大豆根际溶磷真菌的筛选、复配及包埋固定化回用效果
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摘要
为获得具高效溶磷能力的溶磷真菌,本研究从大豆根际土壤中分离筛选出5株具有溶解Ca_3(PO_4)_2、AlPO_4、FePO_4及磷矿粉能力的高效菌株,分别命名为Z2、Z3、Z8、T4和Y2,分析了其对不同磷源的溶解能力,复配了最优溶磷组合同时评价了最优组合固定化回用的效果。结果表明:菌株Z2为黄色蓝状菌(Talaromyces flavus),Z3为绳状篮状菌(Talaromyces funiculosus),Z8为黑曲霉(Aspergilus niger),T4为嗜松蓝状菌(Talaromyces pinophilus),Y2为糙刺蓝状菌(Talaromyces trachyspermus)。培养11 d内,Z2、Z3、Z8、T4及Y2对Ca_3(PO_4)_2的最大溶磷量分别为610.51 mg/mL、674.39 mg/mL、687.10 mg/mL、667.06 mg/mL和629.08 mg/m L,对Al PO_4的最大溶磷量分别为485.76 mg/mL、505.59 mg/mL、559.72 mg/m L、495.97 mg/mL和540.02 mg/mL,对FePO_4的最大溶磷量分别为504.60 mg/mL、511.57 mg/m L、553.21 mg/mL、508.23 mg/m L和520.42 mg/mL以及对磷矿粉的最大溶磷量分别为139.78mg/mL、135.31mg/mL、159.03mg/mL、154.61mg/m L和154.43mg/mL。溶磷过程中,溶磷量与pH值呈现极显著的负相关性,并测得5株真菌释放有机酸含量分别为228.6 mL/L、91.5 mL/L、7 254.1 mL/L、314.4 mL/L和284.6 mL/L。对无拮抗反应的菌株进行随机复配组合,发现菌株Z3、Z8和Y2组成的复配组合溶磷能力最强,其对Ca_3(PO_4)_2溶磷量高达779.94 mg/m L。最优复配组合包埋固定回用至土壤后,土壤中有效磷含量得到明显提升,最高效磷含量可增长至113.5 mg/kg,增长率为127.3%。本研究为微生物溶磷菌肥的开发提供菌种资源,也可为今后的进一步应用研究提供科学依据。
In order to obtain phosphorus-dissolving fungi with high-efficiency phosphorus-dissolving ability,five high-efficiency strains with the ability to dissolve Ca_3(PO_4)_2,AlPO_4,FePO_4and phosphate rock powder were isolated and isolated from soybean rhizosphere soil,respectively named Z2.Z3,Z8,T4 and Y2,the solvency of different phosphorus sources was analyzed,and the optimal phosphorus combination was compounded.The effect of optimal combination and reuse was evaluated.The results showed that the strain Z2 was Talaromyces flavus,Z3was Talaromyces funiculosus,Z8 was Aspergilus niger,T4 was Talaromyces pinophilus,and Y2 was rough.Talaromyces trachyspermus,the maximum phosphorus uptake of Ca_3(PO_4)_2by Z2,Z3,Z8,T4 and Y2 was 610.51 mg/m L,674.39 mg/m L,687.10 mg/mL,667.06 mg/mL and 629.08 mg/mL within 11 days of culture,respectively.The maximum phosphorus solubilization of Al PO_4was 485.76 mg/mL,505.59 mg/mL,559.72 mg/m L,495.97 mg/m L and 540.02 mg/m L,respectively.The maximum phosphorus solubilization of FePO_4was 504.60 mg/m L,511.57 mg/mL,553.21 mg/mL,508.23 mg/mL and 520.42 mg/mL and phosphate rock.The maximum phosphorus solubilization of the powder was 139.78 mg/mL,135.31 mg/m L,159.03 mg/mL,154.61 mg/m L and 154.43 mg/mL respectively.In the process of phosphorus solubilization,the amount of dissolved phosphorus showed a significant negative correlation with the pH value.The organic acid content of the five fungi was 228.6 m L/L,91.5 m L/L,7 254.1 m L/L,314.4 mL/L and 284.6 mL/L.Randomly compounded the strains without antagonistic reaction,it was found that the combination of strains Z3,Z8 and Y2 had the strongest ability to dissolve phosphorus,and the amount of phosphorus dissolved in Ca_3(PO_4)_2was as high as 779.94 mg/mL.After the optimal compounding combination was embedded and fixed to the soil,the available phosphorus content in the soil was significantly improved,and the most efficient phosphorus content could be increased to 113.5 mg/kg,with a growth rate of 127.3%.This study provides strain resources for the development of microbial phosphate solubilizing fertilizer,and can provide scientific basis for further application research in the future.
In order to obtain phosphorus-dissolving fungi with high-efficiency phosphorus-dissolving ability,five high-efficiency strains with the ability to dissolve Ca_3(PO_4)_2,AlPO_4,FePO_4and phosphate rock powder were isolated and isolated from soybean rhizosphere soil,respectively named Z2.Z3,Z8,T4 and Y2,the solvency of different phosphorus sources was analyzed,and the optimal phosphorus combination was compounded.The effect of optimal combination and reuse was evaluated.The results showed that the strain Z2 was Talaromyces flavus,Z3was Talaromyces funiculosus,Z8 was Aspergilus niger,T4 was Talaromyces pinophilus,and Y2 was rough.Talaromyces trachyspermus,the maximum phosphorus uptake of Ca_3(PO_4)_2by Z2,Z3,Z8,T4 and Y2 was 610.51 mg/m L,674.39 mg/m L,687.10 mg/mL,667.06 mg/mL and 629.08 mg/mL within 11 days of culture,respectively.The maximum phosphorus solubilization of Al PO_4was 485.76 mg/mL,505.59 mg/mL,559.72 mg/m L,495.97 mg/m L and 540.02 mg/m L,respectively.The maximum phosphorus solubilization of FePO_4was 504.60 mg/m L,511.57 mg/mL,553.21 mg/mL,508.23 mg/mL and 520.42 mg/mL and phosphate rock.The maximum phosphorus solubilization of the powder was 139.78 mg/mL,135.31 mg/m L,159.03 mg/mL,154.61 mg/m L and 154.43 mg/mL respectively.In the process of phosphorus solubilization,the amount of dissolved phosphorus showed a significant negative correlation with the pH value.The organic acid content of the five fungi was 228.6 m L/L,91.5 m L/L,7 254.1 m L/L,314.4 mL/L and 284.6 mL/L.Randomly compounded the strains without antagonistic reaction,it was found that the combination of strains Z3,Z8 and Y2 had the strongest ability to dissolve phosphorus,and the amount of phosphorus dissolved in Ca_3(PO_4)_2was as high as 779.94 mg/mL.After the optimal compounding combination was embedded and fixed to the soil,the available phosphorus content in the soil was significantly improved,and the most efficient phosphorus content could be increased to 113.5 mg/kg,with a growth rate of 127.3%.This study provides strain resources for the development of microbial phosphate solubilizing fertilizer,and can provide scientific basis for further application research in the future.
引文
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Jain R.,Saxena J.,and Sharma V.,2014,Differential effects of immobilized and free forms of phosphate-solubilizing fungal strains on the growth and phosphorus uptake of mung bean plants,Ann.Microbiol.,64(4):1523-1534
Jiang H.M.,Yin Z.W.,Shi F.C.,Liu C.Y.,and Fan B.Q.,2018,Isolation,identification of a salt-tolerant,phosphate-solubilizing and crop-growth promoting Aspergillus japonicas,Weishengwu Xuebao(Acta Microbiologica Sinica),58(5):862-881(江红梅,殷中伟,史发超,刘彩月,范丙全,2018,一株耐盐日本曲霉的筛选及其溶磷促生作用,微生物学报,58(5):862-881)
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Kaur G.,and Reddy M.S.,2015,Effects of phosphate-solubilizing bacteria,rock phosphate and chemical fertilizers on maize-wheat cropping cycle and economics,Pedosphere,25(3):428-437
Ram H.,Malik S.S.,Dhaliwal S.S.,Kumar D.B.,and Singh Y.,2015,Growth and productivity of wheat affected by phosphorus-solubilizing fungi and phosphorus levels,Plant Soil Environ.,61(3):122-126
Li F.C.,Wang Z.H.,Dai J.,Li Q.,Wang X.,Xue C.,Liu H.,and He G.,2015,Fate of nitrogen from green manure,straw,and fertilizer applied to wheat under different summer fallow management strategies in dryland,Biol.Fertil.Soils,51(7):769-780
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Lin X.,2018,The promoting effect of phosphate solubilizing bacteria on remediation of cadmium-contaminated saline soil by Suaeda salsa,Thesis for M.S.,Jinan University,Su pervisor:Li Q.S.,pp.12-13(林欣,2018,一株溶磷菌对盐地碱蓬修复盐渍土镉污染的促进效应,硕士学位论文,暨南大学,导师:李取生,pp.12-13)
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Muleta D.,2010,Legume responses to arbuscular mycorrhizal fungi inoculation in sustainable agricultur,In:Khan M.S.,Musarrat J.,and Zaidi A.,(eds.),Microbes for legume improvement,Springer,Vienna,Austria,pp.293-323
Rashid M.,Khalil S.,Ayub N.,Alam S.,and Latif F.,2004,Organic acids production and phosphate solubilization by phosphate solubilizing microorganisms(PSM)under in vitro conditions,Pakistan J.Biol.Sci.,7(2):187-196
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Schoumans O.F.,Chardon W.J.,Bechmann M.E.,Gascuel-Odoux C.,Hofman G.,Kronvang B.,Rubaek G.H.,Ulén B.,and Dorioz J.M.,2014,Mitigation options to reduce phosphorus losses from the agricultural sector and improve surface water quality:A review,Sci.Total Environ.,468-469:1255-1266
Shao K.,2017,Screening and identification of phosphorous microorganism and preparation of microcapsule fertilizer,Thesis for M.S.,Suzhou University of Science and Technology,Supervisors:Qiu Y.X.,and Wang Y.L.,pp.50-53(邵锴,2017,高效溶磷菌的筛选、鉴定及微胶囊菌肥制备,硕士学位论文,苏州科技大学,导师:邱业先,王友良,pp.50-53)
Singh H.,and Reddy M.S.,2011,Effect of inoculation with phosphate solubilizing fungus on growth and nutrient uptake of wheat and maize plants fertilized with rock phosphate in alkaline soils,Eur.J.Soil Biol.,47(1):30-34
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de Oliveira Mendes G.,de Freitas A.L.M.,Pereira O.L.,da Silva I.R.,Vassilev N.B.,and Costa M.D.,2014,Mechanisms of phosphate solubilization by fungal isolates when exposed to different P sources,Ann.Microbiol.,64(1):239-249
Elias F.,Woyessa D.,and Muleta D.,2016,Phosphate solubilization potential of rhizosphere fungi isolated from plants in jimma zone,southwest ethiopia,Int.J.Microbiol.,2016(3):1-11
Elsas J.D.V.,Chiurazzi M.,Mallon C.A.,Elhottova D.,Kristufek V.,Salles J.F.,and Salles J.F.,2012,Microbial diversity determines the invasion of soil by a bacterial pathogen,Proc.Natl.Acad.Sci.USA,109(4):1159-1164
Fan Y.H.,Wang J.,Liu X.H.,Zhang L.Y.,and Li X.B.,2015,Isolation,identification and fermentation optimization of a halotolerant strain with phosphate-solubilizing activity,Turang Tongbao(Chinese Journal of Soil Science),46(2):362-367(范延辉,王君,刘雪红,张丽燕,李小彬,2015,一株耐盐解磷真菌的筛选、鉴定及其发酵优化,土壤通报,46(2):362-367)
Garcia-Conesa M.T.,Crepin V.F.,Goldson A.J.,Williamson G.,Cummings N.J.,Connerton I.F.,Faulds C.B.,Kroon P.A.,2004,The feruloyl esterase system of Talaromyces stipitatus:production of three discrete feruloyl esterases,including a novel enzyme,Ts FaeC,with a broad substrate specificity,J.Biotechnol.,108(3):227-241
Gong X.,Wang J.H.,Guan J.F.,Yang X.C.,and Chen D.C.,2014,Impact of reclaimed water irrigation on soil chemical properties and culturable microorganisms,Huanjing Kexue(Environmental Science),35(9):3572-3579(龚雪,王继华,关健飞,杨雪辰,陈黛慈,2014,再生水灌溉对土壤化学性质及可培养微生物的影响,环境科学,35(9):3572-3579)
Illmer P.,and Schinner F.,1995,Solubilization of inorganic calcium phosphates-solubilization mechanisms,Soil Biol.Biochem.,27(3):257-263
Jain R.,Saxena J.,and Sharma V.,2010,The evaluation of free and encapsulated Aspergillus awamori for phosphate solubilization in fermentation and soil-plant system,Appl.Soil Ecol.,46(1):90-94
Jain R.,Saxena J.,and Sharma V.,2012,Solubilization of inorganic phosphates by Aspergillus awamori S19 isolated from rhizosphere soil of a semi-arid region,Ann.Microbiol.,62(2):725-735
Jain R.,Saxena J.,and Sharma V.,2014,Differential effects of immobilized and free forms of phosphate-solubilizing fungal strains on the growth and phosphorus uptake of mung bean plants,Ann.Microbiol.,64(4):1523-1534
Jiang H.M.,Yin Z.W.,Shi F.C.,Liu C.Y.,and Fan B.Q.,2018,Isolation,identification of a salt-tolerant,phosphate-solubilizing and crop-growth promoting Aspergillus japonicas,Weishengwu Xuebao(Acta Microbiologica Sinica),58(5):862-881(江红梅,殷中伟,史发超,刘彩月,范丙全,2018,一株耐盐日本曲霉的筛选及其溶磷促生作用,微生物学报,58(5):862-881)
Juarez Jimenez B.,Reboleiro Rivas P.,Gonzalez L.J.,Pesciaroli C.,Barghini P.,and Fenice M.,2012,Immobilization of Delftia tsuruhatensis in macro-porous cellulose and biodegradation of phenolic compounds in repeated batch process,J.Biotechnol.,157(1):148-153
Kaur G.,and Reddy M.S.,2015,Effects of phosphate-solubilizing bacteria,rock phosphate and chemical fertilizers on maize-wheat cropping cycle and economics,Pedosphere,25(3):428-437
Ram H.,Malik S.S.,Dhaliwal S.S.,Kumar D.B.,and Singh Y.,2015,Growth and productivity of wheat affected by phosphorus-solubilizing fungi and phosphorus levels,Plant Soil Environ.,61(3):122-126
Li F.C.,Wang Z.H.,Dai J.,Li Q.,Wang X.,Xue C.,Liu H.,and He G.,2015,Fate of nitrogen from green manure,straw,and fertilizer applied to wheat under different summer fallow management strategies in dryland,Biol.Fertil.Soils,51(7):769-780
Li S.P.,2017,Biological characteristics of two strains of Trichoderma spp.isolated from rice rhizosphere and their biocontrol potential research,Thesis for M.S.,Huazhong Agricultural University,Supervisor:Xie J.T.,pp.44-46(李松鹏,2017,两株水稻根际木霉菌株生物学特性及生防潜能研究,硕士学位论文,华中农业大学,导师:谢甲涛,pp.44-46)
Lin X.,2018,The promoting effect of phosphate solubilizing bacteria on remediation of cadmium-contaminated saline soil by Suaeda salsa,Thesis for M.S.,Jinan University,Su pervisor:Li Q.S.,pp.12-13(林欣,2018,一株溶磷菌对盐地碱蓬修复盐渍土镉污染的促进效应,硕士学位论文,暨南大学,导师:李取生,pp.12-13)
Mendes G.D.O.,Freitas A.L.M.,Pereira O.L.,Silva I.R.,Vassilev N.B.,and Costa M.D.,2014,Mechanisms of phosphate solubilization by fungal isolates when exposed to different Psources,Ann.Microbiol.,64(1):239-249
Muleta D.,2010,Legume responses to arbuscular mycorrhizal fungi inoculation in sustainable agricultur,In:Khan M.S.,Musarrat J.,and Zaidi A.,(eds.),Microbes for legume improvement,Springer,Vienna,Austria,pp.293-323
Rashid M.,Khalil S.,Ayub N.,Alam S.,and Latif F.,2004,Organic acids production and phosphate solubilization by phosphate solubilizing microorganisms(PSM)under in vitro conditions,Pakistan J.Biol.Sci.,7(2):187-196
Scervino J.M.,Mesa M.P.,Mo仵nica I.D.,Recchi M.,Moreno S.,and Godeas A.,2013,Soil fungal isolates produce different organic acid patterns involved in phosphate salts solubilization,Biol.Fertil.Soils,49(6):779-779
Schoumans O.F.,Chardon W.J.,Bechmann M.E.,Gascuel-Odoux C.,Hofman G.,Kronvang B.,Rubaek G.H.,Ulén B.,and Dorioz J.M.,2014,Mitigation options to reduce phosphorus losses from the agricultural sector and improve surface water quality:A review,Sci.Total Environ.,468-469:1255-1266
Shao K.,2017,Screening and identification of phosphorous microorganism and preparation of microcapsule fertilizer,Thesis for M.S.,Suzhou University of Science and Technology,Supervisors:Qiu Y.X.,and Wang Y.L.,pp.50-53(邵锴,2017,高效溶磷菌的筛选、鉴定及微胶囊菌肥制备,硕士学位论文,苏州科技大学,导师:邱业先,王友良,pp.50-53)
Singh H.,and Reddy M.S.,2011,Effect of inoculation with phosphate solubilizing fungus on growth and nutrient uptake of wheat and maize plants fertilized with rock phosphate in alkaline soils,Eur.J.Soil Biol.,47(1):30-34
Smith D.R.,King K.W.,Johnson L.,Francesconi W.,Richards P.,Baker D.,and Sharpley A.N.,2015,Surface runoff and tile drainage transport of phosphorus in the midwestern United States,J.Environ.Qual.,44(2):495
Sperber J.,1958,The incidence of apatite-solubilizing organisms in the rhizosphere and soil,Aust.J.Agr.Res.,9(6):778-781
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