尾矿土中解磷细菌的筛选及对镉的耐受研究
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摘要
为了筛选出具有较好的有解磷效果的细菌,以尾矿土为材料进行筛选,分离纯化具有高效解磷作用和高耐受重金属的细菌菌株,并对其进行分子鉴定。分离得到136株细菌,筛选出4株解磷效果较好的菌株。对这4株细菌进行分子鉴定,通过16S rRNA测序分析,4株细菌属于芽孢杆菌属(Bacillus)。从4株细菌中进一步优化,筛选出解磷效果最好的菌株WDN-5,分析WDN-5菌株对重金属镉的耐受性。研究结果表明:WDN-5在发酵第五天的解磷效果最好,溶磷量达到77.14μg/mL;WND-5对镉的致死浓度为300mg/L,对镉的耐受性较高;在50—300 mg/L的镉胁迫下,随着镉浓度增加,WDN-5的溶磷量逐渐降低。
In order to study the phosphorus-dissolving bacteria from magnetite tailings,the experiment was done with V-Ti magnetite tailings as material. Phosphorus-dissolving experiment was performed on the 136 bacteria from V-Ti magnetite mine tailings in the lab. Four bacteria showing high phosphate-soiubilizing activity were selected in this study,and then the 16 S rRNA genes were sequenced and analyzed. These four bacteria were identified as the genus of Bacillus by phylogenetic analysis. The WDN-5 with highest phosphate-soiubilizing activity was selected from the four phosphate-soiubilizing bacteria by the test of optimum phosphate-soiubilizing time,and its heavey metal resistance and phosphate-soiubilizing effect under the cadmium stress were analyzed. As the result,WDN-5 showed best phosphate-soiubilizing effect with 77. 14μg/ml of solubilizing phosphate at the fifth culturing day. The lethal concentration of cadmium against WDN-5 was 300 mg/L,which indicated that WDN-5 showed high resistance activity against cadmium. Under the stress of 50-300 mg/L,the phosphate-soiubilizing content of WDN-5 gradually decreased with cadmium concentration increasing.
In order to study the phosphorus-dissolving bacteria from magnetite tailings,the experiment was done with V-Ti magnetite tailings as material. Phosphorus-dissolving experiment was performed on the 136 bacteria from V-Ti magnetite mine tailings in the lab. Four bacteria showing high phosphate-soiubilizing activity were selected in this study,and then the 16 S rRNA genes were sequenced and analyzed. These four bacteria were identified as the genus of Bacillus by phylogenetic analysis. The WDN-5 with highest phosphate-soiubilizing activity was selected from the four phosphate-soiubilizing bacteria by the test of optimum phosphate-soiubilizing time,and its heavey metal resistance and phosphate-soiubilizing effect under the cadmium stress were analyzed. As the result,WDN-5 showed best phosphate-soiubilizing effect with 77. 14μg/ml of solubilizing phosphate at the fifth culturing day. The lethal concentration of cadmium against WDN-5 was 300 mg/L,which indicated that WDN-5 showed high resistance activity against cadmium. Under the stress of 50-300 mg/L,the phosphate-soiubilizing content of WDN-5 gradually decreased with cadmium concentration increasing.
引文
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[20]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.
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[23]李剑峰,师尚礼,张淑卿.不同Ca3(PO4)2含量及菌种保存温度下SL01菌株的解磷及生长能力[J].中国生态农业学报,2010,18(1)∶94-97.
[24]钟宇舟.四川盆地大豆根瘤内生细菌分离鉴定及促生效果研究[D].雅安:四川农业大学硕士学位论文,2016.
[25]亢涵,藏春月,焦洋,等.高效除磷菌P7的生长特征及除磷性能研究[J].沈阳建筑大学学报(自然科学版),2017,(5)∶945-952.
[2]谭晓娟.攀枝花钒钛矿区植物对钛的富集特性研究[J].四川环境,2012,31(2)∶46-51.
[3]滕彦国,倪师军,张成江.攀枝花矿业基地矿区环境现状与环境恢复的途径[J].矿业安全与环保,2001,28(3)∶28-30.
[4]庹先国,徐争启,滕彦国,等.攀枝花钒钛磁铁矿区土壤重金属地球化学特征及污染评价[J].矿物岩石地球化学通报,2007,26(2)∶127-131.
[5]魏远,顾红波,薛亮,等.矿山废弃地土地复垦与生态恢复研究进展[J].中国水土保持科学,2012,10(2)∶107-114.
[6]徐翔.建设攀枝花地区矿业生态工业园的可行性分析[J].矿产保护与利用,2010,(4)∶15-17.
[7]张溪,周爱国,甘义群,等.金属矿山土壤重金属污染生物修复研究进展[J].环境科学与技术,2010,33(3)∶106-112.
[8]姜丽新,徐秀茹.尾矿库对周边环境的影响及废弃地再利用[J].辽宁工程技术大学学报,2011,30(s1)∶8-10.
[9]刘惠欣,李富平.矿区尾矿土壤生物修复技术[J].环境科技,2008,21(a01)∶139-141.
[10]康贻军,程洁,梅丽娟,等.植物根际促生菌作用机制研究进展[J].应用生态学报,2010,21(1)∶232-238.
[11]周德超.氮、磷、钾在植物体中的主要生理作用及植物对养分的吸收[J].生物学通报,1983,(5)∶9-10.
[12]李剑峰,张淑卿,师尚礼,等.解磷根瘤菌液体培养基类型、浓度及透气条件的比较[J].草原与草坪,2010,30(1)∶28-32.
[13]张宝贵,李贵桐.土壤生物在土壤磷有效化中的作用[J].土壤学报,1998,(1)∶102-111.
[14]Brookes P C,Powlson D S,Jenkinson D S.Phosphorus in the Soil Microbial Biomass[J].Soil Biology&Biochemistry,1984,16(2)∶169-175.
[15]韩文星,姚拓,梁启鹏,等.PGPR菌肥对燕麦根系性状影响的研究[J].草原与草坪,2008,(4)∶1-4.
[16]田江.尾矿废弃地土壤中解磷菌的筛选、鉴定和特性研究[D].北京:北京林业大学硕士学位论文,2014.
[17]陈廷伟.解磷巨大芽胞杆菌分类名称、形态特征及解磷性能述评[J].中国土壤与肥料,2005,(1)∶7-9.
[18]Reddy M S,Kumar S,Babita K,et al.Biosolubilization of Poorly Soluble Rock Phosphates by Aspergillus Tubingensis and Aspergillus Niger[J].Bioresource Technology,2002,84(2)∶187-189.
[19]李显刚,姚拓,王小利,等.一株分离自葛藤根际高效溶磷细菌特性研究及菌株鉴定[J].中国土壤与肥料,2012,(2)∶87-91.
[20]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.
[21]Pea A,Rmn K,Jwb S.Inorganic Phosphate Solubilization by Two Penicillium Species in Solution Culture and Soil[J].Soil Biology&Biochemistry,1988,20(4)∶459-464.
[22]Illmer P,Schinner F.Solubilization of Inorganic Calcium Phosphates—solubilization Mechanisms[J].Soil Biology&Biochemistry,1995,27(3)∶257-263.
[23]李剑峰,师尚礼,张淑卿.不同Ca3(PO4)2含量及菌种保存温度下SL01菌株的解磷及生长能力[J].中国生态农业学报,2010,18(1)∶94-97.
[24]钟宇舟.四川盆地大豆根瘤内生细菌分离鉴定及促生效果研究[D].雅安:四川农业大学硕士学位论文,2016.
[25]亢涵,藏春月,焦洋,等.高效除磷菌P7的生长特征及除磷性能研究[J].沈阳建筑大学学报(自然科学版),2017,(5)∶945-952.