洋葱假单胞菌抗汞质粒的研究

用一种改良的方法提取洋葱假单胞菌的抗汞质粒,经过电泳分析,该质粒分子量约1.21×107D,23kb,把纯化的质粒直接转化至大肠杆菌E.coliDH5中,在含100μg/mL的氨苄青霉素和30μg/mL HgCl2的肉汤平板上筛选转化子的抗汞菌株,从生长测定表明,含有抗汞质粒的E.coliDH5α转化株可在含60μg/mL HgCl2的培养基中生长良好,且有较强的去汞功能。质粒经限制性内切酶BamHI、EcoRI、PastI分别酶切,均无酶切位点,且大肠杆菌E.coliDH5α能稳定保存上述质粒。     

一株高效解磷细菌的筛选及其在红壤性水稻土中的施用效果

解磷菌能使土壤中被活性铁、铝等吸附固定的难溶性或不溶性的非有效态磷转化为易于被植物吸收利用的有效态磷,从而大大提高磷肥的利用率.通过对江西鹰潭红壤分离筛选,获得一株性状稳定的高效解磷细菌T4.经鉴定,菌株T4为伯克霍尔德菌属(Burkholderia sp.);T4溶解AlPO4、磷矿粉的能力均比较高,分别为334.2 mg L-1、193.1mg L-1;研究了各种理化因子对T4解磷能力的影响,确定了T4的最佳培养条件为乳糖15 g L-1,KNO3 1.0 g L-1,pH为7.0,温度为30℃,在该条件下T4溶解AlPO4的量为806.3 mg L-1.在江西鹰潭红壤性水稻土的施用试验表明,将菌株T4制成微生物菌剂施用于水稻田可起到减施化肥的作用.图6表2参28     

越南伯克霍尔德菌降解水中微囊藻毒素-LR

探讨利用越南伯克霍尔德菌(Burkholderia Vietnamiensis)降解微囊藻毒素-LR(MC-LR)的可能性和降解机理。结果表明,在1 mg/L的MC-LR溶液中投加体积分数为5%菌株,初始pH为5,温度为30℃的好氧条件下,经过48 h培养后对MC-LR的降解效率达到97.6%。首先,降解溶液的液相色谱分析结果发现,MC-LR在238 nm的特征峰消失。其次,动力学研究发现降解过程符合一级动力学反应方程。最后,SEM、FTIR表征技术对菌株降解前后样品进行分析发现微生物形态和降解产物。因此,Burkholderia Vietnamiensis可能将水中的MC-LR作为碳、氮源利用从而将其降解,Burkholderia Vietnamiensis作为生物降解MC-LR的一种途径是有效的。     

2,2',4,4'-四溴联苯醚高效好氧降解菌的鉴定及其降解路径

2,2',4,4'-四溴联苯醚(BDE-47)是环境和生物体中普遍检出且生物毒性较大的一种多溴联苯醚同系物.本研究从广东省贵屿镇电子垃圾拆解厂周边采集的农田土壤中分离出一株BDE-47的高效好氧降解菌GYP1,考察了其对BDE-47的降解性能、降解路径及不同的环境因素对菌株降解BDE-47的影响.根据菌株形态特征、16S r DNA基因序列及系统发育树分析,将该菌鉴定为伯克霍尔德菌(Burkholderia cepacia).结果表明,菌株GYP1在好氧条件下能够利用BDE-47为唯一碳源生长,在BDE-47初始质量浓度为1 mg·L~(-1)、温度为30℃、摇床转速为150r·min~(-1)的条件下避光培养4 d,该菌株对BDE-47的降解率可达到82.4%.菌株GYP1在15~35℃和p H=4.0~7.0的环境条件范围内对BDE-47均能保持良好的降解性能,但外加蔗糖、酵母粉、联苯醚等碳源会明显抑制其对BDE-47的降解.在菌株GYP1对BDE-47的降解过程中检测到6-OH-BDE-47、5-OH-BDE-47、4'-OH-BDE-17、2'-OH-BDE-3这4种羟基多溴联苯醚及2,4-二溴苯酚,证明菌株GYP1对BDE-47的好氧降解机理主要是羟基化过程.     

伯克氏菌对水稻种子萌发及初生幼苗耐镉性的影响

在无菌条件下研究了伯克氏菌对镉胁迫下水稻种子萌发及初生幼苗耐镉性的影响。结果表明：伯克氏菌D54能在含500mg·L^-1Cd的培养基中正常生长,显示出极强的耐镉能力。在50 mg·L^-1的镉胁迫下,接种伯克氏菌能显著提高水稻种子的萌发率,但在其他镉处理下,接种伯克氏菌对种子萌发无显著影响。接种伯克氏菌对水稻种子的发芽指数没有影响,但在10 mg·L^-1Cd处理下,接种伯克氏菌能显著提高水稻种子的活力指数和初生幼苗的根长、根表面积、根尖总数和耐性系数。接种伯克氏菌对初生幼苗的芽鲜重无影响,但能显著提高根鲜重。在50 mg·L^-1和100 mg·L^-1的高浓度镉处理下,接种伯克氏菌对根系生长没有影响。     

Tween-80和H2O2对Burkholderia cepacia降解油脂的影响

油脂废水是一种重要的水污染源。研究了表面活性剂和H₂O₂对洋葱伯克霍尔德氏菌(Burkholderia cepacia)降解油脂的影响。结果表明：适量的表面活性剂和H₂O₂促进油脂的降解。表面活性剂可以增强油脂在水相中的溶解度，提高油脂在水中的乳化程度从而增加其生物可利用性。当Tween-80浓度600mg/L时，油脂降解率达到91%。低浓度H₂O₂不仅能刺激微生物生长，而且使培养液中的pH值保持相对稳定，提高了微生物对油脂的降解率。H₂O₂在浓度为400mg/L时，能使油脂降解率达到90%。     

Biodegradation of carbofuran in sequencing batch reactor augmented with immobilised Burkholderia cepacia PCL3 on corncob

The performance of sequencing batch reactors (SBRs) augmented with immobilised Burkholderia cepacia PCL3 on corncob for biodegradation of carbofuran in basal salt medium (BSM) was studied. A 2.0-L SBR with a working volume of 1.5 L was operated for a total cycle of 48 h, consisting of 1.0 h fill phase, 46 h react phase and 1.0 h decant phase. The initial pH of the feed medium was 7.0. Air was fed into the reactor at a controlled flow rate of 600 mL·min ^?1. The effect of hydraulic retention time (HRT) (14 to 6 days) on carbofuran-degradation efficiency was investigated at a carbofuran concentration in the feed medium of 20 mg·L ^?1. The shortest HRT resulting in complete degradation of carbofuran was 8 days. At 75% of the optimum HRT (6 days), the effects of biostimulation using organic amendments, i.e. molasses, cassava pulp, rice bran and spent yeast, and the effect of carbofuran concentration in the feed medium (20–80 mg·L ^?1) were investigated. The optimum conditions for SBRs were an initial carbofuran concentration of 40 mg·L ^?1 and 0.1 g·L ^?1 of rice bran as a biostimulated amendment. Complete degradation of carbofuran with a first-order kinetic constant (k ₁) of 0.044 h^?1 was achieved under these optimum conditions.     

Isolation and characterization of a heavy metal-resistant Burkholderia sp. from heavy metal-contaminated paddy field soil and its potential in promoting plant growth and heavy metal accumulation in metal-polluted soil

A heavy metal-resistant bacterial strain was isolated from heavy metal-contaminated soils and identified as Burkholderia sp. J62 based on the 16S rDNA gene sequence analysis. The heavy metal- and antibiotic resistance, heavy metal solubilization of the isolate were investigated. The isolate was also evaluated for promoting plant growth and Pb and Cd uptakes of the plants from heavy metal-contaminated soils in pot experiments. The isolate was found to exhibit different multiple heavy metal and antibiotic resistance characteristics. Atomic absorption spectrometer analysis showed increased bacterial solubilization of lead and cadmium in solution culture and in soils. The isolate produced indole acetic acid, siderophore and 1-aminocyclopropane-1-carboxylate deaminase. The isolate also solubilized inorganic phosphate. Inoculation with the isolate was found to significantly (p < 0.05) increase the biomass of maize and tomato plants. Increase in tissue Pb and Cd contents varied from 38% to 192% and from 5% to 191% in inoculated plants growing in heavy metal-contaminated soils compared to the uninoculated control, respectively. These results show that heavy metal-solubilizing and plant growth promoting bacteria are important for plant growth and heavy metal uptake which may provide a new microbial enhanced-phytoremediation of metal-polluted soils.     

Biodegradation of s-triazine herbicide atrazine by Enterobacter cloacae and Burkholderia cepacia sp. from long-term treated sugarcane-cultivated soils in Kenya

In this study soils from sugarcane-cultivated fields were screened for bacterial species capable of atrazine (6-chloro-N²-ethyl-N⁴-isopropyl-1,3,5-triazine-2,4-diamine) degradation due to long exposure of the soils to this herbicide. To enrich for atrazine degraders, Minimal Salt Medium containing atrazine as the sole N source and glucose as the C source was inoculated with soils impacted with this herbicide and incubated. Bacterial growth was monitored by measuring optical density. The degradation of atrazine was followed by measuring residual atrazine in liquid cultures over a given time period by high performance liquid chromatography. Bacterial strains isolated from the enrichment cultures were characterized by biochemical tests and identified by 16S rRNA gene sequencing. Two bacterial strains coded ISL 8 and ISL 15 isolated from two different fields were shown to have 94 and 96% 16S rRNA gene sequence similarity to Burkholderia cepacia respectively. Another bacterial sp., ISL 14 was closely related to Enterobacter cloacae with a 96% 16S rRNA gene sequence similarity. There was not much difference between the extents of atrazine degradation by the enrichment cultures with communities (79–82% applied amount) from which pure strains were isolated and the pure strains themselves in liquid cultures that showed a degradation of 53–83% of applied amount. The study showed existence of bacterial strains in different sugarcane-cultivated fields which can use atrazine as a nitrogen source. The bacterial strains isolated can be used to enhance the degradation of atrazine in contaminated soils where atrazine is still considered to be recalcitrant.     

菲降解菌的特性及其降解酶纯化研究

研究考察了一株伯克霍尔德氏菌(Burkholderia sp.AFF)的基本特性,并对降解过程中的邻苯二酚2,3-双加氧酶进行了初步分离纯化。菌株AFF对链霉素和四环素具有抗性;当培养基中NaCl的浓度>1.5%时,菌株的生长降解能力受到抑制。菌株AFF生长与降解菲的最佳条件:温度30℃,pH=7.0,摇床转速为150r/min,接种量10%。菌株AFF的细胞粗提物具有邻苯二酚2,3-双加氧酶活性,用阴离子层析交换柱对其进行纯化,对收集到的具有酶活的组分进行SDS-PAGE分析,结果表明目的蛋白分子量在31kDa和43kDa之间。经过一步纯化后,比酶活提高33%,蛋白的回收率为27.6%。