Application of a compound microbial agent in improving nitrogen availability in peanut rhizosphere soil

Abstract

This invention discloses the application of a compound microbial agent in improving nitrogen availability in peanut rhizosphere soil, belonging to the field of microbial technology. This invention is the first to apply a compound microbial agent composed of *Arthrobacter oryzae* (ARZ1) and *Sphingomonas melonis* (SPZ1) to peanut cultivation. The two bacteria in this compound microbial agent work synergistically to significantly promote the increase of total nitrogen, nitrate nitrogen, and ammonia nitrogen content in peanut rhizosphere soil, especially increasing the number of pods, pod weight, and kernel weight per peanut plant. Application during peanut cultivation can significantly increase peanut yield.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of microbial technology, and particularly relates to application of a compound microbial agent in improving nitrogen availability in rhizosphere soil of peanuts. BACKGROUND

[0002] China is one of the countries with the largest arable land in the world. According to statistics, the average utilization rate of nitrogen fertilizer in farmland in China is only 33% in the season, and the nitrogen availability is low. In recent years, the annual loss of urea alone reaches more than 20 million tons. At the same time, low nitrogen fertilizer utilization rate also causes problems such as resource waste, environmental pollution, and decline in crop yield and quality, which seriously restricts the sustainable development of agriculture.

[0003] Peanut is a major oil crop and economic crop in China, and also an important source of export earnings. The total yield of peanuts ranks first among oil crops, and the oil yield is higher than that of other oil crops, which plays a key role in ensuring food and oil security in China and improving the dietary structure. At the same time, peanut rhizobia play a crucial role in nitrogen fixation. These rhizobia can form a symbiotic relationship with the root system of peanuts, and through this symbiotic relationship, they can convert atmospheric nitrogen into nitrogen compounds that can be directly absorbed and utilized by plants. However, excessive nitrogen fertilizer can inhibit the nitrogen fixation of rhizobia, causing a "nitrogen repression" effect. However, when the supply of nitrogen, that is, the nitrogen availability, during the growth of peanuts is synchronized with the demand for nutrients during the growth of peanuts, the utilization rate of nitrogen can be significantly improved. This means that if the nutrient supply can be synchronized with the demand of peanuts through reasonable measures, sufficient nitrogen can be provided in a timely manner during the critical period of peanut growth to meet the needs of different growth and development stages of peanuts, which not only can improve the utilization efficiency of nitrogen, reduce the waste of nitrogen and environmental pollution, but also can promote the healthy growth of peanuts, improve yield and quality.

[0004] Microorganisms are key factors in regulating soil nutrient turnover. If effective microbial agents can be screened to promote nitrogen availability in rhizosphere soil of peanuts and improve nitrogen utilization rate of peanuts, it will have important significance for promoting high-yield and high-quality production of peanuts and sustainable development. SUMMARY

[0005] In view of the deficiencies of the prior art, the purpose of the present application is to provide a compound microbial agent for improving nitrogen availability in rhizosphere soil of peanuts and its application in peanut production.

[0006] In order to achieve the above technical purpose, the present inventors explore the effects of different strains on total nitrogen, ammonia nitrogen, nitrate nitrogen, single plant fruit number, single plant fruit weight and single plant kernel weight in peanut rhizosphere soil through a large number of experiments, and finally obtain the following technical scheme: a plant Arthrobacter oryzae ARZ1, the strain preservation number of which is CCTCC No: M20241883. A Sphingomonas melonis SPZ1, the strain preservation number of which is CCTCC No: M20241884.

[0007] It should be noted that the plant Arthrobacter oryzae ARZ1 used in the present application is isolated from peanut rhizosphere soil, can survive in peanut rhizosphere and soil, grows well in a pH 7.0-9.0 environment, and survives well in a pH 8.0 environment. The strain is identified as plant Arthrobacter oryzae ARZ1 by using morphological characteristics, culture characteristics and physiological and biochemical characteristics of microorganisms, and the strain has been preserved in the China Center for Type Culture Collection on September 2, 2024, with a preservation number of CCTCC No: M20241883.

[0008] The Sphingomonas melonis SPZ1 used in the present application is also isolated from peanut rhizosphere soil, can survive in peanut rhizosphere and soil, grows well in a pH 6.0-9.0 environment, and survives well in a pH 7.0 environment. The strain is identified as Sphingomonas melonis SPZ1 by using morphological characteristics, culture characteristics and physiological and biochemical characteristics of microorganisms, and the strain has been preserved in the China Center for Type Culture Collection on September 2, 2024, with a preservation number of CCTCC No: M20241884.

[0009] The above two strains have a significant positive promoting effect on total nitrogen, ammonia nitrogen, nitrate nitrogen, single plant fruit number, single plant fruit weight and single plant kernel weight in the rhizosphere soil of two peanut varieties, Qinghua No. 6 and Qinghua No. 22, when used alone or in combination, and therefore the present application provides the following new uses:

[0010] (1) Application of ARZ1 strain or SPZ1 strain or complex strain of ARZ1 and SPZ1 in improving total nitrogen content in peanut rhizosphere soil;

[0011] (2) Application of ARZ1 strain or SPZ1 strain or complex strain of ARZ1 and SPZ1 in improving nitrate nitrogen content in peanut rhizosphere soil;

[0012] (3) Application of ARZ1 strain or SPZ1 strain or complex strain of ARZ1 and SPZ1 in increasing the content of ammonia nitrogen in peanut rhizosphere soil;

[0013] (4) Application of ARZ1 strain or SPZ1 strain or complex strain of ARZ1 and SPZ1 in increasing the fruit number of peanut;

[0014] (5) Application of ARZ1 strain or SPZ1 strain or complex strain of ARZ1 and SPZ1 in increasing the fruit weight of peanut;

[0015] (6) Application of ARZ1 strain or SPZ1 strain or complex strain of ARZ1 and SPZ1 in increasing the kernel weight of peanut.

[0016] In the above applications, the ARZ1 strain is Arthrobacter oryzae ARZ1 CCTCC No: M20241883, and the SPZ1 strain is Sphingomonas melonis SPZ1 CCTCC No: M20241884.

[0017] Further preferably, the complex strain is used in a ratio of 1:1 of Sphingomonas melonis SPZ1 and Arthrobacter oryzae ARZ1.

[0018] It should be noted that the present inventors accidentally found that Sphingomonas melonis SPZ1, Arthrobacter oryzae ARZ1 and the complex bacterial liquid of the two strains have a significant effect on increasing the total nitrogen in peanut rhizosphere soil, and can significantly increase the content of ammonia nitrogen and nitrate nitrogen in peanut rhizosphere soil, and can significantly increase the fruit number, fruit weight and kernel weight of single peanut plant. Based on this finding, the present inventors prepared the two strains into a complex microbial agent by inoculating in nitrogen-fixing culture solution for fermentation.

[0019] The complex microbial agent can be prepared by the following method: mixing Sphingomonas melonis SPZ1 bacterial liquid and Arthrobacter oryzae ARZ1 bacterial liquid cultured to logarithmic growth phase, and adding a protective agent to the mixed bacterial liquid, adjusting the viable bacterial count to 1.0×10 10cfu / mL, pH 6-9; the protective agent is composed of 90-120 g / L skim milk powder, 25-35 mL / L glycerol, 90-120 g / L malt dextrin, 120-180 g / L honey locust sugar, and 8-13 g / L sodium L-glutamate.

[0020] The fermentation culture method of Sphingomonas melonis SPZ1 is as follows: Sphingomonas melonis SPZ1 is inoculated into beef extract proteose peptone culture solution at an inoculation amount of 4%-10%, and incubated at 25-27°C for 24-72 hours; after centrifugal washing, a mixed solution containing 0.5 g / L of Yeast Extract, 20 g / L of Mannitol, 0.2 g / L of K2HPO4, 0.8 g / L of KH2PO4, 0.2 g / L of MgSO4·7H2O, 0.1 g / L of CaSO4·2H2O, 0.01 g / L of FeCl3, 0.002 g / L of Na2MoO4·2H2O, and 15 g / L of Agar is added to each bacterial precipitate, pH is adjusted to 6-9, and the viable count of Sphingomonas melonis is adjusted to 1.0×10 10 cfu / mL, and then poured and cultured.

[0021] The fermentation culture method of Arthrobacter oryzae ARZ1 is as follows: Arthrobacter oryzae ARZ1 is inoculated into beef extract proteose peptone culture solution at an inoculation amount of 4%-10%, and incubated at 25-27°C for 24-72 hours; after centrifugal washing, a mixed solution containing 0.5 g / L of Yeast Extract, 20 g / L of Mannitol, 0.2 g / L of K2HPO4, 0.8 g / L of KH2PO4, 0.2 g / L of MgSO4·7H2O, 0.1 g / L of CaSO4·2H2O, 0.01 g / L of FeCl3, 0.002 g / L of Na2MoO4·2H2O, and 15 g / L of Agar is added to each bacterial precipitate, pH is adjusted to 6-9, and the viable count of Arthrobacter oryzae is adjusted to 1.0×10 10 cfu / mL, and then poured and cultured.

[0022] Further preferably, the complex strain is prepared into a product with a total viable count of 1.0×10 10compound microbial agent of 1.0*109cfu / mL; the first application is in the flowering stage of peanut, and the dosage is 2.5-3.5L / 667m 2 ; the second application is in the pod setting stage of peanut, and the dosage is 2.5-3.5L / 667m 2 .

[0023] Further preferably, if the artificial application mode is adopted, the water is diluted and then sprayed on the roots of peanuts by using an agricultural sprayer; if the unmanned aerial vehicle spraying is used, the microbial solution is directly diluted with water and placed in a pesticide tank and sprayed before rain.

[0024] Further preferably, when the peanuts are in the film mulching planting mode, the microbial solution is drip irrigated under the film, and the irrigation depth is 8-10cm to wet the soil.

[0025] Further preferably, no matter which microbial agent application mode is selected, attention should be paid to the fact that the microbial solution and the pesticide cannot be used together, and the interval between the two is more than 2 days.

[0026] Compared with the prior art, the active bacteria in the compound microbial agent are composed of Arthrobacter oryzae ARZ1 and Sphingomonas melonis SPZ1, and the two bacteria in the compound microbial agent have obvious promoting effects on improving the total nitrogen, nitrate nitrogen and ammonium nitrogen contents of the rhizosphere soil of peanuts through synergistic action, especially can improve the fruit number, fruit weight and kernel weight of a single peanut plant, and can greatly improve the yield of peanuts when applied in the process of peanut planting. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 : plate culture and scanning electron microscope image of Arthrobacter oryzae ARZ1;

[0028] Figure 2 : plate culture and scanning electron microscope image of Sphingomonas melonis SPZ1;

[0029] Figure 3 : influence of microbial agent application of each group on total nitrogen of peanut soil;

[0030] Figure 4 : influence of microbial agent application of each group on nitrate nitrogen of peanut soil;

[0031] Figure 5 : influence of microbial agent application of each group on ammonium nitrogen of peanut soil;

[0032] Figure 6 : influence of microbial agent application of each group on fruit number of a single peanut plant;

[0033] Figure 7 Effect of each group of microbial agent application on peanut single plant fruit weight;

[0034] Figure 8 Effect of each group of microbial agent application on peanut single plant kernel weight;

[0035] wherein, Figures 3-8 Different letters in the table represent significant differences (p<0.05) between different treatments in the same group. DETAILED DESCRIPTION

[0036] The above content of the present application will be further described in the form of examples, but it should not be understood that the scope of the above subject matter of the present application is limited to the following examples only, and any technology realized based on the above content of the present application belongs to the scope of the present application. In addition, the experimental methods in the following examples are all conventional methods unless otherwise specified.

[0037] Example 1: Screening and isolation of ARZ1 and SPZ1 strains

[0038] (1) The discovery, screening, isolation and identification process of the strain. Based on the screening of corn and peanut intercropping specific root exudates addition test, the inventors found that root exudate addition can increase the nitrogen content of peanut rhizosphere soil and stimulate the nitrogen absorption of peanut plants. Therefore, we speculate that the addition of root exudates reassembles the soil microbial community, thereby promoting the absorption and utilization of soil nitrogen by peanuts. On this basis, we made a suspension of the peanut rhizosphere soil sample under the above root exudate addition treatment, coated the bacterial liquid on the bacterial culture medium, and carried out strain isolation and purification. The 16S rRNA sequence of the isolated strain was obtained by using high-throughput sequencing technology, and the 16S rRNA sequence was compared with the nr database of NCBI. The sequence clustering analysis was carried out by comparison degree and similarity, and the phylogenetic tree was constructed. The isolated and purified strain was compared with known strains, and comprehensive analysis was carried out in terms of morphology, physiology and biochemistry, etc. to determine the bacterial species. In order to further verify the function of the strain, the isolated and purified strain was used to carry out peanut cultivation addition test.

[0039] (2) Microbial properties. Arthrobacter oryzae ARZ1 colonies are white, round, wet ( Figure 1 ), can grow on nitrogen fixation medium, the suitable growth pH is 6-9, and the temperature is 25-27℃. Sphingomonas melonis SPZ1 colonies are yellow, round, small and wet, smooth and opaque, with neat edges and no halo ( Figure 2 ), can grow on nitrogen fixation medium, the suitable growth pH is 6-9, and the temperature is 25-27℃.

[0040] Example 2: Preparation of ARZ1, SPZ1 inoculants and complex inoculants

[0041] (1) Arthrobacter oryzae ARZ1 was inoculated into beef extract peptone culture solution (beef extract 3 g / L, peptone 10 g / L, Nacl 5 g / L, agar 15 g / L) at an inoculation amount of 4%-10%, and incubated at 25-27°C for 24-72 hours. After centrifugal washing, 0.5 g / L of Yeast Extract, 20 g / L of Mannitol, 0.2 g / L of K2HPO4, 0.8 g / L of KH2PO4, 0.2 g / L of MgSO4·7H2O, 0.1 g / L of CaSO4·2H2O, 0.01 g / L of FeCl3, 0.002 g / L of Na2MoO4·2H2O, and 15 g / L of Agar were added to the bacterial precipitate, and the pH was adjusted to 6-9. The viable count of Arthrobacter oryzae ARZ1 was adjusted to about 1.0 x 10 10 cfu / mL, and mixed uniformly. The suspension was then poured into the culture solution and incubated. The specific operation method was as follows: the Arthrobacter oryzae ARZ1 suspension was diluted and added to sterile nitrogen-fixing culture medium. After solidification, the culture was incubated upside down. Single colonies were taken and suspended repeatedly to obtain pure culture.

[0042] (2) Sphingomonas melonis SPZ1 was inoculated into beef extract peptone culture solution (beef extract 3 g / L, peptone 10 g / L, Nacl 5 g / L, agar 15 g / L) at an inoculation amount of 4%-10%, and incubated at 25-27°C for 24-72 hours. After centrifugal washing, 0.5 g / L of Yeast Extract, 20 g / L of Mannitol, 0.2 g / L of K2HPO4, 0.8 g / L of KH2PO4, 0.2 g / L of MgSO4·7H2O, 0.1 g / L of CaSO4·2H2O, 0.01 g / L of FeCl3, 0.002 g / L of Na2MoO4·2H2O, and 15 g / L of Agar were added to the bacterial precipitate, and the pH was adjusted to 6-9. The viable count of Sphingomonas melonis SPZ1 was adjusted to about 1.0 x 10 10cfu / mL, mixed and then poured into culture; the specific operation method of pouring culture is: after diluting Sphingomonas melonis SPZ1 suspension, adding it into sterile nitrogen fixation medium, inverting culture after solidification, single strain proliferation to form colonies, taking single colony to prepare suspension, repeating multiple times to obtain pure culture.

[0043] (3) Preparation of protective agent, composition: 100 g / L skimmed milk powder, 30 mL / L glycerol, 100 g / L malt dextrin, 150 g / L mucuna sugar and 10 g / L L-glutamic acid sodium, pH 6-9.

[0044] (4) Adding the protective agent prepared in step (3) to the plant Arthrobacter ARZ1 bacterial liquid obtained in step (1), adjusting the viable count to about 1.0×10 10 cfu / mL, to obtain ARZ1 bacterial agent. Adding the protective agent prepared in step (3) to the Sphingomonas melonis SPZ1 bacterial liquid obtained in step (2), adjusting the viable count to about 1.0×10 10 cfu / mL, to obtain SPZ1 bacterial agent.

[0045] Mixing the bacterial liquids obtained in steps (1) and (2) according to the viable count ratio of Sphingomonas melonis SPZ1: plant Arthrobacter ARZ1 being 1:1, and adding the protective agent prepared in step (3) to the mixed bacterial liquid, adjusting the total effective viable count to about 1.0×10 10 cfu / mL, to obtain a composite bacterial agent.

[0046] Example 3: Test of ARZ1 bacterial agent in pot peanut planting

[0047] Test type: pot experiment

[0048] Test site: Qingdao Agricultural University (Qingdao, China)

[0049] Test time: May 2023-September 2023

[0050] Test crop: peanuts (variety Qinghua No. 6, Qinghua No. 22)

[0051] Test soil: sterilized soil (soil pH 6.70, organic matter 11.92 g / kg, alkali hydrolysis nitrogen 99.86 mg / kg, available phosphorus 35.22 mg / kg, available potassium 89.79 mg / kg).

[0052] Test groups: CK is blank control, ARZ1-F and ARZ1-T are treated with plant Arthrobacter ARZ1 bacterial agent, each treatment has 3 repeats, and they are arranged randomly in groups.

[0053] Test design: PVC pots were used for potting, with an inner diameter of 16 cm and a pot height of 23.5 cm. Bacterial agent addition was performed 25 days after peanut seeding (mixing the bacterial solution with 200 mL water and pouring it onto the peanut roots), and sampling was performed at the harvest period after bacterial solution addition.

[0054] Amount and method: ARZ1-F was applied with 0.05 mL / pot of ARZ1 bacterial agent prepared in Example 2, ARZ1-T was applied with 0.1 mL / pot of ARZ1 bacterial agent prepared in Example 2, the bacterial solution was diluted to 200 mL respectively and poured onto the peanut roots, and the CK was 200 mL of blank water solution.

[0055] Table 1 Soil total nitrogen content under treatment of plant Arthrobacter ARZ1

[0056]

[0057] Note: Different letters represent significant differences between different treatments in the same group (p < 0.05).

[0058] From the test results in Table 1, it can be seen that ARZ1-T treatment has a significant effect on the improvement of soil total nitrogen content of two peanut varieties. The soil total nitrogen content of Qinghua No. 6 under ARZ1-T treatment is significantly increased by 352.08% and 124.54% compared with CK and ARZ1-F respectively. The soil total nitrogen content of Qinghua No. 22 under ARZ1-T treatment is significantly increased by 116.35% and 125.00% compared with CK and ARZ1-F respectively. In summary, plant Arthrobacter ARZ1 (Arthrobacter oryzae) ARZ1-T treatment can significantly improve the total nitrogen content in the rhizosphere of peanuts.

[0059] Example 4: Test of SPZ1 bacterial agent in pot-grown peanut planting

[0060] Test type: potting test

[0061] Test site: Qingdao Agricultural University (Qingdao, China)

[0062] Test time: May 2023-September 2023

[0063] Test crop: peanuts (varieties Qinghua No. 6 and Qinghua No. 22)

[0064] Test soil: sterilized soil (soil pH 6.70, organic matter 11.92 g / kg, alkali hydrolyzable nitrogen 99.86 mg / kg, available phosphorus 35.22 mg / kg, available potassium 89.79 mg / kg).

[0065] Test groups: CK is the blank control, SPZ1-F and SPZ1-T are both treated with bacterial agent of guaia sphingomonas SPZ1, each treatment has 3 repeats, and the arrangement is random block.

[0066] Test design: PVC pots were used for potting, with an inner diameter of 16 cm and a pot height of 23.5 cm. Bacterial agent addition was performed 25 days after peanut seeding (mix the bacterial solution with 200 ml water and pour it onto the peanut roots), and sampling was performed at the harvest period after bacterial solution addition.

[0067] Usage and method: SPZ1-F was applied with 0.05 mL of SPZ1 bacterial agent prepared in Example 2 per pot, and SPZ1-T was applied with 0.1 mL of SPZ1 bacterial agent prepared in Example 2 per pot. The bacterial solution was diluted to 200 mL, respectively, and poured onto the peanut roots. The CK was 200 mL of blank water solution.

[0068] Table 2 Soil total nitrogen content under Sphingomonas melonis SPZ1 treatment

[0069] Note: Different letters represent significant differences between different treatments in the same group (p < 0.05).

[0070] From the test results in Table 2, it can be seen that SPZ1-T treatment has a significant effect on the improvement of soil total nitrogen content of the two varieties. The soil total nitrogen content of Qinghua No. 6 under SPZ1-T treatment was significantly increased by 69.00% and 53.64% compared with CK and SPZ1-F, respectively. The soil total nitrogen content of Qinghua No. 22 under SPZ1-T treatment was significantly increased by 27.88% and 12.71% compared with CK and SPZ1-F, respectively. In summary, Sphingomonas melonis SPZ1-T treatment can significantly improve the soil total nitrogen content of peanut plants.

[0071] Example 5: Test of compound bacterial agent in field peanut planting

[0072] Test site: Pingdu City, Qingdao City, Shandong Province

[0073] Test time: May 2024-September 2024

[0074] Test crop: peanuts (varieties Qinghua No. 6 and Qinghua No. 22)

[0075] Test soil: The soil is sand slurry black soil (soil pH is 7.06, organic matter is 16.18 g / kg, alkali-hydrolyzable nitrogen is 77.76 mg / kg, available phosphorus is 45.58 mg / kg, and available potassium is 78.69 mg / kg).

[0076] Test groups:

[0077] CK is a blank water solution control,

[0078] ARZ1-F group was applied with ARZ bacterial agent prepared in Example 2 (total number of living bacteria was about 1.0 x 10 10 cfu / mL) at an application amount of 1.5 L per 667 m 2 ;

[0079] ARZ1-T group was applied with ARZ bacterial agent prepared in Example 2 (total number of living bacteria was about 1.0 x 10 10 cfu / mL) at an application amount of 3 L per 667 m 2 ;

[0080] SPZ1-F group was applied with SPZ1 bacterial agent prepared in Example 2 (total number of living bacteria was about 1.0 x 10 10 cfu / mL) at an application amount of 1.5 L per 667 m 2 ;

[0081] SPZ1-T group was applied with SPZ1 bacterial agent prepared in Example 2 (total number of living bacteria was about 1.0 x 10 10 cfu / mL) at an application amount of 3 L per 667 m 2 ;

[0082] ST+AT group was applied with the compound bacterial agent prepared in Example 2, i.e. mixing the gua sphingomonas SPZ1 and the plant arthrobacter ARZ1 in a ratio of 1:1 (total number of living bacteria was about 1.0 x 10 10 cfu / mL) at an application amount of 3 L per 667 m 2 ;

[0083] Application amount and method: the first application, at the beginning of peanut flowering, the bacterial agent amount designed in each test group mentioned above was diluted to 15 L with clean water, and then sprayed on the peanut roots by artificial application using an agricultural sprayer; the second application, the amount and application method were the same as the first application. It should be noted that the bacterial solution should not be used together with pesticides, and the two should be used more than 2 days apart.

[0084] Planting management: the peanuts were planted by covering the ridges, with a ridge distance of 90 cm, 2 rows per ridge, a small row distance of 35 cm on the ridge, and a hole distance of 11 cm (2 seeds per hole). Compound fertilizer (N-P2O5-K2O: 15-15-15) was applied at 50 kg per mu, and no additional fertilizer was applied in the later period.

[0085] Test results:

[0086] 1) Effect on total nitrogen in peanut soil

[0087] Through Figure 3The experimental results show that, compared with CK and four single strain groups, the complex strain ST+AT treatment can significantly improve the soil total nitrogen content of two peanut varieties. Among them, compared with the control group CK, the soil total nitrogen of Qinghua No. 6 is significantly increased by 442.29%, and the soil total nitrogen of Qinghua No. 22 is significantly increased by 179%. Statistical analysis shows that the soil total nitrogen content under the treatment of complex strain ST+AT is significantly or extremely significantly higher than that of single strain addition.

[0088] 2) Effect on soil nitrate nitrogen of peanut

[0089] Through Figure 4 The experimental results show that, compared with CK and four single strain groups, the complex strain ST+AT treatment can significantly improve the soil total nitrogen content of two peanut varieties. Among them, compared with the control group CK, the soil total nitrogen of Qinghua No. 6 is significantly increased by 442.29%, and the soil total nitrogen of Qinghua No. 22 is significantly increased by 179%. Statistical analysis shows that the soil total nitrogen content under the treatment of complex strain ST+AT is significantly or extremely significantly higher than that of single strain addition.

[0090] 3) Effect on soil ammonium nitrogen of peanut

[0091] Through Figure 5 The experimental results show that, compared with CK and four single strain groups, the complex strain ST+AT treatment can significantly improve the soil total nitrogen content of two peanut varieties. Among them, compared with the control group CK, the soil total nitrogen of Qinghua No. 6 is significantly increased by 442.29%, and the soil total nitrogen of Qinghua No. 22 is significantly increased by 179%. Statistical analysis shows that the soil total nitrogen content under the treatment of complex strain ST+AT is significantly or extremely significantly higher than that of single strain addition.

[0092] 4) Effect on the number of single plant results of peanut

[0093] Through Figure 6 The experimental results show that, compared with CK and four single strain groups, the complex strain ST+AT treatment can significantly improve the soil total nitrogen content of two peanut varieties. Among them, compared with the control group CK, the soil total nitrogen of Qinghua No. 6 is significantly increased by 442.29%, and the soil total nitrogen of Qinghua No. 22 is significantly increased by 179%. Statistical analysis shows that the soil total nitrogen content under the treatment of complex strain ST+AT is significantly or extremely significantly higher than that of single strain addition.

[0094] 5) Effect on the single plant fruit weight of peanut

[0095] Through Figure 7The results of the comparison of the test results show that, compared with CK and four single strain groups, the compound strain ST+AT treatment can significantly improve the single plant fruit weight of two peanut varieties. Among them, compared with the control group CK, under the treatment of compound strain ST+AT, the single plant fruit weight of Qinghua No. 6 peanut is significantly increased by 69.41%, and the single plant fruit weight of Qinghua No. 22 peanut is significantly increased by 97.53%. Statistical analysis shows that the single plant fruit weight of peanuts treated with compound strain ST+AT is significantly or extremely significantly higher than that treated with single strain.

[0096] 6) Effect on peanut single plant kernel weight

[0097] Through Figure 8 The results of the comparison of the test results show that, compared with CK and four single strain groups, the compound strain ST+AT treatment can significantly improve the single plant fruit weight of two peanut varieties. Among them, compared with the control group CK, under the treatment of compound strain ST+AT, the single plant fruit weight of Qinghua No. 6 peanut is significantly increased by 69.41%, and the single plant fruit weight of Qinghua No. 22 peanut is significantly increased by 97.53%. Statistical analysis shows that the single plant fruit weight of peanuts treated with compound strain ST+AT is significantly or extremely significantly higher than that treated with single strain.

[0098] In summary, through statistical comparison, compared with single strain and blank control, the content of total nitrogen, nitrate nitrogen and ammonium nitrogen in soil and the results number, fruit weight and kernel weight of single plant peanut treated with compound inoculant ST+AT are significantly improved. This shows that the compound inoculant prepared by Sphingomonas melonis SPZ1 and Arthrobacteroryzae ARZ1 (the total number of effective viable bacteria is about 1.0×10 10 cfu / mL) can significantly improve the content of total nitrogen, nitrate nitrogen and ammonium nitrogen in peanut rhizosphere soil, and can significantly improve the results number, fruit weight and kernel weight of peanuts. Applying it to the process of peanut planting can improve the nitrogen availability and yield of peanuts in the rhizosphere.

Claims

1. Application of ARZ1 strain or SPZ1 strain or complex strain of ARZ1 and SPZ1 in improving total nitrogen content in rhizosphere soil of peanut, wherein the ARZ1 strain is plant Arthrobacter sp. Arthrobacter oryzae ARZ1 CCTCC No: M20241883, and the SPZ1 strain is Pseudomonas sp. Sphingomonas melonis SPZ1 CCTCC No: M20241884.

2. The use of ARZ1 strain or SPZ1 strain or the complex strain of ARZ1 and SPZ1 in improving the nitrate nitrogen content in the rhizosphere soil of peanuts, wherein the ARZ1 strain is Arthrobacter sp. Arthrobacter oryzae ARZ1 CCTCC No: M20241883, and the SPZ1 strain is Sphingomonas sp. Sphingomonas melonis SPZ1 CCTCC No: M20241884.

3. The use of ARZ1 strain or SPZ1 strain or the complex strain of ARZ1 and SPZ1 in increasing the content of ammonia nitrogen in rhizosphere soil of peanuts, wherein the ARZ1 strain is Arthrobacter sp. Arthrobacter oryzae ARZ1 CCTCC No: M20241883, and the SPZ1 strain is Sphingomonas sp. Sphingomonas melonis SPZ1 CCTCC No: M20241884.

4. The use of ARZ1 strain or SPZ1 strain or the complex strain of ARZ1 and SPZ1 in increasing the number of single plant results, wherein the ARZ1 strain is Arthrobacter sp. ARZ1 CCTCC No: M20241883, and the SPZ1 strain is Sphingomonas sp. SPZ1 CCTCC No: M20241884. Arthrobacter oryzae ARZ1 CCTCC No:M20241883, and the SPZ1 strain is Sphingomonas sp. SPZ1 CCTCC No: M20241884. Sphingomonas melonis )SPZ1 CCTCC No:M20241884.

5. The use of ARZ1 strain or SPZ1 strain or the complex strain of ARZ1 and SPZ1 in increasing the single plant fruit weight of peanut, wherein the ARZ1 strain is Arthrobacter sp. ARZ1 CCTCC No: M20241883, and the SPZ1 strain is Sphingomonas sp. SPZ1 CCTCC No: M20241884. Arthrobacter oryzae ARZ1 CCTCC No:M20241883, the SPZ1 strain is Sphingomonas sp. ( Sphingomonas melonis ) SPZ1 CCTCC No: M20241884.

6. The use of ARZ1 strain or SPZ1 strain or the complex strain of ARZ1 and SPZ1 in increasing the single seed weight of peanut, wherein the ARZ1 strain is Arthrobacter sp. ARZ1 CCTCC No: M20241883, and the SPZ1 strain is Sphingomonas sp. SPZ1 CCTCC No: M20241884. Arthrobacter oryzae ARZ1 CCTCC No:M20241883, and the SPZ1 strain is Sphingomonas sp. SPZ1 CCTCC No: M20241884. Sphingomonas melonis )SPZ1 CCTCC No:M20241884.

7. Use according to any one of claims 1 to 6, characterized in that, The aforementioned compound strain, when used, contains Sphingosine monocytogenes of Cucurbita ( Sphingomonas melonis SPZ1 and Plantar Arthrobacter Arthrobacter oryzae The ratio of live bacteria in ARZ1 is 1:

1.

8. Use according to claim 7, characterized in that, The complex strain is prepared into a bacterial liquid with a total number of living bacteria of 1.0×10 10 cfu / mL; the first application is performed at the beginning of the peanut flowering period, with a dosage of 2.5-3.5 L / 667 m 2 ; the second application is performed at the peanut pod setting period, with a dosage of 2.5-3.5 L / 667 m 2 .

9. Use according to claim 7, characterized in that, If artificial application is adopted, after dilution with clean water, agricultural sprayer is used to spray peanut roots; if unmanned aerial vehicle spraying is used, the diluted bacterial solution is directly placed in the pesticide tank and sprayed before rain.

10. Use according to claim 7, characterized in that, When the peanut is planted in a film mulching mode, the bacterial solution is drip irrigated by using the film mulching drip irrigation method, and the irrigation depth is 8-10 cm to wet the soil.

Images