利用CRISPR/Cas9系统敲除斑马鱼bco1基因与bco1l基因

β-胡萝卜素-15,15'-加氧酶(β-carotene-15,15'-momoxygenase 1,bco1)是β-胡萝卜素转化成维生素A过程中的关键酶,bco1与bco1l是编码此酶的主要基因。本实验利用CRISPR/Cas9技术敲除斑马鱼的与β-胡萝卜素-15,15'-加氧酶编码相关的基因bco1与bco1l,以便深入开展对斑马鱼bco1的功能研究。分别在斑马鱼bco1与bco1l基因2号外显子选取sg RNA识别位点,体外转录制备sg RNA并与Cas9 m RNA混合对斑马鱼Ⅰ细胞期受精卵进行显微注射,24 h后收集部分胚胎进行PCR检测并将PCR产物进行单克隆测序确定sg RNA的有效性,构建首建鱼,并在此基础上通过PCR检测、凝胶电泳及测序筛选可遗传突变体。本研究分别获得了bco1基因突变与bco1l基因突变,分析表明这些缺失和插入均可导致编码序列的移码,为研究鱼类胡萝卜素代谢及相关发育过程等后续研究提供了材料。     

Analysis of a broad range of perfluoroalkyl acids in accipiter feathers: method optimization and their occurrence in Nam Co Basin,Tibetan Plateau

Perfluoroalkyl acids (PFAAs) are ubiquitous in the environment. They are prone to accumulate in organisms and have raised public attention in recent decades. Feather samples have been successfully applied as nondestructive indicators for several contaminants. However, a sophisticated analytical method for determining PFAAs in feathers is still lacking. In the present study, a series of conditions, such as the use of the solid-phase extraction cartridge type and extraction/digestion methods, were optimized for the analysis of 13 PFAAs in feathers. According to the spiked recoveries, a weak-anion exchange cartridge was chosen and the methanol was selected as the extraction solvent. In the present study, an optimized pretreatment procedure combined with high-performance liquid chromatography–electrospray ionization–tandem mass spectrometric (HPLC–ESI–MS/MS) method was established for the determination of PFAAs in feathers. The recoveries and method detection limits of the PFAAs ranged from 71 to 120% and 0.16 to 0.54 ng/g, respectively. Finally, 13 PFAAs in four accipiter feather samples from Nam Co Basin, Tibetan Plateau, were analyzed, indicating that PFOS was the predominant PFAA in accipiter feathers, with an average of 4.67 ng/g, followed by the short-chain PFAAs, PFBS and PFBA, with averages of 1.91 and 1.39 ng/g, respectively. These results partly indicated the current situation of PFAA pollution in the Nam Co Basin, especially the existence of short-chain PFAAs in this region.     

固定化真菌对铅离子生物吸附的研究

采用多种固定化材料包埋枝孢霉属(Cladosporium sp.),结果表明:以一定比例混合的海藻酸钠-明胶包埋的枝孢霉小球对Pb2+具有最佳吸附效果。通过正交试验确定了固定化最优操作条件,最优条件为:菌量为10g/L,包埋剂量为1.5∶1.5,CaCl2浓度为4%,钙化时间为4h。并考察了不同因素如接触反应时间,溶液的pH,温度对生物吸附的影响。结果表明:平衡吸附的时间为3h左右,最适pH为4.0,在15℃~45℃的温度范围内,吸附量有缓慢增加,在一定的浓度范围(30~700mg/L)内,生物吸附随浓度的增加而增加,生物吸附过程较好地符合Langmuir模型。     

活性污泥法处理低浓度含磷废水的试验研究

对比研究了厌氧—好氧交替条件下(AAA工艺)活性污泥对三组磷浓度不同的模拟生活废水的除磷效果。试验结果表明,活性污泥对磷浓度为6mg/L左右的废水处理效果最好,磷去除率可达97%;对磷浓度为16.66mg/L和2mg/L左右废水的磷去除率则分别为78.4%和75.3%;并探讨了活性污泥法的除磷机理。     

苏南丘陵地区森林土壤酶活性季节变化

研究了苏南丘陵区不同林分土壤养分、酶活性的剖面分布以及酶活性的季节动态变化特征。研究结果表明,研究区土壤偏酸性,土壤中TN质量分数较高,有机质、TK、AK质量分数中等,TP、AP质量分数较低,土壤TN、有机质、AK、AP的质量分数及蔗糖酶、脲酶、H2O2酶、磷酸酶活性随着土壤深度的增加而逐渐降低。土壤酶活性在不同季节也存在差异,蔗糖酶活性的季节变化表现为：秋季〉夏季〉春季〉冬季,脲酶活性为：夏季〉春季〉秋季〉冬季,毛竹林土壤的磷酸酶活性为：秋季〉春季〉夏季〉冬季,其它3种林分土壤的磷酸酶活性为：秋季〉夏季〉春季〉冬季,H2O2酶活性在马尾松、麻栎、毛竹林土壤内表现为：春季〉秋季〉夏季〉冬季,在杉木内林土壤内表现为：春季〉夏季〉秋季〉冬季。在α=0.01或α=0.05水平下,土壤酶活性与土壤TN、AP、AK、有机质及重元素Pb、Cu、Zn、Cr、Ni呈显著或极显著相关,与土壤容重呈显著或极显著负相关,冬季蔗糖酶与其它3个季节的蔗糖酶呈现极显著或显著正相关;秋季磷酸酶与其它3个季节的磷酸酶呈现极显著或显著正相关;夏季脲酶与其它3个季节的脲酶呈现极显著正相关;夏季H2O2酶与其它3个季节的H2O2酶呈现极显著正相关。     

金沙江干热河谷侵蚀陡坡植被恢复对土壤质量的影响

植被恢复关系到金沙江干热河谷陡坡冲沟侵蚀的发育,土壤质量的演化.文章以元谋干热河谷侵蚀陡坡为例,通过测定植被恢复地与侵蚀裸地的土壤质量,探讨了植被恢复对侵蚀陡坡土壤质量的影响.结果表明:侵蚀陡坡植被恢复有效地防止了土壤侵蚀的发生.与侵蚀裸地相比,植被恢复地土壤剖面层次保持完整,母质层以上有明显的A、B层,土壤厚度变异小且厚度大,物理性粘粒含量高,基本保持着母质为粘土层的变性燥红土质地粘重特征.植被恢复在3年多时间内对土壤紧实度尚没有起到明显改善作用.土壤容重依然偏大.植被恢复对雨季陡坡土壤水分的改善效果好,而在旱季改善效果不好.雨季,植被恢复地水分含量高,不同坡位土壤水分差异不明显.旱季,植被恢复地与侵蚀裸地土壤水分状况相似.两种类型的坡地土壤水分含量都很低,不同坡位土壤水分差异明显,而且土壤水分含量沿向下坡方向降低.植被恢复减弱了陡坡土壤侵蚀带来的有机质流失,促进了有机质的积累.因此,针对目前植被恢复的不足,采取有效的土壤改良措施,保土保水,提高土壤水分储量,减小土壤水分的时空差异,同时降低土壤紧实度,是今后侵蚀陡坡植被恢复值得考虑的一项工作.     

纳米二氧化钛对小鼠肝、肾细胞DNA的损伤

为探讨纳米TiO2对肝、肾细胞DNA的损伤效应,实验制备了锐钛矿型纳米TiO2颗粒(20～100nm),并采用不同浓度的纳米TiO(20、0.1、0.2、0.4、0.8mg·mL-1)对25只昆明雄性小鼠进行染毒,5d后用单细胞凝胶电泳技术检测小鼠肝、肾细胞DNA的损伤程度.结果表明,随着纳米TiO2染毒浓度的升高,小鼠肝、肾细胞尾部DNA百分率(Tail DNA%)和尾矩(Tail Moment)均呈逐渐升高趋势;对于肝细胞,0.1mg·mL-1组Tail DNA%及Tail Moment与对照组无显著差异(p>0.05),≥0.2mg·mL-1组则显著升高(p<0.05或p<0.01);与肝细胞相比,肾细胞Tai lDNA%及Tail Moment变化更为显著,0.1mg·mL-1组即与对照组存在极显著差异(p<0.01).以上结果表明,纳米TiO2染毒可导致小鼠肝、肾细胞DNA损伤,且随染毒浓度的增加,损伤逐渐加重,呈一定的剂量-效应关系;与肝细胞相比,肾细胞对纳米TiO2更为敏感,较低浓度染毒即可造成DNA损伤.     

流态化TiO_2光催化降解甲苯的实验

在自制的流化床反应器中,以实验室制备的TiO2/硅胶为催化剂,对甲苯、空气混合气进行光催化降解,探讨了初始浓度、紫外光强度、操作气速和床层高度等因素对甲苯降解率的影响。实验结果表明:在低浓度范围内(20~55mg/m3),甲苯降解率不随浓度变化,高达100%,但浓度进一步增加后降解率下降;甲苯降解率随紫外光强度的增加而提高,但提幅不大;甲苯初始浓度越低,维持最高降解率的时间越长;操作气速存在最佳值3·98cm/s,达到此值时降解效果最好;甲苯降解率随床层高度(静态高度)的增加而线性提高。     

Arsenic accumulation in the hyperaccumulator Chinese brake and its utilization potential for phytoremediation

The unique property of arsenic hyperaccumulation by the newly discovered Chinese brake (Pteris vittata L.) fern is of great significance in the phytoremediation of arsenic-contaminated soils. The objectives of this study were to (i) examine arsenic accumulation characterized by its distribution pattern in Chinese brake, and (ii) assess the phytoextraction potential of the plant. Young ferns with five or six fronds were transferred to an arsenic-contaminated soil containing 98 mg As kg-1 and grown for 20 wk in a greenhouse. At harvest, the Chinese brake produced a total dry biomass of 18 g plant-1. Arsenic concentration in the fronds was 6000 mg kg-1 dry mass after 8 wk of transplanting, and it increased to 7230 mg kg-1 after 20 wk with a bioconcentration factor (ratio of plant arsenic concentration to water-soluble arsenic in soil) of 1450 and a translocation factor (ratio of arsenic concentration in shoot to that in root) of 24. The arsenic concentrations increased as the fronds aged, with the old fronds accumulating as much as 13,800 mg As kg-1. Most (approximately 90%) of the arsenic taken up by the Chinese brake was transported to the fronds, with the lowest arsenic concentrations in roots. About 26% of the initial soil arsenic was removed by the plant after 20 wk of transplanting. Our data suggest that the arsenic hyperaccumulating property of the Chinese brake could be exploited on a large scale to remediate arsenic contaminated soils.     

Future solutions for the treatment and disposal of hazardous wastes in China

The current status of the treatment and disposal of hazardous wastes in China is summarized on the basis of the results of the Declaring and Registration Project initiated nationally in 1995. A principle framework for the sound management of hazardous wastes is proposed, which includes three levels of technical solutions. Large-scale enterprises are encouraged to recycle, to treat, and to dispose of wastes by means of constructing facilities, and to have their extra capacities available to the public for a reasonable fee. Municipal governments, provincial governments, and the Central Government are to plan and construct centralized facilities to recycle, treat, and dispose of wastes. For a solution at the manufacturing level, recycling is identified as the main approach. Centralized facilities at the municipal level will mainly focus on special wastes that are unsuitable to transport and store, such as hospital waste, and for the technical solution at this level, incineration and recycling are identified as the main approaches. For the technical solution at the provincial and national levels, landfill and incineration are identified as the main approaches. Based on this principle and the current available data on hazardous wastes, a preliminary plan for the spatial distribution of cross-provincial centralized treatment and disposal facilities of hazardous wastes is presented. The construction of approximately nine cross-provincial comprehensive facilities is proposed. A priority list for the construction of these planned facilities is also presented.