一株高效解磷真菌新菌株的筛选鉴定及解磷特性
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摘要
从辽宁省辽中县多年耕种的日光温室番茄根际土壤中筛选出一株解磷真菌,通过菌落形态特征和ITS rDNA序列对比,鉴定该菌株为草酸青霉菌的一株新菌株,将其命名为PSF1.该菌株能利用葡萄糖、蔗糖、乳糖、半乳糖、可溶性淀粉等多种碳源和硫酸铵、氯化铵、硝酸铵、硝酸钾、尿素等多种氮源进行生长代谢并表现出较强的解磷能力,在C/N 10∶1～60∶1、初始pH 7～8的条件下生长情况较好且解磷能力较高.该菌株有很强的产酸能力,在培养过程中培养液pH由7.00～7.50下降至2.06～4.87;在4种磷源培养液中的最高解磷量分别为磷酸三钙(869.62 mg·L~(-1))>磷矿粉(233.56 mg·L~(-1))>磷酸铝(44.77 mg·L~(-1))>磷酸铁(28.42 mg·L~(-1)).Pearson相关分析表明,菌株在磷酸三钙、磷矿粉和磷酸铁培养液中的解磷量与pH的变化之间呈极显著负相关关系,在磷酸铝培养液中无显著相关关系.菌株PSF1解磷能力强,生长条件广,推测其在土壤中有较强的解磷能力.
A phosphate solubilizing fungus was isolated from the rhizosphere of tomato in greenhouse in Liaozhong County, Liaoning Province, Northeast China. The strain was identified as a new strain of Penicillium oxalicum by morphological characteristics and ITS rDNA sequence comparison, and then was named PSF1. Strain PSF1 could utilize various carbon sources such as glucose, sucrose, lactose, galactose, soluble starch and nitrogen sources such as ammonium sulfate, ammonium chloride, ammonium nitrate, potassium nitrate, urea for growth and metabolism, with an efficient phosphate solubilizing capacity. It grew well and had a high ability of phosphate solubilization under the conditions of C/N 10:1-60:1 and initial pH 7-8. Strain PSF1 had strong acid production ability, with the pH of culture mediums decreasing from 7.00-7.50 to 2.06-4.87 during the culture process. The highest phosphate solubilizing capacity in four phosphorus sources mediums was tricalcium phosphate(869.62 mg·L~(-1)) > phosphate rock power(233.56 mg·L~(-1)) > aluminum phosphate(44.77 mg·L~(-1)) > iron phosphate(28.42 mg·L~(-1)). Results from Pearson correlation analysis showed that there were significant negative correlations between the changes of phosphate solubili-zing capacity and pH in tricalcium phosphate, phosphate rock power and iron phosphate mediums, but no significant correlation in aluminum phosphate medium. Strain PSF1 had strong phosphate solu-bilizing capacity and wide growing conditions, thus would have strong phosphate solubilizing capacity in soil.
A phosphate solubilizing fungus was isolated from the rhizosphere of tomato in greenhouse in Liaozhong County, Liaoning Province, Northeast China. The strain was identified as a new strain of Penicillium oxalicum by morphological characteristics and ITS rDNA sequence comparison, and then was named PSF1. Strain PSF1 could utilize various carbon sources such as glucose, sucrose, lactose, galactose, soluble starch and nitrogen sources such as ammonium sulfate, ammonium chloride, ammonium nitrate, potassium nitrate, urea for growth and metabolism, with an efficient phosphate solubilizing capacity. It grew well and had a high ability of phosphate solubilization under the conditions of C/N 10:1-60:1 and initial pH 7-8. Strain PSF1 had strong acid production ability, with the pH of culture mediums decreasing from 7.00-7.50 to 2.06-4.87 during the culture process. The highest phosphate solubilizing capacity in four phosphorus sources mediums was tricalcium phosphate(869.62 mg·L~(-1)) > phosphate rock power(233.56 mg·L~(-1)) > aluminum phosphate(44.77 mg·L~(-1)) > iron phosphate(28.42 mg·L~(-1)). Results from Pearson correlation analysis showed that there were significant negative correlations between the changes of phosphate solubili-zing capacity and pH in tricalcium phosphate, phosphate rock power and iron phosphate mediums, but no significant correlation in aluminum phosphate medium. Strain PSF1 had strong phosphate solu-bilizing capacity and wide growing conditions, thus would have strong phosphate solubilizing capacity in soil.
引文
[1] Lu R-K (鲁如坤).The phosphorus level of soil and environmental protection of water body.Phosphate & Compound Fertilizer (磷肥与复肥),2003,18(1):4-8 (in Chinese)
[2] Huang C-Y (黄昌勇).Pedology.3rd Ed.Beijing:China Agriculture Press,2010 (in Chinese)
[3] Kucey RMN.Phosphate-solublizing bacteria and fungi in various cultivated and virgin alberta soils.Canadian Journal of Soil Science,1983,63:671-678
[4] Zhao X-R (赵小蓉),Lin Q-M (林启美).Research progress of phosphate solubilizing microorganisms.Soil and Fertilizer Sciences in China (土壤肥料),2001(3):7-11 (in Chinese)
[5] Chen Z (陈哲),Wu M-N (吴敏娜),Qin H-L (秦红灵),et al.Advances in research on molecular mechanisms of phosphate solubilizing microorganisms in soil.Acta Pedologica Sinica (土壤学报),2009,46(5):925-931 (in Chinese)
[6] Jiang H-H (姜焕焕),Qi P-S (祁佩时),Wang T (王通),et al.Isolation,identification and biological identification of phosphate solubilizing fungi.Chinese Journal of Soil Science (土壤通报),2018,49(4):856-861 (in Chinese)
[7] Li X-P (李学平),Xie W-J (谢文军),Fan Y-H (范延辉).Phosphate-solubilizing ability of Aspergillus tubi-ngensis and its effects on growth of wheat in seedling stage.Bulletin of Soil and Water Conservation (水土保持通报),2017,37(1):93-96 (in Chinese)
[8] Qiao Z-W (乔志伟),Hong J-P (洪坚平),Xie Y-H (谢英荷),et al.Screening,identification and dissolving characteristics of a strong phosphorus solubilizing fungi in calcareous soil.Chinese Journal of Applied and Environmental Biology (应用与环境生物学报),2013,19(5):873-877 (in Chinese)
[9] Chen J (陈俊),Lu J-K (陆俊锟),Kang L-H (康丽华),et al.Primary identification,capability of phosphate-solubilization and optimization of medium of some microorganism from mangrove.Microbiology China (微生物学通报),2009,36(8):1183-1188 (in Chinese)
[10] Wang L-J (王莉晶),Gao X-R (高晓蓉),Lyu J (吕军),et al.Isolation and identification of phosphate solu-bilizing fungus C2’ and its effect on phosphorus solubilization in soil.Chinese Journal of Soil Science (土壤通报),2009,40(4):771-775 (in Chinese)
[11] Kang Y-J (康贻军),Hu J (胡健),Shan J (单君),et al.Solubilization capacity of insoluble phosphates and its mechanism by two phosphate solubilizing fungi (PSF).Microbiology China (微生物学通报),2006,33(5):22-27 (in Chinese)
[12] Asea PEA,Kucey RMN,Stewart JWB.Inorganic phosphate solubilization by two Penicillium species in solution culture and soil.Soil Biology and Biochemistry,1988,20:459-464
[13] Zhao X-R (赵小蓉),Lin Q-M (林启美),Li B-G (李保国).The relationship between rock phosphate solubilization and pH and organic acid production of microorganisms.Journal of Microbiology (微生物学杂志),2003,23(3):5-7 (in Chinese)
[14] Lin Q-M (林启美),Wang H (王华),Zhao X-R (赵小蓉),et al.Capacity of some bacteria and fungi in dissolving phosphate rock.Microbiology China (微生物学通报),2001,28(2):26-30 (in Chinese)
[15] Narsian V,Patel HH.Aspergillus aculeatus as a rock phosphate solubilizer.Soil Biology and Biochemistry,2000,32:559-565
[16] Fan B-Q (范丙全),Jin J-Y (金继运),Ge C (葛诚).Isolation of Penicillium oxalicum and its effect on solubilization of insoluble phosphate under different conditions.Scientia Agricultura Sinica (中国农业科学),2002,35(5):525-530 (in Chinese)
[17] Reyes I,Bernier L,Simard RR,et al.Effect of nitrogen source on the solubilization of different inorganic phosphates by an isolate of Penicillium rugulosum and two UV-induced mutants.FEMS Microbiology Ecology,1999,28:281-290
[18] Zhao X-R (赵小蓉),Lin Q-M (林启美),Li B-G (李保国).Effect of C,N sources and C/N ratio on the solu-bilization of rock phosphate by some microorganisms.Journal of Plant Nutrition and Fertilizers (植物营养与肥料学报),2002,8(2):197-204 (in Chinese)
[19] Liu X-F (刘晓芳),Huang X-D (黄晓东),Kong J (孔健).Effect of C,N sources and C/N ratio on the phosphate solubilization of Aspergillus niger ML2 and ML4.Journal of Shandong University (山东大学学报),2005,40(2):121-124 (in Chinese)
[20] Hu B-W (胡百文).Research on Screening of Phosphate-Solubilizing Bacteria and Their Ecological Characteristics.Master Thesis.Dalian:Dalian Ocean Univer-sity,2014 (in Chinese)
[21] Scervino JM,Papinutti VL,Godoy MS,et al.Medium pH,carbon and nitrogen concentrations modulate the phosphate solubilization efficiency of Penicillium purpurogenum through organic acid production.Journal of Applied Microbiology,2011,110:1215-1223
[22] Beckie HJ,Schlechte D,Moulin AP,et al.Response of alfalfa to inoculation with Penicillium bilaii (Provide).Canadian Journal of Plant Science,1998,78:91-102
[23] Zhang J (张健).Molecular Mechanism of Phosphate Solubilization to Phosphate Deficient Stress in Penicil-lium oxalicum BK.Master Thesis.Dalian:Dalian University of Technology,2014 (in Chinese)
[24] Zhao X-R (赵小蓉),Lin Q-M (林启美),Li B-G (李保国).Preliminary study on solubilizing ability of phosphate solubilizing bacteria to 4 insoluble phosphate species.Acta Microbiologica Sinica (微生物学报),2002,42(2):236-241 (in Chinese)
[25] Shi F-C (史发超),Yin Z-W(殷中伟),Jiang H-M (江红梅),et al.Screening and identification of a phosphate solubilizing fungus and its effect on phosphorus solubilization and growth promotion.Acta Microbiologica Sinica (微生物学报),2014,54(11):1333-1343 (in Chinese)
[26] Xue D (薛冬),Huang X-D (黄向东),Yang R-X (杨瑞先),et al.Screening and phosphate-solubilizing characteristics of phosphate-solubilizing actinomycetes in rhizosphere of tree peony.Chinese Journal of Applied Ecology (应用生态学报),2018,29(5):1645-1652 (in Chinese)
[27] Whitelaw MA,Harden TJ,Bender GL.Plant growth promotion of wheat inoculated with Penicillium radicum sp.Nov.Australian Journal of Soil Research,1997,35:291-300
[28] Wang D (王丹),Zhan J (詹婧),Sun Q-Y (孙庆业).Phosphate solubilization of Aureobasidium pullulan F4 and its mechanism.Chinese Journal of Applied Ecology (应用生态学报),2014,25(7):2079-2084 (in Chinese)
[29] Li W-H (李文红),Shi J-Y (施积炎).Isolation,purification and phosphate-solubilizing capability of phosphorous bacteria in West Lake sediment.Chinese Journal of Applied Ecology (应用生态学报),2006,17(11):2112-2116 (in Chinese)
[30] Hu S (胡山),Niu S-Q (牛世全),Long Y (龙洋),et al.Identification of an efficient phosphorus solu-bilizing bacteria from saline-alkali soil in the Hexi Corridor and optimization of its condition.Microbiology China (微生物学通报),2017,44(2):358-365 (in Chinese)
[31] Qiao Z-W (乔志伟),Hong J-P (洪坚平),Xie Y-H (谢英荷),et al.Screening,identification and phosphate-solubilizing characteristics of Rahnella sp.phosphate-solubilizing bacteria in calcareous soil.Chinese Journal of Applied Ecology (应用生态学报),2013,24(8):2294-2300 (in Chinese)
[32] Wang J-J (王俊娟),Yan A-H (阎爱华),Wang W (王薇),et al.Screening,identification and phosphate-solubilizing characteristics of phosphate-solubili-zing bacteria strain D2 (Pantoea sp.) in rhizosphere of Pinus tabuliformis in iron tailings yard.Chinese Journal of Applied Ecology (应用生态学报),2016,27(11):3705-3711 (in Chinese)
[33] He M-X (贺梦醒),Gao Y (高毅),Hu Z-X (胡正雪),et al.Screening,identification and phosphate solubilizing capability of phosphate-solubilizing bacterial strain B25.Chinese Journal of Applied Ecology (应用生态学报),2012,23(1):235-239 (in Chinese)
[34] Lu LL,Shu YQ.Effect of different carbon sources on phosphate solubilizing fungi isolated from phosphate mines.International Conference on Computer Distributed Controland Intelligent Environmental Monitoring,Changsha,China,2011:1124-1127
[35] Whitelaw MA,Harden TJ,Helyar KR.Phosphate solubilisation in solution culture by the soil fungus Penicil-lium radicum.Soil Biology and Biochemistry,1999,31:655-665
[36] BajPai PD,Sundrara Rao WVB.Phoshpate solubilizing baeteira.Ⅰ.Solubilization of phosphate in liquid culture by selected bacteria as affected by different pH values.Soil Science and Plant Nutrition,1971,17:41-43
[37] Xiao CQ,Chi RA,Hu LH.Solubilization of aluminum phosphate by specific Penicillium spp.Journal of Central South University,2013,20:2109-2114
[38] Zheng X-L (郑肖兰),He C-P (贺春萍),Li R (李锐),et al.Isolation,identification and test of biological property of a phosphorus-solubilizing fungus in rhizosphere of Piper nigrum L.Chinese Journal of Tropical Crops (热带作物学报),2012,33(10):1861-1868 (in Chinese)
[2] Huang C-Y (黄昌勇).Pedology.3rd Ed.Beijing:China Agriculture Press,2010 (in Chinese)
[3] Kucey RMN.Phosphate-solublizing bacteria and fungi in various cultivated and virgin alberta soils.Canadian Journal of Soil Science,1983,63:671-678
[4] Zhao X-R (赵小蓉),Lin Q-M (林启美).Research progress of phosphate solubilizing microorganisms.Soil and Fertilizer Sciences in China (土壤肥料),2001(3):7-11 (in Chinese)
[5] Chen Z (陈哲),Wu M-N (吴敏娜),Qin H-L (秦红灵),et al.Advances in research on molecular mechanisms of phosphate solubilizing microorganisms in soil.Acta Pedologica Sinica (土壤学报),2009,46(5):925-931 (in Chinese)
[6] Jiang H-H (姜焕焕),Qi P-S (祁佩时),Wang T (王通),et al.Isolation,identification and biological identification of phosphate solubilizing fungi.Chinese Journal of Soil Science (土壤通报),2018,49(4):856-861 (in Chinese)
[7] Li X-P (李学平),Xie W-J (谢文军),Fan Y-H (范延辉).Phosphate-solubilizing ability of Aspergillus tubi-ngensis and its effects on growth of wheat in seedling stage.Bulletin of Soil and Water Conservation (水土保持通报),2017,37(1):93-96 (in Chinese)
[8] Qiao Z-W (乔志伟),Hong J-P (洪坚平),Xie Y-H (谢英荷),et al.Screening,identification and dissolving characteristics of a strong phosphorus solubilizing fungi in calcareous soil.Chinese Journal of Applied and Environmental Biology (应用与环境生物学报),2013,19(5):873-877 (in Chinese)
[9] Chen J (陈俊),Lu J-K (陆俊锟),Kang L-H (康丽华),et al.Primary identification,capability of phosphate-solubilization and optimization of medium of some microorganism from mangrove.Microbiology China (微生物学通报),2009,36(8):1183-1188 (in Chinese)
[10] Wang L-J (王莉晶),Gao X-R (高晓蓉),Lyu J (吕军),et al.Isolation and identification of phosphate solu-bilizing fungus C2’ and its effect on phosphorus solubilization in soil.Chinese Journal of Soil Science (土壤通报),2009,40(4):771-775 (in Chinese)
[11] Kang Y-J (康贻军),Hu J (胡健),Shan J (单君),et al.Solubilization capacity of insoluble phosphates and its mechanism by two phosphate solubilizing fungi (PSF).Microbiology China (微生物学通报),2006,33(5):22-27 (in Chinese)
[12] Asea PEA,Kucey RMN,Stewart JWB.Inorganic phosphate solubilization by two Penicillium species in solution culture and soil.Soil Biology and Biochemistry,1988,20:459-464
[13] Zhao X-R (赵小蓉),Lin Q-M (林启美),Li B-G (李保国).The relationship between rock phosphate solubilization and pH and organic acid production of microorganisms.Journal of Microbiology (微生物学杂志),2003,23(3):5-7 (in Chinese)
[14] Lin Q-M (林启美),Wang H (王华),Zhao X-R (赵小蓉),et al.Capacity of some bacteria and fungi in dissolving phosphate rock.Microbiology China (微生物学通报),2001,28(2):26-30 (in Chinese)
[15] Narsian V,Patel HH.Aspergillus aculeatus as a rock phosphate solubilizer.Soil Biology and Biochemistry,2000,32:559-565
[16] Fan B-Q (范丙全),Jin J-Y (金继运),Ge C (葛诚).Isolation of Penicillium oxalicum and its effect on solubilization of insoluble phosphate under different conditions.Scientia Agricultura Sinica (中国农业科学),2002,35(5):525-530 (in Chinese)
[17] Reyes I,Bernier L,Simard RR,et al.Effect of nitrogen source on the solubilization of different inorganic phosphates by an isolate of Penicillium rugulosum and two UV-induced mutants.FEMS Microbiology Ecology,1999,28:281-290
[18] Zhao X-R (赵小蓉),Lin Q-M (林启美),Li B-G (李保国).Effect of C,N sources and C/N ratio on the solu-bilization of rock phosphate by some microorganisms.Journal of Plant Nutrition and Fertilizers (植物营养与肥料学报),2002,8(2):197-204 (in Chinese)
[19] Liu X-F (刘晓芳),Huang X-D (黄晓东),Kong J (孔健).Effect of C,N sources and C/N ratio on the phosphate solubilization of Aspergillus niger ML2 and ML4.Journal of Shandong University (山东大学学报),2005,40(2):121-124 (in Chinese)
[20] Hu B-W (胡百文).Research on Screening of Phosphate-Solubilizing Bacteria and Their Ecological Characteristics.Master Thesis.Dalian:Dalian Ocean Univer-sity,2014 (in Chinese)
[21] Scervino JM,Papinutti VL,Godoy MS,et al.Medium pH,carbon and nitrogen concentrations modulate the phosphate solubilization efficiency of Penicillium purpurogenum through organic acid production.Journal of Applied Microbiology,2011,110:1215-1223
[22] Beckie HJ,Schlechte D,Moulin AP,et al.Response of alfalfa to inoculation with Penicillium bilaii (Provide).Canadian Journal of Plant Science,1998,78:91-102
[23] Zhang J (张健).Molecular Mechanism of Phosphate Solubilization to Phosphate Deficient Stress in Penicil-lium oxalicum BK.Master Thesis.Dalian:Dalian University of Technology,2014 (in Chinese)
[24] Zhao X-R (赵小蓉),Lin Q-M (林启美),Li B-G (李保国).Preliminary study on solubilizing ability of phosphate solubilizing bacteria to 4 insoluble phosphate species.Acta Microbiologica Sinica (微生物学报),2002,42(2):236-241 (in Chinese)
[25] Shi F-C (史发超),Yin Z-W(殷中伟),Jiang H-M (江红梅),et al.Screening and identification of a phosphate solubilizing fungus and its effect on phosphorus solubilization and growth promotion.Acta Microbiologica Sinica (微生物学报),2014,54(11):1333-1343 (in Chinese)
[26] Xue D (薛冬),Huang X-D (黄向东),Yang R-X (杨瑞先),et al.Screening and phosphate-solubilizing characteristics of phosphate-solubilizing actinomycetes in rhizosphere of tree peony.Chinese Journal of Applied Ecology (应用生态学报),2018,29(5):1645-1652 (in Chinese)
[27] Whitelaw MA,Harden TJ,Bender GL.Plant growth promotion of wheat inoculated with Penicillium radicum sp.Nov.Australian Journal of Soil Research,1997,35:291-300
[28] Wang D (王丹),Zhan J (詹婧),Sun Q-Y (孙庆业).Phosphate solubilization of Aureobasidium pullulan F4 and its mechanism.Chinese Journal of Applied Ecology (应用生态学报),2014,25(7):2079-2084 (in Chinese)
[29] Li W-H (李文红),Shi J-Y (施积炎).Isolation,purification and phosphate-solubilizing capability of phosphorous bacteria in West Lake sediment.Chinese Journal of Applied Ecology (应用生态学报),2006,17(11):2112-2116 (in Chinese)
[30] Hu S (胡山),Niu S-Q (牛世全),Long Y (龙洋),et al.Identification of an efficient phosphorus solu-bilizing bacteria from saline-alkali soil in the Hexi Corridor and optimization of its condition.Microbiology China (微生物学通报),2017,44(2):358-365 (in Chinese)
[31] Qiao Z-W (乔志伟),Hong J-P (洪坚平),Xie Y-H (谢英荷),et al.Screening,identification and phosphate-solubilizing characteristics of Rahnella sp.phosphate-solubilizing bacteria in calcareous soil.Chinese Journal of Applied Ecology (应用生态学报),2013,24(8):2294-2300 (in Chinese)
[32] Wang J-J (王俊娟),Yan A-H (阎爱华),Wang W (王薇),et al.Screening,identification and phosphate-solubilizing characteristics of phosphate-solubili-zing bacteria strain D2 (Pantoea sp.) in rhizosphere of Pinus tabuliformis in iron tailings yard.Chinese Journal of Applied Ecology (应用生态学报),2016,27(11):3705-3711 (in Chinese)
[33] He M-X (贺梦醒),Gao Y (高毅),Hu Z-X (胡正雪),et al.Screening,identification and phosphate solubilizing capability of phosphate-solubilizing bacterial strain B25.Chinese Journal of Applied Ecology (应用生态学报),2012,23(1):235-239 (in Chinese)
[34] Lu LL,Shu YQ.Effect of different carbon sources on phosphate solubilizing fungi isolated from phosphate mines.International Conference on Computer Distributed Controland Intelligent Environmental Monitoring,Changsha,China,2011:1124-1127
[35] Whitelaw MA,Harden TJ,Helyar KR.Phosphate solubilisation in solution culture by the soil fungus Penicil-lium radicum.Soil Biology and Biochemistry,1999,31:655-665
[36] BajPai PD,Sundrara Rao WVB.Phoshpate solubilizing baeteira.Ⅰ.Solubilization of phosphate in liquid culture by selected bacteria as affected by different pH values.Soil Science and Plant Nutrition,1971,17:41-43
[37] Xiao CQ,Chi RA,Hu LH.Solubilization of aluminum phosphate by specific Penicillium spp.Journal of Central South University,2013,20:2109-2114
[38] Zheng X-L (郑肖兰),He C-P (贺春萍),Li R (李锐),et al.Isolation,identification and test of biological property of a phosphorus-solubilizing fungus in rhizosphere of Piper nigrum L.Chinese Journal of Tropical Crops (热带作物学报),2012,33(10):1861-1868 (in Chinese)