山东茶园土壤高活性解钾细菌的筛选鉴定及肥效研究
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摘要
为了筛选到适合山东茶园土壤环境的解钾菌株,以提高土壤钾素的高效转化效率,本研究经过菌株富集、分离、纯化等步骤,从山东茶园土壤中分离出9株解钾细菌。通过有效钾含量比较和菌株分类,筛选出1株活性最强菌株K2,经鉴定K2菌株为枯草芽孢杆菌(Bacillus subtilis)。发酵试验表明,菌株K2最适宜生长的条件为pH值6.0,温度35℃,以麦芽糖、淀粉、玉米粉为碳源,酵母膏为氮源。施用该菌剂后土壤速效钾和速效磷含量比对照最大分别提高28.40%和28.49%。茶叶产量最大提高36.30%,同时茶叶中氨基酸含量显著增加,酚氨比值降低,有利于茶叶品质的提升。菌株K2可能是一株茶园土壤高效解钾细菌,可作为后续进一步研究茶园专用微生物菌剂的参考。
The potassium bacterium suitable for soil environment in Shandong tea garden was screened to improve the transformation efficiency of soil potassium. By the experimental procedure of enrichment culture and microbial purification, 9 potassium bacterium strains were obtained from the soil of tea garden in Shandong Province. According to the activity comparison and taxonomy of strains, a strain with the highest activity was selected. The strain K2 was identified as Bacillus subtilis. Meanwhile, the effects of growth environment of the strains were measured. The results showed that the optimum p H and temperature for K2 were 6.0 and 35 ℃. It was suitable for K2 to ferment with maltose, starch and corn flour as the carbon source, and with yeast extract as the nitrogen source. After applying the bacteria agent, the content s of available potassium and phosphorus in tea garden soil increased by 28.40% and 28.49% respectively. Meanwhile the tea yield increased by 36.30%. The amino acid contents in tea were also significantly increased, and the polyphenols/amino acids decreased, which benefit tea quality. The K2 may be a high efficient potassium bacterium in tea garden soil, which can be used as a reference for further research on special microbial inoculum in tea garden.
The potassium bacterium suitable for soil environment in Shandong tea garden was screened to improve the transformation efficiency of soil potassium. By the experimental procedure of enrichment culture and microbial purification, 9 potassium bacterium strains were obtained from the soil of tea garden in Shandong Province. According to the activity comparison and taxonomy of strains, a strain with the highest activity was selected. The strain K2 was identified as Bacillus subtilis. Meanwhile, the effects of growth environment of the strains were measured. The results showed that the optimum p H and temperature for K2 were 6.0 and 35 ℃. It was suitable for K2 to ferment with maltose, starch and corn flour as the carbon source, and with yeast extract as the nitrogen source. After applying the bacteria agent, the content s of available potassium and phosphorus in tea garden soil increased by 28.40% and 28.49% respectively. Meanwhile the tea yield increased by 36.30%. The amino acid contents in tea were also significantly increased, and the polyphenols/amino acids decreased, which benefit tea quality. The K2 may be a high efficient potassium bacterium in tea garden soil, which can be used as a reference for further research on special microbial inoculum in tea garden.
引文
[1]万广华,李涛,王国华.山东省土壤钾素含量及分布研究[J].山东农业科学,2000(3):31-33.
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[5]蒋宝贵.解磷解钾自生固氮菌的分离筛选及鉴定[J].华中农业大学学报,2005,24(1):43-48.
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[8]阳洁,江院,王晓甜,等.几株高效溶磷解钾药用稻内生固氮菌的筛选与鉴定[J].农业生物技术学报,2016,24(2):186-195.
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[10]Basak BB,Biswas DR.Co-inoculation of potassium solubilizing and nitrogen fixing bacteria on solubilization of waste mica and their effect on growth promotion and nutrient acquisition by a forage crop[J].Biology and Fertility of Soils,2010,46:641-648.
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[13]韩晓阳,李智,张丽霞,等.茶园土壤高活性固氮菌的筛选鉴定及接种效果初步研究[J].茶叶科学,2014,34(5):497-505.
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[15]Weisburgw G,Bams M,Elletier D A.16s Ribosomal DNA amplification for phylogenetic study[J].J Bacterial,1991,173(2):697-703.
[16]Martin K?nneke,Anne E Bernhard,JoséR de la Torre,et al.Isolation of an autotrophic ammonia-oxidizing marine archaeon[J].Nature,2005,437:543-546.
[17]Buchanan R E,Gibbons N E.伯杰细菌鉴定手册[M].北京:科学出版社,1984:329-339.
[18]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:138-140.
[19]鲍士旦.土壤农化分析[M].北京:中国农业出版社,2000.
[20]麻瑞阳,张爱民,惠小双,等.高效解磷解钾菌NX-11菌株的分离筛选,鉴定及最佳培养条件的确定[J].华北农学报,2013,28(2):202-208.
[21]肖胜.不同种类解钾菌对烟叶化学成分及经济性状的影响[J].安徽农业科学,2017,45(3):46-48.
[22]顾杰,孟丽媛,孙乐妮,等.水稻根系固氮菌促生特性评价[J].安徽农业大学学报,2012,39(3):426-429.
[23]孙建光,胡海燕,刘君,等.农田环境中固氮菌的促生潜能与分布特点[J].中国农业科学,2012,45(8):1532-1544.
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[26]杨红武,李帆,唐春闺,等.溶磷解钾微生物在植烟土壤中的应用[J].湖南农业科学,2014,20:31-32.
[27]张爱民,李乃康,赵钢勇,等.土壤中解磷、解钾微生物研究进展[J].河北大学学报(自然科学版),2015,35(4):442-448.
[28]燕红,于彩莲,包鑫,等.高效解磷兼解钾活性菌株分离筛选的初步研究[J].扬州大学学报(农业与生命科学版),2016,37(1):81-90.
[29]Ma Y,Prasad MN,Rajkumar M,et al.Plant growth promoting rhizobacteria and endophytes accelerate phytoremediation of metalliferous soils[J].Biotechnol Adv,2011,29(2):248-258.
[30]Rajkumar M,Ae N,Prasad MN,et al.Potential of siderophore-producing bacteria for improving heavy metal phytoextraction[J].Trends Biotechnol,2010,28(3):142-149.
[31]韩丽珍,邓兆辉,朱春艳,等.茶树根际促生菌的筛选与促生特性的研究[J].山地农业生物学报,2016,35(1):51-56.
[2]刘钦普,林振山,周亮.山东省化肥使用时空分异及潜在环境风险评价[J].农业工程学报,2015,31(7):208-214.
[3]Huang P M,Wang M K,Chiu C Y.Soil mineral-organic matter-microbe interactions:impact on biogeochemical processes and biodiversity in soil[J].Pedobiologia,2005,49(6):609-635.
[4]张爱民,李乃康,赵钢勇,等.土壤中解磷、解钾微生物研究进展[J].河北大学学报(自然科学版),2015,35(4):442-448.
[5]蒋宝贵.解磷解钾自生固氮菌的分离筛选及鉴定[J].华中农业大学学报,2005,24(1):43-48.
[6]饶正华,林启美.解钾菌与解磷菌及固氮菌的相互作用[J].生态学杂志,2002,21(2):71-73.
[7]李新新,高新新,陈星,等.一株高效解钾菌的筛选、鉴定及发酵条件的优化[J].土壤学报,2014,51(2):382-388.
[8]阳洁,江院,王晓甜,等.几株高效溶磷解钾药用稻内生固氮菌的筛选与鉴定[J].农业生物技术学报,2016,24(2):186-195.
[9]Basak BB,Biswas DR.Influence of potassium solubilizing microorganism(Bacillus mucilaginosus)and waste mica on potassium uptake dynamics by sudan grass(Sorghum vulgare Pers.)grown under two Alfisols[J].Plant and Soil,2009,317:235-255.
[10]Basak BB,Biswas DR.Co-inoculation of potassium solubilizing and nitrogen fixing bacteria on solubilization of waste mica and their effect on growth promotion and nutrient acquisition by a forage crop[J].Biology and Fertility of Soils,2010,46:641-648.
[11]赵斌,何绍江.微生物学实验[M].北京:科学出版社,2002:215.
[12]Laemmli U K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature,1970,227:680-685.
[13]韩晓阳,李智,张丽霞,等.茶园土壤高活性固氮菌的筛选鉴定及接种效果初步研究[J].茶叶科学,2014,34(5):497-505.
[14]Tan Z Y,Kan F L,Peng G X,et al.Rhizobium yanglingense sp.nov.,isolated from arid and semi-arid regions in China[J].Int J Syst Evol Microbiol,2001,51:909-914.
[15]Weisburgw G,Bams M,Elletier D A.16s Ribosomal DNA amplification for phylogenetic study[J].J Bacterial,1991,173(2):697-703.
[16]Martin K?nneke,Anne E Bernhard,JoséR de la Torre,et al.Isolation of an autotrophic ammonia-oxidizing marine archaeon[J].Nature,2005,437:543-546.
[17]Buchanan R E,Gibbons N E.伯杰细菌鉴定手册[M].北京:科学出版社,1984:329-339.
[18]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:138-140.
[19]鲍士旦.土壤农化分析[M].北京:中国农业出版社,2000.
[20]麻瑞阳,张爱民,惠小双,等.高效解磷解钾菌NX-11菌株的分离筛选,鉴定及最佳培养条件的确定[J].华北农学报,2013,28(2):202-208.
[21]肖胜.不同种类解钾菌对烟叶化学成分及经济性状的影响[J].安徽农业科学,2017,45(3):46-48.
[22]顾杰,孟丽媛,孙乐妮,等.水稻根系固氮菌促生特性评价[J].安徽农业大学学报,2012,39(3):426-429.
[23]孙建光,胡海燕,刘君,等.农田环境中固氮菌的促生潜能与分布特点[J].中国农业科学,2012,45(8):1532-1544.
[24]孙海新.茶树根际促生细菌的研究[D].山东:山东农业大学,2006.
[25]梁成华,魏丽萍,罗磊.土壤固钾与释钾机制研究进展.地球科学进展[J].2002,17(5):679-684.
[26]杨红武,李帆,唐春闺,等.溶磷解钾微生物在植烟土壤中的应用[J].湖南农业科学,2014,20:31-32.
[27]张爱民,李乃康,赵钢勇,等.土壤中解磷、解钾微生物研究进展[J].河北大学学报(自然科学版),2015,35(4):442-448.
[28]燕红,于彩莲,包鑫,等.高效解磷兼解钾活性菌株分离筛选的初步研究[J].扬州大学学报(农业与生命科学版),2016,37(1):81-90.
[29]Ma Y,Prasad MN,Rajkumar M,et al.Plant growth promoting rhizobacteria and endophytes accelerate phytoremediation of metalliferous soils[J].Biotechnol Adv,2011,29(2):248-258.
[30]Rajkumar M,Ae N,Prasad MN,et al.Potential of siderophore-producing bacteria for improving heavy metal phytoextraction[J].Trends Biotechnol,2010,28(3):142-149.
[31]韩丽珍,邓兆辉,朱春艳,等.茶树根际促生菌的筛选与促生特性的研究[J].山地农业生物学报,2016,35(1):51-56.