玉米叶绿体表达载体的构建及AsRED基因原核表达

从玉米幼嫩叶片中提取玉米叶绿体基因DNA,通过PCR克隆出叶绿体同源重组片段trnA和trnI、叶绿体特异性启动子Prrn以及终止子psbA.构建玉米叶绿体表达载体pBAIRTARED,含有一个人工操纵子,其中,筛选标记基因aadA和红色荧光蛋白报告基因AsRED处于Prrn启动子和psbA终止子控制.将构建的载体转化大肠杆菌BL21（DE3）,观测到重组细胞呈现红色,表明构建的载体可以用于玉米叶绿体转化以及表达报告基因.     

长双歧杆菌NCC2705高效转化系统的建立及GFP表达

目的:建立长双歧杆菌NCC2705高效稳定的电转化系统,并以其为宿主表达绿色荧光蛋白(GFP),构建一个稳定的带报告基因的表达载体。方法:从双歧杆菌克隆载体pDG7中切取其在双歧杆菌中复制所必需的pMB1复制子插入质粒pUC19的多克隆位点区,并将gfp基因在双歧杆菌中进行表达;设计双歧杆菌电击转化体系,研究在不同电击电压条件下的电转效率。结果:首先构建了大肠杆菌-长双歧杆菌穿梭质粒pUB1和带有gfp基因的表达质粒pUB2,用电击法将之转化至双歧杆菌,当细菌生长到D600nm约为0.5时制备感受态细胞,在电容25μF、电阻200Ω、电压12.5 kV/cm的电击条件下进行完整质粒转化,可得到较高的转化率。结论:构建了以长双歧杆菌NCC2705为宿主的高效稳定的表达系统,为进一步研究益生菌的分子生物学和功能特性提供了基础。     

The Importance of Defining 'Forest': Tropical Forest Degradation, Deforestation, Long-term Phase Shifts, and Further Transitions

While research continues on the causes, consequences, and rates of deforestation and forest degradation in the tropics, there is little agreement about what exactly is being lost, what we want back, and to whom the 'we' refers. Particularly unsettling is that many analyses and well-intended actions are implemented in fogs of ambiguity surrounding definitions of the term 'forest'—a problem that is not solely semantic; with development of markets for biomass carbon, vegetation classification exercises take on new relevance. For example, according to the basic implementation guidelines of the Kyoto Protocol, closed canopy natural forest could be replaced by monoclonal plantations of genetically engineered exotic tree species and no deforestation would have occurred. Following these same guidelines, carbon credits for afforestation could be available for planting trees in species-rich savannas; these new plantations would count towards a country moving towards the 'forest transition,' the point at which there is no net 'forest' loss. Such obvious conflicts between biodiversity conservation and carbon sequestration might be avoided if 'forest' was clearly defined and if other vegetation types and other ecosystem values were explicitly recognized. While acknowledging that no one approach to vegetation classification is likely to satisfy all users at all scales, we present an approach that recognizes the importance of species composition, reflects the utility of land-cover characteristics that are identifiable via remote sensing, and acknowledges that many sorts of forest degradation do not reduce carbon stocks ( e.g ., defaunation) or canopy cover ( e.g ., over-harvesting of understory nontimber forest products).     

Effects of grazing intensity on soil carbon stocks following deforestation of a Hawaiian dry tropical forest

The effects of forest-to-pasture conversion on soil carbon (C) stocks depend on a combination of climatic and management factors, but factors that relate to grazing intensity are perhaps the least understood. To understand the long-term impact of grazing in converted pastures, methods are needed that accurately measure the impact of grazing on recent plant inputs to soil C in a variety of pasture management and climate settings. Here, we present an analysis from Hawai'i of changes in vegetation structure and soil organic carbon (SOC) along gradients of grazing intensity and elevation in pastures converted from dry tropical forest 100 years ago. We used hyperspectral remote sensing of photosynthetic vegetation, nonphotosynthetic vegetation (NPV) and exposed substrate to understand the effects of grazing on plant litter cover, thus, estimating recent plant inputs to soils (the NPV component). Forest-to-pasture conversion caused a shift from C3 to C4 plant physiology, thus the δ ¹³C method was used in soil cores to measure the fraction of SOC accumulated from pasture vegetation sources following land conversion. SOC decreased in pasture by 5–9 kg C m⁻², depending upon grazing intensity. SOC derived from C3 (forest) sources was constant across the grazing gradient, indicating that the observed variation in SOC was attributable to changes in C inputs following deforestation. Soil C stocks were also reduced in pastures relative to forest soils. We found that long-term grazing lowers SOC following Hawaiian forest-to-pasture conversion, and that these changes are larger in magnitude that those occurring with elevation (climate). Further we demonstrate a relationship between remotely sensed measurements of surface litter and field SOC measurements, allowing for regional analysis of pasture condition and C storage where limited field data are available.     

Bt基因甜菜叶绿体转化载体构建及毒蛋白表达

利用植物叶绿体基因组在进化中高度保守的特点,根据烟草、菠菜、水稻叶绿体基因组全序列资料设计合成引物,PCR扩增并克隆了甜菜叶绿体两个重要功能基因rbcL和atpB(GenBank登录号分别为DQ067450和DQ067451),并以其作为定点整合外源基因的同源重组片段,构建了Bt基因CryIAc甜菜叶绿体定点转化载体pSKARBt,酶切鉴定表明:所构建载体符合预期设计。对克隆菌菌体总蛋白进行了生物杀虫试验,结果表明:Bt基因CryIAc能够在叶绿体特异性启动子及终止子的调控下表达,并对二龄末甘蓝夜蛾有很强的毒杀作用。该载体构建对培育甜菜高抗虫品种具有重要应用价值。叶绿体转化及后续工作正在进行中。     

Leaf lifespan and lifetime carbon balance of individual leaves in a stand of an annual herb, Xanthium canadense

Leaf lifespan in response to resource availability has been documented in many studies, but it still remains uncertain what determines the timing of leaf shedding. Here, we evaluate the lifetime carbon (C) balance of a leaf in a canopy as influenced by nitrogen (N) availability. Stands of Xanthium canadense were established with high-nitrogen (HN) and low-nitrogen (LN) treatments and temporal changes of C gain of individual leaves were investigated with a canopy photosynthesis model. Daily C gain of a leaf was maximal early in its development and subsequently declined. Daily C gain at shedding was nearly zero in HN, while it was still positive in LN. Sensitivity analyses showed that the decline in the daily C gain resulted primarily from the reduction in light level in HN and by the reduction in leaf N in LN. Smaller leaf size in LN than in HN led to higher light levels in the canopy, which helped leaves of the LN stand maintain for a longer period. These results suggest that the mechanism by which leaf lifespan is determined changes depending on the availability of the resource that is most limiting to plant growth.     

Niche construction,human behavior,and the adaptive‐lag hypothesis

Niche construction is the process whereby organisms modify selective environments, thereby affecting evolution. The niche‐construction perspective is particularly relevant to researchers using evolutionary methods to interpret human behavior and society. On the basis of niche‐construction theory, we argue against the hypothesis that modern humans experience an atypically large adaptive lag. We stress that humans construct their world largely to suit themselves and frequently buffer adaptive lag through cultural niche construction. Where they are unable to do that, natural selection of genes rapidly ensues. Our argument has implications for evolutionary psychology and human behavioral ecology, and suggests that the methods of the latter are potentially applicable to all human societies, even postindustrial ones.     

Environmental Enhancement Through Use of Recycled Aggregate Concrete in a Two-Stage Mixing Approach

As a high priority of waste management and recycling by the Hong Kong government, Recycled Aggregate (RA) has been used in various construction applications, mainly as sub-grade, roadwork, and unbound materials. However, higher-grade applications are rare. The major barrier encountered is the variation of quality within RA, which causes lower strength, resulted from crystallization of Recycled Aggregate Concrete (RAC). Therefore, the objective of this study is to examine the crystallization of RAC in a Two-Stage Mixing Approach. Following are the five areas of interest: (i) investigate the waste problems in construction activities; (ii) examine the crystal development on the hydration of cement paste; (iii) develop a two-stage mixing approach (TSMA) for improving the performance of RAC; (iv) explore the crystallization of TSMA in comparison with the Normal Mixing Approach (NMA) through use of Differential Scanning Calorimetry (DSC); and (v) verify the results obtained from DSC analysis with those obtained from compressive strength testing. This study adopted 0, 20, and 100% RA substitution in virgin aggregate and measured by DSC and compressive strength on both TSMA and NMA. TSMA uses only half the water for mixing, forming a thin layer of cement slurry on the surface of RA that will permeate into the porous old cement mortar and fill old cracks and voids in the pre-mix process. The results from DSC analysis clearly demonstrated that TSMA can give a better crystallization of CaO·SiO₂·H₂O [CSH] and Ca(OH)₂[CH]. The optimal situation occurs on 20% RA substitution in virgin aggregate, balancing the advantages of each, a finding supported by the results from compressive strength testing. Therefore, TSMA is a superior methodology and opens a wider application for the use of RAC.     

The effect of population structure on the adaptive radiation of microbial populations evolving in spatially structured environments

Spatial structure is thought to be an important factor influencing the emergence and maintenance of genetic diversity. Previous studies have demonstrated that environmental heterogeneity, provided by spatial structure, leads to adaptive radiation of populations. In the present study, we investigate not only the impact of environmental heterogeneity on adaptive radiation, but also of population fragmentation and niche construction. Replicate populations founded by a single genotype of Escherichia coli were allowed to evolve for 900 generations by serial transfer in either a homogeneous environment, or a spatially structured environment that was either kept intact or destroyed with each daily transfer. Only populations evolving in the structured environment with intact population structure diversified: clones are significantly divergent in sugar catabolism, and show frequency-dependent fitness interactions indicative of stable coexistence. These findings demonstrate an important role for population fragmentation, a consequence of population structure in spatially structured environments, on the diversification of populations.     

贵州省城镇建设用地扩展的时空演变特征

选择贵州省的85个城镇为研究区,运用1973年1∶5万地形图,1986、1995、2000和2007年4期高分辨率遥感影像,选取城镇扩展动态度指数、城镇扩展贡献率指数和城镇扩展强度指数3个指标,构建城镇综合扩展程度指数模型,定量分析了1973—2007年4个时期贵州省城镇建设用地扩展的总体特征和时空分异特征。结果表明:从1973—2007年,贵州省城镇建设用地呈持续增长态势,城镇建设用地面积共增加17550.8915hm2,其中2000—2007年贵州省城镇建设用地扩展面积最大,动态度、贡献率和强度指数均最大;研究区建设用地扩展具有明显的空间分异特征,1973—1986年,城镇建设用地扩展主要集中在省会城市贵阳市,1986—1995年,地级城市所在城镇扩展面积和强度快速提高,1995—2000年,交通沿线城镇扩展速度加快,2000—2007年,贵州省城镇扩展强度和综合扩展程度不仅在贵阳市和地级城市较快,而且在区位和自然条件良好的县级城镇也迅速提高。