柠条根际产铁载体促生菌的分离鉴定及其促生特性
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摘要
以柠条根际土壤为试材,采用CAS(铬天青S)选择性培养基分离筛选具有产铁载体功能的根际促生菌株,通过16S r DNA序列分析进行菌株鉴定,并测定菌株解磷、固氮、产IAA(吲哚乙酸)和ACC(1-氨基环丙烷-1-羧酸)脱氨酶等促生活性,通过盆栽试验分析验证菌株Fe11对柠条幼苗生长的影响。结果表明:共分离出14株产铁载体根际促生菌,其中6株菌具备较强的产铁载体能力。所有菌株的16S r DNA序列同源性与假单胞菌达到99%~100%,假单胞菌属细菌是柠条根际产铁载体的优势菌群。多数菌株兼具2种以上促生活性,菌株Fe9、Fe11和Fe13表现出多种促生潜能。回接试验证实,接种菌株Fe11使柠条幼苗的株高、根长、叶片数、鲜质量和干质量分别增长17.25%、6.68%、17.75%、42.51%和20.67%,表明菌株Fell对柠条的生长具有良好的促进效果。
Rhizobacteria of producing siderophore were isolated from rhizosphere soil of Caragana spp.grown in desert grassland with CAS( chrome azurol S) medium. The diversity of the strains were analysed based on the 1 6 S r DNA sequences. The ability of phosphate dissolving,nitrogen fixation,indoleacetic acid( IAA) and ACC deaminase production were evaluated. The effect on promoting growth of Caragana spp.seedlings were tested by inoculating experiment with strain Fe11. The results showed that total of 14 strains of siderophore-producing were isolated from rhizosphere soil of Caragana spp.,and 6 of them had strong ability to produce siderophore. The 16 S r DNA sequences of all strains had 99%-100%similarity with that of Pseudomonas,which was dominant siderophore-producing rhizobacteria of Caragana spp.Most of the strains had more than two kinds of growth-promoting characteristics.Among them,strains Fe9,Fe11 and Fe13 showed a diverse growth-promoting potential.The results of inoculation experiment demonstrated that strain Fe11 had obvious growth-promoting effect on Caragana spp. seedlings. The plant height,root length,leaf number,fresh weight and dry weight were increased by 17. 25%,6. 68%,17. 75%,42. 51% and 20. 67%,respectively.
Rhizobacteria of producing siderophore were isolated from rhizosphere soil of Caragana spp.grown in desert grassland with CAS( chrome azurol S) medium. The diversity of the strains were analysed based on the 1 6 S r DNA sequences. The ability of phosphate dissolving,nitrogen fixation,indoleacetic acid( IAA) and ACC deaminase production were evaluated. The effect on promoting growth of Caragana spp.seedlings were tested by inoculating experiment with strain Fe11. The results showed that total of 14 strains of siderophore-producing were isolated from rhizosphere soil of Caragana spp.,and 6 of them had strong ability to produce siderophore. The 16 S r DNA sequences of all strains had 99%-100%similarity with that of Pseudomonas,which was dominant siderophore-producing rhizobacteria of Caragana spp.Most of the strains had more than two kinds of growth-promoting characteristics.Among them,strains Fe9,Fe11 and Fe13 showed a diverse growth-promoting potential.The results of inoculation experiment demonstrated that strain Fe11 had obvious growth-promoting effect on Caragana spp. seedlings. The plant height,root length,leaf number,fresh weight and dry weight were increased by 17. 25%,6. 68%,17. 75%,42. 51% and 20. 67%,respectively.
引文
[1]BUTLER J L,WILLIAMS M A,BOTTOMLEY P J,et al.Microbial community dynamics associated with rhizosphere carbon flow[J].Applied and Environmental Microbiology,2003,69:6793-6800.
[2]艾超,孙静文,王秀斌,等.植物根际沉积与土壤微生物关系研究进展[J].植物营养与肥料学报,2015,21(5):1343-1351.
[3]张东艳,刘晔,吴越,等.花生根际产IAA菌的筛选鉴定及其效应研究[J].中国油料作物学报,2016,38(1):104-110.
[4]ZHANG L Z,FAN J J,NI W,et al.Isolation of phosphate solubilizing fungus(Aspergillus niger)from Caragana rhizosphere and its potential for phosphate solubilization[J].Acta Ecologica Sinica,2011,31(24):7571-7578.
[5]ELENA L C,VINCENZA A.Improvement of Brassica napus growth under cadium stress by eadmium-resistant rhizobaeteria[J].Soil Biology&Bioehemistry,2008,40:74-84.
[6]张晖,宋圆圆,吕顺,等.香蕉根际促生菌的抑菌活性及对作物生长的促进作用[J].华南农业大学学报,2015,36(3):65-70.
[7]PIROMYOU P,BURANABANYAT B,TANTASAWAT,et al.Effect of plant growth promoting rhizobacteria(PGPR)inoculation on microbial community structure in rhizosphere of forage corn cultivated in Thailand[J].European Journal Soil Biololgy,2011,47(1):44-54.
[8]PAYNE S M.Detection,isolation,and characterization of siderophores[J].Methods Enzymol,1994,235:329-344.
[9]金崇伟,俞雪辉,郑绍建.微生物在植物铁营养中的潜在作用[J].植物营养与肥料学报,2005,11(5):688-695.
[10]梅新兰,闪安琪,蒋益,等.适应玉米的溶磷细菌筛选及其对玉米生长的影响[J].土壤学报,2016,53(2):502-509.
[11]荣良燕,姚拓,赵桂琴,等.产铁载体PGPR菌筛选及其对病原菌的拮抗作用[J].植物保护,2011,37(1):59-64.
[12]关林婧,马成仓.21世纪锦鸡儿属植物研究进展[J].草地学报,2014,22(4):697-705.
[13]赵翔,谢志雄,陈绍兴,等.适合高产铁载体细菌筛选、检测体系的改进与探析[J].微生物学通报,2006,33(6):95-98.
[14]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.
[15]滕松山,刘艳萍,赵蕾.具ACC脱氨酶活性的碱蓬内生细菌的分离、鉴定及其生物学特性[J].微生物学报,2010,50(11):1503-1509.
[16]PENROSE D M,GLICK B R.Methods for isolating and characterizing ACC deaminase-containing plant growth-promoting rhizobacteria[J].Plant Physiology,2003,118:10-15.
[17]孙红启.铁载体和铁离子对细菌生长过程的影响[D].济南:山东大学,2008.
[18]朱彭玲,杜秉海,丁延芹,等.新疆棉花根际土壤铁载体产生菌的遗传多样性及系统发育研究[J].中国农业科学,2009,42(5):1568-1574.
[19]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.
[20]谭石勇,易永健,汪洪鹰,等.苎麻促生菌的筛选、鉴定及其促生效应[J].微生物学通报,2015,42(3):525-533.
[21]王小兵,叶赛克,汪晓丽,等.江苏沿海滩涂植物根际促生菌的筛选[J].扬州大学学报(农业与生命科学),2016,37(2):81-86.
[22]赵燕娜,廖超英,李晓明.毛乌素沙地4种固沙植物根际与非根际土壤生物学特性[J].干旱区研究,2015,32(4):680-686.
[23]REQUENA N,PEREZ-SOLIS E,AZCN-AGUILAR C,et al.Management of indigenous plant-microbe symbioses aids restoration of desertified ecosystems[J].Applied and Environmental Microbiology,2001,67(2):495-498.
[2]艾超,孙静文,王秀斌,等.植物根际沉积与土壤微生物关系研究进展[J].植物营养与肥料学报,2015,21(5):1343-1351.
[3]张东艳,刘晔,吴越,等.花生根际产IAA菌的筛选鉴定及其效应研究[J].中国油料作物学报,2016,38(1):104-110.
[4]ZHANG L Z,FAN J J,NI W,et al.Isolation of phosphate solubilizing fungus(Aspergillus niger)from Caragana rhizosphere and its potential for phosphate solubilization[J].Acta Ecologica Sinica,2011,31(24):7571-7578.
[5]ELENA L C,VINCENZA A.Improvement of Brassica napus growth under cadium stress by eadmium-resistant rhizobaeteria[J].Soil Biology&Bioehemistry,2008,40:74-84.
[6]张晖,宋圆圆,吕顺,等.香蕉根际促生菌的抑菌活性及对作物生长的促进作用[J].华南农业大学学报,2015,36(3):65-70.
[7]PIROMYOU P,BURANABANYAT B,TANTASAWAT,et al.Effect of plant growth promoting rhizobacteria(PGPR)inoculation on microbial community structure in rhizosphere of forage corn cultivated in Thailand[J].European Journal Soil Biololgy,2011,47(1):44-54.
[8]PAYNE S M.Detection,isolation,and characterization of siderophores[J].Methods Enzymol,1994,235:329-344.
[9]金崇伟,俞雪辉,郑绍建.微生物在植物铁营养中的潜在作用[J].植物营养与肥料学报,2005,11(5):688-695.
[10]梅新兰,闪安琪,蒋益,等.适应玉米的溶磷细菌筛选及其对玉米生长的影响[J].土壤学报,2016,53(2):502-509.
[11]荣良燕,姚拓,赵桂琴,等.产铁载体PGPR菌筛选及其对病原菌的拮抗作用[J].植物保护,2011,37(1):59-64.
[12]关林婧,马成仓.21世纪锦鸡儿属植物研究进展[J].草地学报,2014,22(4):697-705.
[13]赵翔,谢志雄,陈绍兴,等.适合高产铁载体细菌筛选、检测体系的改进与探析[J].微生物学通报,2006,33(6):95-98.
[14]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.
[15]滕松山,刘艳萍,赵蕾.具ACC脱氨酶活性的碱蓬内生细菌的分离、鉴定及其生物学特性[J].微生物学报,2010,50(11):1503-1509.
[16]PENROSE D M,GLICK B R.Methods for isolating and characterizing ACC deaminase-containing plant growth-promoting rhizobacteria[J].Plant Physiology,2003,118:10-15.
[17]孙红启.铁载体和铁离子对细菌生长过程的影响[D].济南:山东大学,2008.
[18]朱彭玲,杜秉海,丁延芹,等.新疆棉花根际土壤铁载体产生菌的遗传多样性及系统发育研究[J].中国农业科学,2009,42(5):1568-1574.
[19]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.
[20]谭石勇,易永健,汪洪鹰,等.苎麻促生菌的筛选、鉴定及其促生效应[J].微生物学通报,2015,42(3):525-533.
[21]王小兵,叶赛克,汪晓丽,等.江苏沿海滩涂植物根际促生菌的筛选[J].扬州大学学报(农业与生命科学),2016,37(2):81-86.
[22]赵燕娜,廖超英,李晓明.毛乌素沙地4种固沙植物根际与非根际土壤生物学特性[J].干旱区研究,2015,32(4):680-686.
[23]REQUENA N,PEREZ-SOLIS E,AZCN-AGUILAR C,et al.Management of indigenous plant-microbe symbioses aids restoration of desertified ecosystems[J].Applied and Environmental Microbiology,2001,67(2):495-498.