| In this paper, using the plate separation method isolation phosphate solubilizing bacteria in Shanxi of calcareous soil, through indoor culture, pot experiment and field experiment to research the combination of phosphate solubilizing bacteria influence on the reclaimed soil, The main conclusions are as follows:1. Using phosphorus bacteria selective medium isolate and screening 440 soil samples of 44 counties in 10 cities in Shanxi province. Initial screened phosphate solubilizing bacteria 147 strains, and let the 147 strains of phosphate solubilizing bacteria inoculate in three calcium phosphate liquid medium, further screened 25 strains bacteria which phosphate solubilizing capacity were greater than 200mg/L After 5 times of purification, there were 12 strains of phosphorus phosphorus solubilizing ability significantly decreased or loss phosphorus solubilizing ability, and eventually selected 13 bacterial strains which were genetic stability and phosphorus solubilizing amount is greater than 200mg/L in three of calcium phosphate liquid culture medium; Identified 13 strains of dissolved phosphorus bacteria through morphological observation, physiological and biochemical identification and 16sRAN sequencing results. Among them,13 strains were belong to Enterobacter sp; 3 strains were belong to Rahnella bacteria; 1 strain was belong to Waxy Bacillus; 2 strains were belong to fluorescent Pseudomonas. Phosphate solubilization capacity of 13 strains of phosphate solubilizing bacteria in the three calcium phosphate liquid medium is 296.5 mg/L-563.5mg/L, in phosphorus slag liquid culture medium the ability of phosphorus solubilizing is 5.35 mg/L-18.49mg/L, in liquid iron phosphate medium phosphate solubilization capacity is 38.33 mg/L-173.63mg/L, in aluminum phosphate liquid medium is 32.03 mg/L-72.07mg/L.2. The maximum value of available phosphorus content in tricalcium phosphate, aluminum phosphate and iron phosphate nutrient solution of soluble phosphorus umaine bacterium W25 respectively were 403.4、110、216.6mg/L. Available phosphorus content in culture and solution pH show significant correlation (p<0.05); Umaine type bacteria W25 had stronger buffering capacity in cultivating 2-4 d, Buffering capacity of umaine type bacteria W25 begined to weaken from cultivating 5 d, after cultivating 7 d W25 basically lost buffering capacity after cultivating 7 d; W25 had the highest ability to dissolve with glucose as carbon source and ammonium nitrate as the nitrogen source. Culture available phosphorus content in tricalcium phosphate liquid medium was 427.1mg/L, The order on the use of carbon of Umaine type bacteria W25 sequentially was glucose> lactose> sucrose> mannitol> starch, and the order on the use of nitrogen sequentially was ammonium nitrate> chloride> sulfate> nitrite> nitrate.3. The concentration of carbon and nitrogen in the medium had a deeper effect on dissolved phosphorus ability of Rahnella.sp W25. Appropriate NO3-/NH4+ was beneficial to the growth and dissolving phosphorus capacity of the strain. Compared with ammonium sulfate as the only nitrogen resource, the capacity of dissolving phosphorus of the strain respectively increased 6.27% and 6.31% when NO3-/NH4+ was 25/75 and 50/50.The capacity of dissolving phosphorus of Rahnella.sp W25 and the species of the organic acids were dissimilar in different culture of carbon, nitrogen and phosphorus. The capacity of dissolving phosphorus of the strain was related to the species and content of the organic acids.4. Phosphorus bacteria Bacillus cereus、Larne type bacteria, Pseudomonas 1,2, and a combination of each other were not antagonistic relationship, available phosphorus content of Larne type bacteria+Pseudomonasl+Pseudomonas2, in culture medium of tricalcium phosphate was 609.10mg/L, from the different considerations of the ability to dissolve tricalcium phosphate with combinations of phosphorus bacteria, we determined the best combination of phosphorus bacteria was Larne type bacteria +Pseudomonasl+Pseudomonas2; The order on the use of carbon of Larne type bacteria+Pseudomonasl+Pseudomonas2 sequentially was glucose> mannitol> sucrose> maltose> starch, the order on the use of nitrogen sequentially was ammonium> potassium> nitrate> urea> ammonium chloride; The growth and phosphate solubilization capacity of Larne type bacteria+Pseudomonasl+ Pseudomonas2 changed with different carbon sources, nitrogen sources, initial pH of culture medium and bacterial inoculum, the best growth conditions of Larne type bacteria+Pseudomonas 1+Pseudomonas2 was glucose 15g/L, ammonium sulfate 0.13g/L, initial pH of 7, bacterial inoculum 3%, the order on all factors of the impact on the growth of Larne type bacteria+Pseudomonasl+Pseudomonas2 sequentially was glucose> ammonium sulfate> the bacterial inoculum> pH; Culture medium formula of the best dissolved phosphorus ability of Larne type bacteria+Pseudomonas 1+Pseudomonas2 was glucose 15g/L, ammonium sulfate 0.67g/L, initial pH of 7, bacterial inoculum 1%, the order on all factors of the impact on soluble phosphorus ability of Larne type bacteria+Pseudomonas 1+Pseudomonas2 sequentially was glucose> pH> the bacterial inoculum> ammonium sulfate, on the best condition, soluble phosphorus ability was 664.29mg/L, significantly increased 55.19 mg/L than ordinary dissolved phosphorus bacteria fermentation medium.5. The number of reclaimed soil phosphorus bacteria reached a maximum after cultivated 7d with different phosphorus bacteria treatment, was 6.5×108, was 65 times,29.5 times,9.3 times,5.4 times of phosphorus bacteria、phosphate bacteria+urinary, phosphate bacteria+glucose, phosphorus bacteria+glucose+urine treatment respectively, the number of soil phosphorus bacteria still kept 3.3×104(CFU/g fresh soil) after cultivated 60d with different phosphorus bacteria treatment, phosphorus bacteria and glucose, urea and manures applied into the reclamation soil, has significantly promoting effect on number of reclaimed soil phosphorus bacteria;On reclamation soil phosphorus bacteria、glucose、urea and manures mixed use, could significantly improve the content of available phosphorus, the content with phosphorus bacteria+glucose+urine+manures treatment was 16.63mg/kg、183.17mg/kg higher than manures、bacteria+glucose treatment respectively; Reclamation soil alkaline phosphorus content with phosphorus bacteria+glucose+urine+manures treatment、phosphorus bacteria+glucose+urine treatment、 phosphorus bacteria treatment were significantly improve 9.56、6.54、2.22 times than phosphorus bacteria treatment respectively, soil acid phosphorus content with phosphorus bacteria+ glucose+urine+manures treatment was 10.33mg/kg, significantly difference with other phosphorus bacteria treatment. Phosphorus bacteria treatment compared with the blank treatment, the maximum soil phosphorus uptake reduced 64.1 mg/kg, adsorption constant reduced 0.008, maximum buffer capacity reduced 10.58 mL/g, desorption phosphorus content increases, the average desorption rate increased by 2.03%, phosphorus bacteria* glucose、urea and manures mixed use treatment has bigger influence on soil adsorption and desorption than phosphorus bacteria treatment, increased 54.95mg/kg, adsorption constant reduced 0.019, maximum buffer capacity reduced 16.93 mL/g, desorption phosphorus content increases, the average desorption rate increased by 20.52%.6 In the whole growth period of rape of the potted rapeseed experiments, among different fertilizer treatments, phosphorus bacteria fertilizer and calcium phosphate on reclamation soil available phosphorus content increased the most effectively, phosphorus bacteria fertilizer plus tricalcium phosphate fertilizer treatment on reclamation soil available p content were higher than other treatments, available phosphorus content with soil phosphorus bacteria fertilizer+ tricalcium phosphate processing in seedling stage increased 0.29-1.61 times than other treatments, Medium-term increased 0.08-3.76 times more than other treatments, harvest time increased 0.40- 4.45 times than other treatments. In the seedling stage and harvest time, among different fertilizer treatments, Ca2-P、Cag-P、Al-P、Fe-P content with phosphorus bacteria fertilizer+ tricalcium phosphate treatment was the highest. In the seedling stage, the soil Ca2-P、Ca8-P、 Al-P and Fe-P content of phosphorus bacteria fertilizer+ tricalcium phosphate treatment were 23.2-43.4 mg/kg,62.3-127.3 mg/kg,32.2-133.1 mg/kg,4.840.1 mg/kg higher than other treatments respectively. In rape harvest time, soil Ca2-P, Ca8-P, Al-P and Fe-P content of phosphorus bacteria fertilizer+ tricalcium phosphate processing were 5.3 to 20.1 mg/kg,63.95 to 97.83 mg/kg,23.9 to 74.3 mg/kg,3.1 to 34.9 mg/kg higher than other treatments respectively. In rape harvest time, compared with seedling stage, reclamation soil Ca10-P content with blank test, organic fertilizer, phosphorus bacteria fertilizer, phosphorus bacteria fertilizer+ tricalcium phosphate, organic fertilizer+ tricalcium phosphate fertilizer, phosphorus bacteria fertilizer+ ground phosphate rock, organic fertilizer+ ground phosphate rock, decreased respectively 0.5%,1.5%,5.4%,5.3%,5.4%,2.8%,1.7%, In harvest time, reclamation soil O- P content with different fertilizer treatments has no change compared with seedling stage.Olsen-P and Ca2-P were significantly positively correlated (P< 0.01), Olsen P and Ca8-P, Al-P, Fe-P have significant correlation (P< 0.05), and have no correlation with O-P, Ca10-P, Olsen P and Ca10-P does not have relevance.Reclamation soil alkaline phosphatase activity increased by 4.54%-118.24% with different fertilizer treatments than the blank test. Soil invertase activity increased by 12.66%-12.66% than the blank test, Soil urease activity increased by 77.01%-77.01% than the blank test, in the soil phosphorus bacteria fertilizer processing, the three kinds of enzyme activity is higher than other processing. Different fertilizer treatments compare to blank test treatment, rapeseed production increased by 16.2%-16.2%, phosphorus bacteria fertilizer+ tricalcium phosphate processing rapeseed yield was 101.6 g/pot, increased by 7.8% to 52.8% than other fertilizer treatments7.1n the field of corn test with different fertilizer treatments, reclamation soil microbial nitrogen content with the phosphorus bacteria fertilizer processing is largest, increased 6.6 times,3.4 times,1.2 times than the blank experiment, regular fertilizing respectively. Soil available p, available k, phosphatase, invertase, urease content was highest with the treatment of phosphorus bacteria fertilizer, Phosphorus bacteria fertilizer treatment, available soil phosphorus higher than other processing, increased by 0.38 to 4.51 times, soil available potassium increased by 17.6% to 64.2%, the soil phosphatase content increased by 20.3% to 35.5%, soil urease content increased by 40.1%-169.7%, soil invertase content increased by 21.1%-90.5%, Applying phosphorus bacteria fertilizer can obviously improve reclaimed soil nutrients and biological activity; In different processing, the order on corn production sequentially was phosphorus bacteria fertilizer> conventional fertilization> matrix blank, Using phosphorus bacteria fertilizer processing significantly increase corn production, was535 kg/mu, compared with the matrix fertilization and regular fertilizing increased by 8.1%,11.1% respectively, on the reclamation soil applying phosphorus bacteria fertilizer can significantly increase the maize yield. |
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