Nanomanipulation of Single RNA Molecules by Optical Tweezers

A large portion of the human genome is transcribed but not translated. In this post genomic era, regulatory functions of RNA have been shown to be increasingly important. As RNA function often depends on its ability to adopt alternative structures, it is difficult to predict RNA three-dimensional structures directly from sequence. Single-molecule approaches show potentials to solve the problem of RNA structural polymorphism by monitoring molecular structures one molecule at a time. This work presents a method to precisely manipulate the folding and structure of single RNA molecules using optical tweezers. First, methods to synthesize molecules suitable for single-molecule mechanical work are described. Next, various calibration procedures to ensure the proper operations of the optical tweezers are discussed. Next, various experiments are explained. To demonstrate the utility of the technique, results of mechanically unfolding RNA hairpins and a single RNA kissing complex are used as evidence. In these examples, the nanomanipulation technique was used to study folding of each structural domain, including secondary and tertiary, independently. Lastly, the limitations and future applications of the method are discussed.     

Use of Label-free Optical Biosensors to Detect Modulation of Potassium Channels by G-protein Coupled Receptors

Ion channels control the electrical properties of neurons and other excitable cell types by selectively allowing ions to flow through the plasma membrane¹. To regulate neuronal excitability, the biophysical properties of ion channels are modified by signaling proteins and molecules, which often bind to the channels themselves to form a heteromeric channel complex^2,3. Traditional assays examining the interaction between channels and regulatory proteins require exogenous labels that can potentially alter the protein''s behavior and decrease the physiological relevance of the target, while providing little information on the time course of interactions in living cells. Optical biosensors, such as the X-BODY Biosciences BIND Scanner system, use a novel label-free technology, resonance wavelength grating (RWG) optical biosensors, to detect changes in resonant reflected light near the biosensor. This assay allows the detection of the relative change in mass within the bottom portion of living cells adherent to the biosensor surface resulting from ligand induced changes in cell adhesion and spreading, toxicity, proliferation, and changes in protein-protein interactions near the plasma membrane. RWG optical biosensors have been used to detect changes in mass near the plasma membrane of cells following activation of G protein-coupled receptors (GPCRs), receptor tyrosine kinases, and other cell surface receptors. Ligand-induced changes in ion channel-protein interactions can also be studied using this assay. In this paper, we will describe the experimental procedure used to detect the modulation of Slack-B sodium-activated potassium (K_Na) channels by GPCRs.     

Complete genome sequence of the bacteriochlorophyll a-containing Roseibacterium elongatum type strain (DSM 19469T), a representative of the Roseobacter group isolated from Australian coast sand

Roseibacterium elongatum Suzuki et al. 2006 is a pink-pigmented and bacteriochlorophyll a-producing representative of the Roseobacter group within the alphaproteobacterial family Rhodobacteraceae. Representatives of the marine ‘Roseobacter group’ were found to be abundant in the ocean and play an important role in global and biogeochemical processes. In the present study we describe the features of R. elongatum strain OCh 323^T together with its genome sequence and annotation. The 3,555,102 bp long genome consists of one circular chromosome with no extrachromosomal elements and is one of the smallest known Roseobacter genomes. It contains 3,540 protein-coding genes and 59 RNA genes. Genome analysis revealed the presence of a photosynthetic gene cluster, which putatively enables a photoheterotrophic lifestyle. Gene sequences associated with quorum sensing, motility, surface attachment, and thiosulfate and carbon monoxide oxidation could be detected. The genome was sequenced as part of the activities of the Transregional Collaborative Research Centre 51 (TRR51) funded by the German Research Foundation (DFG).     

人为干扰对大兴安岭北坡兴安落叶松林粗木质残体的影响

比较了兴安落叶松天然林和两种不同干扰类型兴安落叶松林(一次干扰林、二次干扰林)之间活立木蓄积、粗木质残体(CWD)蓄积和组成的差异.结果表明：天然林、一次干扰林和二次干扰林的活立木蓄积量分别为161.6、138.3和114.8 m³·hm^-2,粗木质残体的蓄积量分别为69.77、36.64和32.61 m³·hm^-2.天然林粗木质残体大部分径级在20～40 cm,其中倒木、枯立木分别占总材积的72%和28%;一次干扰林和二次干扰林粗木质残体大部分径级在10～30 cm,其中倒木、枯立木和伐桩分别占各自总材积的70%、14%、16%和57%、15%、28%.人为干扰造成兴安落叶松林粗木质残体蓄积减少,改变了粗木质残体的组成.     

北京市农业景观生态与美学质量空间评价

选择了重点反映农业景观生态与美学功能的生态系统功能、自然性、开阔与多样性、污染概率和整洁度等10个单一空间显性指数,基于田间调查和专家系统赋予其不同权重,构建了农业景观质量综合评价指标,并基于地理信息系统支持下的土地利用数据和利用遥感获得的植被指数,对北京市农业景观质量及其空间差异进行了评价.结果表明：北京市农业景观质量的区域差别较大,城区及小城镇边缘的农业景观质量最差,近郊外围及远郊区较好;北京市农业景观的优劣主要与地形和人为压力有关.基于空间信息技术和景观指数,在大尺度上构建的综合指标体系,对评价农业景观质量的空间差异具有重要意义.     

基于高光谱遥感的小麦叶干重和叶面积指数监测

生物量和叶面积指数(LAI)是描述作物长势的重要参数, 叶干重和LAI的实时动态监测对小麦(Triticum aestivum)生长诊断和管理调控具有重要意义。为分析多种高光谱参数估算小麦叶干重和LAI的效果, 建立小麦叶干重和LAI的定量监测模型, 该研究连续3年采用不同小麦品种进行不同施氮水平的大田试验, 于小麦不同生育期采集田间冠层高光谱数据并测定叶片叶干重和LAI。试验结果显示, 小麦叶干重和LAI随施氮水平的提高而增加, 随生育进程呈单峰动态变化模式。小麦叶干重和LAI与光谱反射率间相关性较好的区域主要位于红光波段(590~710 nm, r<-0.60)和近红外波段(745~1 130 nm, r>0.69)。对于不同试验条件下的叶干重和LAI, 可以使用统一的光谱参数进行定量反演, 其中基于RVI (810, 560)、FD755、GM1、SARVI (MSS)和TC3等光谱参数的方程拟合效果较好。经不同年际独立试验数据的检验表明, 以参数RVI (810, 560)、GM1、SARVI (MSS)、PSSRb、(R_750-800/R_695-740) -1、VOG2和MSR705为变量建立的叶干重和LAI监测模型均给出较好的检验结果。因此, 利用关键特征光谱参数可以有效地评价小麦叶片生长状况, 尤其是光谱参数RVI (810, 560)、GM1和SARVI (MSS)可以对不同条件下小麦叶干重和LAI进行准确可靠的监测。     

猪CAST基因多态性与肌纤维组织学特性及屠宰性状的相关性分析

采用3种限制性核酸内切酶对45头金皮(金华×皮特兰)F₂代猪钙蛋白酶抑制蛋白基因(CAST)的第6外显子和第7外显子之间的片段进行了PCR-RFLP分析, 结果显示在第6内含子内均检测到MspⅠ、HinfⅠ和RsaⅠ酶切多态性。根据酶切结果将CAST基因分为3种基因型(AACCEE、BBDDFF、ABCDEF), 其基因型频率分别为0.1778, 0.2222, 0.6000。对背最长肌做连续性石蜡切片, 分别采用苏木精-伊红和肌球蛋白重链免疫组织化学方法染色并拍照, ImageJ软件统计肌纤维横截面积、密度、直径及MHCⅠ型肌纤维的比例。采用SPSS程序分析了CAST不同基因型对金皮F₂代猪肌纤维组织学特性和屠宰性状的影响。结果表明: BBDDFF基因型个体的肌纤维横截面积和眼肌面积显著地高于ABCDEF基因型个体的肌纤维横截面积和眼肌面积(P < 0.05)。     

Influence of potassium fertilization and foliar application of zinc and phosphorus on growth,yield components,yield and fiber properties of Egyptian cotton (Gossypium barbadense L.)

Aims Supplying optimal quantities of mineral nutrients to growing crop plants is one way to improve crop yields. Nutrients need to be used rationally in order to avoid a negative ecological impact and undesirable effects on the sustainability of agricultural production systems. Excessive application of nutrients also affects the farmer's economy. In order to calculate the amount of fertilizer to be applied to crops, it is necessary to develop recommendation programmers that adjust nutrient rates to crop requirements.Methods Experiments in two successive seasons were conducted to investigate the effect of K fertilization and foliar application of Zn and P on yield and fiber properties of cotton cv. Giza 86. Potassium (0.0 and 47.4 kg of K ha^-1) was soil applied, while chelated zinc (0.0 and 57.6 g of Zn ha^-1, applied twice at 70 and 85 days after sowing 'DAS') and phosphorus (0.0, 576, 1?152 and 1?728 g of P ha^-1, applied twice at 80 and 95 DAS) were applied to the foliage.Important findings Dry matter yield, total chlorophyll concentration, K, Zn and P uptake per plant, number of opened bolls per plant, boll weight, seed index, lint index, seed cotton yield per plant, seed cotton and lint yield ha^-1 and earliness of harvest increased with the application of K, Zn and P. Treatments generally had no significant effect on lint percentage and fiber properties, with exceptions, for micronaire reading and flat bundle strength, and uniformity ratio, where the mean values of these characters were significantly increased over the untreated control by applying K, and for the micronaire reading in the first season, when applying P at 1?728 g ha^-1, and uniformity ratio in the second season, when applying P at 1?152 and 1?728 g ha^-1, where the mean values of these characters were significantly increased over the untreated control by applying P. Under the conditions of this study, applying K fertilization at 47.4 kg ha^-1 combined with spraying cotton plants with zinc at 57.6 g ha^-1 and also with P at 1?728 g ha^-1 improved growth and yield of Egyptian cotton.     

遥感数据支持下不同地表覆盖的区域蒸散

将地表能量平衡系统(SEBS)扩展成遥感日蒸散估算模型,利用MODIS遥感数据估算了黄淮海地区的区域蒸散,并在地理信息系统的支持下分析了不同地表覆盖下的区域蒸散统计分布特征.在缺乏各地表覆盖类型相应蒸散量实测值进行对比的情况下,以2001年4月17日估算的日蒸散量为例,通过各地表覆盖类型日蒸散量间的相互对比分析表明,SEBS估算的区域日蒸散量具有一定的合理性.分析结果表明：在黄淮海地区,荒地具有最低的蒸散量;森林、灌木、草地等地表覆盖类型具有中等的蒸散量;而水体、湿地以及耕地具有较高的蒸散量.可能由于包含绿地和水面,城镇用地的蒸散量也较高.土壤含水量的空间差异性导致森林、灌木、草地和耕地等地表覆盖类型的蒸散量具有明显的空间差异性.耕地蒸散量的空间差异性可以为制定合理、高效的农田灌溉计划提供指示作用.SEBS遥感日蒸散模型的局限性在于有可能低估水体和湿地等地表覆盖类型的蒸散量.     

上海九段沙互花米草种群动态遥感研究

该研究应用遥感技术(RS),结合地理信息系统(GIS)、全球定位系统(GPS)和野外实地调查,对长江口新生沙洲九段沙互花米草(Spartina alterniflora)引种7年来的种群扩散过程进行了调查分析。通过不同时相遥感卫星影像的解译,结合现场调查,分析了外来种互花米草种群自1997年引种后7年期间的种群扩散动态过程。至2004年,互花米草群落已从种植的100 hm²扩展到1 014 hm²。互花米草种群扩散过程可分为3个阶段:1997年的成活定居期;1998至2000年的滞缓期;2000年以后的快速扩散期。互花米草种群的增长速度远高于土著种芦苇(Phragmites australis),显示了互花米草极强的扩散能力、竞争优势和更广的生态幅。研究结果表明,3S技术可应用于植被分类、群落动态和生物入侵动态监测。3S技术具有宏观、快速、动态和综合的优势,可为外来植物的定量调查、分布格局和扩散动态研究提供有效的手段,为滩涂资源生物多样性保护和可持续开发利用提供科学依据。