Parsimony using MEGA5.1 software, and the sequence homology was analyzed. The results are shown in Figures 2-4 , according to the 16S sequence analysis results, Bacteria 10 was named Cupriavidus basilensis-10 (CB-10), i.e., the rhizosphere Cupriavidus basilensis-10 of the present embodiment.

[0047] Example 2

[0048] This embodiment provides a phosphorus solubility test of *Basella cordata-10* prepared in the rhizosphere based on Example 1. The specific steps are as follows:

[0049] (1) Preparation of potassium antimony tartrate solution: Weigh 0.5g of potassium antimony tartrate [K(SbO)C4H4O6], dissolve it in 100mL of pure water to prepare a 0.5% potassium antimony tartrate solution.

[0050] (2) Preparation of molybdenum-antimony mixed solution: Take 10g of ammonium molybdate [(NH4)6Mo7O 24 Dissolve [·4H2O] in 450mL of pure water, slowly add 153mL of concentrated sulfuric acid while stirring. Then add 100mL of 0.5% potassium antimony tartrate solution, and finally add water to make up to 1L. Shake well and store in a brown bottle.

[0051] (3) Preparation of molybdenum-antimony anti-color development agent: Before use (i.e., prepare on the same day), weigh 1.5g of L-ascorbic acid, dissolve it in 100mL of molybdenum-antimony mixture, mix well, and the shelf life is 24h.

[0052] (4) 100 μL of *Basella alba*-10 culture was added to 50 mL of inorganic phosphorus liquid medium (culture group). Both the culture group and the control group were cultured at 37℃ and 160 rpm for 3 days with shaking to obtain the fermentation broth. 10 mL of the fermentation broth was then transferred to a centrifuge tube and centrifuged at 5000 rpm for 10 min. The supernatant was collected, and 5 mL of the supernatant was transferred to a 50 mL volumetric flask and diluted to 20 mL with sterile distilled water. Three drops of 2,6-dinitrophenol indicator were slowly added, followed by the addition of 10% sodium hydroxide solution or 10% dilute sulfuric acid solution to adjust the pH until the solution turned slightly yellow. Finally, 5 mL of molybdenum-antimony anti-chromic agent was added, shaken well, and allowed to stand at room temperature for 30 min. The absorbance was then measured at 680 nm to calculate the available phosphorus content. The result for the culture group was 0.30 mg·L⁻¹. -1 The result for the control group was 0.07 mg·L⁻¹. -1 See results Figure 5 .

[0053] pass Figure 5 It can be seen that the available phosphorus content in the rhizosphere *Basella aurea*-10 supplement group was 4.31 times that of the control group (P<0.05), indicating that *Basella aurea*-10 has a strong phosphorus-solubilizing ability.

[0054] Example 3

[0055] This embodiment provides a method for preparing a fungal agent based on *Basellium clavatum*-10 from the rhizosphere prepared in Example 1, comprising the following steps:

[0056] (1) The rhizosphere Cupriavidus basilensis-10 stored at -80°C was activated on R2A medium;

[0057] (2) A single colony of the rhizosphere Cupriavidus basilensis-10 was picked and inoculated into a conical flask containing 20 mL of R2A liquid medium, and cultured at a temperature of 28°C and a rotation speed of 200 rpm / min for 10 h. When the OD value of the bacterial solution was 0.7, a rhizosphere Cupriavidus basilensis-10 inoculum was obtained. 600

[0058] Example 4

[0059] This example provides an effect test of the rhizosphere Cupriavidus basilensis-10 prepared based on Example 1, which is to prove that the rhizosphere Cupriavidus basilensis-10 can promote the growth of the C. fortunei seedlings, increase the pigment content of the C. fortunei seedling leaves, regulate the soluble substance content of the C. fortunei seedling leaves and roots, and increase the effective phosphorus content of the C. fortunei seedling leaves. Specifically,

[0060] (1) The C. fortunei seeds (picked from the Hunan Provincial Botanical Garden) were sown in 50-hole forest nursery hole trays, with 5 seeds per hole, and placed in a greenhouse for germination. After the germinated seedlings grew to about 10 cm, they were transplanted.

[0061] (2) Ten seedlings with similar growth vigor were selected and transferred to plastic flowerpots containing 750 g of nutrient soil (diameter x height of 19.7 cm x 20.5 cm), with one seedling per pot. The potted seedlings were transferred to the C. fortunei experimental base on the third floor of the School of Life and Environmental Sciences of Central South University of Forestry and Technology for growth, with attention paid to watering management. When the C. fortunei seedlings grew to the four-leaf stage (about 40 cm), a rhizosphere Cupriavidus basilensis-10 inoculation experiment was conducted.

[0062] (3) Five of the 10 C. fortunei seedlings were randomly selected and inoculated with 25 mL of rhizosphere Cupriavidus basilensis-10 using the root irrigation method, labeled as the CB-10 group. The remaining 5 C. fortunei seedlings were inoculated with 25 mL of R2A liquid medium as a control group, labeled as the CK group. The C. fortunei seedlings in the CK and CB-10 groups were cultured for 40 d. See Figure 10 .

[0063] 1. Rhizosphere Cupriavidus basilensis-10 promotes the growth of C. fortunei seedlings

[0064] ​The average plant height and ground diameter of the CB-10 group of S. chenmoui seedlings were 47.9 cm and 6.80 mm, respectively, and the average plant height and ground diameter of the CK group of S. chenmoui seedlings were 34.6 cm and 4.93 mm, respectively, which were 38.44% and 37.91% higher than those of the CK group of S. chenmoui seedlings (P<0.05), see Table 1. Figure 6 A and Figure 6 B, the average total fresh weight, leaf fresh weight, stem fresh weight, and root fresh weight of the CB-10 group of S. chenmoui seedlings were 62.54 g, 27.52 g, 7.20 g, and 21.72 g, respectively, and the average total fresh weight, leaf fresh weight, stem fresh weight, and root fresh weight of the CK group of S. chenmoui seedlings were 25.98 g, 11.41 g, 3.30 g, and 9.21 g, respectively, which were 1.40 times, 1.41 times, 1.18 times, and 1.36 times higher than those of the CK group of S. chenmoui seedlings (P<0.05), see Table 2. Figure 6 C- Figure 6 F. At the same time, the average total root length and thick root number of the CB-10 group of S. chenmoui seedlings were 21.67 cm and 13, respectively, and the average total root length and thick root number of the CK group of S. chenmoui seedlings were 19.88 cm and 4, respectively, which were 0.09 times and 2.25 times higher than those of the CK group of S. chenmoui seedlings, see Table 3. Figure 6 G and Figure 6 H. The above results show that the rhizosphere Cupriavidus basilensis-10 can promote the growth of S. chenmoui seedlings.

[0065] 2. Effect of rhizosphere Cupriavidus basilensis-10 on the pigment content of S. chenmoui seedling leaves

[0066] After 40 days of culture, the average chlorophyll a, chlorophyll b, carotenoid, and total chlorophyll in the leaves of the CB-10 group of S. chenmoui seedlings were 6.62 mg / g, 3.55 mg / g, 1.38 mg / g, and 10.18 mg / g, respectively, and the average chlorophyll a, chlorophyll b, carotenoid, and total chlorophyll in the leaves of the CK group of S. chenmoui seedlings were 5.03 mg / g, 2.95 mg / g, 0.74 mg / g, and 7.99 mg / g, respectively, which were 31.62%, 20.34%, 87.99%, and 27.45% higher than those of the CK group of S. chenmoui seedlings, and there were significant differences between the two groups (P<0.05), see Table 4. Figure 7 The above results show that the rhizosphere Cupriavidus basilensis-10 can increase the pigment content of S. chenmoui seedling leaves, and it is speculated that the rhizosphere Cupriavidus basilensis-10 promotes the growth of S. chenmoui seedlings by increasing the content of photosynthetic pigments.

[0067] 3. Test on rhizosphere Cupriavidus basilensis-10 regulating soluble substance content in leaves and roots of P. davidiana seedlings

[0068] After 40 days of culture, the average soluble sugar content in the roots and leaves of P. davidiana seedlings in the CB-10 group was 0.15 mg / g and 0.28 mg / g, respectively, and the average soluble sugar content in the roots and leaves of P. davidiana seedlings in the CK group was 0.12 mg / g and 0.15 mg / g, respectively, which was increased by 25.00% and 86.67% (P<0.05) compared with the average soluble sugar content in the roots and leaves of P. davidiana seedlings in the CK group, see Figure 8 A and Figure 8 B. At the same time, the average fructose content in the roots and leaves of P. davidiana seedlings in the CB-10 group was 48.83 μg / g and 63.63 μg / g, respectively, and the average fructose content in the roots and leaves of P. davidiana seedlings in the CK group was 37.86 μg / g and 67.36 μg / g, respectively, wherein the average fructose content in the roots was increased by 28.99% (P<0.05) compared with the average fructose content in the roots of P. davidiana seedlings in the CK group, and the average fructose content in the leaves was decreased by 5.58% compared with the average fructose content in the leaves of P. davidiana seedlings in the CK group, but there was no significant difference between the two groups, see Figure 8 C and Figure 8 D. The above results show that rhizosphere Cupriavidus basilensis-10 promotes the synthesis and accumulation of soluble sugar in the roots and leaves of P. davidiana seedlings, and promotes the accumulation of fructose in the roots.

[0069] 4. Test on rhizosphere Cupriavidus basilensis-10 increasing effective phosphorus content in leaves of P. davidiana seedlings

[0070] After 40 days of culture, the average effective phosphorus content in the leaves of P. davidiana seedlings in the CB-10 group was 1.29 mg / g, and the average effective phosphorus content in the leaves of P. davidiana seedlings in the CK group was 0.79 mg / g, which was increased by 63.72% (P<0.05) compared with the average effective phosphorus content in the leaves of P. davidiana seedlings in the CK group, see Figure 9 A. At the same time, the average effective phosphorus content in the roots of P. davidiana seedlings in the CB-10 group was 0.31 mg / g, and the average effective phosphorus content in the roots of P. davidiana seedlings in the CK group was 0.40 mg / g, which was decreased by 22.24% compared with the average effective phosphorus content in the roots of P. davidiana seedlings in the CK group, but there was no significant difference between the two groups, see Figure 9 B. The above results show that rhizosphere Cupriavidus basilensis-10 significantly promotes the accumulation of effective phosphorus in the leaves of P. davidiana seedlings, and has no significant effect on the effective phosphorus content in the roots.

[0071] Although the principles of the present application have been described in connection with the preferred embodiments thereof with reference to the drawings, it should be understood that the application is not limited to the construction and arrangements of the preferred embodiments as set forth above and above. The skilled in the art will appreciate that various adaptations and modifications of the preferred embodiments described above can be accomplished using equivalent means, without departing from the scope of the present application. Accordingly, the application is not limited to the specific embodiments described above, but only by the scope of the appended claims.

Claims

1. A rhizosphere Cupriavidus basilensis-10, characterized in that, The rhizosphere Cupriavidus basilensis-10 has a preservation number of GDMCC No: 65501.

2. A rhizosphere inoculant of Cupriavidus basilensis-10, characterized in that, The rhizosphere Cupriavidus basilensis-10 is prepared by the method of claim 1.

3. A method for preparing the rhizosphere bacterial inoculant of Cupriavidus basilensis-10 according to claim 2, characterized by, The method comprises the following steps: When the OD of the bacterial liquid is 0.7, the inoculum of Cupriavidus basilensis-10 is obtained. 600 When the OD of the bacterial liquid is 0.7, the inoculum of Cupriavidus basilensis-10 is obtained.

4. The method of preparing the inoculant of Cupriavidus basilensis-10 according to claim 3, characterized in that, The temperature of the culture is 25-30℃, the time is 8-12h, the rotation speed is 150-250rpm; and / or, the culture medium is R2A liquid medium.

5. Application of the rhizosphere Cupriavidus basilensis-10 of claim 1 or the rhizosphere Cupriavidus basilensis-10 inoculant of claim 2 in promoting the growth of P. fortunei seedlings.

6. Use according to claim 5, characterized in that, The indexes of promoting the growth of P. fortunei seedlings include the plant height, ground diameter, total fresh weight, leaf fresh weight, stem fresh weight, root fresh weight, total root length and thick root number of P. fortunei seedlings; The method for promoting the growth of P. fortunei seedlings by the rhizosphere Cupriavidus basilensis-10 inoculant is as follows: 20-30mL of the rhizosphere Cupriavidus basilensis-10 inoculant is inoculated to each P. fortunei seedling growing to four-leaf stage by the method of root irrigation.

7. Application of the rhizosphere Cupriavidus basilensis-10 of claim 1 or the rhizosphere Cupriavidus basilensis-10 inoculant of claim 2 in increasing the pigment content of P. fortunei seedling leaves.

8. Application of the rhizosphere Cupriavidus basilensis-10 of claim 1 or the rhizosphere Cupriavidus basilensis-10 inoculant of claim 2 in adjusting the soluble substance content of P. fortunei seedling leaves and roots.

9. Application of the rhizosphere Cupriavidus basilensis-10 of claim 1 or the rhizosphere Cupriavidus basilensis-10 inoculant of claim 2 in increasing the effective phosphorus content of P. fortunei seedling leaves.

Images