牡丹根际溶磷放线菌的筛选及其溶磷特性
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摘要
通过从牡丹根际土壤中分离筛选溶磷放线菌,得到一株具有较强溶磷能力的菌株PSPSA1,根据形态特征、生理生化特性以及16S rDNA序列分析对菌株进行鉴定,并研究其溶磷遗传稳定性及溶磷特性.菌株PSPSA1被鉴定为白网链霉菌,具有较好的溶磷遗传稳定性.在不同磷源培养液中溶磷量依次为磷酸钙(158.5 mg·L~(-1))>磷酸铝(139.9 mg·L~(-1))>磷酸铁(127.7 mg·L~(-1))>卵磷脂(45.6 mg·L~(-1)),在无机磷培养液中的溶磷量均与pH呈现显著负相关性,在有机磷培养液中的溶磷量与pH没有显著相关性.在不同碳源条件下的溶磷量依次为乳糖>葡糖糖>麦芽糖>果糖>蔗糖>淀粉>纤维素,在不同氮源条件下的溶磷量依次为蛋白胨>硝酸铵>硫酸铵>硝酸钾>尿素,以葡萄糖为碳源、蛋白胨为氮源时,菌株的溶磷量最高可达202.6 mg·L~(-1).土培60 d,单施菌株土壤有效磷含量比对照增加68.2%,菌株与有机肥混施土壤有效磷含量比单施有机肥增加76.7%.表明菌株PSPSA1能够溶解多种难溶磷,在土壤中溶磷效果显著,与有机肥混施其溶磷能力明显提高,有望成为高效生物磷肥的优良菌种.
A strain PSPSA1 with strong phosphate-solubilizing ability was isolated from the rhizosphere of tree peony. This strain was identified based on morphological characteristics,physiological and biochemical experiments,and 16 S rDNA sequence analysis. Furthermore,its genetic stability and phosphate-solubilizing characteristics were investigated. Strain PSPSA1 was identified as the actinomycete Streptomyces albireticuli and showed high genetic stability. The order of phosphate solubilization capacity( PSC) in different phosphorus-source culture media was calcium phosphate( 158.5 mg·L~(-1)) > aluminum phosphate( 139.9 mg·L~(-1)) > ferric phosphate( 127.7 mg·L~(-1)) >lecithin( 45.6 mg·L~(-1)). The PSC and pH were significantly negatively correlated in inorganic phosphorus solution,but were not correlated in organic phosphorus solution. When using different carbon sources,PSC was in the order of lactose > glucose > maltose > fructose > sucrose > starch >cellulose. For different nitrogen sources,PSC was in the order of peptone > ammonium nitrate >ammonium sulfate > potassium nitrate > urea. Using glucose as a carbon source and peptone as a nitrogen source,strain PSPSA1 achieved the highest PSC of 202.6 mg·L~(-1). After incubated for 60 days,the available phosphorus content in soil with a single application of strain PSPSA1 increased by 68.2% compared with control,and the available phosphorus content in soil with mixed application of strain PSPSA1 and organic fertilizer increased by 76.7% compared with a single application of organic fertilizer. The results indicated that the strain PSPSA1 had a strong ability to dissolve insoluble phosphorus and its mixed application with organic fertilizer improved the phosphorus solubilization. Thus,PSPSA1 is a promising microbial resource for the production of high-efficiency biological phosphorus fertilizer.
A strain PSPSA1 with strong phosphate-solubilizing ability was isolated from the rhizosphere of tree peony. This strain was identified based on morphological characteristics,physiological and biochemical experiments,and 16 S rDNA sequence analysis. Furthermore,its genetic stability and phosphate-solubilizing characteristics were investigated. Strain PSPSA1 was identified as the actinomycete Streptomyces albireticuli and showed high genetic stability. The order of phosphate solubilization capacity( PSC) in different phosphorus-source culture media was calcium phosphate( 158.5 mg·L~(-1)) > aluminum phosphate( 139.9 mg·L~(-1)) > ferric phosphate( 127.7 mg·L~(-1)) >lecithin( 45.6 mg·L~(-1)). The PSC and pH were significantly negatively correlated in inorganic phosphorus solution,but were not correlated in organic phosphorus solution. When using different carbon sources,PSC was in the order of lactose > glucose > maltose > fructose > sucrose > starch >cellulose. For different nitrogen sources,PSC was in the order of peptone > ammonium nitrate >ammonium sulfate > potassium nitrate > urea. Using glucose as a carbon source and peptone as a nitrogen source,strain PSPSA1 achieved the highest PSC of 202.6 mg·L~(-1). After incubated for 60 days,the available phosphorus content in soil with a single application of strain PSPSA1 increased by 68.2% compared with control,and the available phosphorus content in soil with mixed application of strain PSPSA1 and organic fertilizer increased by 76.7% compared with a single application of organic fertilizer. The results indicated that the strain PSPSA1 had a strong ability to dissolve insoluble phosphorus and its mixed application with organic fertilizer improved the phosphorus solubilization. Thus,PSPSA1 is a promising microbial resource for the production of high-efficiency biological phosphorus fertilizer.
引文
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[14]Planting Management,Ministry of Agriculture of the People’s Republic of China(中华人民共和国农业部种植业管理司).General Technical Requirements for Production Strain Quality of Microbial Fertilizer(NY/T1847-2010).Beijing:China Standards Press,2011:4-5(in Chinese)
[15]Chen Y-F(陈宇丰),Ke C-L(柯春亮),Zhou D-B(周登博),et al.Screening,identification and potassium-dissolving characteristics of potassium-dissolving actinomycete in banana rhizosphere soil.Biotechnology Bulletin(生物技术通报),2015,31(6):129-137(in Chinese)
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[17]Bao S-D(鲍士旦).Agricultural and Chemistry Analysis of Soil.Third Edition.Beijing:China Agriculture Press,2000:81-83(in Chinese)
[18]Marcela FC,Angelica Q,Christian D,et al.Evaluation of actinomycete strains for key traits related with plant growth promotion and mycorrhiza helping activities.Applied Soil Ecology,2010,45:209-217
[19]Farhat MB,Boukhris I,Chouayekh H.Mineral phosphate solubilization by Streptomyces sp.CTM396 involves the excretion of gluconic acid and is stimulated by humic acids.FEMS Microbiology Letters,2015,362:1-8
[20]Li Y-E(李玉娥),Yao T(姚拓),Zhu Y(朱颖),et al.Isolation and characteristics of phosphorussolubilizing bacteria in rhizosphere of alfalfa and sainfoin in Lanzhou.Chinese Journal of Grassland(中国草地学报),2009,31(1):45-51(in Chinese)
[21]Liu W-G(刘文干),He Y-Q(何园球),Zhang K(张坤),et al.Isolation,identification and characterization of a strain of phosphate-solubilizing bacteria from red soil.Acta Microbiologica Sinica(微生物学报),2012,52(3):326-333(in Chinese)
[22]Li H-Y(李海云),Niu S-Q(牛世全),Kong W-B(孔维宝),et al.Screening and identification of a phosphate solubilizing strain isolated from pig manure compost and determination of its phosphate solubilizing capacity.Acta Scientiae Circumstantiae(环境科学学报),2015,35(5):1464-1470(in Chinese)
[23]Narsian V,Patel HH.Aspergillus aculeatus as a rock phosphate solubilizer.Soil Biology and Biochemistry,2000,32:559-565
[24]Zhao X-R(赵小蓉),Lin Q-M(林启美),Li B-G(李保国),et al.The solubilization of four insoluble phosphates by some microorganisms.Acta Microbiologica Sinica(微生物学报),2002,42(2):236-241(in Chinese)
[25]Lu R-X(陆瑞霞),Wang X-L(王小利),Li X-G(李显刚),et al.Capability of dissolving phosphate and characteristics of phosphate-dissolving bacteria in rhizosphere of Astragalus bhotanensis in Guizhou.Chinese Journal of Grassland(中国草地学报),2012,34(4):101-108(in Chinese)
[26]Park JH,Bolan N,Megharaj M,et al.Isolation of phosphate solubilizing bacteria and their potential for lead immobilization in soil.Journal of Hazardous Materials,2011,185:829-836
[27]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)
[28]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 p H and organic acid production of microorganisms.Journal of Microbiology(微生物学杂志),2003,23(3):5-7(in Chinese)
[29]Ahuja A,Ghosh SB,D'souza SF.Isolation of a starch utilizing,phosphate solubilizing fungus on buffered medium and its characterization.Bioresource Technology,2007,98:3408-3411
[30]Lindsay WL.Chemical Equilibria in Soils.Caldwell,NJ:The Blackburn Press,2001
[31]Wei M(魏猛),Zhang A-J(张爱君),Zhuge Y-P(诸葛玉平),et al.Effects of long-term fertilization on soil fertility in yellow fluvo-aquic soil.Chinese Journal of Applied Ecology(应用生态学报),2017,28(3):838-846(in Chinese)
[32]Qi R,Li J,Lin Z,et al.Temperature effects on soil organic carbon,soil labile organic carbon fractions,and soil enzyme activities under long-term fertilization regimes.Applied Soil Ecology,2016,102:36-45
[2]Sharma SB,Sayyed RZ,Trivedi MH,et al.Phosphate solubilizing microbes:Sustainable approach formanaging phosphorus deficiency in agricultural soils.Springer Plus,2013,2:587-601
[3]Wang T(王同),Kong L-Y(孔令雅),Jiao J-G(焦加国),et al.Screeing of phosphate-solubilizing bacteria in red soil and their acting mechanisms.Acta Pedologica Sinica(土壤学报),2014,51(2):373-380(in Chinese)
[4]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-solubilizing 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)
[5]Liu Z,Li YC,Zhang S,et al.Characterization of phosphate-solubilizing bacteria isolated from calcareous soils.Applied Soil Ecology,2015,96:217-224
[6]Gong M-B(龚明波),Fan B-Q(范丙全),Jin Z-G(金振国),et al.Screening and application of phosphate-dissolving microorganism suitable for corn production.Acta Microbiologica Sinica(微生物学报),2010,50(12):1619-1625(in Chinese)
[7]Hamdali H,Bouizgarne B,Hafidi M,et al.Screening for rock phosphate solubilizing Actinomycetes from Moroccan phosphate mines.Applied Soil Ecology,2008,38:12-19
[8]Liliensiek AK,Thakuria D,Clipson N.Influences of plant species composition,fertilisation and Lolium perenne ingression on soil microbial community structure in three Irish grasslands.Microbial Ecology,2012,63:509-521
[9]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)
[10]Xue D,Huang XD.Changes in soil microbial community structure with planting years and cultivars of tree peony(Paeonia suffruticosa).World Journal of Microbiology and Biotechnology,2014,2:389-397
[11]Huang XD,Xue D,Xue L.Changes in microbial biomass,activity,functional diversity,and enzyme activity in tree peony(Paeonia suffruticosa)garden soils.HortScience,2014,11:1408-1413
[12]Wang Z(王圳),Zhang J-C(张金池),Cui X-X(崔晓晓),et al.Effects of VA mycorrhiza on rhizosphere nutrient of the herb community in Karst area.Ecology and Environmental Sciences(生态环境学报),2010,19(7):1574-1577(in Chinese)
[13]Yao H-Y(姚槐应),Huang C-Y(黄昌勇).Soil Microbial Ecology and Its Experimental Techniques.Beijing:Science Press,2007(in Chinese)
[14]Planting Management,Ministry of Agriculture of the People’s Republic of China(中华人民共和国农业部种植业管理司).General Technical Requirements for Production Strain Quality of Microbial Fertilizer(NY/T1847-2010).Beijing:China Standards Press,2011:4-5(in Chinese)
[15]Chen Y-F(陈宇丰),Ke C-L(柯春亮),Zhou D-B(周登博),et al.Screening,identification and potassium-dissolving characteristics of potassium-dissolving actinomycete in banana rhizosphere soil.Biotechnology Bulletin(生物技术通报),2015,31(6):129-137(in Chinese)
[16]Actinomycetes Classification Group,Institute of Microbiology,Chinese Academy of Sciences(中国科学院微生物研究所放线菌分类组).Manual of Determinative Streptomyces.Beijing:Science Press,1975:13-16(in Chinese)
[17]Bao S-D(鲍士旦).Agricultural and Chemistry Analysis of Soil.Third Edition.Beijing:China Agriculture Press,2000:81-83(in Chinese)
[18]Marcela FC,Angelica Q,Christian D,et al.Evaluation of actinomycete strains for key traits related with plant growth promotion and mycorrhiza helping activities.Applied Soil Ecology,2010,45:209-217
[19]Farhat MB,Boukhris I,Chouayekh H.Mineral phosphate solubilization by Streptomyces sp.CTM396 involves the excretion of gluconic acid and is stimulated by humic acids.FEMS Microbiology Letters,2015,362:1-8
[20]Li Y-E(李玉娥),Yao T(姚拓),Zhu Y(朱颖),et al.Isolation and characteristics of phosphorussolubilizing bacteria in rhizosphere of alfalfa and sainfoin in Lanzhou.Chinese Journal of Grassland(中国草地学报),2009,31(1):45-51(in Chinese)
[21]Liu W-G(刘文干),He Y-Q(何园球),Zhang K(张坤),et al.Isolation,identification and characterization of a strain of phosphate-solubilizing bacteria from red soil.Acta Microbiologica Sinica(微生物学报),2012,52(3):326-333(in Chinese)
[22]Li H-Y(李海云),Niu S-Q(牛世全),Kong W-B(孔维宝),et al.Screening and identification of a phosphate solubilizing strain isolated from pig manure compost and determination of its phosphate solubilizing capacity.Acta Scientiae Circumstantiae(环境科学学报),2015,35(5):1464-1470(in Chinese)
[23]Narsian V,Patel HH.Aspergillus aculeatus as a rock phosphate solubilizer.Soil Biology and Biochemistry,2000,32:559-565
[24]Zhao X-R(赵小蓉),Lin Q-M(林启美),Li B-G(李保国),et al.The solubilization of four insoluble phosphates by some microorganisms.Acta Microbiologica Sinica(微生物学报),2002,42(2):236-241(in Chinese)
[25]Lu R-X(陆瑞霞),Wang X-L(王小利),Li X-G(李显刚),et al.Capability of dissolving phosphate and characteristics of phosphate-dissolving bacteria in rhizosphere of Astragalus bhotanensis in Guizhou.Chinese Journal of Grassland(中国草地学报),2012,34(4):101-108(in Chinese)
[26]Park JH,Bolan N,Megharaj M,et al.Isolation of phosphate solubilizing bacteria and their potential for lead immobilization in soil.Journal of Hazardous Materials,2011,185:829-836
[27]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)
[28]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 p H and organic acid production of microorganisms.Journal of Microbiology(微生物学杂志),2003,23(3):5-7(in Chinese)
[29]Ahuja A,Ghosh SB,D'souza SF.Isolation of a starch utilizing,phosphate solubilizing fungus on buffered medium and its characterization.Bioresource Technology,2007,98:3408-3411
[30]Lindsay WL.Chemical Equilibria in Soils.Caldwell,NJ:The Blackburn Press,2001
[31]Wei M(魏猛),Zhang A-J(张爱君),Zhuge Y-P(诸葛玉平),et al.Effects of long-term fertilization on soil fertility in yellow fluvo-aquic soil.Chinese Journal of Applied Ecology(应用生态学报),2017,28(3):838-846(in Chinese)
[32]Qi R,Li J,Lin Z,et al.Temperature effects on soil organic carbon,soil labile organic carbon fractions,and soil enzyme activities under long-term fertilization regimes.Applied Soil Ecology,2016,102:36-45