Biodegradability of Vegetation-Derived Dissolved Organic Carbon in a Cool Temperate Ombrotrophic Bog

Dissolved organic carbon (DOC) plays a key role in the peatland carbon balance and serves numerous ecological and chemical functions including acting as a microbial substrate. In this study, we quantify the concentration, biodegradability, and intrinsic properties of DOC obtained from peat, fresh material, and litter from nine species of ombrotrophic bog vegetation. Potential biodegradability was assessed by incubating vegetation extracts for 28 days in the dark and measuring percent DOC loss as the fraction of biodegradable DOC (%BDOC) while DOC properties were characterized using UV–Vis absorbance and fluorescence measurements. The mean initial DOC concentration extracted differed significantly among species (P < 0.05) and was significantly higher in fresh material, 217 ± 259 mg DOC l^?1, than either litter or peat extracts with mean concentrations of 82.1 ± 117 mg DOC l^?1 and 12.7 ± 1.0 mg DOC l^?1, respectively (P < 0.05). %BDOC also differed significantly among species (P < 0.05) and ranged from 52 to 73% in fresh cuttings with the greatest fraction observed in S. magellanicum; 22–46% in litter; and 24% in peat. The majority of variability (82.5%) in BDOC was explained by initial absorbance at 254 nm and total dissolved nitrogen concentration which was further resolved into significant non-linear relationships between %BDOC and both humic-like and protein-like DOC fractions (P < 0.05). Our results highlight the extremely heterogeneous nature of the surface vegetation-derived DOC input in peatlands and stress the importance of vegetation species in peatland ecosystem function.     

Insect herbivory, organic matter deposition and effects on belowground organic matter fluxes in a central European oak forest

Apart from the forest floor, the canopy of forested ecosystems functions as the second most important source for dissolved and particulate fractions of organic and inorganic C and N compounds. However, under mass outbreak situations of insect herbivores this flux path of organic matter is considerably intensified clearly exceeding C and N fluxes from the forest floor. In this paper we report on herbivore-altered C and N fluxes from the canopy to the forest floor and effects on forest floor nutrient fluxes during severe defoliating herbivory of the winter moth (Operophtera brumata) and the mottled umber moth (Eranis defoliaria) in an oak forest in Germany. Over the course of 6.5 months we followed the C and N fluxes with bulk deposition, throughfall solution, insect frass deposits (green-fall together with insect faeces) and with forest floor solution in an 117-yr-old oak (Quercus petraea) forest. Compared to the control, herbivore defoliation significantly enhanced throughfall inputs of total and dissolved organic carbon and nitrogen by a factor of 3 and 2.5 (for TOC and DOC), and by 1.4 and 1.3 times (for TNb and DNb), respectively. Frass plus green-fall C and N fluxes peaked in May with 592 kg C?ha^?1 and 33.5 kg N?ha^?1 representing 79.6% (for C) and 78.3% (for N) of the total C and N input over 2.5 months. The quantitative and qualitative C and N input via faeces and litter deposition significantly differ between the insect affected and non-affected site. However, the C and N fluxes with throughfall did not significantly correlate with forest floor leachates. In this context, forest floor fluxes of TOC, DOC and NO₃-N were significantly lower at the infested site compared to the control, whereas fluxes of NH₄-N together with DON were significantly higher. The study demonstrates the importance of linking the population and associated frass dynamics of herbivorous insects with the cycling of nutrients and organic matter in forest ecosystems, highlighting the remarkable alterations in the timing, amounts and nature of organic matter dynamics on the ecosystem level. Consequently, the ecology of phytophagous insects allows partly to explain temporal-spatial alterations in nutrient cycling and thus ecosystem functioning.     

Carbon Dioxide and Methane Fluxes From Tree Stems,Coarse Woody Debris,and Soils in an Upland Temperate Forest

Forest soils and canopies are major components of ecosystem CO₂ and CH₄ fluxes. In contrast, less is known about coarse woody debris and living tree stems, both of which function as active surfaces for CO₂ and CH₄ fluxes. We measured CO₂ and CH₄ fluxes from soils, coarse woody debris, and tree stems over the growing season in an upland temperate forest. Soils were CO₂ sources (4.58 ± 2.46 µmol m^?2 s^?1, mean ± 1 SD) and net sinks of CH₄ (?2.17 ± 1.60 nmol m^?2 s^?1). Coarse woody debris was a CO₂ source (4.23 ± 3.42 µmol m^?2 s^?1) and net CH₄ sink, but with large uncertainty (?0.27 ± 1.04 nmol m^?2 s^?1) and with substantial differences depending on wood decay status. Stems were CO₂ sources (1.93 ± 1.63 µmol m^?2 s^?1), but also net CH₄ sources (up to 0.98 nmol m^?2 s^?1), with a mean of 0.11 ± 0.21 nmol m^?2 s^?1 and significant differences depending on tree species. Stems of N. sylvatica, F. grandifolia, and L. tulipifera consistently emitted CH₄, whereas stems of A. rubrum, B. lenta, and Q. spp. were intermittent sources. Coarse woody debris and stems accounted for 35% of total measured CO₂ fluxes, whereas CH₄ emissions from living stems offset net soil and CWD CH₄ uptake by 3.5%. Our results demonstrate the importance of CH₄ emissions from living stems in upland forests and the need to consider multiple forest components to understand and interpret ecosystem CO₂ and CH₄ dynamics.     

Impact of elevated CO<Subscript>2</Subscript> in <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis> rooted stem cuttings inoculated with <Emphasis Type="Italic">Frankia</Emphasis>

Impact of different levels of elevated CO ₂ on the activity of Frankia (Nitrogen-fixing actinomycete) in Casuarina equisetifolia rooted stem cuttings has been studied to understand the relationship between C. equisetifolia, Frankia and CO₂. The stem cuttings of C. equietifolia were collected and treated with 2000 ppm of Indole Butyric Acid (IBA) for rooting. Thus vegetative propagated rooted stem cuttings of C. equisetifolia were inoculated with Frankia and placed in the Open top chambers (OTC) with elevated CO₂ facilities. These planting stocks were maintained in the OTC for 12 months under different levels of elevated CO₂ (ambient control, 600 ppm, 900 ppm). After 12 months, the nodule numbers, bio mass, growth, and photosynthesis of C. equisetifolia rooted stem cuttings inoculated with Frankia were improved under 600 ppm of CO₂. The rooted stem cuttings of C. equisetifolia inoculated with Frankia showed a higher number of nodules under 900 ppm of CO₂ and cuttings without Frankia inoculation exhibited poor growth. Tissue Nitrogen (N) content was also higher under 900 ppm of CO₂ than ambient control and 600 ppm levels. The photosynthetic rate was higher (17.8 μ mol CO₂ m^?2 s^?1) in 900 ppm of CO₂ than in 600 ppm (13.2 μ mol CO₂ m^?2 s^?1) and ambient control (8.3 μ mol CO₂ m^?2 s^?1). This study showed that Frankia can improve growth, N fixation and photosynthesis of C. equietifolia rooted stem cuttings under extreme elevated CO₂ level conditions (900 ppm).     

Oxygen transfer as a tool for fine-tuning recombinant protein production by <Emphasis Type="Italic">Pichia pastoris</Emphasis> under glyceraldehyde-3-phosphate dehydrogenase promoter

Effects of oxygen transfer on recombinant protein production by Pichia pastoris under glyceraldehyde-3-phosphate dehydrogenase promoter were investigated. Recombinant glucose isomerase was chosen as the model protein. Two groups of oxygen transfer strategies were applied, one of which was based on constant oxygen transfer rate where aeration rate was Q _O/V = 3 and 10 vvm, and agitation rate was N = 900 min^?1; while the other one was based on constant dissolved oxygen concentrations, C _DO = 5, 10, 15, 20 and 40 % in the fermentation broth, by using predetermined exponential glucose feeding with μ _o = 0.15 h^?1. The highest cell concentration was obtained as 44 g L^?1 at t = 9 h of the glucose fed-batch phase at C _DO = 20 % operation while the highest volumetric and specific enzyme activities were obtained as 4440 U L^?1 and 126 U g^?1 cell, respectively at C _DO = 15 % operation. Investigation of specific enzyme activities revealed that keeping C _DO at 15 % was more advantageous with an expense of relatively higher by-product formation and lower specific cell growth rate. For this strategy, the highest oxygen transfer coefficient and oxygen uptake rate were K _L a = 0.045 s^?1 and OUR = 8.91 mmol m^?3 s^?1, respectively.     

The nitrite reductase activity of horse heart carboxymethylated-cytochrome <Emphasis Type="Italic">c</Emphasis> is modulated by cardiolipin

Horse heart carboxymethylated cytc (CM-cytc) displays myoglobin-like properties. Here, the effect of cardiolipin (CL) liposomes on the nitrite reductase activity of ferrous CM-cytc [CM-cytc-Fe(II)], in the presence of sodium dithionite, is reported between pH 5.5 and 7.6, at 20.0 °C. Cytc-Fe(II) displays a very low value of the apparent second-order rate constant for the NO₂ ^?-mediated conversion of cytc-Fe(II) to cytc-Fe(II)-NO [k _on = (7.3 ± 0.7) × 10^?2 M^?1 s^?1; at pH 7.4], whereas the value of k _on for NO₂ ^? reduction by CM-cytc-Fe(II) is 1.1 ± 0.2 M^?1 s^?1 (at pH 7.4). CL facilitates the NO₂ ^?-mediated nitrosylation of CM-cytc-Fe(II) in a dose-dependent manner, the value of k _on for the NO₂ ^?-mediated conversion of CL–CM-cytc-Fe(II) to CL–CM-cytc-Fe(II)-NO (5.6 ± 0.6 M^?1 s^?1; at pH 7.4) being slightly higher than that for the NO₂ ^?-mediated conversion of CL–cytc-Fe(II) to CL–cytc-Fe(II)-NO (2.6 ± 0.3 M^?1 s^?1; at pH 7.4). The apparent affinity of CL for CM-cytc-Fe(II) is essentially pH independent, the average value of B being (1.3 ± 0.3) × 10^?6 M. In the absence and presence of CL liposomes, the nitrite reductase activity of CM-cytc-Fe(II) increases linearly on lowering pH and the values of the slope of the linear fittings of Log k _on versus pH are ?1.05 ± 0.07 and ?1.03 ± 0.03, respectively, reflecting the involvement of one proton for the formation of the transient ferric form, NO, and OH^?. These results indicate that Met80 carboxymethylation and CL binding cooperate in the stabilization of the highly reactive heme-Fe atom of CL–CM-cytc.     

Characterization of alcohol dehydrogenase from <Emphasis Type="Italic">Kangiella koreensis</Emphasis> and its application to production of all-<Emphasis Type="Italic">trans</Emphasis>-retinol

A recombinant alcohol dehydrogenase (ADH) from Kangiella koreensis was purified as a 40 kDa dimer with a specific activity of 21.3 nmol min^?1 mg^?1, a K _m of 1.8 μM, and a k _cat of 1.7 min^?1 for all-trans-retinal using NADH as cofactor. The enzyme showed activity for all-trans-retinol using NAD ⁺ as a cofactor. The reaction conditions for all-trans-retinol production were optimal at pH 6.5 and 60 °C, 2 g enzyme l^?1, and 2,200 mg all-trans-retinal l^?1 in the presence of 5 % (v/v) methanol, 1 % (w/v) hydroquinone, and 10 mM NADH. Under optimized conditions, the ADH produced 600 mg all-trans-retinol l^?1 after 3 h, with a conversion yield of 27.3 % (w/w) and a productivity of 200 mg l^?1 h^?1. This is the first report of the characterization of a bacterial ADH for all-trans-retinal and the biotechnological production of all-trans-retinol using ADH.     

<Emphasis Type="Italic">Pseudomonas</Emphasis> spp. increases root biomass and tropane alkaloid yields in transgenic hairy roots of <Emphasis Type="Italic">Datura</Emphasis> spp.

Transgenic hairy roots of Datura spp., established using strain A4 of Agrobacterium rhizogenes, are genetically stable and produce high levels of tropane alkaloids. To increase biomass and tropane alkaloid content of this plant tissue, four Pseudomonas strains, Pseudomonas fluorescens P64, P66, C7R12, and Pseudomonas putida PP01 were assayed as biotic elicitors on transgenic hairy roots of Datura stramonium, Datura tatula, and Datura innoxia. Alkaloids were extracted from dried biomass, and hyoscyamine and scopolamine were quantified using liquid chromatography-tandem mass spectrometry analysis. D. stramonium and D. innoxia biomass production was stimulated by all Pseudomonas spp. strains after a 5-d treatment. All strains of P. fluorescens increased hyoscyamine yields compared to untreated cultures after both 5 and 10 d of treatment. Hyoscyamine yields were highest in D. tatula cultures exposed to a 5-d treatment with C7R12 (16.633 + 0.456 mg g^?1 dry weight, a 431% increase) although the highest yield increases compared to the control were observed in D. stramonium cultures exposed to strains P64 (511% increase) and C7R12 (583% increase) for 10 d. D. innoxia showed the highest scopolamine yields after elicitation with P. fluorescens strains P64 for 5 d (0.653 + 0.021 mg g^?1 dry weight, a 265% increase) and P66 for 5 and 10 d (5 d, 0.754 + 0.0.031 mg g^?1 dry weight, a 321% increase; 10 d 0.634 + 0.046 mg g^?1 dry weight, a 277% increase). These results show that the Pseudomonas strains studied here can positively and significantly affect biomass and the yields of hyoscyamine and scopolamine from transgenic roots of the three Datura species.     

Single point mutations enhance activity of <Emphasis Type="Italic">cis</Emphasis>-epoxysuccinate hydrolase

Objectives

To enhance activity of cis-epoxysuccinate hydrolase from Klebsiella sp. BK-58 for converting cis-epoxysuccinate to tartrate.

Results

By semi-saturation mutagenesis, all the mutants of the six important conserved residues almost completely lost activity. Then random mutation by error-prone PCR and high throughput screening were further performed to screen higher activity enzyme. We obtained a positive mutant F10D after screening 6000 mutations. Saturation mutagenesis on residues Phe10 showed that most of mutants exhibited higher activity than the wild-type, and the highest mutant was F10Q with activity of 812 U mg^?1 (k _cat /K _m , 9.8 ± 0.1 mM^?1 s^?1), which was 230 % higher than that of wild-type enzyme 355 U mg^?1 (k _cat /K _m , 5.3 ± 0.1 mM^?1 s^?1). However, the thermostability of the mutant F10Q slightly decreased.

Conclusions

The catalytic activity of a cis-epoxysuccinate hydrolase was efficient improved by a single mutation F10Q and Phe10 might play an important role in the catalysis.

     

Zonal distribution of coarse woody debris and its contribution to net primary production in a secondary mangrove forest

Coarse woody debris (CWD) plays an important role in long-term carbon storage in forest ecosystems. However, few studies have examined CWD in mangrove forests. A secondary mangrove forest on an estuary of the Trat River showed different structures along vegetation zones ranging from the river’s edge to inland parts of the forest (the Sonneratia–Avicennia, Avicennia, Rhizophora, and Xylocarpus zones, respectively). The mass distribution of CWD stock in downed wood and standing dead trees along these vegetation zones was evaluated. Most of the CWD stock in the Sonneratia–Avicennia and Avicennia zones was found in downed wood, while it mainly accumulated in standing dead trees in the Rhizophora and Xylocarpus zones. The total mass of CWD stock that accumulated in each zone ranged from 1.56–8.39 t ha^?1, depending on the forest structure and inundation regimes. The annual woody debris flux in each zone was calculated by summing the necromass (excluding foliage) of dead trees and coarse litter from 2010 to 2013. The average woody debris flux was 5.4 t ha^?1 year^?1, and its zonal variation principally depended on the necromass production that resulted from forest succession, high tree-density, and lightning. Over all the zones, the above- and below-ground net primary production (ANPP and BNPP, respectively) was estimated at 18.0 and 3.6 t ha^?1 year^?1, respectively. The magnitude of BNPP and its contribution to the NPP was markedly increased when fine root production was taken into consideration. The contribution of the woody debris flux without root necromass to the ANPP ranged from 12 to 28%.     

Non-native liana, <Emphasis Type="Italic">Euonymus fortunei</Emphasis>, associated with increased soil nutrients,unique bacterial communities,and faster decomposition rate

Invasive plants have wide-ranging impacts on native systems including reducing native plant richness and altering soil chemistry, microbes, and nutrient cycling. Increasingly, these effects are found to linger long after removal of the invader. We examined how soil chemistry, bacterial communities, and litter decomposition varied with cover of Euonymus fortunei, an invasive evergreen liana, in two central Kentucky deciduous forests. In one forest, E. fortunei invaded in the late 1990s but invasion remained patchy and we paired invaded and uninvaded plots to examine the associations between E. fortunei cover and our response variables. In the second forest, E. fortunei had completely invaded the forest by 2005; areas where it had been selectively removed by 2010 were paired with an adjacent invaded plot. Where E. fortunei had patchily invaded, E. fortunei patches had up to 3.5× nitrogen, 2.7× carbon, and 1.9× more labile glomalin in soils than uninvaded plots, whereas there were no differences in soil characteristics between invaded and removal plots. In the patchily invaded forest, bacterial community composition varied among invaded and non-invaded plots, whereas bacterial communities did not vary among invaded and removal plots. Finally, E. fortunei leaf litter decomposed faster (k = 4.91 year^?1) than the native liana (k = 3.77 year^?1), Vitis vulpina; decomposition of both E. fortunei and V. vulpina was faster in invaded (k = 7.10 year^?1) than removal plots (k = 4.77 year^?1). Our findings suggest that E. fortunei invasion increases the rate of leaf litter decomposition via high-quality litter, alters the decomposition environment, and shifts in the soil biotic communities associated with a dense mat of wintercreeper. Land managers with limited resources should target the densest mats for the greatest restoration potential and remove wintercreeper patches before they establish dense mats.     

A saponin isolated from <Emphasis Type="Italic">Agapanthus africanus</Emphasis> differentially induces apoplastic peroxidase activity in wheat and displays fungicidal properties

A spirostane with an attached trisaccharide, (25R)-5α-spirostane-2α,3β,5α-triol 3-O-(O-α-l-rhamnopyranosyl-(1 → 2)-O-(β-d-galactopyranosyl-(1 → 3))-β-d-glucopyranoside), was isolated and identified from the aerial parts of Agapanthus africanus by activity-guided fractionation. Fungicidal properties of the crude extract, semi-purified fractions as well as the purified active saponin from A. africanus were screened in vitro against Fusarium oxysporum. At a concentration of 1 mg mL^?1, the crude extract and semi-purified ethyl acetate and dichloromethane fractions showed significant antifungal activity. The purified saponin inhibited the in vitro mycelial growth of F. oxysporum completely (100 %) at a concentration of 125 µg mL^?1. Furthermore, to verify previously observed induced resistance by crude extracts of A. africanus towards leaf rust, intercellular PR-protein activity was determined in wheat seedlings following foliar application of the purified saponin at 100 µg mL^?1. In vitro peroxidase enzyme activity increased significantly (60 %) in wheat seedlings 48 h after treatment with the purified saponin, demonstrating its role as an elicitor to activate a defence reaction in wheat.     

Responses of Tree Transpiration and Growth to Seasonal Rainfall Redistribution in a Subtropical Evergreen Broad-Leaved Forest

Precipitation changes such as more frequent drought and altered precipitation seasonality may impose substantial impacts on the structure and functioning of forest ecosystems. A better understanding of tree responses to precipitation changes can provide fundamental information for the conservation and management of forests under future climate regimes. We conducted a 2-year seasonal rainfall redistribution experiment to assess the responses of tree transpiration and growth to manipulated precipitation changes in a subtropical evergreen broad-leaved forest. Three precipitation treatments were administered including a drier dry season and wetter wet season treatment (DD), an extended dry season and wetter wet season treatment (ED), and an ambient control treatment, with the total amount of annual rainfall being kept the same among the three treatments. Our results showed that the DD and ED treatments reduced daily transpiration of Schima superba by 8–16 and 13–25%, respectively. The ED treatment also reduced the DBH increment of larger S. superba individuals. In contrast, neither treatment showed obvious effects on the transpiration and DBH increment of another dominant species Michelia macclurei. However, the transpiration of both species showed clear inter-annual differences between the 2 years with contrasting annual rainfall (2094 vs 1582 mm). S. superba had a lower transpiration-to-precipitation ratio (T/P) compared to M. macclurei and showed decreased sensitivities to total solar radiation and vapor pressure deficit under the DD and ED treatments. These results indicate the deep-rooted S. superba may be suppressed with a lower ability to obtain water and assimilate carbon compared to the shallow-rooted M. macclurei under the precipitation seasonality changes, which could potentially cause shifts in species dominance within the forest community.     

Shrub Expansion of <Emphasis Type="Italic">Alnus viridis</Emphasis> Drives Former Montane Grassland into Nitrogen Saturation

The N₂-fixing shrub Alnus viridis is currently encroaching on montane grasslands in the Alps as a result of reduced land management and complete abandonment. Alnus introduces large amounts of nitrogen (N) into these formerly N-poor grasslands and restricts the succession to montane forests. We studied pools and fluxes of N and the associated C pools in pastures (controls) and adjacent Alnus shrublands at two elevations (1650 versus 1950 m a.s.l.) in three valleys in the Swiss central Alps. The total N and C pools stored in 50-year-old Alnus shrubland did not exceed those in adjacent pastures with a total of approximately 610 g N m^?2 in phytomass plus soil (down to 30 cm) at both elevations. In Alnus stands, reduced soil N pools balanced the gain in phytomass N pools, a likely result of a faster turnover of soil N. The soil solution under Alnus was continuously enriched with nitrate, with a total N leaching of 0.79 g N m^?2 season^?1 (June–October) under 50-year-old stands at both elevations and the highest flux of 1.76 g N m^?2 season^?1 in 25-year-old shrubland at low elevation, clearly indicating an excess of available N in Alnus shrubland. In contrast, N leaching across all pastures was close to zero (0.08 g N m^?2) throughout the season. At the catchment scale, streamlet water showed increased nitrate concentrations with typical flushing peaks in spring and autumn, provided more than one fifth of the catchment area was covered by Alnus shrubs. We conclude that the expansion of Alnus rapidly converts centuries-old, N-poor grassland into N saturated shrubland, irrespective of elevation, and it reduces the C storage potential of the landscape because the Alnus dominance constrains re-establishment of a natural montane forest.     

Integrated control of tobacco black shank by combined use of riboflavin and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> strain Tpb55

We investigated the effect of riboflavin on the biocontrol activity of Bacillus subtilis Tpb55 against Phytophthora nicotianae (Pn), which causes tobacco black shank. Riboflavin (0.2 mg ml^?1) significantly improved the biocontrol activity of Tpb55 (2.0 × 10⁸ cfu ml^?1). Riboflavin (0.02–0.5 mg ml^?1) alone could not significantly inhibit Pn growth. However, it enhanced the B. subtilis population, both in vitro and in tobacco roots and significantly increased the activity of defense enzymes, peroxidase, catalase, superoxide dismutase, and β-1,3-glucanase, in the roots of B. subtilis-treated tobacco seedlings. Our results indicate that riboflavin can stimulate the growth of B. subtilis Tpb55 and induce resistance to Pn in tobacco plants. These findings should boost the prospects for practical application of B. subtilis Tpb55 as a biocontrol agent against black shank of tobacco.     

Fluxes and concentrations of dissolved organic carbon and nitrogen – a synthesis for temperate forests

Dissolved organic carbon (DOC) and nitrogen (DON)represent an important part of the C and N cycles inforest ecosystems. Little is known about the controlson fluxes and concentrations of these compounds insoils under field conditions. Here we compiledpublished data on concentrations and fluxes of DOC andDON from 42 case studies in forest ecosystems of thetemperate zone in order to evaluate controls on alarger temporal and spatial scale. The focus was onannual fluxes and concentrations in throughfall,forest floor leachates and soil solutions. In allcompartments considered, concentrations and fluxesdiffered widely between the sites. Highestconcentrations of DOC and DON were generally observedin forest floor leachates and in A horizons. Highestfluxes occurred in forest floor leachates. The fluxesof DOC and DON in forest floor leachates increasedwith increasing annual precipitation and were alsopositively related to DOC and DON fluxes withthroughfall. Variation in throughfall fluxes couldexplain 46% and 65% of the variation in DOC and DONfluxes from the forest floor, respectively. No generaldifference in DOC and DON concentrations and fluxes inforest floor leachates was found when comparingconiferous and hardwood sites. Concentrations of DOCin forest floor leachates were positively correlatedto the pH of the forest floor. Furthermore, there wasno relationship between organic C and N stocks, soilC/N, litterfall or mineral N inputs and concentrationsand fluxes of DOC and DON in forest floor leachates.Including all compartments, fluxes of DOC and DON werehighly correlated. Ratios of DOC to DON calculatedfrom fluxes from the forest floor were independent ofthe amount of annual precipitation, pointing to asimilar response of DOC and DON to precipitationconditions. A decrease in the ratio of DOC to DON withsoil depth as observed on a plot-scale, was notconfirmed by data analysis on a large scale. Thecontrols observed on annual fluxes and concentrationsof DON and DOC at regional scale differed from thosereported for smaller time and space scales.     

Phycoremediation potential,physiological, and biochemical response of <Emphasis Type="Italic">Amphora subtropica</Emphasis> and <Emphasis Type="Italic">Dunaliella</Emphasis> sp. to nickel pollution

Metal pollution can produce many biological effects on aquatic environments. The marine diatom Amphora subtropica and the green alga Dunaliella sp. possess a high metal absorption capacity. Nickel (Ni) removal by living cells of A. subtropica and Dunaliella sp. was tested in cultures exposed to different Ni concentrations (100, 200, 300, and 500 mg L^?1). The amount of Ni removed by the microalgae increased with the time of exposure and the initial Ni concentration in the medium. The metal, which was mainly removed by bioadsorption to Dunaliella sp. cell surfaces (93.63% of total Ni (for 500 mg Ni L^?1) and by bioaccumulation (80.82% of total Ni (for 300 mg Ni L^?1) into Amphora subtropica cells, also inhibited growth. Exposure to Ni drastically reduced the carbohydrate and protein concentrations and increased total lipids from 6.3 to 43.1 pg cell^?1, phenolics 0.092 to 0.257 mg GAE g^?1 (Fw), and carotenoid content, from 0.08 to 0.59 mg g^?1 (Fw), in A. subtropica. In Dunaliella sp., total lipids increased from 26.1 to 65.3 pg cell^?1, phenolics from 0.084 to 0.289 mg GAE g^?1 (Fw), and carotenoid content from 0.41 to 0.97 mg g^?1 (Fw). These compounds had an important role in protecting the algae against ROS generated by Ni. In order to cope with Ni stress shown by the increase of TBARS level, enzymatic (SOD, CAT, and GPx) ROS scavenging mechanisms were induced.     

Enhancing carotenoid production in <Emphasis Type="Italic">Rhodotorula mucilaginosa</Emphasis> KC8 by combining mutation and metabolic engineering

Rhodotorula mucilaginosa has been considered as a potential industrial yeast due to its unicellular and fast-growing characteristics, and its ability to produce carotenoids, including torularhodin. However, its low total carotenoid production limits its commercial application. In this study, mutation breeding and metabolic engineering were employed to enhance carotenoid production in the R. mucilaginosa strain KC8. After chemical–physical mutagenesis, R. mucilaginosa K4 with a 67% greater concentration of carotenoids (14.47 ± 0.06 mg L^?1) than R. mucilaginosa KC8 (8.67 ± 0.07 mg L^?1) was obtained. To further enhance carotenoid production, gene HMG1 encoding the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase was introduced from another yeast, Saccharomyces cerevisiae, and overexpressed in R. mucilaginosa K4. The carotenoid production of HMG1-gene-overexpression transformant G1 reached 16.98 mg L^?1. To relieve the feedback inhibition of ergosterol, and to down-regulate ergosterol synthesis, ketoconazole, an ergosterol synthesis inhibitor, was added at a concentration of 28 mg L^?1. The carotenoid production of the transformant G1 reached 19.14 ± 0.09 mg L^?1, which was 121% higher than in R. mucilaginosa KC8. This suggests that a combination of chemical–physical mutagenesis, overexpression of the HMG1 gene, and adding ketoconazole is an effective strategy to improve carotenoid production.     

Effects of both substrate and nitrogen and phosphorus fertilizer on <Emphasis Type="Italic">Sphagnum palustre</Emphasis> growth in subtropical high-mountain regions and implications for peatland recovery

Human activities have recently caused severe destruction of Sphagnum wetlands in subtropical high-mountain regions, calling for urgent efforts to restore Sphagnum wetlands. Through a greenhouse experiment in western Hubei, China, we studied the effects of different substrate types (peat and mountain soil) and different levels of nitrogen (N) (0, 2, 4, 6, 10 g m^?2 year^?1) and phosphorus (P) (0, 0.2, 0.5, 1, 2 g m^?2 year^?1) on the growth of Sphagnum palustre, which was evaluated by four growth indicators: length growth, number of capitula, coverage change and biomass. We aimed to determine the optimal nutrient conditions for S. palustre growth, which would contribute to the rapid colonization and restoration of Sphagnum wetlands. The results showed that the different substrates significantly influenced S. palustre growth. Compared with those of peat, the acidic properties of the local yellow brown soil in the subtropical high-mountain regions were more favorable for S. palustre growth. As N addition increased, the four growth indicators responded inconsistently to the different substrates. While the number of capitula markedly increased, the other three indicators significantly decreased in the mountain soil or exhibited no definitive changes in the peat. The addition of P markedly promoted S. palustre growth in both substrates. However, a threshold for P fertilization existed; the highest productivity occurred at P additions of 0.2 and 0.5 g m^?2 year^?1 in the peat and mountain soil, respectively. The N and P contents in the capitula increased in parallel as the N and P fertilization rates increased, suggesting that these nutrients were absorbed proportionately and were used during the growth of S. palustre.     

<Emphasis Type="Italic">Plectosphaerella cucumerina</Emphasis> as a bioherbicide for <Emphasis Type="Italic">Cirsium arvense</Emphasis>: proof of concept

Plectosphaerella cucumerina (Lindf.) W. Gams was evaluated as a bioherbicide for Cirsium arvense L. (Scop.) using a Canadian and a New Zealand isolate. Both isolates defoliated C. arvense when applied at 10¹³ conidia ha^?1 in water volumes ranging from 250 to 6400 l ha^?1 with a rapid decline in effect with declining conidial dose. Repeat application and the addition of the adjuvant Pulse^® penetrant to the conidial suspension increased the disease severity in C. arvense. Maximum disease occurred at 20 °C with a 48 h post-application dew period. The experiments demonstrate that P. cucumerina can defoliate C. arvense under the environmental conditions of temperate pastures where the weed is problematic. The results also show that modifications to formulation and strategic application may reduce the 48 h dew period requirement and risk to non-target species respectively, supporting the conclusion that the fungus has potential as a bioherbicide for C. arvense.