核酸技术在立枯丝核菌分类与生态学研究中的应用

立枯丝核菌是一种传播广、危害大的土传病原菌,对其展开的研究已涉及各个方面并逐渐深入,近几年借助于核酸技术对其所展开的研究更成为热点。对核酸技术在立枯丝核菌的分类学研究(主要包括G Cmol%含量测定、核酸杂交、各种DNA指纹技术、特异PCR扩增、测序DNA的序列分析)、监测群体动态变化的生态学研究(实时荧光PCR技术)中的应用作一简要的介绍。     

植物功能基因组研究进展

随着植物基因组计划的深入,植物基因组学研究的重点已经转变为基因组功能的研究,即利用基因组序列的信息和高通量的系统分析技术,在基因组水平研究植物结构和组织与植物功能在细胞、有机体和进化上的关系.对功能基因组学研究的内容、方法以及最新研究进展进行了综述.     

匍匐茎草本植物形态可塑性、整合作用与觅食行为研究进展

综述了匍匐茎型克隆植物在形态可塑性、整合作用及觅食行为方面的研究进展。资源斑块性分布是生境异质性的特征之一,适应于异质性生境,匍匐茎植物对环境资源表现了一系列可塑性反应。本文着重从匍匐茎植物对光、水、肥的可塑性反应及其整合作用以及觅食行为等方面的研究进行总结分析,以期对匍匐茎型克隆植物进行更广泛深入的研究。     

Prevalence and evolution of drug resistance HIV-1 variants in Henan, China

To understand the prevalence and evolution of drug resistant HIV strains in Henan China after the implementation of free antiretroviral therapy for AIDS patients. 45 drug naive AIDS patients, 118 AIDS patients who received three months antiretroviral therapy and 124 AIDS patients who received six months antiretroviral treatment were recruited in the southern part of Henan province. Information on general condition, antiretroviral medicines, adherence and clinical syndromes were collected by face to face interview. Meanwhile, 14ml EDTA anticoagulant blood was drawn. CD4/CD8 T cell count, viral load and genotypic drug resistance were tested. The rates of clinical improvement were 55.1% and 50.8% respectively three months and six months after antiretroviral therapy. The mean CD4 cell count after antiretroviral therapy was significantly higher than in drug naive patients. The prevalence rate of drug resistant HIV strains were 13.9%, 45.4% and 62.7% in drug naive patients, three month treatment patients and six month treatment patients, respectively.The number of resistance mutation codons and the frequency of mutations increased significantly with continued antiretroviral therapy. The mutation sites were primarily at the 103, 106 and 215 codons in the three-month treatment group and they increased to 15 codon mutations in the six-month treatment group. From this result, the evolution of drug resistant strains was inferred to begin with the high level NNRTI resistant strain, and then develop low level resistant strains to NRTIs. The HIV strains with high level resistance to NVP and low level resistance to AZT and DDI were highly prevalent because of the AZT DDI NVP combination therapy. These HIV strains were also cross resistant to DLV, EFV, DDC and D4T. Poor adherence to therapy was believed to be the main reason for the emergence and prevalence of drug resistant HIV strains. The prevalence of drug resistant HIV strains was increased with the continuation of antiretroviral therapy in the southern part of Henan province. Measures, that could promote high level adherence,provide new drugs and change ART regimens in failing patients, should be implemented as soon as possible.     

Impacts of population growth and economic development on the nitrogen cycle in Asia

Asia is the major consumer of fertilizer nitrogen and energy in the world, and consequently shares a considerable proportion of the world creation of reactive nitrogen (Nr). However, if estimated on per capita basis, Asia is characterized by a lower arable land area, fertilizer nitrogen consumption, energy consumption, and gross domestic product, as well as lower daily protein intake. To meet the increasing needs for food and energy for the growing population combined with the improvement of living standards, Nr will inevitably increase. The present study estimates the creation of Nr and the emissions of various N compounds into environment in Asia currently and in 2030. In comparison with the world averages, the lower fertilizer nitrogen and energy use efficiencies, and the lower use of animal wastes for agriculture imply that there is potential for moderating the increase in Nr and its impacts on the environment. Strategies for moderating the increase are discussed.     

人工栽培铁皮石斛菌根的细胞学研究

从细胞学水平上研究了人工栽培铁皮石斛的菌根特征。结果表明:真菌与铁皮石斛根成功地形成了菌根,真菌菌丝穿透根被细胞,并在根被的某个细胞中定殖,然后经由外皮层的通道细胞进入皮层薄壁细胞;皮层薄壁细胞层正是真菌与植物体相互作用的活跃部位;真菌侵染大多数的皮层薄壁细胞,并不能侵染内皮层及中柱细胞,从而使根仍然保持生活力。     

应用DGGE研究微生物群落时的常见问题分析

变性梯度凝胶电泳(DGGE)是通过核酸片段对微生物群落进行研究,可以监测未培养细菌及其功能基因,被广泛地应用于微生物群落多样性和动态分析,并成为微生物分子生态学研究中的重要手段之一。文中论述了DGGE操作过程中遇到的常见问题,并提出了相应的解决方法。全面分析了样品预处理过程和PCR扩增效果对DGGE分析的影响,探讨了DGGE图谱的优化过程和图谱分析方法,并对DGGE的应用前景进行了综述。     

利用FRET技术在活细胞内研究红景天甙对Aβ25-35诱导PCI2细胞凋亡的抑制作用

为研究红景天甙（salidroside）对β淀粉样肽25-35（β amyloid peptide25-35,Aβ25-35）诱导PC12细胞凋亡的抑制作用,采用Cell Counting Kit-8（CCK-8）分析细胞的存活率,通过光镜检测细胞形态并配以Hoechst染色检测细胞核固缩,利用荧光共振能量转移（fluorescence resonance energy transfer,FRET）技术在单个活细胞中检测caspase-3和caspase-8活性的动态变化。结果表明,红景天甙可剂量依赖性抑制Aβ25-35引起的细胞凋亡,提高细胞的存活率;红景天甙对caspase-3的活性有明显的抑制作用,而且Aβ25-35诱导细胞凋亡不依赖于caspase-8的激活。这些结果提示抑制caspase-3的活性是红景天甙抑制Aβ25-35诱导PC12细胞凋亡的机制之一。     

滇池试验围隔内不同形态铁浓度的变化与物化因子的关系

在蓝藻水华形成以后,通过围隔实验,从2003年6月份到10月份定期采样测定水体中的pH、溶解氧(DO)、水温、总铁、亚铁、过滤性铁(<0.45μm)和可溶性磷的浓度,研究物化因子对不同形态铁浓度变化的影响。实验结果表明,蓝藻水华优势种微囊藻在pH 7—9和水温17.5—20.5℃的条件下,生长旺盛,消耗了大量的亚铁,使亚铁浓度大幅度下降;溶解氧和磷酸盐对亚铁浓度没有显著影响;在水华蓝藻严重发生的条件下,水体中的总铁和过滤性铁浓度没有显著意义的变化,而亚铁浓度的变化与水华蓝藻的种群密度成显著负相关(r=-0.8391,P<0.05)。     

黄瓜膨胀素的重组表达及活性分析

目的:提高纤维素的酶水解效率和开发高效的纤维素酶水解过程。方法:采用RT-PCR方法从黄瓜胚轴细胞中分离了膨胀素S1的cDNA,并使之与毕赤酵母表达质粒pPICZ(A连接,形成重组质粒pPICZ(A-exs1。通过电转化方法,用质粒pPICZ(A-exs1转化巴氏毕赤酵母GS115,得到重组菌株P.pastoris-exs1。在该重组菌株中,膨胀素的基因通过同源重组整合在毕赤酵母的染色体上,并处于毕赤酵母甲醇氧化酶启动子的下游。重组菌株P.pastoris-exs1在甲醇诱导下可合成并分泌膨胀素。结果:培养上清液没有纤维素酶活性,但具有破坏滤纸纤维素结晶结构的能力。培养上清液与里氏木霉纤维素酶等量混合后,可使纤维素酶的滤纸酶活力提高50%。结论:采用巴氏毕赤酵母GS115重组成功表达了黄瓜膨胀素,其表达产物可以促进纤维素酶对滤纸的水解。