高效解磷细菌菌株CT45-1的鉴定及其对烟草的促生作用
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摘要
使用以Ca_3(PO_4)_2为唯一磷源的NBRIP液体培养基,对一株来源于潍坊烟区根际土壤的高效解磷细菌菌株CT45-1的解磷能力进行定量测定分析,并利用分子生物学方法对该菌株进行鉴定;通过盆栽试验,了解菌株CT45-1对烟株的促生作用。定量分析结果表明,随着培养时间的增加,接种菌株CT45-1的液体培养基中可溶性磷含量先增加后降低并趋于稳定,振荡培养48 h后,培养液中可溶性磷含量最高,为246.99 mg/L;培养过程中pH值先降低后升高,然后趋于稳定,48 h时pH值最小(3.17)。经recA序列分析,将CT45-1鉴定为新洋葱伯克霍尔德氏菌(Burkholderia cenocepacia)。盆栽试验结果表明,不同处理浓度的CT45-1菌液处理烟株后,其株高、鲜重均显著高于对照,植株全磷含量分别比对照高30.28%、22.40%。
The NBRIP liquid medium with Ca_3(PO_4)_2 as the sole phosphorus source was used to quantitatively analyze the phosphate-dissolving ability of a high-efficiency phosphate-dissolving bacterial strain CT45-1 separated from the rhizosphere soil of Weifang tobacco-growing area. The strain was identified by molecular biological method, and its promoting-effect on tobacco growth was studied by pot experiment. The results of quantitative analysis showed that, with the increase of culture time, the soluble phosphorus content in the liquid medium with inoculated strain CT45-1 increased firstly, then decreased and tended to be stable. After 48 hours of shaking culture, the soluble phosphorus in the culture solution had the highest content as 246.99 mg/L. The pH value decreased firstly, then rose and stabilized during the culture. The pH value as 3.17 was minimum at the 48 th h. The strain CT45-1 was identified as a new Burkholderia cenocepacia by recA sequence analysis. The results of pot experiment showed that the plant height and fresh weight of tobacco treated by two concentrations of CT45-1 bacterial solution were significantly higher than those of the control, and the total phosphorus content of the two treatments were 30.28% and 22.40% higher than that of the control.
The NBRIP liquid medium with Ca_3(PO_4)_2 as the sole phosphorus source was used to quantitatively analyze the phosphate-dissolving ability of a high-efficiency phosphate-dissolving bacterial strain CT45-1 separated from the rhizosphere soil of Weifang tobacco-growing area. The strain was identified by molecular biological method, and its promoting-effect on tobacco growth was studied by pot experiment. The results of quantitative analysis showed that, with the increase of culture time, the soluble phosphorus content in the liquid medium with inoculated strain CT45-1 increased firstly, then decreased and tended to be stable. After 48 hours of shaking culture, the soluble phosphorus in the culture solution had the highest content as 246.99 mg/L. The pH value decreased firstly, then rose and stabilized during the culture. The pH value as 3.17 was minimum at the 48 th h. The strain CT45-1 was identified as a new Burkholderia cenocepacia by recA sequence analysis. The results of pot experiment showed that the plant height and fresh weight of tobacco treated by two concentrations of CT45-1 bacterial solution were significantly higher than those of the control, and the total phosphorus content of the two treatments were 30.28% and 22.40% higher than that of the control.
引文
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[19] 任嘉红,刘辉,吴晓蕙,等.南方红豆杉根际溶无机磷细菌的筛选、鉴定及其促生效果[J].微生物学报,2012,53 (3):294-303.
[20] 陶涛,叶明,刘冬,等.无机解磷细菌的筛选、鉴定及其溶磷能力研究[J].合肥工业大学学报(自然科学版),2011,34(2):304-308.
[21] 陈俊,陆俊锟,康丽华,等.红树林溶磷菌的初步鉴定、溶磷能力测定及其优化培养[J].微生物学通报,2009,36 (8):1183-1188.
[22] Hameeda B,Harini G,Rupela O P,et al.Growth promotion of maize by phosphate-solubilizing bacteria isolated from composts and macrofauna[J].Microbiological Research,2008,163(2):234-242.
[23] 王婧.桔黄假单胞菌JD37菌株对植物的促生作用及其微生物肥料的研制[D].上海:上海师范大学,2012.
[24] Yu X,Liu X,Zhu T H,et al.Isolation and characterization of phosphate solubilizing bacteria from walnut and their effect on growth and phosphorus mobilization[J].Biology and Fertility of Soils,2011,47(4):437-446.
[25] 程宝森,房玉林,刘延琳,等.渭北旱塬葡萄根际解磷细菌的筛选及其对葡萄的促生效应[J].西北农业学报,2009,18(4):185-190.
[2] 邵宗臣,赵美芝.土壤中积累态磷活化动力学的研究I.有机质的影响[J].土壤学报,2002,39(3):318-325.
[3] 余旋,朱天辉,刘旭.不同解磷菌剂对美国山核桃根际微生物和酶活性的影响[J].林业科学,2012,48(2):117-123.
[4] 贺梦醒,高毅,胡正雪,等.解磷菌株B25的筛选、鉴定及其解磷能力[J].应用生态学报,2012,23(1):235-239.
[5] 韩烁,夏冬亮,李潞滨,等.毛竹根部解磷细菌的筛选及多样性研究[J].河北农业大学学报,2010,33(2):26-31.
[6] 胡晓峰.溶磷菌的筛选、溶磷条件优化及对玉米的促生作用研究[D].南京:南京农业大学,2010.
[7] Chen Y P,Rekha P D,Arun A B,et al.Phosphate solubilizing bacteria from subtropical soil and their tricalcium phosphate solubilizing abilities[J].Applied Soil Ecology,2006,34(1):33-41.
[8] 林启美,赵海英,赵小蓉.4株溶磷细菌和真菌溶解磷矿粉的特性[J].微生物学通报,2002,29(6):24-28.
[9] 贺梦醒,高毅,胡正雪,等.解磷菌株B25的筛选、鉴定及其解磷能力[J].应用生态学报,2012,23(1):235-239.
[10] 崔邢,张亮,林勇明,等.不同土壤条件下解磷菌处理对巨尾桉土壤有效磷含量的影响[J].应用与环境生物学报,2015,21(4):740-746.
[11] Ahemad M,Kibret M.Mechanisms and applications of plant growth promoting rhizobacteria:current perspective[J].Journal of King Saud University-Science,2014,26(1):1-20.
[12] Li Y,Liu X,Hao T,et al.Colonization and maize growth promotion induced by phosphate solubilizing bacterial isolates[J].International Journal of Molecular Sciences,2017,18(7):1253-1268.
[13] 鲁如坤.土壤农业化学分析方法[M].北京:中国农业科学技术出版社,2000:66-185.
[14] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:169.
[15] Mahenthiralingam E,Bischof J,Byrne S K,et al.DNA-based diagnostic approaches for identification of Burkholderia cepacia complex,Burkholderia vietnamiensis,Burkholderia multivorans,Burkholderia stabilis,and Burkholderia cepacia genomovars Ⅰ and Ⅲ[J].Journal Clinical Microbiology,2000,38(9):3165-3173.
[16] Molla M A Z,Chowdhury A A,Islam A,et al.Microbial mineralization of organic phosphate in soil[J].Plant and Soil,1984,78(3):393-399.
[17] Illmer P,Schinner F.Solubilization of inorganic calcium phosphates-solubilization mechanisms[J].Soil Biology and Biochemistry,1995,27(3):257-263.
[18] Oliveira C A,Alves V M C,Marriel I E,et al.Phosphate solubilizing microorganisms isolated from rhizosphere of maize cultivated in an oxisol of the Brazilian Cerrado Biome[J].Soil Biology and Biochemistry,2008,41(9):1782-1787.
[19] 任嘉红,刘辉,吴晓蕙,等.南方红豆杉根际溶无机磷细菌的筛选、鉴定及其促生效果[J].微生物学报,2012,53 (3):294-303.
[20] 陶涛,叶明,刘冬,等.无机解磷细菌的筛选、鉴定及其溶磷能力研究[J].合肥工业大学学报(自然科学版),2011,34(2):304-308.
[21] 陈俊,陆俊锟,康丽华,等.红树林溶磷菌的初步鉴定、溶磷能力测定及其优化培养[J].微生物学通报,2009,36 (8):1183-1188.
[22] Hameeda B,Harini G,Rupela O P,et al.Growth promotion of maize by phosphate-solubilizing bacteria isolated from composts and macrofauna[J].Microbiological Research,2008,163(2):234-242.
[23] 王婧.桔黄假单胞菌JD37菌株对植物的促生作用及其微生物肥料的研制[D].上海:上海师范大学,2012.
[24] Yu X,Liu X,Zhu T H,et al.Isolation and characterization of phosphate solubilizing bacteria from walnut and their effect on growth and phosphorus mobilization[J].Biology and Fertility of Soils,2011,47(4):437-446.
[25] 程宝森,房玉林,刘延琳,等.渭北旱塬葡萄根际解磷细菌的筛选及其对葡萄的促生效应[J].西北农业学报,2009,18(4):185-190.
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