电转化法将luxAB基因导入毒死蜱降解茵β菌株的研究

采用电转化法将带发光酶基因luxAB的质粒pTR102导人毒死蜱降解菌β菌株,研究了细胞生长状态、电压强度、电击时间和质粒浓度对转化效率的影响,并对转化子的生理特性进行研究.实验结果表明,以处于对数生长早期的菌液制备感受态细胞,在质粒浓度16.5μg·mL-1、电压2.5kV,电击时间3ms条件下的转化率最高,可达2....     

利用PCR-DGGE溯源典型农村塘坝饮用水中的污染

以华南地区典型农村塘坝饮用水为研究对象,采用基于大肠杆菌的两种特异性基因(β-D-葡萄糖苷酶编码基因uidA和膜外周磷通道蛋白编码基因phoE)的群体差异分析(PCR-DGGE),对饮用水中的污染情况进行了溯源研究.结果表明,采用富集培养的混合菌体DNA作为模板扩增大肠杆菌特异性基因更加实用.uidA和phoE的引物均...     

Archaeal community structure along a gradient of petroleum contamination in saline-alkali soil

The impact of petroleum on archaeal community in salinealkali soils was investigated, which will expand the knowledge of the archaeal population involved in the natural attenuation of hydrocarbons in extreme environments.     

Expression of sulfur uptake assimilation-related genes in response to cadmium, bensulfuron-methyl and their co-contamination in rice roots

The responses of sulfur （S） uptake assimilation-related genes＇ expression in roots of two rice cultivars to cadmium （Cd）, bensulfuron-methyl （BSM） and their co-contamination （Cd＋BSM） were investigated by gene-chip microarray analysis and quantitative real-time PCR （QRT-PCR） technology. Treatments of Cd and Cd＋BSM induced expression of sulfate transporter and permease genes, and promoted sulfate uptake in rice roots. Cd＋BSM could alleviate Cd toxicity to cv. Fengmeizhan seedlings, probably due to Cd＋BSM promoting greater S absorption by seedlings. Cd and Cd＋BSM induced expression of sulfate assimilation-related genes, and thus activated the sulfur assimilation pathway. Cd and Cd＋BSM induced expression of phytochelatin synthase and metallothionein genes, and induced expression of glutathione S-transferases （GSTs）, glutathione synthase （GS） and S- containing antioxidation enzyme genes, which detoxified Cd2＋. It is suggested that （to cope with the toxicity of Cd, BSM and their co-contamination） the S uptake and assimilation pathway was activated in rice roots by increased expression of related genes, thus enhancing the supply of organic S for synthesis of Cd or BSM resistance-related substances.     

Community structure and elevational diversity patterns of soil Acidobacteria

Acidobacteria is one of the most dominant and abundant phyla in soil,and was believed to have a wide range of metabolic and genetic functions. Relatively little is known about its community structure and elevational diversity patterns. We selected four elevation gradients from 1000 to 2800 m with typical vegetation types of the northern slope of Shennongjia Mountain in central China. The vegetation types were evergreen broadleaved forest,deciduous broadleaved forest,coniferous forest and sub-alpine shrubs. We analyzed the soil acidobacterial community composition,elevational patterns and the relationship between Acidobacteria subdivisions and soil enzyme activities by using the 16 S rRNA meta-sequencing technique and multivariate statistical analysis. The result found that 19 known subdivisions as well as an unclassified phylotype were presented in these forest sites,and Subdivision 6 has the highest number of detectable operational taxonomic units(OTUs). A significant single peak distribution pattern(P 0.05) between the OTU number and the elevation was observed. The Jaccard and Bray–Curtis index analysis showed that the soil Acidobacteria compositional similarity significantly decreased(P 0.01) with the increase in elevation distance. Mantel test analysis showed the most of the soil Acidobacteria subdivisions had the significant relationship(P 0.01) with different soil enzymes. Therefore,soil Acidobacteria may be involved in different ecosystem functions in global elemental cycles. Partial Mantel tests and CCA analysis showed that soil pH,soil temperature and plant diversity may be the key factors in shaping the soil Acidobacterial community structure.     

星肋小环藻、铜绿微囊藻春化作用基因控制机理

实验证实了星肋小环藻、铜绿微囊藻的春化作用后,其暴发控制机理成为研究重点。该文通过分析单细胞生物基因操作原理、遗传物质转录复制控制、温度对蛋白质的作用和酶原激活过程等,研究微藻春化作用基因控制机理,提出基因控制模型:激活基因-操纵基因连锁的基因操纵模型,由基因操纵子、调节基因和激活基因组成。该模型阐释2种藻类春末夏初大规模暴发机理:春季,调节基因产生的调节蛋白原被调节蛋白激活酶激活产生的调节蛋白与操纵基因结合,启动结构基因转录合成DNA合成酶系,DNA复制完成,藻体裂殖(繁殖)群体增长;夏季,高温下温度敏感的调节蛋白激活酶变性,使调节蛋白原激活产生变构的调节蛋白,同时与操纵基因和激活基因结合,激活基因不能产生调节蛋白激活酶激活调节蛋白原产生激活蛋白,这时铜绿微囊藻进入生长被抑制状态,星肋小环藻进入高温休眠状态;进入秋季,因之前无低温出现使热变性的激活酶复性,激活产生正常的调节蛋白与操纵基因结合,启动结构基因复制繁殖所需DNA酶系,尽管秋季温光适宜,星肋小环藻不会生长活动,铜绿微囊藻可生长但生长速度低于春末夏初时的生长速度;进入冬季,低温使热变性的激活酶复性,等待来年春暖激活调节蛋白原,启动新一年的藻生长。     

污水处理厂中红霉素抗药性基因的污染特征及选择性因子

采用固相萃取-高效液相色谱串联质谱(SPE-HPLC-MS/MS)检测分析了上海某污水处理厂中12种不同药品和个人护理品(pharmaceutical and personal care products,PPCPs)的含量水平和分布特征,采用实时荧光定量PCR方法定量检测分析了7种红霉素抗药性基因(ERY-ARGs)在污水处理厂中的分布和丰度变化.结果表明,在污水处理厂中共检出5种PPCPs(包括磺胺甲唑、红霉素、四环素、卡马西平和三氯生),浓度范围分别为24.5~38.7、47.5~49.2、43.1~85.4、2.5~3.9和423.2~8 973.3ng·L-1.两段A/O生物处理工艺对三氯生具有较好的去除效果,但对其余4种PPCPs的去除效果较差.与此同时,目标ERY-ARGs在污水处理厂中均被检出,在原水中的丰度达9.28×103(erm A)~1.83×108(ere A)copies·L-1,虽然两段A/O工艺能够显著降低污水中的ERY-ARGs(1.19 log~3.97 log),但其在最终处理出水中的浓度仍然较高.通过相关性分析可知,ERY-ARGs与红霉素和三氯生之间均具有显著相关性(P<0.05),表明污水处理厂中红霉素对ERY-ARGs的产生和传播具有重要影响,而三氯生对ERY-ARGs可能存在交叉选择性作用.     

南海水域不同深度非光合微生物的固碳潜能及其对不同电子供体的响应

通过对南海水域多个区域不同深度的海水分别以H2、Na2S2O3、Na NO2为电子供体经过一定时间的驯化培养后,测定其中非光合微生物的固碳潜力,并统计南海水域不同深度非光合微生物在不同电子供体条件下固碳潜力的差异性,最后结合不同深度海水的主要固碳基因丰度差异,分析南海水域不同深度海洋非光合微生物在不同电子供体条件下固碳潜能差异的原因.结果发现以Na NO2为电子供体时,海洋非光合微生物固碳能力普遍较低,各深度之间没有显著差异;以H2为电子供体时,表层海水中的非光合微生物的固碳潜力显著高于深层海水中的;而以Na2S2O3为电子供体时,深层海水中的非光合微生物的固碳潜力显著高于表层海水中的.基因分析结果表明,固碳基因cbb L在表层海水中的丰度高于深层海水,而cbb M基因在深层海水中的丰度高于表层海水.硫细菌大多以拥有cbb M基因为主,而氢细菌大多以拥有cbb L基因为主.因此不同海洋深度非光合微生物对不同电子供体响应的差异性可能和优势菌群结构的差异有关.海洋表层和深层溶解氧、无机碳含量的差异是导致菌群结构差异,乃至固碳潜力差异的重要原因.     

3种常规消毒方法对磺胺类抗性基因削减效果的比较

近年来,抗生素抗性基因(antibiotic resistance genes,ARGs)对人类和环境的潜在风险引起了广泛关注.本研究从1家城市污水处理厂二级出水中分离出两株磺胺类抗性细菌,控制浓度投加到灭菌后的出水中,分别进行氯消毒、紫外消毒、臭氧消毒处理,采用菌落计数法、q PCR法对消毒后目标细菌及磺胺类抗性基因(sulⅠ、sulⅡ)进行分析比较.同时采用"消毒+DNase I酶"实验研究磺胺类抗性基因在各消毒过程中的行为特征.结果表明,3种消毒均有效削减细菌浓度,但对抗性基因削减程度不高.结合"消毒+DNase I酶"实验分析,氯消毒削减磺胺类抗性基因与微生物量的降低有关;紫外消毒能直接破坏抗性基因;而臭氧消毒在大量削减细菌量的同时,抗性基因进入到胞外环境中,其潜在环境风险不容忽视.     

零价铁对污泥高温厌氧消化过程中四环素抗性基因及第一类整合子的消减影响

为研究污泥高温厌氧消化过程中零价铁(Fe0)对抗生素抗性基因的消减影响,采用荧光定量PCR方法定量检测了7种四环素抗性基因(TC-ARGs,包括tetA、tetC、tetG、tetM、tetO、tetW和tetX)及第一类整合子int I1基因在厌氧消化过程中的丰度变化,同时检测分析了挥发性脂肪酸(VFAs)的质量浓度变化,并探讨了目标基因与VFAs之间的相关关系.结果表明,适量Fe0(如0.10 g·g~(-1),以Fe0/VSS计)不仅可以强化污泥厌氧消化进程,显著提高总VFAs和乙酸的产生量,而且能够增强TCARGs和int I1基因的消减效果.尽管如此,过量Fe0(如1.17 g·g~(-1),以Fe0/VSS计)对污泥高温厌氧消化过程中TC-ARGs和int I1基因消减的增强效果并不显著,推测可能是由于基因水平转移的发生所导致的.通过相关性分析可知,TC-ARGs(除tetO基因外)和int I1基因与乙酸之间存在显著负相关性,表明乙酸可能对TC-ARGs和int I1基因的消减具有促进作用.