一种耐铀植物促生菌的筛选及促生特性研究
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摘要
采用富集培养法和LB平板法从铀尾矿污染区蓼科植物酸模根部中筛选和分离一株具有较强铀耐受能力的细菌菌株,通过特征反应和平均吸光度法分析其生长特性和不同培养条件下植物促生特性。结果表明:该菌株可在铀浓度350 mg/kg(土)条件下生长。具有较强的分泌IAA能力,普通条件下24 h产IAA 40.21 mg/L,最佳产IAA条件为温度为35℃,pH 7,转速为150 r/min,氮源为酵母膏,碳源为甘油;兼具ACC脱氨能力,普通条件下24 h产ACC酶活为0.32 U/μg,最佳的ACC酶活条件为温度为30℃,pH 7,转速180 r/min。结合形态学特征,生理生化初步特征和16s rDNA序列确定菌株为木糖氧化无色杆菌。不同铀浓度的盆栽实验接种该菌种能使苜蓿干重分别提高17.9%-110.4%;对铀的富集率分别提高12.2%-180.6%。
A uranium-resistant and plant growth-promoting bacterial strain was isolated from the root of a native Rumex acetosa in auranium mining by enrichment culture and LB plate. Its growth characteristics and the performance of plant growth-promoting under differentexperimental conditions were analyzed by characteristic reaction and average absorbance method. The results showed that the strain grew wellat the uranium concentration of 350 mg/kg,it produced indole-3-acetic acid(IAA)and IAA reached 40.21 mg/L in 24 h at normal condition.The optimal condition for IAA production was the temperature 30℃,pH 7,rotating speed 150 r/min,yeast extract as nitrogen source,andglycerol as carbon source. The strain also had deaminase capacity of ACC,and ACC enzymatic activity was 0.32 U/μg for 24 h at normalcondition. The optimal condition for ACC enzyme activity was temperature 35℃,pH 7,and rotating speed 180 r/min. The strain was identifiedas Achromobacter xylosoxidans while combined morphological characteristic with physiological and biochemical preliminary characteristics and16 s rDNA sequence. Dry biomass of Medicago sativa increased by 17.9%-110.4% and U was enriched by 12.2%-180.6% under different Uconcentration.
A uranium-resistant and plant growth-promoting bacterial strain was isolated from the root of a native Rumex acetosa in auranium mining by enrichment culture and LB plate. Its growth characteristics and the performance of plant growth-promoting under differentexperimental conditions were analyzed by characteristic reaction and average absorbance method. The results showed that the strain grew wellat the uranium concentration of 350 mg/kg,it produced indole-3-acetic acid(IAA)and IAA reached 40.21 mg/L in 24 h at normal condition.The optimal condition for IAA production was the temperature 30℃,pH 7,rotating speed 150 r/min,yeast extract as nitrogen source,andglycerol as carbon source. The strain also had deaminase capacity of ACC,and ACC enzymatic activity was 0.32 U/μg for 24 h at normalcondition. The optimal condition for ACC enzyme activity was temperature 35℃,pH 7,and rotating speed 180 r/min. The strain was identifiedas Achromobacter xylosoxidans while combined morphological characteristic with physiological and biochemical preliminary characteristics and16 s rDNA sequence. Dry biomass of Medicago sativa increased by 17.9%-110.4% and U was enriched by 12.2%-180.6% under different Uconcentration.
引文
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[25]徐文思,姜瑛,李引,等.一株植物促生菌的筛选、鉴定及其对花生的促生效应研究[J].土壤, 2014, 46(1):119-125.
[26]蒙渊.产铁载体和脱氨酶的氢氧化细菌筛选及促生作用研究[D].西安:西北大学, 2011.
[27]沈萍,刘维红,闫淑珍,等. XG32菌株产ACC脱氢酶的培养条件和酶活影响因素[J].南京师大学报:自然科学版,2008, 31(1):104-108.
[28]罗胜联.一种促进植物生长和重金属污染土壤修复的内生菌及应用:中国, 201010609930. 9[P]. 2011-08-24.
[29]Zhang XC, Lin L, et al. Colonization and modulation ofhost growthand metal uptake by endophytic bacteria of Sedum alfredii[J].International Journal of Phytoremediation, 2013, 15(1):51-64.
[30]Muhammad TA, Muhammad N, Muhammad I, et al. Bacterialendophytes enhance phytostabilizationin soils contaminated withuranium and lead[J]. International Journal of Phytoremediation,2017, 19(755).
[31]Peralta-Videa JR, Rose GDl, Gonzalez JH, et al. Effect of thegrowth stage on the heavy metal tolerance of alfalfa plant[J].Advances in Environmental Research, 2004, 8(3):679-685.
[32]唐永金,罗学刚,曾峰.不同植物对高浓度铀胁迫的响应与铀富集植物筛选[J].核农学报, 2013, 27(12):1920-1926.
[33]金星,罗学刚,唐永金.外源硼酸对铀胁迫下四季豆生理以及铀富集的影响[J].中国农学通报, 2016, 32(33):175-181.
[2]秦茂钊.铀矿山废渣治理方法研究[D].重庆:重庆大学,2003.
[3]张洪,杨亚新,等.湖南某退役铀矿山水环境重金属污染现状评价[J].有色金属(矿山部分), 2014(3):41-44.
[4]Kuppusamy S, Palanisami T, Megharaj M, et al. In-situ remediationapproachesforthemanagementofcontaminatedsites:acomprehensive overview[J]. Rev Environ Contam Toxicol, 2016,236(18):1-115.
[5]陈亚刚,陈雪梅,张玉刚,等.微生物抗重金属的生理机制[J].生物技术通报, 2009(10):60-65.
[6]陈可,胡南,陈威,等.两种植物根际促生菌对博落回抗干旱及富集铀性能的增强作用研究[J].环境科学学报, 2018, 38(10):4142-4149.
[7]何琳燕,李娅,刘涛,等.龙葵根际和内生Cd抗性细菌的筛选及其生物学特性[J].生态与农村环境学报, 2011, 27:83-88.
[8]李艳梅,王琼瑶,涂卫国.镍胁迫下产铁载体细菌对花生的促生性[J].微生物学通报, 2017, 44(8):1882-1890.
[9]Dell’Amico E, Cavalca L, et al. Analysis of rhizobacterial communities in perennial Graminaceae from polluted water meadow soil,and screening of metal-resistant, potentially plant growth-promotingbacteria[J]. FEMS Microbiol Ecol, 2005, 52:153-162.
[10]Glickmann E, Dessaux Y. A critical examination of the specificityof the Salkowski reagent for indolic compounds producedbyphytopathogenic bacteria[J]. Appl Environ Microbiol, 1995, 619(2):793-796.
[11]李振东,陈秀蓉,李鹏,等.珠芽蓼内生菌Z5产IAA和抑菌能力测定及其鉴定[J].草业学报, 2007, 19(2):61-68.
[12]张国壮.产脱氨酶根际促生菌的分离鉴定及其接种对小麦乙烯代谢和生长的效应[D].杨凌:西北农林科技大学, 2014.
[13]Dong X, Cai M. Identificationmanual of common bacteriasystem[M]. Beijing:Science Press, 2001.
[14]杨圣,罗学刚,李波,等.重金属富集植物纤维素降解菌株的分离鉴定[J].安全与环境学报, 2016, 16(2):218-225.
[15]苏峰丙,罗学刚,唐永金,等.不同含磷化合物固化修复铀污染土壤的研究[J].核农学报, 2017, 32(2):407-415.
[16]Glick BR. Using soil bacteria to facilitate phytoremediation[J].Biotechnol Adv, 2010, 28(3):367-374.
[17]Ying M, Mani R, Helena F. Inoculation of plant growth promotingbacterium Achromobacter xylosoxidans strain Ax10 for theimprovement of copperphytoextraction by Brassica juncea[J].Journal of Environmental Management, 2009, 9:831-837.
[18]Junkang G, Jie C. Effect of Cd-tolerant plant growth-promotingrhizobiumon plant growth and Cd uptake by Lolium multiflorumLam. and Glycine max(L.)Merr. in Cd-contaminated soil[J].Plant Soil, 2013, 375(2):205-214.
[19]Kadhimi AA, Alhasnawi AN, Mohamad A, et al. Tissue cultureand some of the factors affecting them and the micropropagation ofstrawberry[J]. Life Sci J, 2014, 11:484-493.
[20]潘风山,陈宝,等.一株镉超积累植物东南景天特异内生细菌的筛选及鉴定[J].环境科学学报, 2014, 34(2):449-456.
[21]Govarthanan M, Lee KJ, Cho M, et al. Significance of autochthonous Bacillus sp. KK1 on biomineralization of lead in mine tailings[J].Chemosphere, 2013, 90:2267-2272.
[22]李交昆,余黄,曾伟民,等.根际促生菌强化植物修复重金属污染土壤的研究进展[J].生命科学, 2017, 29(5):434-442.
[23]吉云秀.含脱氨酶分离及提高植物抗逆性[D].大连:大连海事大学, 2007.
[24]RajkumarM,AeN,FreitasH.Endophyticbacteriaandtheirpotential to enhance heavy metal phytoextraction[J].Chemosphere, 2009, 77(2):153-160.
[25]徐文思,姜瑛,李引,等.一株植物促生菌的筛选、鉴定及其对花生的促生效应研究[J].土壤, 2014, 46(1):119-125.
[26]蒙渊.产铁载体和脱氨酶的氢氧化细菌筛选及促生作用研究[D].西安:西北大学, 2011.
[27]沈萍,刘维红,闫淑珍,等. XG32菌株产ACC脱氢酶的培养条件和酶活影响因素[J].南京师大学报:自然科学版,2008, 31(1):104-108.
[28]罗胜联.一种促进植物生长和重金属污染土壤修复的内生菌及应用:中国, 201010609930. 9[P]. 2011-08-24.
[29]Zhang XC, Lin L, et al. Colonization and modulation ofhost growthand metal uptake by endophytic bacteria of Sedum alfredii[J].International Journal of Phytoremediation, 2013, 15(1):51-64.
[30]Muhammad TA, Muhammad N, Muhammad I, et al. Bacterialendophytes enhance phytostabilizationin soils contaminated withuranium and lead[J]. International Journal of Phytoremediation,2017, 19(755).
[31]Peralta-Videa JR, Rose GDl, Gonzalez JH, et al. Effect of thegrowth stage on the heavy metal tolerance of alfalfa plant[J].Advances in Environmental Research, 2004, 8(3):679-685.
[32]唐永金,罗学刚,曾峰.不同植物对高浓度铀胁迫的响应与铀富集植物筛选[J].核农学报, 2013, 27(12):1920-1926.
[33]金星,罗学刚,唐永金.外源硼酸对铀胁迫下四季豆生理以及铀富集的影响[J].中国农学通报, 2016, 32(33):175-181.
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