牛蒡根际土壤解磷菌的筛选、鉴定及生长特性
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摘要
为了提高牛蒡根际土壤的有效磷含量,以促进牛蒡的生长,采用解磷圈法及磷钼蓝比色法,从牛蒡根际土壤中筛选具有较高解磷能力的细菌,并对其进行鉴定,研究其生长特性。结果表明,筛选获得1株高效解磷菌JL-1,其在无机磷液体培养基发酵2 d,解磷量为50. 4μg/m L。对其进行形态学观察、生理生化鉴定及16S r DNA序列分析,确定其为Acinetobacter indicus,此菌株在温度26～37℃、pH值6. 5～8. 0条件下,生长较好,最适温度为30℃,最适p H值为7. 0;可以耐受NaCl质量浓度为60 g/L,生长最适NaCl质量浓度为10 g/L。
To increase the effective phosphate concentration in Arctium lappa rhizosphere soil and promote the growth of Arctium lappa,phosphate-solubilizing circle method and phosphomolybdenum blue colorimetry were applied to isolate phosphate-solubilizing bacteria,which was then identified,and the growth characteristics of which was also studied. The results showed that a strain of phosphate-solubilizing bacteria named JL-1 was screened,2 d after fermentation in liquid inorganic phosphorus medium,the content of solubilizing phosphate reached 50. 4 μg/m L. The strain JL-1 was identified as Acinetobacter indicus through morphologic characteristics,physiological and biochemical characteristics and 16 S r DNA sequence analysis. The better temperature and pH value range for growth of Acinetobacter indicus JL-1 were 26—37 ℃ and 6. 5—8. 0,and the best temperature,p H value for growth of JL-1 were 30 ℃,7. 0,respectively. The NaCl tolerance concentration of JL-1 was 60 g/L,and the optimal NaCl concentration for growth of JL-1 was 10 g/L.
To increase the effective phosphate concentration in Arctium lappa rhizosphere soil and promote the growth of Arctium lappa,phosphate-solubilizing circle method and phosphomolybdenum blue colorimetry were applied to isolate phosphate-solubilizing bacteria,which was then identified,and the growth characteristics of which was also studied. The results showed that a strain of phosphate-solubilizing bacteria named JL-1 was screened,2 d after fermentation in liquid inorganic phosphorus medium,the content of solubilizing phosphate reached 50. 4 μg/m L. The strain JL-1 was identified as Acinetobacter indicus through morphologic characteristics,physiological and biochemical characteristics and 16 S r DNA sequence analysis. The better temperature and pH value range for growth of Acinetobacter indicus JL-1 were 26—37 ℃ and 6. 5—8. 0,and the best temperature,p H value for growth of JL-1 were 30 ℃,7. 0,respectively. The NaCl tolerance concentration of JL-1 was 60 g/L,and the optimal NaCl concentration for growth of JL-1 was 10 g/L.
引文
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[22]郝晶,洪坚平,谢英荷,等.石灰性土壤磷细菌的分离、筛选及解磷效果[J].山西农业科学,2005,33(4):56-59.
[23]巩文峰,邢瑜琪,卓玛曲措,等.一株色季拉山长鞭红景天根际溶磷菌的分离、鉴定及其低温适应性分析[J].南方农业学报,2018,49(2):280-286.
[24]代志,高俊明.兼具解磷解钾功能生防菌分离鉴定及效果评价[J].山西农业科学,2018,46(4):627-633.
[25]杨慧,范丙全,龚明波,等.一株新的溶磷草生欧文氏菌的分离、鉴定及其溶磷效果的初步研究[J].微生物学报,2008,48(1):51-56.
[26]陈倩颖.解有机磷细菌的分离鉴定及其解磷特性研究[D].合肥:安徽农业大学,2009.
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[28]布坎南R E,吉本斯N E.伯杰细菌鉴定手册[M]//中国科学院生物研究所《伯杰细菌鉴定手册》翻译组. 8版.北京:科学出版社,1984.
[29] Cheng H R,Jiang N. Extremely rapid extraction of DNA from bacteria and yeasts[J]. Biotechnology Letters,2006,28(1):55-59.
[30] Karen M K,Sherry L H,Douglas C N. Novel,attached,sulfur-oxidizing bacteria at shallow hydrothermal vents possess vacuoles not involved in respiratory nitrate accumulation[J]. Applied and Environmental Microbiology,2004,70:7487-7496.
[31]杨天学,席北斗,栗越妍,等.耐高温解无机磷菌解磷条件的优化[J].环境科学研究,2009,22(3):36-40.
[32]刘玉华,王慧,胡晓珂.不动杆菌属(Acinetobacter)细菌降解石油烃的研究进展[J].微生物学通报,2016,43(7):1579-1589.
[33]张慧智,史学正,于东升,等.中国土壤温度的季节性变化及其区域分异研究[J].土壤学报,2009,46(2):227-234.
[2] Eftekhari S G H,Falah N A R,Ali A G,et al. Effect of phosphate solubilizing bacteria and phosphate fertilizers on rice growth parameters[J]. Iranian Journal of Soil Research,2009,23(2):229-238.
[3] Rodríguez H,Fraga R. Phosphate solubilizing bacteria and their role in plant growth promotion[J]. Biotechnology Advances,1999,17(4/5):319-339.
[4] Zeng Q W,Wu X Q,Wen X Y. Identification and characterization of the rhizosphere phosphate-solubilizing bacterium Pseudomonas frederiksbergensis,JW-SD2,and its plant growth-promoting effects on poplar seedlings[J].Annals of Microbiology,2017,67(3):219-230.
[5]郑少玲,陈琼贤,谭炽强,等.解磷细菌对难溶性磷的有效化作用[J].华南农业大学学报,2007,28(4):38-41.
[6] Vassilev N,Eichler-L9bermann B,Vassileva M. Stress-tolerant P-solubilizing Microorganisms[J]. Applied Microbiology and Biotechnology,2012,95(4):851.
[7] Hayat R,Ali S,Amara U,et al. Soil beneficial bacteria and their role in plant growth promotion:A review[J].Annals of Microbiology,2010,60(4):579-598.
[8] Kuhad R C,Singh S,Lata,et al. Phosphate-solubilizing microorganisms[J]. Soil Biology,2011,108:65-84.
[9]戴沈艳,贺云举,申卫收,等.一株高效解磷细菌的紫外诱变选育及其在红壤稻田施用效果[J].生态环境学报,2010,19(7):1646-1652.
[10] Richardson A E. Prospects for using soil microorganisms to improve the acquisition of phosphorus by plants[J].Australian Journal of Plant Physiology,2001,28(9):897-906.
[11]王海东,吴文华,王印,等.牛蒡成分提取、活性研究及加工利用概述[J].食品工业,2014,35(4):164-167.
[12]常慧萍,邢文会,夏铁骑,等.根际促生细菌的筛选及其对小麦幼苗的促生作用[J].河南农业科学,2016,45(12):52-57.
[13]郎敬.大豆根际解磷菌分离及其定殖分析[D].哈尔滨:东北林业大学,2014.
[14]夏觅真,马忠友,曹媛媛,等.棉花根际固氮菌、解磷菌及解钾菌的相互作用[J].中国微生态学杂志,2010,22(2):102-105.
[15]孙婷婷,陈晏,樊剑波,等.土壤悬液培养法研究长期施肥下花生根际解磷菌溶磷特性[J].土壤学报,2017,54(1):227-236.
[16]刘小玉,付登强,贾效成,等.油茶根际土壤解磷菌的筛选、鉴定及培养条件[J].西南农业学报,2016,29(11):2637-2642.
[17]杨艳华,李俊州,臧睿,等.茶树根际土壤解磷细菌的筛选及解磷活性分析[J].河南农业科学,2014,43(9):60-65.
[18]万兵兵,刘晔,吴越,等.烟草根际解磷解钾菌的筛选鉴定及应用效果研究[J].河南农业科学,2016,45(9):46-51.
[19]李海峰,张月阳,曹健,等.耐寡营养高效解磷菌株XMT-5的分离鉴定及解磷特性[J].河南农业科学,2017,46(8):67-71.
[20]屈建航,张璐洁,符运会,等. 1株高效溶磷细菌的分离、鉴定和溶磷能力研究[J].河南农业科学,2018,47(3):55-58,91.
[21]张淑红.兼溶5种难溶磷的溶磷细菌筛选及其对花生的促生作用[J].河南农业科学,2015,44(11):58-61,72.
[22]郝晶,洪坚平,谢英荷,等.石灰性土壤磷细菌的分离、筛选及解磷效果[J].山西农业科学,2005,33(4):56-59.
[23]巩文峰,邢瑜琪,卓玛曲措,等.一株色季拉山长鞭红景天根际溶磷菌的分离、鉴定及其低温适应性分析[J].南方农业学报,2018,49(2):280-286.
[24]代志,高俊明.兼具解磷解钾功能生防菌分离鉴定及效果评价[J].山西农业科学,2018,46(4):627-633.
[25]杨慧,范丙全,龚明波,等.一株新的溶磷草生欧文氏菌的分离、鉴定及其溶磷效果的初步研究[J].微生物学报,2008,48(1):51-56.
[26]陈倩颖.解有机磷细菌的分离鉴定及其解磷特性研究[D].合肥:安徽农业大学,2009.
[27]黄亚东,时小艳.微生物实验技术[M].北京:中国轻工业出版社,2012.
[28]布坎南R E,吉本斯N E.伯杰细菌鉴定手册[M]//中国科学院生物研究所《伯杰细菌鉴定手册》翻译组. 8版.北京:科学出版社,1984.
[29] Cheng H R,Jiang N. Extremely rapid extraction of DNA from bacteria and yeasts[J]. Biotechnology Letters,2006,28(1):55-59.
[30] Karen M K,Sherry L H,Douglas C N. Novel,attached,sulfur-oxidizing bacteria at shallow hydrothermal vents possess vacuoles not involved in respiratory nitrate accumulation[J]. Applied and Environmental Microbiology,2004,70:7487-7496.
[31]杨天学,席北斗,栗越妍,等.耐高温解无机磷菌解磷条件的优化[J].环境科学研究,2009,22(3):36-40.
[32]刘玉华,王慧,胡晓珂.不动杆菌属(Acinetobacter)细菌降解石油烃的研究进展[J].微生物学通报,2016,43(7):1579-1589.
[33]张慧智,史学正,于东升,等.中国土壤温度的季节性变化及其区域分异研究[J].土壤学报,2009,46(2):227-234.