典型滨岸草地生态系统碳稳定同位素组成及分布特征

通过测定上海市青浦区东风港百慕大、白花三叶草、高羊茅和白茅等4种典型滨岸草本植物各组织以及不同垂直深度土壤有机质δ¹³C值,对滨岸草地生态系统的植物-土壤碳稳定同位素特征进行了分析.结果表明: 白花三叶草、高羊茅属于C3植物,百慕大、白茅属于C4植物,其茎叶、凋落物和根系各组织间δ¹³C值无显著差异.C3和C4植物样带表层土壤有机质δ¹³C值随着土壤深度递增而呈现截然不同的变化特征,这与样带本底δ¹³C值以及碳稳定同位素分馏效应有关,同时还受植物根系分布深度的影响.植物输入是土壤有机碳(SOC)的最主要来源,植物有机体δ¹³C组成对土壤有机质δ¹³C值有直接影响,植物各组分δ¹³C值与土壤有机质δ¹³C值均存在极显著相关.4种草本植物样带SOC含量与δ¹³C值均呈极显著相关,其中,C3植物样带SOC含量与δ¹³C值呈线性负相关,C4植物样带SOC含量与δ¹³C值呈线性正相关.     

Soil water use strategy of dominant species in typical natural and planted shrubs in loess hilly region

Aims Artemisia gmelinii is a dominant specie naturally established after abandonment of cultivated lands in the Loess Plateau, and Caragana korshinskii is one of the main planted shrub species to control soil erosion. Improved understanding of water use strategies of these two species is of great significance to evaluate the sustainable development of the Loess Plateau under the trend of climate warming and increasing drought events.
Methods Stable oxygen-18 isotope was used to determine seasonal variations in the water sources of native A. gmelinii communities established after abandonment of cultivated lands for 7 and 30 years and planted C. korshinskii after 30 years. The contributions of soil water from different depths to water uptake were estimated by the MixSIR Bayesian mixing model. The geometric mean regression method was used to fit the line of precipitation to get the local meteoric water line (LWML).
Important findings The stable hydrogen isotope rate (δD) and stable oxygen isotope rate (δ¹⁸O) of soil water and xylem water plotted to the right side of the LWML, indicating that the isotopic compositions of soil water were enriched due to evaporation. The native A. gmelinii communities established after abandonment of cultivated lands for 7 years and planted C. korshinskii after 30 years showed plasticity in switching water sources from different soil layers, extracting water from shallow soil (0-40 cm) when soil water was available, but deeper soil (40-80 cm) when shallow soil water was dry. In contrast, A. gmelinii growing in site after cultivation abandonment for 30 years mainly relied on water from the surface soil (0-10 cm) throughout the growing season. Our results suggest that the ability of A. gmelinii to compete for soil water reduces with aging of the community while the planted C. korshinskii will have competitive advantage under the condition of increasing frequency of drought events in the future.     

环境微生物群落功能研究的新方法和新策略

微生物群落在驱动生物地球化学循环中扮演着重要角色,传统的研究方法可对微生物群落进行遗传结构的解析,但不能有效地与功能研究耦联.概述了近年发展起来的基于核酸和蛋白质水平的分子生物学新方法--环境mRNA 和 rRNA同时荧光原位杂交(FISH)、寡核苷酸微阵列技术(Oligonucleotide Microarray)、 稳定性同位素联合宏基因组学(SIP-enabled Metagenomics)和环境蛋白质组学(Metaproteomics)在环境微生物群落功能研究中的应用,并且对其发展趋势进行了分析和展望.     

辽东楤木光合和蒸腾作用对光照和土壤水分的响应过程

应用CIRAS-2型便携式光合作用系统,测定了不同土壤含水量下辽东楤木光合作用与蒸腾作用的光响应过程,探讨了辽东楤木对光照环境和土壤水分的适应性.结果表明:辽东楤木的净光合速率(P_n)、蒸腾速率(T_r)和叶片水分利用效率(WUE)对光量子通量密度(PFD)的响应过程不同;在强光范围内(PFD 800～1 800 μmol·m^-2·s^-1),随着光强增加,辽东楤木的P_n变化较小,而T_r逐渐减小,WUE明显提高.辽东楤木的光饱和点(LSP)和光补偿点(LCP)分别在800和30 μmol·m^-2·s^-1左右,且受土壤含水量变化的影响较小;但其光合量子效率(Ф)和暗呼吸速率(R_d)受土壤含水量变化的影响较大.辽东楤木的P_n和WUE对土壤含水量的变化有明显的阈值响应,其高效用水的土壤相对含水量(RWC)在44％～79％;在此范围内,辽东楤木能同时获得较高的光合速率和水分利用效率.     

利用CRISPR/Cas9n double nick系统构建人DNAH2基因敲除的U2OS细胞株

基于CRISPR/Cas9n double nick技术构建人DNAH2(Homo sapiens dynein,axonemal,heavy chain 2)基因敲除的U2OS稳定细胞株,旨在研究DNAH2基因的生物学功能。首先设计并合成A、B两个sg RNA(Single guide RNA)以及各自的互补链,退火连接形成DNAH2 sg RNA-A、B双链,再分别与带有BbsⅠ粘性末端的p X462线性载体相连,形成p X462-DNAH2-A、p X462-DNAH2-B重组真核表达质粒。质粒共转染至U2OS细胞后,加入嘌呤霉素,以有限稀释法获得阳性单克隆细胞株,再以蛋白印迹实验检测DNAH2蛋白的表达,最后通过PCR-基因测序技术分析突变特点。结果显示A、B sg RNA双链成功插入p X462载体,U2OS-DNAH2-KO单克隆细胞株中DNAH2蛋白不表达,DNAH2基因发生移码突变,从而证实利用CRISPR/Cas9n double nick系统成功构建人DNAH2基因敲除的U2OS稳定细胞株,为研究DNAH2基因提供有利工具。     

Isotope labeling to determine the dynamics of metabolic response in CHO cell perfusion bioreactors using MALDI‐TOF‐MS

The steady‐state operation of Chinese hamster ovary (CHO) cells in perfusion bioreactors requires the equilibration of reactor dynamics and cell metabolism. Accordingly, in this work we investigate the transient cellular response to changes in its environment and their interactions with the bioreactor hydrodynamics. This is done in a benchtop perfusion bioreactor using MALDI‐TOF MS through isotope labeling of complex intracellular nucleotides (ATP, UTP) and nucleotide sugars (UDP‐Hex, UDP‐HexNAc). By switching to a ¹³C₆ glucose containing feed media during constant operation at 20 × 10⁶ cells and a perfusion rate of 1 reactor volume per day, isotopic steady state was studied. A step change to the ¹³C₆ glucose medium in spin tubes allowed the determination of characteristic times for the intracellular turnover of unlabeled metabolites pools, (≤0.56 days), which were confirmed in the bioreactor. On the other hand, it is shown that the reactor residence time (1 day) and characteristic time for glucose uptake (0.33 days), representative of the bioreactor dynamics, delayed the consumption of ¹³C₆ glucose in the bioreactor and thus the intracellular ¹³C enrichment. The proposed experimental approach allowed the decoupling of bioreactor hydrodynamics and intrinsic dynamics of cell metabolism in response to a change in the cell culture environment. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1630–1639, 2017     

Strategic analysis in evolutionary genetics and the theory of games

This paper is written in memory of John Maynard Smith. In a brief survey it discusses essential aspects of how game theory in biology relates to its counterpart in economics, the major transition in game theory initiated by Maynard Smith, the discrepancies between genetic and phenotypic models in evolutionary biology, and a balanced way of reconciling these models. In addition, the paper discusses modern problems in understanding games at the genetic level using the examples of conflict between endosymbionts and their hosts, and the molecular interactions between parasites and the mammalian immune system.     

Effects of plant harvesting and nutrient enrichment on phytoplankton community structure in a shallow urban lake

The utility of shallow water bodies in urban environments is frequently compromised either by dense beds of submerged plants or cyanobacterial blooms associated with nutrient enrichment. Although submerged plants are often harvested to facilitate recreational uses, this activity may alter the phytoplankton community, which in turn, also may restrict the use of the lake. We tested whether (i) plant harvesting reduced the abundance of flagellate algae and increased the abundance of cyanobacteria, and (ii) whether increasing levels of nutrient enrichment caused shifts in the dominance of heterocytous cyanobacteria, non-heterocytous cyanobacteria and Chlorophyta, in a shallow urban lake in Southern Australia as has been observed for shallow Danish lakes in previous studies. These predictions were tested with large (3000 l), replicated mesocosms in a warm, highly productive, shallow lake densely colonised by the submerged angiosperm, Vallisnaria americana Michaux. The heterokont algae, Chlorophyta, Cyanobacteria and Cryptophyta were the most numerous algal divisions in the lake. The Euglenophyta, although uncommon in early summer, became more abundant towards the end of summer. The Dinophyta and Charophyta were rare. The abundance of the heterokont algae and Euglenophyta was significantly reduced by plant harvesting even after plants had partially re-established 18 weeks after initial harvesting. The decline in the Euglenophyta in response to plant harvesting is consistent with earlier findings, that the relative abundance of flagellate algae tends to be greater in the presence of submerged plants. Contrary to our prediction, we found that the Cyanobacteria did not increase in response to plant harvesting, however the response may be altered under higher nutrient levels. Algal responses to nutrient enrichment in the presence of dense V. americana plants generally followed the patterns observed in shallow Danish lakes despite the large differences in climatic conditions. Both studies found that the abundance of heterocytous cyanobacteria declined at higher levels of nutrient enrichment, whereas non-heterocytous cyanobacteria and chlorophytes increased.     

Northern map turtles (Graptemys geographica) derive energy from the pelagic pathway through predation on zebra mussels (Dreissena polymorpha)

1. Zebra mussels ( Dreissena polymorpha ) derive their energy from the pelagic energy pathway by filtering plankton. Because zebra mussels occur in high densities in littoral habitats, they potentially constitute an important trophic link between littoral consumers and pelagic energy sources. Northern map turtles ( Graptemys geographica ) are widespread in North America and consume zebra mussels.
2. We used stable isotopes analyses to quantify the flow of energy from the pelagic pathway to northern map turtles and to infer the contribution of zebra mussels to map turtle biomass. We then built a bioenergetic model to estimate the annual intake of zebra mussels by northern map turtles in Lake Opinicon, Ontario, Canada.
3. Stable isotopes analyses indicated that zebra mussels constitute between 0% and 14% of the diet of males and between 4% and 36% of the diet of females. Assuming that zebra mussels account for all of the pelagic contribution, we estimated that map turtles consume 3200 kg of zebra mussels annually. Because female map turtles are much larger than males and consume more zebra mussels, they are responsible for 95% of the zebra mussel biomass ingested annually.
4. The pelagic pathway supports an important part of the standing crop biomass of map turtles in Lake Opinicon. We highlight the importance of freshwater turtles in lake ecosystems. Unravelling the trophic interactions mediated by freshwater turtles will lead to a more integrated picture of lake ecosystems.