Initial Copper Stress Strengthens the Resistance of Soil Microorganisms to a Subsequent Copper Stress

To improve the prediction of essential ecosystem functioning under future environmental disturbances, it is of significance to identify responses of soil microorganisms to environmental stresses. In this study, we collected polluted soil samples from field plots with eight copper levels ranging from 0 to 3,200 mg Cu kg^?1 soil. Then, the soils with 0 and 3,200 mg Cu kg^?1 were selected to construct a microcosm experiment. Four treatments were set up including Cu0-C and Cu3200-C without further Cu addition, and Cu0-A and Cu3200-A with addition of 57.5 mg Cu kg^?1 soil. We measured substrate-induced respiration (SIR) and potential nitrification rate (PNR). Furthermore, the abundance of bacterial, archaeal 16S rRNA genes, ammonia-oxidizing bacteria and archaea amoA genes were determined through quantitative PCR. The soil microbial communities were investigated by terminal restriction fragment length polymorphism (T-RFLP). For the field samples, the SIR and PNR as well as the abundance of soil microorganisms varied significantly between eight copper levels. Soil microbial communities highly differed between the low and high copper stress. In the microcosm experiment, the PNR and SIR both recovered while the abundance of soil microorganisms varied irregularly during the 90-day incubation. The differences of microbial communities measured by pairwise Bray–Curtis dissimilarities between Cu0-A and Cu0-C on day 0 were significantly higher after subsequent stress than before. However, the differences of microbial communities between Cu3200-A and Cu3200-C on day 0 changed little between after subsequent stress and before. Therefore, initial copper stress could increase the resistance of soil microorganisms to subsequent copper stress.     

Response of Wild Type of Arabidopsis thaliana to Copper Stress

Wild type of Arabidopsis thaliana plants were cultivated hydroponically in Hoagland and Arnon nutrient solution and treated with copper (5 – 100 µM) for 2, 4, 7 and 14 d. A progressive decrease of the root length and biomass was observed at increasing Cu concentration in the nutrient solution. Roots accumulated higher amounts of Cu than shoots at all Cu treatments. Changes of cell and chloroplast ultrastructure of Cu-treated plants were also observed. Cu application did not induce formation of Cu-phytochelatin complexes. Changes in glutathione and glutathione disulfide content observed in roots and shoots of Cu-treated plants suggest their participation in amelioration of metal-induced oxidative stress.     

Soil Microbial Community Response to Land Use Change in an Agricultural Landscape of Western Kenya

Tropical agroecosystems are subject to degradation processes such as losses in soil carbon, nutrient depletion, and reduced water holding capacity that occur rapidly resulting in a reduction in soil fertility that can be difficult to reverse. In this research, a polyphasic methodology has been used to investigate changes in microbial community structure and function in a series of tropical soils in western Kenya. These soils have different land usage with both wooded and agricultural soils at Kakamega and Ochinga, whereas at Ochinga, Leuro, Teso, and Ugunja a replicated field experiment compared traditional continuous maize cropping against an improved N-fixing fallow system. For all sites, principal component analysis of 16S rRNA gene denaturing gradient gel electrophoresis (DGGE) profiles revealed that soil type was the key determinant of total bacterial community structure, with secondary variation found between wooded and agricultural soils. Similarly, phospholipid fatty acid (PLFA) analysis also separated wooded from agricultural soils, primarily on the basis of higher abundance of monounsaturated fatty acids, anteiso- and iso-branched fatty acids, and methyl-branched fatty acids in the wooded soils. At Kakamega and Ochinga wooded soils had between five 5 and 10-fold higher levels of soil carbon and microbial biomass carbon than agricultural soils from the same location, whereas total enzyme activities were also lower in the agricultural sites. Soils with woody vegetation had a lower percentage of phosphatase activity and higher cellulase and chitinase activities than the agricultural soils. BIOLOG analysis showed woodland soils to have the greatest substrate diversity. Throughout the study the two functional indicators (enzyme activity and BIOLOG), however, showed lower specificity with respect to soil type and land usage than did the compositional indicators (DGGE and PLFA). In the field experiment comparing two types of maize cropping, both the maize yields and total microbial biomass were found to increase with the fallow system. Moreover, 16S rRNA gene and PLFA analyses revealed shifts in the total microbial community in response to the different management regimes, indicating that deliberate management of soils can have considerable impact on microbial community structure and function in tropical soils.     

紫鸭跖草根组织应答铜胁迫的转录组分析

紫鸭跖草是一种高耐铜的超积累植物,本研究首次应用RNA-Seq技术对其转录组进行分析。通过全长转录组分析组装了紫鸭跖草耐铜相关候选基因,通过转录组分析,共获得82 471条N50长度为2 299 bp的高质量unigene,为紫鸭跖草的进一步研究提供了丰富的数据。对照组(CK)、300 μmol/L胁迫组(CT1)和1 000 μmol/L胁迫组(CT2)的测序数据已保存在NCBI的SRA数据库中,登录号为SAMN11265427。CT1相比于CK,共有5 028条unigene在根组织中有显著性差异表达,约占全部unigene的6.10%,其中富集上调和下调的基因分别为3 138和1 890条;CT2相比于CT1,根中共有6 813个unigene富集差异表达,占所有unigene的8.26%。其中富集上调和下调的基因分别为2 555和4 258。随机选取10个基因进行qRT-PCR荧光定量分析,结果与Illumina测序数据一致,验证了基因数据差异表达的有效性。上述实验从分子水平上为研究铜胁迫下铜耐受的分子机制提供了理论依据。     

Assessing Genotypic Diversity and Symbiotic Efficiency of Five Rhizobial Legume Interactions Under Cadium Stress for Soil Phytoremediation

In the framework of soil phytoremediation using local legume plants coupled with their native root-nodulating bacteria to increase forage yields and preserve contaminated soils in arid regions of Tunisia, we investigated the diversity of bacteria from root nodules of Lathyrus sativus, Lens culinaris, Medicago marina, M. truncatula, and M. minima and the symbiotic efficiency of these five legume symbiosis under Cadmium stress. Fifty bacterial strains were characterized using physiological and biochemical features such heavy metals resistant, and PCR-RFLP of 16S rDNA. Taxonomically, the isolates nodulating L. sativus, and L. culinaris are species within the genera Rhizobium and the ones associated to Medicago sp, within the genera Sinorhizobium. The results revealed also that the cadmium tolerance of the different legumes-rhizobia interaction was as follows: M. minima<M. truncatula<M. marina<L. sativus<L. culinaris indicating that the effect of Cadmium on root nodulation and biomass production is more deleterious on M. minima-S. meliloti and M. truncatula-S. meliloti than in other symbiosis. Knowledge on genetic and functional diversity of M. marina, L. sativus and L. culinaris microsymbiotes is very useful for inoculant strain selection and can be selected to develop inoculants for soil phytoremediation.     

蛋白质组学在植物水分胁迫应答中的应用研究进展

水分胁迫是影响植物生长发育的主要生长因子。通过蛋白质组学技术可对水分胁迫下植物差异变化的蛋白和基因进行挖掘,在研究植物抗旱生理机制方面意义重大。总结了植物蛋白质组学的基本方法与关键技术,同时从光合与碳代谢相关蛋白、抗氧化系统、渗透调节蛋白、热激蛋白、胚胎发育晚期丰富蛋白、转录因子等方面综述了近几年国际上在植物水分胁迫蛋白质组研究方面的进展,并展望了今后蛋白质组学技术发展的方向。     

Laboratory Measure of an Ecosystem Response to a Sustained Stress

A new method is described for measuring environmental stress through the use of the duckweed (Lemna minor) rhizosphere.     

Melanin Production by a Filamentous Soil Fungus in Response to Copper and Localization of Copper Sulfide by Sulfide-Silver Staining

Gaeumannomyces graminis var. graminis, a filamentous soil ascomycete, exhibited enhanced cell wall melanin accumulation when exposed to as little as 0.01 mM CuSO(inf4) in minimal broth culture. Because its synthesis was inhibited by tricyclazole, the melanin produced in response to copper was dihydroxynaphthalene melanin. An additional hyphal cell wall layer was visualized by electron microscopy when hyphae were grown in the presence of copper and fixed by cryotechniques. This electron-dense layer was between the outer cell wall and the inner chitin layer and doubled the total wall thickness. In copper-grown cells that were also treated with tricyclazole, this electron-dense layer was absent. Atomic absorption spectroscopy demonstrated that up to 3.5 mg of Cu per g of fungal mycelium was adsorbed or taken up by hyphae grown in 0.06 mM CuSO(inf4). A method for silver enhancement was developed to determine the cellular location of CuS. CuS was present in cell walls and septa of copper-grown hyphae. Electron microscopy of silver-stained cells suggested that CuS was associated with the melanin layer of cell walls.     

Soil Carbon Storage Response to Temperature: an Hypothesis

Recently, global and some regional observations of soil carbonstocks and turnover times have implied that warming may notdeplete soil carbon as much as predicted by ecosystem models.The proposed explanation is that microbial respiration of carbonin ‘old’ mineral pools is accelerated less by warmingthan ecosystem models currently assume. Data on the sensitivityof soil respiration to temperature are currently conflicting.An alternative or additional explanation is that warming increasesthe rate of physico-chemical processes which transfer organiccarbon to ‘protected’, more stable, soil carbonpools. These processes include adsorption reactions, some ofwhich are known to have positive activation energies. Theoretically,physico-chemical reactions may be expected to respond more towarming than enzyme-mediated microbial reactions. A simple analyticalmodel and a complex multi-pool soil carbon model are presented,which separate transfers between pools due to physico-chemicalreactions from those associated with microbial respiration.In the short-term, warming depletes soil carbon. But in thelong-term, carbon losses by accelerated microbial respirationare offset by increases in carbon input to the soil (net production)and any acceleration of soil physico-chemical ‘stabilization’reactions. In the models, if net production rates are increasedin response to notional warming by a factor of 1.3, and microbialrespiration (in all pools) by 1.5, then soil carbon at equilibriumremains unchanged if physico-chemical reactions are acceleratedby a factor of about 2.2 (50% more than microbial reactions).Equilibrium soil carbon increases if physico-chemical reactionsare over 50% more sensitive to warming than soil respiration.Copyright 2001 Annals of Botany Company Soil organic matter, carbon, respiration, temperature, stabilization, decomposition, model     

燕麦植株生长及叶片生理活性对土壤镉胁迫的响应

该研究以燕麦品种‘燕科2号’(籽粒镉积累量高,Yh)和‘200919 7 1’(籽粒镉积累量低,Yl)为材料,采用盆栽试验,在土壤Cd胁迫条件下测定两种类型燕麦植株生长、叶片光合生理参数及抗氧化酶活性的变化,探讨两品种对Cd胁迫的响应差异。结果表明：(1)Cd胁迫条件下,燕麦品种Yh和Yl植株生长、气体交换参数及叶绿素荧光参数均受到抑制,并以单株干重、叶片净光合速率(P_n)、光化学淬灭系数(qP)表现最为突出,Yh的降幅分别为38.2%、35.0%、14.7%,Yl降幅分别为40.8%、57.1%、27.3%。(2)镉胁迫均诱导增强了两个类型燕麦品种叶片抗氧化酶活性,尤其是SOD活性得到显著增强,Yh和Yl的增幅分别为41.9%和44.9%; Cd胁迫也显著提高了MDA含量,Yh和Yl的增幅分别达到22.2%和18.1%。研究发现,在Cd胁迫条件下,与低Cd积累燕麦品种相比,高Cd积累燕麦品种具有较高的生物量和抗氧化酶活性,以及较强的净光合速率和优异的叶绿素荧光特性,从而表现出较强的耐Cd性。     

An Asymmetric Respiratory Response Occurring when Fluoride and Copper Ions are Applied Jointly to Chlorella vulgaris

When a cell suspension of Chlorella vulgaris is treated with a mixture of copper and fluoride ions, the respiration almost ceases even though the two ions individually have lit t le inhibitory effect. If instead of adding the ions simultaneously, the cells are subjected to a period of pretreatment with copper sulphate before sodium fluoride the inhibition of respiration becomes more severe as the pretreatment time is lengthened, except at copper levels so high that inhibition is almost complete at all lengths of pretreatment. When, however, pretreatment is with fluoride ions, the inhibition becomes less as the pretreatment time is lengthened, eventually disappearing when the time interval is 90 min or more. Measurements of the amount of copper in the cells indicate that this asymmetry of response cannot be ascribed to differences in the rate at which copper is taken up. It is suggested that fluoride blocks the main respiratory pathway and that copper probably blocks the hexose monophosphate shunt.     

Characteristics of Woodland Rhizobial Populations from Surface- and Deep-Soil Environments of the Sonoran Desert

A collection of 74 rhizobial isolates recovered from nodules of the desert woody legumes Prosopis glandulosa, Psorothamnus spinosus, and Acacia constricta were characterized by using 61 nutritional and biochemical tests. We compared isolates from A. constricta and Prosopis glandulosa and tested the hypothesis that the rhizobia from a deep-phreatic rooting zone of a Prosopis woodland in the Sonoran Desert of southern California were phenetically distinct from rhizobia from surface soils. Cluster analysis identified four major homogeneous groups. The first phenon contained slow-growing (SG) Prosopis rhizobia from surface and deep-phreatic-soil environments. These isolates grew poorly on most of the media used in the study, probably because of their requirement for a high medium pH. The second group of isolates primarily contained SG Prosopis rhizobia from the deep-phreatic rooting environment and included two fast-growing (FG) Psorothamnus rhizobia. These isolates were nutritionally versatile and grew over a broad pH range. The third major phenon was composed mainly of FG Prosopis rhizobia from surface and dry subsurface soils. While these isolates used a restricted range of carbohydrates (including sucrose) as sole carbon sources, they showed better growth on a range of organic acids as sole carbon sources and amino acids as sole carbon and nitrogen sources than did other isolates in the study. They grew better at 36°C than at 26°C. The FG Acacia rhizobia from surface-soil environments formed a final major phenon that was distinct from the Prosopis isolates. They produced very high absorbance readings on all of the carbohydrates tested except sucrose, grew poorly on many of the other substrates tested, and preferred a 36 to a 26°C incubation temperature. The surface populations of Prosopis rhizobia required a higher pH for growth and, under the conditions used in this study, were less tolerant of low solute potential and high growth temperature than were phreatic-soil isolates. SG Prosopis rhizobia from phreatic and surface soils were physiologically distinct, suggesting adaptation to their respective soil environments.     

Comparative Response of Alfalfa to Pratylenchus penetrans Populations

Four populations of Pratylenchus penetrans did not differ (P > 0.05) in their virulence or reproductive capability on Lahontan alfalfa. There was a negative relationship (r = -0 .7 9 ) between plant survival and nematode inocula densities at 26 ± 3 C in the greenhouse. All plants survived at an inoculum level (Pi) of 1 nematode/cm³ soil, whereas survival rates were 50 to 55% at 20 nematodes/cm³ soil. Alfalfa shoot and root weights were negatively correlated (r = - 0.87; P < 0.05) with nematode inoculum densities. Plant shoot weight reductions ranged from 13 % at Pi 1 nematode/cm³ soil to 69% for Pi 20 nematodes/cm³ soil, whereas root weight reductions ranged from 17% for Pi 1 nematode/cm³ soil to 75% for Pi 20 nematodes/cm³ soil. Maximum and minimum nematode reproduction (Pf/Pi) for the P. penetrans populations were 26.7 and 6.2 for Pi 1 and 20 nematodes/cm³ soil, respectively. There were negative correlations between nematode inoculum densities and plant survival (r = 0.84), and soil temperature and plant survival (r = -0 .7 8 ). Nematode reproduction was positively correlated to root weight (r = 0.89).     

Adaptation of Denitrifying Populations to Low Soil pH

Natural denitrification rates and activities of denitrifying enzymes were measured in an agricultural soil which had a 20-year past history of low pH (pH ca. 4) due to fertilization with acid-generating ammonium salts. The soil adjacent to this site had been limed and had a pH of ca. 6.0. Natural denitrification rates of these areas were of similar magnitude: 158 ng of N g⁻¹ of soil day⁻¹ for the acid soil and 390 ng of N g⁻¹ of soil day⁻¹ at the neutral site. Estimates of in situ denitrifying enzyme activity were higher in the neutral soil, but substantial enzyme activity was also detected in the acid soil. Rates of nitrous oxide reduction were very low, even when NO₃⁻ and NO₂⁻ were undetectable, and were ca. 400 times lower than the rates of N₂O production from NO₃⁻. Denitrification rates measured in slurries of the acid and neutral soil showed distinctly different pH optima (pH 3.9 and pH 6.3) which were near the pH values of the two soils. This suggests that an acid-tolerant denitrifying population had been selected during the 20-year period of low pH.     

Barn Owl Productivity Response to Variability of Vole Populations

We studied the response of the barn owl annual productivity to the common vole population numbers and variability to test the effects of environmental stochasticity on their life histories. Current theory predicts that temporal environmental variability can affect long-term nonlinear responses (e.g., production of young) both positively and negatively, depending on the shape of the relationship between the response and environmental variables. At the level of the Czech Republic, we examined the shape of the relationship between the annual sum of fledglings (annual productivity) and vole numbers in both non-detrended and detrended data. At the districts’ level, we explored whether the degree of synchrony (measured by the correlation coefficient) and the strength of the productivity response increase (measured by the regression coefficient) in areas with higher vole population variability measured by the s-index. We found that the owls’ annual productivity increased linearly with vole numbers in the Czech Republic. Furthermore, based on district data, we also found that synchrony between dynamics in owls’ reproductive output and vole numbers increased with vole population variability. However, the strength of the response was not affected by the vole population variability. Additionally, we have shown that detrending remarkably increases the Taylor’s exponent b relating variance to mean in vole time series, thereby reversing the relationship between the coefficient of variation and the mean. This shift was not responsible for the increased synchrony with vole population variability. Instead, we suggest that higher synchrony could result from high food specialization of owls on the common vole in areas with highly fluctuating vole populations.     

Decadal Response of Arctic Freshwaters to Burgeoning Goose Populations

The Arctic is faced with rapid climatic changes, but in some areas, drastic changes in the abundance of herbivores represent an even greater agent of change. Increasing goose populations, especially midcontinent lesser snow geese (Chen caerulescens), have led to an extensive loss of vegetation in terrestrial habitats in the Arctic through heavy grazing and destructive foraging. Our aim was to evaluate the effect of geese on the freshwater systems in their Arctic breeding grounds. We sampled the water chemistry of lakes and ponds across a major goose breeding area in the Eastern Canadian Arctic and compared results to samples taken 13 years earlier to determine whether the changes in water chemistry, if evident, were consistent with effects of geese or of climate. Our results suggest that nutrient loadings have increased while most other parameters associated with the underlying geology and hydrology of the region have stayed in a similar range as a decade ago. The most significant changes were linked to nitrogen and phosphorus; phosphorus concentrations doubled between 2001/2002 and 2015, with the highest levels and greatest changes observed for wetlands inside versus outside of goose breeding areas. Our results suggest that geese are most strongly affecting nutrient loads in freshwaters inside breeding areas, which show evidence of ornithological eutrophication. Nutrient changes of this magnitude, especially in typically oligotrophic Arctic lakes, can have profound consequences on ecosystem structure and function and demonstrate how burgeoning waterfowl populations can act as a vector of rapid environmental change in Arctic freshwaters.     

Structural Changes of Cell Wall and Lignifying Enzymes Modulations in Bean Roots in Response to Copper Stress

Fourteen-day-old bean seedlings were cultured in nutrient solution containing Cu²⁺ ions at various concentrations (50 and 75 μM of CuSO₄) for 3 days. This excess of copper induced a reduction in the water volume absorbed by the plants. Moreover, this reduction was accompanied by an increase of the amount of copper taken up by the roots. Analysis by native gel electrophoresis of cell wall peroxidase activities in the roots revealed a stimulation of two anionic isoforms (A₂ and A₃) under cupric stress conditions. Moreover, the activity of phenylalanine ammonia lyase (EC. 4.3.1.5), which plays an important role in plant defense, was enhanced. Copper-treated bean roots showed modifications in the cell walls of various tissues. Label for lignin, callose, and suberin with berberine-aniline blue revealed abnormal cell wall thickenings in the endodermis, the phloem, and the xylem, especially in plants treated with 75 μM CuSO₄.