一个RNAi载体上反向重复片段的测序策略

相邻的反向重复DNA片段有形成单链内二级结构的倾向,属于一种测序困难的DNA模板。解决RNAi载体插入的反向重复片段的测序问题,为该类载体正确性的测序验证奠定基础。采用常规分子克隆方法构建表达小麦TaATG2串联反向重复片段的RNAi载体,设计2种策略对经菌落PCR初步鉴定的载体进行测序验证:一种是以完整的载体质粒为模板进行测序;另一种是先对载体进行酶切处理,切除反向重复片段中的一个后对保留另一个片段的线性载体进行测序。结果表明,第一种测序策略受到串联反向重复片段形成的单链内部二级结构的影响,测序信号在反向重复片段处出现衰减或乱峰,无法读取序列。第二种测序策略排除了2个反向重复片段之间的干扰,保留在载体上的片段测序信号清晰,序列准确。采用酶切切除一个片段后进行测序的方法,经过2次酶切和2次测序可以有效地对载体上的2个反向重复片段分别进行序列测定,进而确认构建载体的正确性。     

基于多尺度协同的长沙市生态网络构建与层级优化

城市化背景下人类与自然环境的矛盾呈现出多尺度、层级化特征，而传统生态网络的构建方式较少考虑不同尺度下生态要素的关系，无法从区域落实到中心城区，难以形成系统性的解决方案。研究在综合梳理各尺度生态网络构建方法的基础上，以长沙市为例，基于形态学空间格局分析（Morphological Spatial Pattern Analysis，MSPA）、景观连通性原理和生态斑块重要性评价识别生态源地，并通过多层级生态阻力面的确定，综合运用最小费用路径（Least-cost path method，LCP）、电路理论、层级传导理论、尺度嵌套等方法对市域、都市区、中心城区的生态网络进行了协同构建和层级优化，最后基于不同尺度生态网络的特点应用并落实到多层级的国土空间规划体系中。研究结果表明：（1）长沙市域生态网络和都市区生态网络具有较好的层级嵌套特征；共识别两尺度生态叠合源地14个、生态叠合廊道15条，主要通过中心城区内的湘江、浏阳河和捞刀河部分河段与外围生态绿圈相衔接，形成"外环内楔"的空间格局。（2）确定市域重要廊道、市域潜在廊道、生态叠合廊道、都市区重要廊道、都市区潜在廊道的核心保护面积共501.14 km²，并提取位于生态廊道核心保护区范围内的生态夹点和生态障碍点，以进一步落实生态保护修复策略。（3）得到具有重要生态连通功能的中心城区生态绿道长度441.2 km，生态修复单元56个，并结合生态阻力值划分为5级进行针对性修复。（4）基于不同尺度生态网络的衔接、嵌套，最终构建"市域总体生态安全格局-都市区城市生态空间发展格局-以城市绿道为基础的中心城区生态修复单元"，并与不同层级的国土空间规划体系相对应。研究结果将为以大城市为中心的跨尺度生态系统修复和生态安全格局构建提供科学参考。     

Developmental costs of rapid growth in a damselfly

Abstract.  1. Developmental costs of rapid growth in terms of increased fluctuating asymmetry are expected to contribute to the widespread occurrence of growth rates below the physiological maximum, but have rarely been demonstrated. Here, these costs are studied for the first time in an invertebrate, the damselfly Lestes viridis , using a rearing experiment where early- and late-hatched larvae of both sexes were reared at decreasing or permanent water levels.
2. Late-hatched animals were more asymmetrical than early-hatched animals except for males in the drying treatment. Also, females were more asymmetrical than males except in early-hatched animals in the drying treatment.
3. The data presented suggest that in females but not in males treatment groups with higher growth rates have more asymmetrical wings. However, at the individual level no relationship between growth rate and asymmetry was present.
4. Possible reasons why the suggested trade-off between growth and developmental instability was not present at the individual level, and at the group level only in females, are discussed.     

An Experimental study of reduced parental effort and future reproductive success in the puffin, Fratercula arctica

1. The puffin, a long-lived seabird, was studied on the Isle of May, East Scotland between 1990 and 1992. During two of these years, parental effort was experimentally decreased by supplementary feeding of young. This aimed to identify inter-year reproductive costs, and show whether they took the form of reduced adult survival, reduced fledging success and/or a reduction in the 'quality' of offspring in the following year.
2. The feeding treatment significantly reduced the daily number of feeds delivered by experimental parents by 67% in 1990 and 87% in 1991.
3. The proportions of experimental and control parents returning to the colony in the year following manipulation did not differ significantly, although in 1991, 2·5 times as many controls (young unfed) as experimental birds (young fed) failed to return.
4. The fledging success of experimental pairs in the year following manipulation (68%) was significantly higher than that of controls (24%).
5. Experimental pairs raised young with significantly higher body condition (Residual Peak Mass) than that of controls in the year following manipulation (1992).
6. Experimental parents did not differ from controls in their body condition (Lipid Reserve Mass) or rate of reserve depletion, either in the year of manipulation or in the following breeding season; hence there was no evidence for a role of the measured component of body condition in the cost mechanism.
7. The study demonstrated inter-year reproductive costs for puffins and supported the hypothesis that long-lived species reduce the 'quality' of their offspring or abandon a breeding attempt rather than compromise their survival and future opportunities to reproduce.     

Towards automation: Radiata pine shoot hedges in vitro

A novel system for in vitro shoot production has been developed whereby shoot hedges are maintained in one vessel. Monthly crops of shoots are produced for rooting. Radiata pine shoot hedges were maintained on Lepoivre (LP) nutrient agar medium for 18 months using a weekly liquid-nutrient replenishment system. In a separate experiment liquid-LP-nutrient replenishment of shoots twice weekly without transfers (D) resulted in better shoot growth and health than monthly transfers to fresh agar medium (B), monthly transfers to fresh agar medium plus aeration twice weekly (C), or no transfers and no liquid nutrient addition (A). Liquid nutrient replenishment twice weekly was better than 2 weekly or 4 weekly replenishment. The percentage of normal waxy (abundant tubular epicuticular wax) shoots harvested monthly increased significantly over the culture period from 41% at the first harvest to 93% at the eight harvest, and remained high at 97% from the ninth to twelfth harvest. The percentage of wet (no tubular epicuticular wax, small amounts of globular epicuticular wax) shoots harvested showed a corresponding decline—from 59%, to 7% at the eighth harvest. Shoots were harvested at a rate of 672/h (1.19 cents/shoot at a labour cost of NZ$8.00/h) and approximately 1100 shoots were produced per square metre of agar surface per month. Initial problems of contamination and crowding were overcome. These results will greatly facilitate progress towards automation of shoot production and reduction of costs of micropropagated trees. An automated system used in combination with other cost-saving techniques or robotics could potentially result in a substantial reduction in costs. This is the first report of a method of culturing shoots as hedges for a period of up to 18 months without manual subculturing.     

Implementation of advanced Optimum Contribution Selection in small-scale breeding schemes: prospects and challenges in Vorderwald cattle

Vorderwald cattle are a regional cattle breed from the Black Forest in south western Germany. In recent decades, commercial breeds have been introgressed to upgrade the breed in performance traits. On one hand, native genetic diversity of the breed should be conserved. On the other hand, moderate rates of genetic gain are needed to satisfy breeders to keep the breed. These goals are antagonistic, since the native proportion of the gene pool is negatively correlated to performance traits and the carriers of introgressed alleles are less related to the population. Thus, a standard Optimum Contribution Selection (OCS) approach would lead to reinforced selection on migrant contributions (MC). Our objective was the development of strategies for practical implementation of an OCS approach to manage the MC and native genetic diversity of regional breeds. Additionally, we examined the organisational efforts and the financial impacts on the breeding scheme of Vorderwald cattle. We chose the advanced Optimum Contribution Selection (aOCS) to manage the breed in stochastic simulations based on real pedigree data. In addition to standard OCS approaches, aOCS facilitates the management of the MC and the rate of inbreeding at native alleles. We examined two aOCS strategies. Both strategies maximised genetic gain, while strategy (I) conserved the MC in the breeding population and strategy (II) reduced the MC at a predefined annual rate. These two approaches were combined with one of three flows of replacement of sires (FoR strategies). Additionally, we compared breeding costs to clarify about the financial impact of implementing aOCS in a young sire breeding scheme. According to our results, conserving the MC in the population led to significantly (P < 0.01) higher genetic gain (1.16 ± 0.13 points/year) than reducing the MC (0.88 ± 0.10 points/year). In simulation scenarios that conserved the MC, the final value of MC was 57.6% ± 0.004, while being constraint to 58.2%. However, reducing the MC is only partially feasible based on pedigree data. Additionally, this study proves that the classical rate of inbreeding can be managed by constraining only the rate of inbreeding at native alleles within the aOCS approach. The financial comparison of the different breeding schemes proved the feasibility of implementing aOCS in Vorderwald cattle. Implementing the modelled breeding scheme would reduce costs by 1.1% compared with the actual scheme. Reduced costs were underpinned by additional genetic gain in superior simulation scenarios compared to expected genetic gain in reality (+4.85%).     

Persistent courtship reduces male and female longevity in captive tsetse flies Glossina morsitans morsitans Westwood (Diptera: Glossinidae)

Where males can increase their mating success by harassing femalesuntil they accept copulation, harassing tactics can be expectedto evolve to a point where they have costs to the longevityof both sexes. By experimentally manipulating the sex ratioin captive groups of tsetse flies Glossina morsitans morsitans,we demonstrated that the longevity of females declines wheresex ratios are biased toward males, while the longevity of malesdeclines where the sex ratio is biased toward females. Neitherirradiation of males nor prevention of copulation by blockingor damaging the external male genitalia increased the longevityof females caged with them, suggesting that female longevitywas reduced by the physical aspects of male harassment ratherthan by components of the ejaculate     

Corporate sustainability for architecture engineering and construction (AEC) organizations: Framework,transition and implication strategies

This research aims to propose a regenerative sustainability framework for AEC organizations, not only to focus on delivering green certified projects, but to encourage a revitalized approach to systematically drive their sustainability initiative. To achieve this aim, the study conducts an extensive global review of sustainable assessment systems in various industries and organizations, such as corporate sustainable reports, green company competitions, and green-renowned stocks. After learning the best practices from other industries, a sustainable development framework for the AEC organizations is developed. The proposed framework constitutes 110 key indicators covering three-dimensions of sustainability—social, environmental, and economic, respectively—and four-elements of corporate development essentials—projects, operations, governance, and stakeholders. By comparing the proposed framework to existing AEC practices, the study identifies the skewed development of corporate sustainability in the AEC industry where much awareness have been given to project-level environmental matters, but the social dimension, such as social stakeholders and social governance, are seriously lacking and need to be prioritized. A case study of a multi-millions construction company in China was investigated to validate this framework in practice. The case analysis justifies the practical value of the proposed framework, and elaborates the future need of regenerative sustainable initiatives for AEC organizations. The study contributes to the development of corporate sustainability theory in the AEC industry, and also provides industrial practitioners and policy makers with a better understanding of the next generation of corporate sustainable performance and implementation strategies.