应用于矿山修复的高效菌株鉴定与溶岩机制:基于增强回归树分析

岩石矿区废弃地水土流失问题极度严峻,微生物对修复该问题造成的退化生境具有重要意义,为探究微生物对岩石矿区生境修复的有效性与作用机制,采用两次定向筛选,获得1株高效产铁载体及吲哚乙酸(IAA)的硅酸盐矿物分解细菌,利用16S rRNA基因序列分析鉴定该细菌为Pseudomonas protegens.通过该细菌对硅酸盐岩的岩石分解实验,测定培养液pH值,K、Al、Si这3种元素的释放量,检测岩石溶蚀的粒径变化,并分析培养液中不同有机酸,氨基酸及多糖的浓度变化,研究Pseudomonas protegens NLX-4对硅酸盐的溶蚀效果,同时引入增强回归树(boosted regression tree analysis,BRT)分析,探究其促溶机制.分析表明,该细菌通过代谢积累一定浓度的酒石酸(777 mg·L~(-1))以及多糖(8.21 g·L~(-1)),对硅酸盐岩产生释钾、释铝、脱硅的作用,从而有效促进岩石的分解.菌株Pseudomonas protegens NLX-4可作为修复废弃矿地被破坏生境的良好菌种资源.     

3株溶藻细菌的分离鉴定及其溶藻效应

从武汉市的池塘分离到3株编号分别为M6,M8和M13的溶藻细菌,对这3株细菌的生理生化特性、溶藻专一性和溶藻原因进行了研究.结果表明:M6,M8和M13分别属于葡萄球菌属(Staphylococcus sp.)、芽孢杆菌属(Bacillus sp.)和节杆菌属(Arthrobacter sp.);它们分别能溶解鲍氏织线藻、念珠藻、鱼腥藻、坑形席藻、铜绿微囊藻、鞘丝藻等多种蓝藻,并且它们的液体溶藻现象较固体溶藻现象明显;3株溶藻细菌培养液的过滤液仍有溶藻效应,说明溶藻原因可能是细菌释放某种化学物质所致.      

Degradation of polycyclic aromatic hydrocarbons by Pseudomonas sp. JM2 isolated from active sewage sludge of chemical plant

It is important to screen strains that can decompose polycyclic aromatic hydrocarbons (PAHs) completely and rapidly with good adaptability for bioremediation in a local area. A bacterial strain JM2, which uses phenanthrene as its sole carbon source, was isolated from the active sewage sludge from a chemical plant in Jilin, China and identified as Pseudomonas based on 16S rDNA gene sequence analysis. Although the optimal growth conditions were determined to be pH 6.0 and 37℃, JM2 showed a broad pH and temperature profile. At pH 4.5 and 9.3, JM2 could degrade more than 40% of fluorene and phenanthrene (50 mg/L each) within 4 days. In addition, when the temperature was as low as 4℃, JM2 could degrade up to 24% fluorene and 12% phenanthrene. This showed the potential for JM2 to be applied in bioremediation over winter or in cold regions. Moreover, a nutrient augmentation study showed that adding formate into media could promote PAH degradation, while the supplement of salicylate had an inhibitive effect. Furthermore, in a metabolic pathway study, salicylate, phthalic acid, and 9-fluorenone were detected during the degradation of fluorene or phenanthrene. In conclusion, Pseudomonas sp. JM2 is a high performance strain in the degradation of fluorene and phenanthrene under extreme pH and temperature conditions. It might be useful in the bioremediation of PAHs.     

苯并噻吩脱硫菌株的筛选及脱硫活性研究

从孤岛油田油浸土样中筛选到1株能降解苯并噻吩(BT)的脱硫菌,经初步鉴定该菌为戈登氏菌属(Gordona sp.).实验证明:该菌能以类似于4S途径脱除BT及其衍生物中的硫,但是不能脱除二苯并噻吩(DBT)及其衍生物中的硫.GC-MS分析表明该途径的终产物为邻羟基苯乙醛或其异构体苯并呋喃.在以BT为唯一硫源的培养基中30℃培养48h,Gordona sp.C-6能降解0.15mmol/L的BT,终产物占发酵培养基中BT加入量的50%,其余BT在有氧培养过程中挥发.通过Matlab拟合曲线确定以邻羟基苯乙酸为标准品进行产物定量检测的方法.     

Microbial reduction of uranium (VI) by Bacillus sp. dwc-2: A macroscopic and spectroscopic study

The microbial reduction of U(VI) by Bacillus sp. dwc-2, isolated from soil in Southwest China, was explored using transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and X-ray absorption near edge spectroscopy (XANES). Our studies indicated that approximately 16.0% of U(VI) at an initial concentration of 100 mg/L uranium nitrate could be reduced by Bacillus sp. dwc-2 at pH 8.2 under anaerobic conditions at room temperature. Additionally, natural organic matter (NOM) played an important role in enhancing the bioreduction of U(VI) by Bacillus sp. dwc-2. XPS results demonstrated that the uranium presented mixed valence states (U(VI) and U(IV)) after bioreduction, which was subsequently confirmed by XANES. Furthermore, the TEM and high resolution transmission electron microscopy (HRTEM) analysis suggested that the reduced uranium was bioaccumulated mainly within the cell and as a crystalline structure on the cell wall. These observations implied that the reduction of uranium may have a significant effect on its fate in the soil environment in which these bacterial strains occur.     

Biodegradation of phenol by free and immobilized Acinetobacter sp. strain PD12

A new phenol-degrading bacterium with high biodegradation activity and high tolerance of phenol, strain PD 12, was isolated from the activated sludge of Tianjin Jizhuangzi Wastewater Treatment Facility in China. This strain was capable of removing 500 mg phenol/L in liquid minimal medium by 99.6% within 9 h and metabolizing phenol at concentrations up to 1100 mg/L. DNA sequencing and homologous analysis of 16S rRNA gene identified PD12 to be an Acinetobacter sp. Polyvinyl alcohol （PVA） was used as a gel matrix to immobilize Acinetobacter sp. strain PDI2 by repeated freezing and thawing. The factors affecting phenol degradation of immobilized cells were investigated, and the results showed that the immobilized cells could tolerate a high phenol level and protected the bacteria against changes in temperature and pH. Storage stability and reusability tests revealed that the phenol degradation functions of immobilized cells were stable after reuse for 50 times or storing at 4℃ for 50 d. These results indicate that immobilized Acinetobacter sp. strain PD 12 possesses a good application potential in the treatment of phenol-containing wastewater.     

1株海洋异养硝化-好氧反硝化菌的分离鉴定及其脱氮特性

以海水为基质,采用传统的微生物分离纯化方法,从海底沉积物中分离筛选得到1株耐盐异养硝化-好氧反硝化细菌y5,经形态、生理生化特性以及16S rRNA基因序列分析,鉴定该菌为克雷伯氏菌(Klebsiella sp.).对其脱氮特性及影响因素进行了研究,结果表明,菌株y5的最佳碳源为柠檬酸三钠,最适p H值为7.0,最适C/N为17.菌株均能以NH4Cl、Na NO2和KNO3为唯一氮源进行反应,36 h的去除率分别为77.07%、64.14%和100%.3种氮源共存时,36 h的去除率达到100%.表明菌株y5在高盐废水中具有独立高效的异养硝化和好氧反硝化作用.     

慢生型花生根瘤菌16S rRNA分析

以２１株分离自四川,云南不同花生产区的慢性型花生根瘤菌和６株参比菌株为材料,进行了１６ＳｒＤＮＡＰＣＲ－ＲＦＬＰ,结果除证实了慢性型花生根瘤菌同慢生型大豆根瘤菌（Ｂｒａｄｙｒｈｉｚｏｂｉｕｍｊａｐｏｎｉｃｕｍ）高度的遗传相似性外,还发现了一些菌株同Ｂｒａｄｙｒｈｉｚｏｉｂｉｕｍｌｉａｏｎｉｎｇｅｎｓｉｓ和Ｂｒａｄｙｒｈｉｚｏｂｉｕｍｅｌｋａｎｉｉ有较高的遗传相似性,菌株Ｓｐｒ１－２的１６ＳｒＲＮＡ     

氯氰菊酯降解菌的筛选鉴定及其降解特性研究

从农药厂废水排放口附近的污泥中分离到1株能降解氯氰菊酯的细菌LQ-3.根据其形态、生理生化特征和16S rDNA(GenBank Accession No.FJ222585)序列分析,将该菌株鉴定为Starkeya sp..LQ-3菌株只能以共代谢方式降解氯氰菊酯,在有酵母粉、蛋白胨、葡萄糖等营养物质存在的条件下,5 d内对20 mg·L-1氯氰菊酯的降解率达到72.1%.LQ-3菌株降解氯氰菊酯的最适温度为30 ℃左右,pH值为7～8.LQ-3菌株还能降解功夫菊酯、甲氰菊酯、联苯菊酯和溴氰菊酯.酶的定域试验表明,LQ-3菌株降解氯氰菊酯的酶属于胞外酶.     

红球菌Chr-9降解吡啶和苯酚的特性

研究了红球菌(Rhodococcus)Chr-9菌株在基础盐培养基中降解吡啶和苯酚的特性,分析了菌株降解苯酚和吡啶间的差异.结果表明,菌株Chr-9能够在72 h内将基础盐培养基中的吡啶(200 mg L-1)和苯酚(200 mg L-1)完全降解,同时利用吡啶和苯酚进行生长.菌株降解吡啶的最适温度为35℃,最适pH为8.0.菌株降解吡啶和苯酚的速度与底物的初始浓度呈负相关;在无其它氮源的基础盐培养基中,菌株能够利用吡啶和苯酚协同生长.图7参12