RNAi转基因大豆品系‘B5C9123-5’的RPA可视化检测技术

为开发RNAi转基因作物的田间可视化鉴定方法,以DNA嵌合染料SYBR Green I为材料,采用设置重组酶聚合酶扩增(Recombinase Polymerase Amplification,RPA)特异性引物的手段,建立了一种快速、低成本、可视化的RPA用于转基因大豆‘B5C9123-5’的检测。结果表明,在靶标不存在时,RPA体系中游离的SYBR Green I染料使溶液呈现较弱的黄色荧光。在靶标存在的情况下,RPA扩增产生的双链DNA与SYBR Green I结合导致溶液从黄色转变为绿色并使荧光迅速增强。通过对RPA扩增时间和温度进行优化,阳性结果可在20min内用肉眼鉴别,检测限为1.00ng/μL。通过设置不同RPA引物,该方法可用于现场快速检测多种RNAi转基因作物。     

苏丹红Ⅰ褪色分光光度法测定环境水体中六价铬

在盐酸介质中,依据六价铬离子(Cr(Ⅵ))氧化苏丹红Ⅰ使其褪色的原理提出了测定Cr(Ⅵ)离子的新方法,确定了最优的测定条件.在485 nm处,Cr(Ⅵ)离子在0.2～2.6 μg/mL范围内符合比尔定律,线性回归方程为A=0.207 0C+0.002 35,相关系数为0.996 0,表观摩尔吸光系数为6.09×104 L/(cm·mol).该方法可用于水样中的微量Cr(Ⅵ)离子的测定,回收率为97.1％～100.3％,RSD小于3.0％.     

Effects of three methods of restraint an in rhesus and african green monkeys intravenous glucose tolerance testing

The intravenous administration of 0.75 gm glucose per kg and the measurement of serum glucose pretest and at 10, 20, 30, 60, 90 and 120 minutes constitute a satisfactory protocol for intravenous glucose tolerance testing of Rhesus (Macaca mulatto) and African Green (Cercopithecus aethiops) monkeys. No significant differences were noted between animals restrained with ketamine hydrochloride and those restrained with sodium pentobarbital, but the African Green males and females and the male Rhesus monkeys yielded significantly different results while being manually restrained.     

猪细小病毒SYBR Green Ⅰ模式实时定量PCR检测方法的建立

根据GenBank猪细小病毒(PPV)的NS1基因序列,设计一对特异性引物,采用SYBR Green Ⅰ随机结合渗入法,建立实时定量PCR检测方法,构建了检测PPV的标准DNA模版,循环阈值(Ct)与标准质粒DNA模板在7.8×101～7.8×108拷贝/μl浓度范围内呈良好的线性关系,相关系数为0.997。该方法用于猪细小病毒的检测具有很高的特异性,其敏感性与常规PCR相比可以提高100倍,可以用于猪细小病毒的快速检测。     

小麦粒重降低原因与防御对策研究

通过试验和调查研究,对１９９８年小麦粒重降低原因进行了分析。结果表明,厄尔尼诺现象造成气候条件的异常是粒重普遍降低的基本原因;麦蚜为害是粒重大幅度降低的重要原因;而品种的综合抗性差以及栽培管理不及时更是其主要原因。重视培养地力,选用综合抗性强的小麦品种,抓好中后期追肥、浇水管理和加强病早防治等,是保证小麦获得高而稳的粒重,实现小麦持续增产增收的战略性措施。     

茶多酚体外氧化产物颜色稳定性及对PC-3细胞生长的影响

以红茶中提取的茶黄素粗提物、绿茶多酚为对照,利用紫外扫描技术、UV-Vis法及MTT法研究了绿茶多酚纳米固定化多酚氧化酶体外氧化产物的颜色稳定性、自由基清除能力和对前列腺癌细胞PC-3生长的抑制作用。在不同pH值条件下,茶黄素粗提物和绿茶多酚纳米固定化酶体外氧化产物的紫外吸收光谱的变化趋势相似,碱性增加,吸光度增加;绿茶多酚、茶黄素粗提物及绿茶多酚纳米固定化酶体外氧化产物对DPPH·自由基均有明显清除作用,绿茶多酚纳米固定化酶体外氧化产物清除效果较绿茶多酚和茶黄素粗提物强;茶黄素粗提物及绿茶多酚纳米固定化酶体外氧化产物对PC-3细胞的生长都有抑制作用,绿茶多酚纳米固定化酶体外氧化产物优于茶黄素粗提物。因此,绿茶多酚纳米碳酸钙固定化酶体外氧化产物中茶黄素保持了红茶中茶黄素所具有的良好生物学活性。     

三元向度下农业生态补偿的金融支持——以绿色信贷为进入路径

法律制度缺位、农村金融抑制、环境金融发展迟滞阻碍了农业生态补偿投融资机制的良性运行。以绿色信贷为路径,从三元主体向度构建农业生态补偿融资体系,发挥政策性金融作用,调动金融机构实施绿色信贷的积极性,创新环境融资方式是问题的解决之道。     

Urban land use types contribute to grassland conservation: The example of Berlin

Urbanisation is an important driver of biodiversity loss, also contributing to habitat loss and fragmentation of grasslands at the urban-rural interface. While urban green spaces are known to include many grassland habitats, it is uncertain to what extent urban land use types harbour grasslands of special conservation interest and whether patch characteristics and connectivity of these differ from grasslands on agricultural land. By relating the city-wide biotope mapping to the land use mapping of Berlin, Germany, we assessed (1) to which specific urban land use types the major grassland biotope types belong, (2) differences in patch characteristics and connectivity, and (3) the conservation value of grassland patches at a typological level by means of their legal protection status. Grasslands cover 5% of Berlin's surface, and 43% of that area is assigned to legally protected grassland types. The majority of legally protected grassland (71%) lies on urban land opposed to 29% on agricultural land. Airports and historic parks, which only cover 2% of land in Berlin, contain one-third of all protected dry grasslands. Wet grassland is more confined to agricultural land. In airports and agricultural areas, grassland patches are larger but of a more complex shape than those in historic parks. In airports, grassland patches show greater connectivity as they are situated in grassland-dominated surroundings. Grassland in historic parks appears to be more vulnerable due to smaller patch sizes and higher fragmentation. The example of Berlin demonstrates that the urban green infrastructure can clearly contribute to grassland conservation and may thus partially compensate for the decline of traditional grasslands in cultural landscapes. It will be important to involve residents and landowners in urban grassland conservation and management because most grassland of special conservation interest (57%) was found outside of conservation areas.     

Cloning and Sequence Analysis of Major Histocompatibility Complex Ⅰ Gene from Goose F1 of Fast-growth Lines

According to the multiple alignments identified major histocompatibility complex Ⅰ (MHC Ⅰ) gene conserved sequence registered in GenBank from the family ducks (Anatidae) anser waterfowl, a pairs of specific primers for the fragments of MHCⅠgene of goose F1 from fast-growth lines were designed and synthesized by Primer Premier 5.0. Using the genome DNA of goose F1 from fast-growth lines, the target gene fragment was obtained by PCR. To conduct sequencing of the fragments of MHCⅠgene of goose F1 from fast-growth lines and make sequence alignment and analysis of protein structure and function by bioinformatics, and research the characteristics of MHCⅠgene of goose F1 and the physicochemical properties of the protein. Bioinformatics was analyzed the nucleic acid data, deduced amino acid sequence and phylogenetic trees. The result of sequence analysis showed that the fragments of MHCⅠgene of goose F1 from fast-growth lines was 1036 bp in length, which coded 96 amino acids polyprotein. The homology were 93% and 83% with MHC Ⅰ gene and coding sequence of Wulong goose in NCBI respectively. There were 72 different bases sequence and 16 amino acids change. There also was higher homology with other poultry, and existed genetic relationship of Siji goose > chickens > ducks.The homology segment sequences corresponding to the fragments of MHCⅠ gene of goose F1 coded 96 amino acids protein, which molecular weight, PI, positively or negatively charged amino acid, estimated half-life, instability index, aliphatic index and average hydrophobicity were 11.342 ku, 5.32, 14, 17, 2.8 h, 34.92, 42.81, -1.066, respectively, and appeared 9 B cell epitopes, but contained no signal peptide. These results indicated that the protein for hydrophilic non-secreted proteins, had the high immunogenicity. In addition, The protein structure study indicated that alpha-helix, beta-sheet, beta-turn and random coil were 31.25%,16.67%, 14.58% and 37.50%, respectively. There existed amino terminal domain and carboxyl terminal domain in the tertiary structure. Therefore, MHC gene had significant difference between species and populations of individuals by the pathogen pressure in environment, and there were the interaction between polymorphism of MHC molecules and the diversity of antigenic peptide. MHC determined the differences of individual susceptibility to disease, and could be treated as a candidate gene for disease resistance.     

沼液在绿色蔬菜中的应用现状

概述了沼液在蔬菜上的应用方式,进一步讨论了沼液在绿色蔬菜生产应用中存在的问题并提出建议,力求使沼液这一有机能源在绿色蔬菜生产中发挥最大作用。