深部热水硫酸盐还原菌微滴数字PCR检测技术的建立与应用

【背景】地下深部存在一个生物圈,深部沉积岩、玄武岩、花岗岩和变质岩等岩性环境的微生物群落已被调查,而地下深部碳酸盐岩岩溶-裂隙热储层微生物群落特征仍然不清。硫酸盐还原菌(sulfate-reducing bacteria,SRB)是地下深部频繁检出的微生物。【目的】建立快速准确定量深部热水硫酸盐还原菌的微滴数字PCR (droplet digital PCR,ddPCR)技术。【方法】以SRB的功能基因dsrB为检测目标,优化SRB ddPCR技术的退火温度,考察其线性范围、敏感性、重复性和特异性,并将该技术用于实际样品的检测。【结果】SRB ddPCR技术的最佳退火温度为54 °C,检测的线性范围为1.1×100?1.1×105 copies/μL-DNA,相关系数R2为0.996,检出限为1 copy/μL-DNA,重复性的相对标准差优于9%,对3种非SRB人工构建的质粒均没有扩增,显示该技术具有很好的线性关系、敏感性、重复性和特异性。利用该技术对冀中地热区深部热水、浅层水和土壤样品进行了检测,平均含量分别为(4.0±8.4)×103 copies/mL、(1.6±3.5)×102 copies/mL和(1.5±1.2)×103 copies/g-dw。与浅层水和土壤相比,深部热水富含SRB菌。【结论】为了提高地下深部生物圈认识和合理开发利用深部热水,建立了一种快速、灵敏、准确的SRB ddPCR检测技术,同时为其他指示菌检测技术的建立提供了参考。     

Osmotic Water Permeability of Vacuolar and Plasma Membranes Isolated from Maize Roots

The method of stopped flow was used to follow the changes in light scattering by the vesicles of plasmalemma and tonoplast isolated from maize (Zea maysL.) roots and treated by osmotic pressure. In both membrane preparations, the rate of the process depended on the osmotic gradient and was described with the simple exponential function. The rate constants derived from these functions were the following: the coefficient of water permeability in the tonoplast (P= 165 ± 7 m/s) exceeded by an order of magnitude the corresponding index for plasmalemma (11 ± 2 m/s). The presence of HgCl₂(1.6 nmol/g membrane protein) decreased the tonoplast water permeability by 80%. Microviscosity studies of the hydrocarbon zone in the isolated membranes by using a fluorescent diphenylhexatriene probe demonstrated that the two membranes do not differ in the phase state of their lipid bilayer. The authors conclude that the observed difference in water permeability does not depend on the state of the lipid phase and probably reflects the dissimilar functional activity of plasmalemma and tonoplast aquaporins.     

Phosphorus loss from land to water: integrating agricultural and environmental management

Phosphorus (P), an essential nutrient for crop and animal production, can accelerate freshwater eutrophication, now one of the most ubiquitous forms of water quality impairment in the developed world. Repeated outbreaks of harmful algal blooms (e.g., Cyanobacteria and Pfiesteria) have increased society's awareness of eutrophication, and the need for solutions. Agriculture is regarded as an important source of P in the environment. Specifically, the concentration of specialized farming systems has led to a transfer of P from areas of grain production to animal production. This has created regional surpluses in P inputs (mineral fertilizer and feed) over outputs (crop and animal produce), built up soil P in excess of crop needs, and increased the loss of P from land to water. Recent research has shown that this loss of P in both surface runoff and subsurface flow originates primarily from small areas within watersheds during a few storms. These areas occur where high soil P, or P application in mineral fertilizer or manure, coincide with high runoff or erosion potential. We argue that the overall goal of efforts to reduce P loss to water should involve balancing P inputs and outputs at farm and watershed levels by optimizing animal feed rations and land application of P as mineral fertilizer and manure. Also, conservation practices should be targeted to relatively small but critical watershed areas for P export.     

Effect of Elevated CO2 on Photosynthesis and Chlorophyll Fluorescence of Rose Plants Grown at High Temperature and High Photosynthetic Photon Flux Density

Gas exchange and chlorophyll (Chl) fluorescence were measured on young mature leaves of rose plants (Rosa hybrida cvs. First Red and Twingo) grown in two near-to-tight greenhouses, one under control ambient CO₂ concentration, AC (355 µmol mol^–1) and one under CO₂ enrichment, EC (700 µmol mol^–1), during four flushes from late June to early November. Supply of water and mineral elements was non-limiting while temperature was allowed to rise freely during daytime. Leaf diffusive conductance was not significantly reduced at EC but net photosynthetic rate increased by more than 100 %. Although the concentration of total non-structural saccharides was substantially higher in the leaves from the greenhouse with EC, _PS2 (quantum efficiency of radiation use) around noon was not significantly reduced at EC indicating that there was no down-regulation of electron transport. Moreover, CO₂ enrichment did not cause any increase in the risk of photo-damage, as estimated by the 1 – q_P parameter. Non-photochemical quenching was even higher in the greenhouse with EC during the two summer flushes, when temperature and photosynthetic photon flux density (PPFD) were the highest. Hence rose photosynthesis benefits strongly from high concentrations of atmospheric CO₂ at both high and moderate temperatures and PPFD.     

Hotspots of biotic compositional change in lakes along vast latitudinal transects in northern Canada

Ecotones mark zones of rapid change in ecological structure at various spatial scales. They are believed to be particularly susceptible to shifts caused by environmental transformation, making them key regions for studying the effects of global change. Here, we explored the variation in assemblage structure of aquatic primary producer and consumer communities across latitudinal transects in northeastern North America (Québec‐Labrador) to identify spatial patterns in biodiversity that indicated the location of transition zones across the landscape. We analyzed species richness and the cumulative rate of compositional change (expressed as beta‐diversity) of diatoms and chironomids to detect any abrupt shifts in the rate of spatial taxonomic turnover. We used principal coordinates analysis to estimate community turnover with latitude, then applied piecewise linear regression to assess the position of ecotones. Statistically significant changes in assemblage composition occurred at 52 and 55°N, corresponding to the transition between closed‐ and open‐crown forest, and to the southern onset of the forest tundra (i.e., the forest limit), respectively. The spatial distribution of ecotones was most strongly related to air temperature for chironomids and to vegetation‐ and soil‐related chemical attributes of lake water for diatoms, including dissolved organic carbon content and water color. Lakes at mid‐ to high‐latitudes currently face pressures from rapidly rising temperatures, accompanied by large increases in organic carbon inputs from their catchments, often leading to browning and its associated effects. The biota at the base of food webs in lakes located in transition zones are disproportionately affected by the cascading effects of these multi‐factorial changes, concurrent with pronounced terrestrial greening observed in these regions. Similar patterns of biotic shifts have been observed along alpine aquatic transects, indicating the potential for widespread restructuring of cold, high‐altitude and high‐latitude freshwater communities due to global change.     

陕北黄土高原植被生态系统水分利用效率气候时滞效应

基于月尺度气温、降水以及MODIS-GPP、MODIS-ET遥感数据,采用基于像元的时滞偏相关分析,揭示了陕北黄土高原植被生态系统水分利用效率气候时滞效应及其影响因素。结果表明:①2000—2014年,受退耕还林草生态工程和气候变化的双重影响,陕北黄土高原WUE呈波动变化趋势,2001—2003年显著下降,2003—2005年、2010—2013年显著上升。受不同植被类型的影响,年内WUE呈急速双峰、缓速双峰和单峰型3种变化状态。②陕北黄土高原WUE对降水存在明显的时滞效应,滞后时间多为3个月,人为扰动较大的I-1、I-2、Ⅱ-2生态区以及植被覆盖度较高的Ⅲ-3生态区WUE对降水的时滞响应程度明显低于其他生态区;而对气温的时滞效应较弱,仅南部的Ⅲ-2、Ⅲ-3生态区WUE对气温存在1—2个月滞后时间。③不同植被类型WUE对降水的时滞效应较明显,荒漠类植被WUE对降水时滞响应程度最强,滞后时间近3个月,针叶林时滞响应程度最弱,滞后时间为2.1个月;但对气温的时滞效应总体较弱,针叶林WUE对气温滞后时间为15 d,其余大多数植被类型WUE对气温不存在滞后时间。④WUE气候时滞效应对植被覆盖度...     

Studying Liposomes by Tritium Bombardment

Bilayer liposomes from a mixture of dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylethanolamine (DPPC:DPPE=8:2, molar ratio) or DPPC labeled with ¹⁴C-DPPC (DPPC:¹⁴C-DPPC) were bombarded with thermally activated tritium atoms. The tritiated liposomes were hydrolyzed by phospholipase C, and the tritium incorporation into different parts of the bilayer along its thickness was determined. The tritium flux attenuation coefficients were calculated for the headgroup (k₁=0.176±0.032 Å^–1) and acylglycerol residue (k₂=0.046±0.004 Å^–1) layers indicating a preferential attenuation of the tritium flux in the headgroup region and relative transparence of the membrane hydrophobic part. The finding is potentially important to apply tritium bombardment for investigation of spatial organization of transmembrane proteins in their native lipid environment.     

Effects of Angular Leaf Spot and Rust on Leaf Gas Exchange and Yield of Common Bean (Phaseolus Vulgaris)

Isolated and interactive effects of angular leaf spot (caused by Phaeoisariopsis griseola) and rust (caused by Uromyces appendiculatus) on leaf gas exchange and yield was studied in common bean (Phaseolus vulgaris L. cv. Carioca) plants. Gas exchange was measured on 37, 44, 51, and 58 d after planting using a portable photosynthesis system. The inoculation of plants with P. griseola (P), U. appendiculatus (U), and the combination of both pathogens (P+U) caused a significant reduction of net photosynthetic rate (P _N) and yield. The reduction of stomatal conductance (g _s), P _N, and yield was higher under P and combination of P+U than under U treatment. By effect of U, the reduction on yield was higher than the reductions on gas exchange parameters. On the treatment P+U, a reduction of 23 % in P _N and a correspondent reduction of 32 % in yield was observed. The interactive effects of the pathogens on yield could be explained in part by the decreases in g _s and in P _N of diseased bean leaves. The combined effect of both diseases on yield and gas exchange parameters suggests an antagonistic interaction.