解磷菌发酵及溶磷条件的研究
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摘要
从新疆盐碱土中分离纯化得到一株高效解磷菌PS-3,为改善盐碱地磷素供应提供菌种资源。采用单因素和正交试验,对菌株PS-3的发酵条件和溶磷能力进行分析。结果表明,菌株PS-3较适发酵条件为温度35℃、pH 7和盐浓度1%,其对碳、氮源的利用顺序依次为葡萄糖>麦芽糖>果糖>蔗糖>乳糖,硝酸铵>硝酸钠>氯化铵>硫酸铵>尿素,菌株PS-3最适溶磷条件为2%葡萄糖、0.01%硝酸铵、温度35℃、初始pH 7.5、盐浓度2 g/L、接种量4%,在此条件下,其对磷酸三钙的溶解量可达100 2.95mg/L。该菌株具有一定耐盐性和良好的溶磷能力,在生物溶磷方面具有较好的应用潜力。
The aim of the present work is to isolate a highly effective phosphate-solubilizing strain PS-3 for improving phosphorus supplyof saline-alkali soil in Xinjiang. The single factor experiment and the orthogonal experiment were used to analyze the fermentation conditionsand the phosphate-dissolving capability of strain PS-3. The results showed that the appropriate fermentation conditions of the strain PS-3 weretemperature 35℃,pH 7 and salt concentration 1%. The order of the strain utilizing carbon and nitrogen was glucose > maltose > fructose >sucrose > lactose,and ammonium nitrate > sodium nitrate > ammonium chloride > ammonium sulfate > urea,respectively. The optimalconditions for the solubilization of strain PS-3 were 2% glucose,0.01% ammonium nitrate,temperature 35℃,initial pH 7.5,salt content 2 g/L,and inoculation amount 4%. The phosphate-solubilizing capability reached the level of 1 002.95 mg/L under this condition. The strain PS-3 istolerant to salt stress and has desired phosphate-solubilizing capacity,and thus has a better potential for microbial inoculants with phosphorussolubilizing ability.
The aim of the present work is to isolate a highly effective phosphate-solubilizing strain PS-3 for improving phosphorus supplyof saline-alkali soil in Xinjiang. The single factor experiment and the orthogonal experiment were used to analyze the fermentation conditionsand the phosphate-dissolving capability of strain PS-3. The results showed that the appropriate fermentation conditions of the strain PS-3 weretemperature 35℃,pH 7 and salt concentration 1%. The order of the strain utilizing carbon and nitrogen was glucose > maltose > fructose >sucrose > lactose,and ammonium nitrate > sodium nitrate > ammonium chloride > ammonium sulfate > urea,respectively. The optimalconditions for the solubilization of strain PS-3 were 2% glucose,0.01% ammonium nitrate,temperature 35℃,initial pH 7.5,salt content 2 g/L,and inoculation amount 4%. The phosphate-solubilizing capability reached the level of 1 002.95 mg/L under this condition. The strain PS-3 istolerant to salt stress and has desired phosphate-solubilizing capacity,and thus has a better potential for microbial inoculants with phosphorussolubilizing ability.
引文
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[2]Srinivasan R, Alagawadi AR, Yandigeri MS, et al. Characterizationof phosphate-solubilizing microorganisms from salt-affected soils ofIndia and their effect on growth of sorghum plants[Sorghum bicolor(L.)Moench][J]. Annals of Microbiology, 2012, 62(1):93-105.
[3]Saurabh Jyoti S, Satinder Kaur B, Yann LB, et al. Potentialapplication of biohydrogen production liquid waste as phosphatesolubilizing agent-a study using soybean Plants[J]. Appl BiochemBiotechnol, 2016, 178:865-875.
[4]鲁如坤,时正元,顾益初.土壤积累态磷研究II.一次大量施磷的产量效应[J].土壤, 1995, 27(6):286-289.
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[6]银婷婷,王敬敬,柳影,等.高效解磷菌的筛选及其促生机制的初步研究[J].生物技术通报, 2015, 31(12):234-242.
[7]Swaby RJ, Sperber J. Phosphate dissolving micro-organisms in therhizosphere of legumes[J]. Nutrition of the Legumes, 1958:289-294.
[8]OchoalozaFJ,ArtiolaJF,MaierRM.Stabilityconstantsforthecomplexationofvariousmetalswitharhamnolipidbiosurfactant[J]. Journal of Environmental Quality, 2001, 30(2):479.
[9]Yi YHW, Ge Y. Exopolysaccharide:a novel important factor inthe microbial dissolution of tricalcium phosphate[J]. World JMicrobiol Biotechnol, 2008, 24(7):1059-1065.
[10]赵小蓉,林启美.微生物解磷的研究进展[J].中国土壤与肥料, 2001(3):7-11.
[11]Chen YP, Rekha PD, Arun AB, et al. Phosphate solubilizingbacteria from subtropical soil and their tricalcium phosphatesolubilizing abilities[J]. Applied Soil Ecology, 2006, 34(1):33-41.
[12]Kundu BS, Nehra K, Yadav R, et al. Biodiversity of phosphatesolubilizing bacteria in rhizosphere of chickpea, mustard and wheatgrown in different regions of Haryana[J]. Indian Journal ofMicrobiology, 2009, 49(2):120-127.
[13]Nautiyal CS. An efficient microbiological growth medium forscreening phosphate solubilizing microorganisms[J]. FemsMicrobiology Letters, 1999, 170(1):265-270.
[14]鲍士旦.土壤农化分析.[M].第3版.北京:中国农业出版社,2000.
[15]Bianco C, Defez R. Improvement of phosphate solubilization andMedicago plant yield by an indole-3-acetic acid-overproducingstrain of Sinorhizobium meliloti[J]. Applied&EnvironmentalMicrobiology, 2010, 76(14):4626.
[16]Mehta P, Walia A, Chauhan A, et al. Plant growth promoting traitsof phosphate-solubilizing rhizobacteria isolated from apple trees inTrans Himalayan region of Himachal Pradesh[J]. Archives ofMicrobiology, 2013, 195(5):357-369.
[17]Bhagwanjee S, Scribante J, Paruk F. Application of potentialphosphate-solubilizing bacteria and organic acids on phosphatesolubilization from phosphate rock in aerobic rice[J]. TheScientific World Journal, 2013,(2013-10-8), 2013, 2013(4):272409.
[18]Khan MS, Zaidi A, Wani PA. Role of phosphate-solubilizingmicroorganisms in sustainable agriculture—A review[J].Agronomy for Sustainable Development, 2007, 27(1):29-43.
[19]江红梅,殷中伟,史发超,等.一株耐盐日本曲霉的筛选及其溶磷促生作用[J].微生物学报, 2018, 58(5):862-881.
[20]向文良,冯玮,郭建华,等.一株解磷中度嗜盐菌的分离鉴定及解磷特性分析[J].微生物学通报, 2009, 36(3):320-327.
[21]胡山,牛世全,龙洋,等.河西走廊盐碱土壤中一株高效溶磷菌的鉴定及条件优化[J].微生物学通报, 2017, 44(2):358-365.
[22]Ahuja A, Ghosh SB, D’Souza SF. Isolation of a starch utilizing,phosphate solubilizing fungus on buffered medium and itscharacterization[J]. Bioresource Technology, 2007, 98(17):3408-3411.
[23]Jain R, Saxena J, Sharma V. Effect of phosphate-solubilizing fungiAspergillus awamori S29 on mungbean(Vigna radiata cv. RMG492)growth[J]. Folia Microbiologica, 2012, 57(6):533-541.
[24]卫星,徐鲁荣,张丹,等.一株耐硝酸盐的巨大芽孢杆菌溶磷特性研究[J].环境科学学报, 2015, 35(7):2052-2058.
[25]黄达明,李倩,管国强,等.一株解磷细菌的筛选、鉴定及其溶磷培养条件的优化[J].生物技术通报, 2015, 31(2):173-178.
[26]李海云,牛世全,孔维宝,等.猪粪堆肥中一株溶磷菌的筛选鉴定及溶磷能力初步测定[J].环境科学学报, 2015, 35(5):1464-1470.
[2]Srinivasan R, Alagawadi AR, Yandigeri MS, et al. Characterizationof phosphate-solubilizing microorganisms from salt-affected soils ofIndia and their effect on growth of sorghum plants[Sorghum bicolor(L.)Moench][J]. Annals of Microbiology, 2012, 62(1):93-105.
[3]Saurabh Jyoti S, Satinder Kaur B, Yann LB, et al. Potentialapplication of biohydrogen production liquid waste as phosphatesolubilizing agent-a study using soybean Plants[J]. Appl BiochemBiotechnol, 2016, 178:865-875.
[4]鲁如坤,时正元,顾益初.土壤积累态磷研究II.一次大量施磷的产量效应[J].土壤, 1995, 27(6):286-289.
[5]Stevenson FJ, Cole MA. Cycles of soils:carbon, nitrogen,phosphorus, sulfur, micronutrients[M]. Wiley:Quarterly Reviewof Biology, 1999.
[6]银婷婷,王敬敬,柳影,等.高效解磷菌的筛选及其促生机制的初步研究[J].生物技术通报, 2015, 31(12):234-242.
[7]Swaby RJ, Sperber J. Phosphate dissolving micro-organisms in therhizosphere of legumes[J]. Nutrition of the Legumes, 1958:289-294.
[8]OchoalozaFJ,ArtiolaJF,MaierRM.Stabilityconstantsforthecomplexationofvariousmetalswitharhamnolipidbiosurfactant[J]. Journal of Environmental Quality, 2001, 30(2):479.
[9]Yi YHW, Ge Y. Exopolysaccharide:a novel important factor inthe microbial dissolution of tricalcium phosphate[J]. World JMicrobiol Biotechnol, 2008, 24(7):1059-1065.
[10]赵小蓉,林启美.微生物解磷的研究进展[J].中国土壤与肥料, 2001(3):7-11.
[11]Chen YP, Rekha PD, Arun AB, et al. Phosphate solubilizingbacteria from subtropical soil and their tricalcium phosphatesolubilizing abilities[J]. Applied Soil Ecology, 2006, 34(1):33-41.
[12]Kundu BS, Nehra K, Yadav R, et al. Biodiversity of phosphatesolubilizing bacteria in rhizosphere of chickpea, mustard and wheatgrown in different regions of Haryana[J]. Indian Journal ofMicrobiology, 2009, 49(2):120-127.
[13]Nautiyal CS. An efficient microbiological growth medium forscreening phosphate solubilizing microorganisms[J]. FemsMicrobiology Letters, 1999, 170(1):265-270.
[14]鲍士旦.土壤农化分析.[M].第3版.北京:中国农业出版社,2000.
[15]Bianco C, Defez R. Improvement of phosphate solubilization andMedicago plant yield by an indole-3-acetic acid-overproducingstrain of Sinorhizobium meliloti[J]. Applied&EnvironmentalMicrobiology, 2010, 76(14):4626.
[16]Mehta P, Walia A, Chauhan A, et al. Plant growth promoting traitsof phosphate-solubilizing rhizobacteria isolated from apple trees inTrans Himalayan region of Himachal Pradesh[J]. Archives ofMicrobiology, 2013, 195(5):357-369.
[17]Bhagwanjee S, Scribante J, Paruk F. Application of potentialphosphate-solubilizing bacteria and organic acids on phosphatesolubilization from phosphate rock in aerobic rice[J]. TheScientific World Journal, 2013,(2013-10-8), 2013, 2013(4):272409.
[18]Khan MS, Zaidi A, Wani PA. Role of phosphate-solubilizingmicroorganisms in sustainable agriculture—A review[J].Agronomy for Sustainable Development, 2007, 27(1):29-43.
[19]江红梅,殷中伟,史发超,等.一株耐盐日本曲霉的筛选及其溶磷促生作用[J].微生物学报, 2018, 58(5):862-881.
[20]向文良,冯玮,郭建华,等.一株解磷中度嗜盐菌的分离鉴定及解磷特性分析[J].微生物学通报, 2009, 36(3):320-327.
[21]胡山,牛世全,龙洋,等.河西走廊盐碱土壤中一株高效溶磷菌的鉴定及条件优化[J].微生物学通报, 2017, 44(2):358-365.
[22]Ahuja A, Ghosh SB, D’Souza SF. Isolation of a starch utilizing,phosphate solubilizing fungus on buffered medium and itscharacterization[J]. Bioresource Technology, 2007, 98(17):3408-3411.
[23]Jain R, Saxena J, Sharma V. Effect of phosphate-solubilizing fungiAspergillus awamori S29 on mungbean(Vigna radiata cv. RMG492)growth[J]. Folia Microbiologica, 2012, 57(6):533-541.
[24]卫星,徐鲁荣,张丹,等.一株耐硝酸盐的巨大芽孢杆菌溶磷特性研究[J].环境科学学报, 2015, 35(7):2052-2058.
[25]黄达明,李倩,管国强,等.一株解磷细菌的筛选、鉴定及其溶磷培养条件的优化[J].生物技术通报, 2015, 31(2):173-178.
[26]李海云,牛世全,孔维宝,等.猪粪堆肥中一株溶磷菌的筛选鉴定及溶磷能力初步测定[J].环境科学学报, 2015, 35(5):1464-1470.