毛竹林钾矿物分解细菌的分离与鉴定
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摘要
为探明钾矿物分解细菌在毛竹林土壤中的分布情况与溶硅解钾性能,并从毛竹林土壤中分离筛选溶硅解钾能力较强的菌株。本研究对以钾长石为唯一钾源的专性培养基分离获得的钾矿物分解细菌,采用硅钼蓝比色法、火焰光度法及Salkowski比色法分别测定菌株的溶硅、解钾及产吲哚乙酸(IAA)能力,并对兼具溶硅与解钾功能菌株进行16S rDNA测序鉴定。结果表明:(1)鞭根根际土壤中的钾矿物分解细菌菌落形成数显著高于篼根根际土壤(P<0.05);(2)从毛竹根际(鞭根、蔸根)土、非根际土中分离出长势良好的钾矿物分解细菌共计102株,其中51株为功能菌株,具有溶硅能力的34株,具有解钾功能的31株,具产IAA能力的5株;兼具溶硅与解钾能力的14株,兼具溶硅、释钾及产IAA的仅有2株;钾矿物分解细菌发酵过程中pH下降值与水溶性硅相对增加量之间呈极显著负相关(P<0.01),与水溶性钾相对增加量之间呈负相关关系。(3) 14株兼具溶硅释钾的钾矿物分解细菌中有9株属于伯克霍尔德菌属(Burkholderia)属,其余5株分别属于假单胞菌属(Pseudomonas)、泛菌属(Pantoea)、肠杆菌属(Enterobacter)、四折叠球菌属(Quadrisphaera)与节细菌属(Arthrobacter) 5个属。钾矿物分解细菌在毛竹林土壤中分布广泛,其中以鞭根根际土壤中数量居多,其分布表现出明显的根系效应;兼具溶硅与解钾能力的钾矿物分解细菌物种多样性较高,以伯克霍尔德菌属为优势类群。
In order to detect the natural distribution and the characteristics of silicon-dissolving and potassium-solubilizing of the mineral potassium-solubilizing bacteria( MPSB) existing in the soil of Phyllostachys edulis forest,the highly efficient bacterial isolates were gained from the soil of non-rhizosphere,rhizosphere of rhizome roots and base roots of P.edulis.The ability of silicondissolving,potassium-solubilizing,and ability to secrete IAA of the bacterial isolates were also studied with the method of siliconmolybdenum blue colorimetry,flame spectrometry and Salkowski colorimetric,respectively.Then,the Si/K-solubilizing bacteria were identified with the method of 16S rDNA sequence analysis.The results showed that:( 1) the number of colony-forming unit( CFU) of MPSB in the rhizosphere soil of bamboo rhizome roots was significantly higher than that in the rhizosphere soil of base root soil( P<0.05).( 2) There were 51 functional strains were screened out from those 102 strains,which including 34 strains of silicate mineral-solubilizing bacteria,31 strains of potassium-solubilizing bacteria and 5 strains of IAA-secreting bacteria.There were 14 strains able to solubilize both silicate and potassium,of which 2 strains can also secrete IAA.The decreases in fermented liquid pH value were significant negative correlation with the water-soluble Si( P<0.01),and negative correlation with water-soluble K.( 3) Among the 14 strains of Si/K-solubilizing bacteria,9 strains belong to the genus Burkholderia,and the other 5 isolates belonged to Pantoea,Pseudomonas,Quadrisphaera,Enterobacter,Arthrobacter,respectively.The mineral potassium-solubilizing bacteria were widely distributed in bamboo soil,especially in rhizosphere soil of bamboo rhizome roots,which was shown obvious effect on root type.The Si/K-solubilizing bacteria in P.edulis forest displayed higher species diversity,and the dominant bacterial group is Burkholderia.
In order to detect the natural distribution and the characteristics of silicon-dissolving and potassium-solubilizing of the mineral potassium-solubilizing bacteria( MPSB) existing in the soil of Phyllostachys edulis forest,the highly efficient bacterial isolates were gained from the soil of non-rhizosphere,rhizosphere of rhizome roots and base roots of P.edulis.The ability of silicondissolving,potassium-solubilizing,and ability to secrete IAA of the bacterial isolates were also studied with the method of siliconmolybdenum blue colorimetry,flame spectrometry and Salkowski colorimetric,respectively.Then,the Si/K-solubilizing bacteria were identified with the method of 16S rDNA sequence analysis.The results showed that:( 1) the number of colony-forming unit( CFU) of MPSB in the rhizosphere soil of bamboo rhizome roots was significantly higher than that in the rhizosphere soil of base root soil( P<0.05).( 2) There were 51 functional strains were screened out from those 102 strains,which including 34 strains of silicate mineral-solubilizing bacteria,31 strains of potassium-solubilizing bacteria and 5 strains of IAA-secreting bacteria.There were 14 strains able to solubilize both silicate and potassium,of which 2 strains can also secrete IAA.The decreases in fermented liquid pH value were significant negative correlation with the water-soluble Si( P<0.01),and negative correlation with water-soluble K.( 3) Among the 14 strains of Si/K-solubilizing bacteria,9 strains belong to the genus Burkholderia,and the other 5 isolates belonged to Pantoea,Pseudomonas,Quadrisphaera,Enterobacter,Arthrobacter,respectively.The mineral potassium-solubilizing bacteria were widely distributed in bamboo soil,especially in rhizosphere soil of bamboo rhizome roots,which was shown obvious effect on root type.The Si/K-solubilizing bacteria in P.edulis forest displayed higher species diversity,and the dominant bacterial group is Burkholderia.
引文
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[14]韩晓阳,周波,董玉惠,等.山东茶园土壤高活性解钾细菌的筛选鉴定及肥效研究[J].茶叶科学,2018,38(1):78-86.
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[24]孟勇,艾文胜,李美群,等.毛竹鞭根区土壤微生物数量及其与土壤养分含量的关系[J].湖南林业科技,2015,42(5):1-5.
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[27]罗娜,周德明,徐睿,等.降香黄檀、檀香根际解钾菌的筛选与活性研究[J].热带作物学报,2016,37(5):964-970.
[28]鞠伟.杨树根际高效解钾细菌的分离筛选与鉴定[D].南京:南京林业大学,2016.
[29]陈易,程永毅,郭涛,等.一株具紫色土亲和性解钾菌的筛选及促生效应[J].西南大学学报(自然科学版),2016,38(5):58-65.
[30]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):381-388.
[31]王鹏.不同季节不同风化程度矿物表生可培养钾矿物分解细菌多样性研究[D].南京:南京农业大学,2011.
[2]赵建诚.毛竹氮素利用规律及影响因子研究[D].北京:中国林业科学研究院,2016.
[3]辉朝茂,杨宇明,郝吉明.论竹子生态环境效益与竹产业可持续发展[J].西南林学院学报,2003,23(4):25-29.
[4]刘广路,范少辉,苏文会,等.施肥时间对毛竹林生产力分配格局及土壤性质的影响[J].东北林业大学学报,2011,39(4):62-66.
[5]楼一平,李艳霞,BUCKINGHAM K.中国竹林可持续经营认证的必要性和可行性研究[J].竹子研究汇刊,2008,27(3):1-7.
[6]陈廷伟.钾细菌[M].北京:农业出版社,1959.
[7]赵飞,盛下放,黄智,等.山东地区钾矿物分解细菌的分离及生物学特性[J].生物多样性,2008,16(6):593-600.
[8]曹建芳.硅酸盐矿物分解细菌的定向筛选及其活化土壤硅的研究[D].南京:南京农业大学,2010.
[9]盛下放,黄为一.硅酸盐细菌NBT菌株释钾条件的研究[J].中国农业科学,2002,35(6):673-677.
[10]林启美,饶正华,孙焱鑫,等.硅酸盐细菌的筛选及其对番茄营养的影响[J].中国农业科学,2002,35(1):59-62.
[11]连宾,SMITH D L,傅平秋.硅酸盐细菌在工农业生产中的应用及其作用机理[J].贵州科学,2000,18(1/2):43-53.
[12]盛下放.硅酸盐细菌在不同生境土壤中的分布[J].土壤,2004,36(1):81-84.
[13]胡桦,陈强,李登煜,等.紫色土硅酸盐细菌的遗传多样性研究[J].土壤学报,2007,44(2):379-383.
[14]韩晓阳,周波,董玉惠,等.山东茶园土壤高活性解钾细菌的筛选鉴定及肥效研究[J].茶叶科学,2018,38(1):78-86.
[15]张晶晶,李建贵,郭艺鹏.新疆核桃根际土壤中解钾菌的分离筛选及鉴定[J].经济林研究,2016,34(2):30-34.
[16]中国科学院南京土壤研究所微生物室.土壤微生物研究法[M].北京:科学出版社,1985.
[17]张小丽.胀袋酱油中产气微生物的分离、鉴定与特性研究[D].杭州:浙江工商大学,2015.
[18]WELCH S A,ULLMAN W J.The effect of organic acids on plagioclase dissolution rates and stoichiometry[J].Geochimica et Cosmochimica Acta,1993,57(12):2 725-2 736.
[19]鲍世旦.土壤农化分析[M].3版.北京:中国农业出版社,2000.
[20]BANO N,MUSARRAT J.Isolation and characterization of phorate degrading soil bacteria of environmental and agronomic significance[J].Letters in Applied Microbiology,2003,36(6):349-353.
[21]HALL T A.Bio Edit:a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT[J].Nucleic Acids Symposium Series,1999,41:95-98.
[22]TAMURA K,STECHER G,PETERSON D,et al.MEGA6:molecular evolutionary genetics analysis version 6.0[J].Molecular Biology and Evolution,2013,30(12):2 725-2 729.
[23]CHAUDHARY D R,SAXENA J,LORENZ N,et al.Microbial profiles of rhizosphere and bulk soil microbial communities of biofuel crops switchgrass(Panicum virgatum L.)and Jatropha(Jatropha curcas L.)[J].Applied and Environmental Soil Science,2012,2012:906 864.
[24]孟勇,艾文胜,李美群,等.毛竹鞭根区土壤微生物数量及其与土壤养分含量的关系[J].湖南林业科技,2015,42(5):1-5.
[25]潘瑞炽.植物生理学[M].6版.北京:高等教育出版社,2008.
[26]盛下放,黄为一,曹晓英.硅酸盐细菌NBT菌株解钾效能及对钾的吸持作用[J].植物营养与肥料学报,2001,7(4):459-466.
[27]罗娜,周德明,徐睿,等.降香黄檀、檀香根际解钾菌的筛选与活性研究[J].热带作物学报,2016,37(5):964-970.
[28]鞠伟.杨树根际高效解钾细菌的分离筛选与鉴定[D].南京:南京林业大学,2016.
[29]陈易,程永毅,郭涛,等.一株具紫色土亲和性解钾菌的筛选及促生效应[J].西南大学学报(自然科学版),2016,38(5):58-65.
[30]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):381-388.
[31]王鹏.不同季节不同风化程度矿物表生可培养钾矿物分解细菌多样性研究[D].南京:南京农业大学,2011.