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1.
Early stages of biofilm succession in a lentic freshwater environment   总被引:1,自引:0,他引:1  
Sekar  R.  Venugopalan  V.P.  Nandakumar  K.  Nair  K.V.K.  Rao  V.N.R. 《Hydrobiologia》2004,512(1-3):97-108
Initial events of biofilms development and succession were studied in a freshwater environment at Kalpakkam, East Coast of India. Biofilms were developed by suspending Perspex (Plexiglass) panels for 15 days at bimonthly intervals from January 1996 to January 1997. Changes in biofilm thickness, biomass, algal density, chlorophyll a concentration and species composition were monitored. The biofilm thickness, biomass, algal density and chlorophyll a concentration increased with biofilms age and colonization was greater during summer (March, May and July) than other months. The initial colonization was mainly composed of Chlorella vulgaris, Chlorococcum humicolo (green algae), Achnanthes minutissima, Cocconeis scutellum, C. placentula (diatoms) and Chroococcus minutus (cyanobacteria) followed by colonial green algae such as Pediastrum tetras, P. boryanumand Coleochaete scutata, cyanobacteria (Gloeocapsa nigrescens), low profile diatoms (Amphora coffeaeformis, Nitzschia amphibia, and Gomphonema parvulum) and long stalked diatoms (Gomphoneis olivaceumand Gomphonema lanceolatum). After the 10th day, the community consisted of filamentous green algae (Klebshormidium subtile, Oedogonium sp., Stigeoclonium tenue and Ulothrix zonata) and cyanobacteria (Calothrix elenkinii, Oscillatoria tenuis and Phormidium tenue). Based on the percentage composition of different groups in the biofilm, three phases of succession could be identified: the first phase was dominated by green algae, the second by diatoms and the third phase by cyanobacteria. Seasonal variation in species composition was observed but the sequence of colonization was similar throughout the study period.  相似文献   

2.
Algalogically pure cultures were isolated from different aquatic ecosystems and identified as representatives of green algae, euglena algae, and cyanobacteria. Isolated cultures and collection strains of microalgae were screened for productivity. Induced mutagenesis (exposure to UV radiation, 254 nm, 40 Erg/mm2) and selection yielded a mutant strain C-2m2 of Chlorella pyrenoidosa notable for a high activity of lipid biosynthesis and accumulation. Optimization of its culture conditions was conducted. It was found that the active lipid accumulation by the cells of C. pyrenoidosa strain C-2m2 occurred when nitrogen concentration in the medium was tenfold reduced (to the level of 0.004 g/L) at an illuminance of 4 klx.  相似文献   

3.
The nanotechnology industry is growing rapidly, leading to concerns about the potential ecological consequences of the release of engineered nanomaterials (ENMs) to the environment. One challenge of assessing the ecological risks of ENMs is the incredible diversity of ENMs currently available and the rapid pace at which new ENMs are being developed. High-throughput screening (HTS) is a popular approach to assessing ENM cytotoxicity that offers the opportunity to rapidly test in parallel a wide range of ENMs at multiple concentrations. However, current HTS approaches generally test one cell type at a time, which limits their ability to predict responses of complex microbial communities. In this study toxicity screening via a HTS platform was used in combination with next generation sequencing (NGS) to assess responses of bacterial communities from two aquatic habitats, Lake Michigan (LM) and the Chicago River (CR), to short-term exposure in their native waters to several commercial TiO2 nanomaterials under simulated solar irradiation. Results demonstrate that bacterial communities from LM and CR differed in their sensitivity to nano-TiO2, with the community from CR being more resistant. NGS analysis revealed that the composition of the bacterial communities from LM and CR were significantly altered by exposure to nano-TiO2, including decreases in overall bacterial diversity, decreases in the relative abundance of Actinomycetales, Sphingobacteriales, Limnohabitans, and Flavobacterium, and a significant increase in Limnobacter. These results suggest that the release of nano-TiO2 to the environment has the potential to alter the composition of aquatic bacterial communities, which could have implications for the stability and function of aquatic ecosystems. The novel combination of HTS and NGS described in this study represents a major advance over current methods for assessing ENM ecotoxicity because the relative toxicities of multiple ENMs to thousands of naturally occurring bacterial species can be assessed simultaneously under environmentally relevant conditions.  相似文献   

4.
5.
Metabolic engineering of cyanobacteria has the advantage that sunlight and CO2 are the sole source of energy and carbon for these organisms. However, as photoautotrophs, cyanobacteria generally lack transporters to move hydrophilic primary metabolites across membranes. To address whether cyanobacteria could be engineered to produce and secrete organic primary metabolites, Synechococcus elongatus PCC7942 was engineered to express genes encoding an invertase and a glucose facilitator, which mediated secretion of glucose and fructose. Similarly, expression of lactate dehydrogenase- and lactate transporter-encoding genes allowed lactate accumulation in the extracellular medium. Expression of the relevant transporter was essential for secretion. Production of these molecules was further improved by expression of additional heterologous enzymes. Sugars secreted by the engineered cyanobacteria could be used to support Escherichia coli growth in the absence of additional nutrient sources. These results indicate that cyanobacteria can be engineered to produce and secrete high-value hydrophilic products.Metabolic engineering of photosynthetic microbes is attractive because of the efficient use of light energy by these organisms and the potential for CO2 mitigation during production (21). Conventional terrestrial plants capture solar energy at low efficiencies (about 0.1 to 0.25% for corn and up to 1% for switchgrass), while fast-growing prokaryotic and eukaryotic microalgal species are about 1 order of magnitude more productive and their photosynthetic efficiencies can be >10% (12, 13). Genetic tools for engineering cyanobacterial species, including Synechococcus elongatus PCC7942 (Synechococcus), can be applied to metabolic engineering (7). For example, Deng and Coleman (8) expressed pyruvate decarboxylase and alcohol dehydrogenase in cyanobacteria to produce small amounts of ethanol, and Atsumi et al. recently described efficient synthesis of isobutanol using a four-step pathway established in Escherichia coli (2).Much attention has been focused on metabolic engineering to produce fuels. However, fuel molecules are generally toxic to microbes even at moderate concentrations. In addition, on a per-photon basis, the actual market value of fuels is at best comparable to, and generally lower than, the market value of other commodity organic compounds, such as sugars, lactic acid, and amino acids. Engineering cyanobacteria to produce and secrete hydrophilic or charged molecules would thus be economically desirable.Commonly used metabolic engineering organisms, such as E. coli and yeast (e.g., Saccharomyces cerevisiae), express a variety of transport systems for exporting waste products as well as importing nutrients. As photoautotrophs, cyanobacteria lack many of the transporters found in these organisms. In addition, while most microbes store energy by pumping protons across the plasma membrane, cyanobacteria store energy by transporting protons across the thylakoid membrane. In fact, cyanobacteria tend to alkalinize their growth medium in both laboratory and natural conditions (4), and thus how effective heterologous transporters can be in metabolic engineering of cyanobacteria is an open question. Here, we investigated whether heterologous transporters belonging to the major facilitator superfamily, in combination with relevant enzymes, could be introduced into cyanobacteria for production and secretion of useful products.  相似文献   

6.
The occurrence of 23 cyanobacterial species, belonging to 9 different genera and 5 cyanobacterial lichen species of 5 different genera on exposed, open rock surfaces of inselbergs and on soil in savannas of the Orinoco lowlands and the Guayana uplands is described. Their distribution patterns and frequency within the different habitats are given. The filamentous procaryotic blue-green algae/cyanobacteria Stigonema ocellatum and Scytonema crassum, together with the unicellular cyanobacterium Gloeocapsa sanguinea were the most frequent species on rocks, whereas the filamentous cyanobacterium, Schizothrix telephoroides, dominated in cyanobacterial mats on the savanna soil. All species showed intensively coloured sheaths, either brown or yellow in the case of Stigonema ocellatum and Scytonema crassum, or red in Gloeocapsa sanguinea and Schizothrix telephoroides. In addition, a number of cyanobacterial lichens occurred.  相似文献   

7.
Nitrate reductase activity from filamentous, heterocyst-forming cyanobacteria showed a biphasic kinetic behavior with respect to nitrate as the variable substrate. Two kinetic components were detected, the first showing a higher affinity for nitrate (Km, 0.05-0.25 mm) and a lower catalytic activity and the second showing a lower affinity for nitrate (Km, 5-25 mm) and a higher (3- to 5-fold) catalytic activity. In contrast, among unicellular cyanobacteria, most representatives studied exhibited a monophasic, Michaelis-Menten kinetic pattern for nitrate reductase activity. Biphasic kinetics remained unchanged with the use of different assay conditions (i.e. cell disruption or permeabilization, two different electron donors) or throughout partial purification of the enzyme.  相似文献   

8.
The pulmonary damage induced by nanosized titanium dioxide (nano-TiO2) is of great concern, but the mechanism of how this damage may be incurred has yet to be elucidated. Here, we examined how multiple genes may be affected by nano-TiO2 exposure to contribute to the observed damage. The results suggest that long-term exposure to nano-TiO2 led to significant increases in inflammatory cells, and levels of lactate dehydrogenase, alkaline phosphate, and total protein, and promoted production of reactive oxygen species and peroxidation of lipid, protein and DNA in mouse lung tissue. We also observed nano-TiO2 deposition in lung tissue via light and confocal Raman microscopy, which in turn led to severe pulmonary inflammation and pneumonocytic apoptosis in mice. Specifically, microarray analysis showed significant alterations in the expression of 847 genes in the nano-TiO2-exposed lung tissues. Of 521 genes with known functions, 361 were up-regulated and 160 down-regulated, which were associated with the immune/inflammatory responses, apoptosis, oxidative stress, the cell cycle, stress responses, cell proliferation, the cytoskeleton, signal transduction, and metabolic processes. Therefore, the application of nano-TiO2 should be carried out cautiously, especially in humans.  相似文献   

9.
Photoautotrophs are capable of consuming high quantities of CO2, yet scant research exists examining the influence of different CO2 concentrations on the growth of freshwater diazotrophic or non-diazotrophic cyanobacteria. In this study, we cultured two cyanobacteria taxa (Dolichospermum circinale and Microcystis aeruginosa) within controlled atmospheric CO2 chambers at pre-industrial, and post-industrial concentrations. Biovolume and chlorophyll a (Chl-a) differed as a consequence of the adjusted CO2 gradients. Significantly higher biovolume measurements were observed in the elevated CO2 treatment for the diazotrophic species in the initial experiment. However, a follow-up experiment, with a corrected culture replenishment regime showed Chl-a measurements were greater for the diazotrophic and non-diazotrophic species in the elevated CO2 treatment. Increasing CO2 presents a risk to already compromised eutrophic and hyper-eutrophic ecosystems, and we reason increasing CO2 concentrations could affect photosynthetic performance and CO2 assimilation of surface dwelling cyanobacteria. Further experimental work is required to establish ecological thresholds for freshwater ecosystems, as pH levels showed a measurable reduction within the elevated CO2 treatments. As cyanobacteria species may respond quite differently to future CO2 concentrations similar comparative studies should be carried out that focus on CO2 dynamics and pH. The findings of the study indicate diazotrophic cyanobacteria growth in particular may benefit from elevated atmospheric CO2 concentrations.  相似文献   

10.
Recent studies have demonstrated nanosized titanium dioxide (nano-TiO2)-induced fertility reduction and ovary injury in animals. To better understand how nano-TiO2 act in mice, female mice were exposed to 2.5, 5, and 10 mg/kg nano-TiO2 by intragastric administration for 90 consecutive days; the ovary injuries, fertility, hormone levels, and inflammation-related or follicular atresia-related cytokine expression were investigated. The results showed that nano-TiO2 was deposited in the ovary, resulting in significant reduction of body weight, relative weight of ovary and fertility, alterations of hematological and serum parameters and sex hormone levels, atretic follicle increases, inflammation, and necrosis. Furthermore, nano-TiO2 exposure resulted in marked increases of insulin-like growth factor-binding protein 2, epidermal growth factor, tumor necrosis factor-α, tissue plasminogen activator, interleukin-1β, interleukin -6, Fas, and FasL expression, and significant decreases of insulin-like growth factor-1, luteinizing hormone receptor, inhibin α, and growth differentiation factor 9 expression in mouse ovary. These findings implied that fertility reduction and ovary injury of mice following exposure to nano-TiO2 may be associated with alteration of inflammation-related or follicular atresia-related cytokine expressions, and humans should take great caution when handling nano-TiO2.  相似文献   

11.
Oceanographic studies have shown that heterotrophic bacteria can protect marine cyanobacteria against oxidative stress caused by hydrogen peroxide (H2O2). Could a similar interspecific protection play a role in freshwater ecosystems? In a series of laboratory experiments and two lake treatments, we demonstrate that freshwater cyanobacteria are sensitive to H2O2 but can be protected by less-sensitive species such as green algae. Our laboratory results show that green algae degrade H2O2 much faster than cyanobacteria. Consequently, the cyanobacterium Microcystis was able to survive at higher H2O2 concentrations in mixtures with the green alga Chlorella than in monoculture. Interestingly, even the lysate of destructed Chlorella was capable to protect Microcystis, indicating a two-component H2O2 degradation system in which Chlorella provided antioxidant enzymes and Microcystis the reductants. The level of interspecific protection provided to Microcystis depended on the density of Chlorella. These findings have implications for the mitigation of toxic cyanobacterial blooms, which threaten the water quality of many eutrophic lakes and reservoirs worldwide. In several lakes, H2O2 has been successfully applied to suppress cyanobacterial blooms. Our results demonstrate that high densities of green algae can interfere with these lake treatments, as they may rapidly degrade the added H2O2 and thereby protect the bloom-forming cyanobacteria.  相似文献   

12.
Qian H  Hu B  Yu S  Pan X  Wu T  Fu Z 《PloS one》2012,7(3):e33347
  相似文献   

13.
Because cyanobacteria are photosynthetic, fast-growing microorganisms that can accumulate sucrose under salt stress, they have a potential application as a sugar source for the biomass-derived production of renewable fuels and chemicals. In the present study, the production of sucrose by the cyanobacteria Synechocystis sp. PCC6803, Synechococcus elongatus PCC7942, and Anabaena sp. PCC7120 was examined. The three species displayed different growth curves and intracellular sucrose accumulation rates in response to NaCl. Synechocystis sp. PCC6803 was used to examine the impact of modifying the metabolic pathway on the levels of sucrose production. The co-overexpression of sps (slr0045), spp (slr0953), and ugp (slr0207) lead to a 2-fold increase in intracellular sucrose accumulation, whereas knockout of ggpS (sll1566) resulted in a 1.5-fold increase in the production of this sugar. When combined, these genetic modifications resulted in a fourfold increase in intracellular sucrose accumulation. To explore methods for optimizing the transport of the intracellular sucrose to the growth medium, the acid-wash technique and the CscB (sucrose permease)-dependent export method were evaluated using Synechocystis sp. PCC6803. Whereas the acid-wash technique proved to be effective, the CscB-dependent export method was not effective. Taken together, these results suggest that using genetic engineering, photosynthetic cyanobacteria can be optimized for efficient sucrose production.  相似文献   

14.
Culture experiments were conducted on ten phytoplankton species to examine their biological and physiological responses during exposure to oil and a combination of oil and dispersant. The species tested included a range of taxa typically found in the Gulf of Mexico such as cyanobacteria, chlorophytes, and diatoms. Cultures were exposed to Macondo surrogate oil using the water accommodated fraction (WAF), and dispersed oil using a chemically enhanced WAF (CEWAF) and diluted CEWAF, to replicate conditions following the Deepwater Horizon spill in the Gulf of Mexico. A range of responses were observed, that could broadly class the algae as either “robust” or “sensitive” to oil and/or dispersant exposure. Robust algae were identified as Synechococcus elongatus, Dunaliella tertiolecta, two pennate diatoms Phaeodactylum tricornutum and Navicula sp., and Skeletonema grethae CCMP775, and were largely unaffected by any of the treatments (no changes to growth rate or time spent in lag phase relative to controls). The rest of the phytoplankton, all centric diatoms, exhibited at least some combination of reduced growth rates or increased lag time in response to oil and/or dispersant exposure. Photophysiology did not have a strong treatment effect, with significant inhibition of photosynthetic efficiency (Fv/Fm) only observed in the CEWAF, if at all. We found that the effects of oil and dispersants on phytoplankton physiology were species‐dependent, and not always detrimental. This has significant implications on how oil spills might impact phytoplankton community structure and bloom dynamics in the Gulf of Mexico, which in turn impacts higher trophic levels.  相似文献   

15.
The factors and processes driving cyanobacterial blooms in eutrophic freshwater ecosystems have been extensively studied in the past decade. A growing number of these studies concern the direct or indirect interactions between cyanobacteria and heterotrophic bacteria. The presence of bacteria that are directly attached or immediately adjacent to cyanobacterial cells suggests that intense nutrient exchanges occur between these microorganisms. In order to determine if there is a specific association between cyanobacteria and bacteria, we compared the bacterial community composition during two cyanobacteria blooms of Anabaena (filamentous and N2-fixing) and Microcystis (colonial and non-N2 fixing) that occurred successively within the same lake. Using high-throughput sequencing, we revealed a clear distinction between associated and free-living communities and between cyanobacterial genera. The interactions between cyanobacteria and bacteria appeared to be based on dissolved organic matter degradation and on N recycling, both for N2-fixing and non N2-fixing cyanobacteria. Thus, the genus and potentially the species of cyanobacteria and its metabolic capacities appeared to select for the bacterial community in the phycosphere.  相似文献   

16.
Biological soil crusts (BSCs) are key components of ecosystem productivity in arid lands and they cover a substantial fraction of the terrestrial surface. In particular, BSC N2-fixation contributes significantly to the nitrogen (N) budget of arid land ecosystems. In mature crusts, N2-fixation is largely attributed to heterocystous cyanobacteria; however, early successional crusts possess few N2-fixing cyanobacteria and this suggests that microorganisms other than cyanobacteria mediate N2-fixation during the critical early stages of BSC development. DNA stable isotope probing with 15N2 revealed that Clostridiaceae and Proteobacteria are the most common microorganisms that assimilate 15N2 in early successional crusts. The Clostridiaceae identified are divergent from previously characterized isolates, though N2-fixation has previously been observed in this family. The Proteobacteria identified share >98.5% small subunit rRNA gene sequence identity with isolates from genera known to possess diazotrophs (for example, Pseudomonas, Klebsiella, Shigella and Ideonella). The low abundance of these heterotrophic diazotrophs in BSCs may explain why they have not been characterized previously. Diazotrophs have a critical role in BSC formation and characterization of these organisms represents a crucial step towards understanding how anthropogenic change will affect the formation and ecological function of BSCs in arid ecosystems.  相似文献   

17.
Inhalation is the most likely exposure route for individuals working with aerosolizable engineered nano-materials (ENM). To properly perform nanoparticle inhalation toxicology studies, the aerosols in a chamber housing the experimental animals must have: 1) a steady concentration maintained at a desired level for the entire exposure period; 2) a homogenous composition free of contaminants; and 3) a stable size distribution with a geometric mean diameter < 200 nm and a geometric standard deviation σg < 2.5 5. The generation of aerosols containing nanoparticles is quite challenging because nanoparticles easily agglomerate. This is largely due to very strong inter-particle forces and the formation of large fractal structures in tens or hundreds of microns in size 6, which are difficult to be broken up. Several common aerosol generators, including nebulizers, fluidized beds, Venturi aspirators and the Wright dust feed, were tested; however, none were able to produce nanoparticle aerosols which satisfy all criteria 5.A whole-body nanoparticle aerosol inhalation exposure system was fabricated, validated and utilized for nano-TiO2 inhalation toxicology studies. Critical components: 1) novel nano-TiO2 aerosol generator; 2) 0.5 m3 whole-body inhalation exposure chamber; and 3) monitor and control system. Nano-TiO2 aerosols generated from bulk dry nano-TiO2 powders (primary diameter of 21 nm, bulk density of 3.8 g/cm3) were delivered into the exposure chamber at a flow rate of 90 LPM (10.8 air changes/hr). Particle size distribution and mass concentration profiles were measured continuously with a scanning mobility particle sizer (SMPS), and an electric low pressure impactor (ELPI). The aerosol mass concentration (C) was verified gravimetrically (mg/m3). The mass (M) of the collected particles was determined as M = (Mpost-Mpre), where Mpreand Mpost are masses of the filter before and after sampling (mg). The mass concentration was calculated as C = M/(Q*t), where Q is sampling flowrate (m3/min), and t is the sampling time (minute). The chamber pressure, temperature, relative humidity (RH), O2 and CO2 concentrations were monitored and controlled continuously. Nano-TiO2 aerosols collected on Nuclepore filters were analyzed with a scanning electron microscope (SEM) and energy dispersive X-ray (EDX) analysis.In summary, we report that the nano-particle aerosols generated and delivered to our exposure chamber have: 1) steady mass concentration; 2) homogenous composition free of contaminants; 3) stable particle size distributions with a count-median aerodynamic diameter of 157 nm during aerosol generation. This system reliably and repeatedly creates test atmospheres that simulate occupational, environmental or domestic ENM aerosol exposures.  相似文献   

18.
《Aquatic Botany》2005,83(2):87-107
Cyanobacterial mats (CBM), complex assemblages of cyanobacteria, bacteria and algae, are important ecosystem components of oligotrophic marshes in limestone-based regions of the Caribbean. We conducted a mesocosm experiment and evaluated the response of CBM to factorial combinations of low, medium and high phosphorus, nitrogen and salinity. Changes in composition of the main species groups of cyanobacteria and algae, primary production, cellular nutrients and enzymatic activities were recorded as response variables. The redundancy analysis with concentrations of P, N and salinity as explanatory variables showed that the primary production of CBM and the amount of phytoplankton expressed as Chl a were best explained by concentration of P, with less significant positive effect of N and a negative effect of salinity. Abundance of green algae and Chroococcales was positively correlated with increasing concentrations of P and N and reached 27.6% and 21.9%, respectively, in high P and high N treatment at the end of experiment. N2-fixation averaged 75 and 175 nmol C2H4 cm−2 min−1, at low nitrogen and medium or high P, respectively, and it was negatively correlated with nitrogen concentration and positively correlated with abundance of a group of heterocytous cyanobacteria from genus Nostoc. At low N concentrations, increasing P concentrations supported higher N2-fixation. Activity of the alkaline phosphatase, APA, was negatively correlated with P and salinity and positively with N. We also found a significant negative correlation between the APA activity and the P content of the mat. At high P and N concentrations, the mats were impacted by grazing, had a tendency to disintegrate and become shaded out by a massive growth of phytoplankton. We confirmed an overall negative effect of nutrient increase on CBM.  相似文献   

19.
The biotransformation of Hg(II) by cyanobacteria was investigated under aerobic and pH-controlled culture conditions. Mercury was supplied as HgCl2 in amounts emulating those found under heavily impacted environmental conditions where bioremediation would be appropriate. The analytical procedures used to measure mercury within the culture solution, including that in the cyanobacterial cells, used reduction under both acid and alkaline conditions in the presence of SnCl2. Acid reduction detected free Hg(II) ions and its complexes, whereas alkaline reduction revealed that meta-cinnabar (β-HgS) constituted the major biotransformed and cellularly associated mercury pool. This was true for all investigated species of cyanobacteria: Limnothrix planctonica (Lemm.), Synechococcus leopoldiensis (Racib.) Komarek, and Phormidium limnetica (Lemm.). From the outset of mercury exposure, there was rapid synthesis of β-HgS and Hg(0); however, the production rate for the latter decreased quickly. Inhibitory studies using dimethylfumarate and iodoacetamide to modify intra- and extracellular thiols, respectively, revealed that the former thiol pool was required for the conversion of Hg(II) into β-HgS. In addition, increasing the temperature enhanced the amount of β-HgS produced, with a concomitant decrease in Hg(0) volatilization. These findings suggest that in the environment, cyanobacteria at the air-water interface could act to convert substantial amounts of Hg(II) into β-HgS. Furthermore, the efficiency of conversion into β-HgS by cyanobacteria may lead to the development of applications in the bioremediation of mercury.  相似文献   

20.
Effect of nano-TiO2 on photochemical reaction of chloroplasts of spinach   总被引:1,自引:0,他引:1  
The effects of nano-TiO2 (rutile) on the photochemical reaction of chloroplasts of spinach were studied. The results showed that when spinach was treated with 0.25% nano-TiO2, the Hill reaction, such as the reduction rate of FeCy, and the rate of evolution oxygen of chloroplasts was accelerated and noncyclic photophosphorylation (nc-PSP) activity of chloroplasts was higher than cyclic photophosphorylation (c-PSP) activity, the chloroplast coupling was improved and activities of Mg2+-ATPase and chloroplast coupling factor I (CF1)-ATPase on the thylakoid membranes were obviously activated. It suggested that photosynthesis promoted by nano-TiO2 might be related to activation of photochemical reaction of chloroplasts of spinach.  相似文献   

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