Stimulation of methane oxidation by CH<Subscript>4</Subscript>-emitting rose chafer larvae in well-aerated grassland soil

In this study, the impact of rose chafer (Cetonia aurata L.) larvae on net and gross methane (CH₄) fluxes in soil from an old permanent grassland site (Giessen, Germany) was investigated. Previous studies at this site suggested the existence of Scarabaeidae larvae-induced “CH₄-emitting hot spots” within the soil profile which may subsequently lead to increased CH₄ oxidation. The net (soil + larvae) and gross (soil and larvae separated) CH₄ fluxes were studied in a 3-month laboratory incubation. Addition of larvae changed the soil from a net sink (?330 ± 11 ng CH₄ kg^?1 h^?1) to a net source (637 ± 205 ng CH₄ kg^?1 h^?1). Supply of plant litter to the soil + larvae incubation jars tended to increase CH₄ emissions which was not significant due to large variability. After 11–13 weeks of incubation, the net soil CH₄ oxidation was significantly stimulated by 13–21% in the treatments containing larvae when these were taken out. Analysis of archaeal 16S rRNA genes revealed that the majority of the obtained clones were closely related to uncultured methanogens from guts of insects and other animals. Other sequences were relative to cultivated species of Methanobrevibacter, Methanoculleus, and Methanosarcina. Hence, Scarabaeidae larvae in soils (i) may represent an underestimated source of CH₄ emissions in aerobic upland soils, (ii) may stimulate gross CH₄ consumption in their direct soil environment, and, thus, (iii) contribute to the spatial heterogeneity often observed in the field with closed-chamber measurements. Long-term CH₄-flux balances may be wrongly assessed when “exceptional” net CH₄ flux rates (due to larvae hot spots) are excluded from data sets.     

Assessing the potential of using biochar in mine rehabilitation under elevated atmospheric CO<Subscript>2</Subscript> concentration

Purpose

Re-establishment of soil nitrogen (N) capital is a priority in mine rehabilitation. We aimed to evaluate the effects of biochar addition on improving mine spoil N pools and the influence of elevated CO₂ concentration on mine rehabilitation.

Materials and methods

We assessed the effects of pinewood biochar, produced at three temperatures (650, 750 and 850 °C, referred as B₆₅₀, B₇₅₀ and B₈₅₀, respectively), on mine spoil total N concentrations with five different plant species, including a tree species (Eucalyptus crebra), N-fixing shrubs (Acacia floribunda and Allocasuarina littoralis) and C₃ and C₄ grasses (Austrodanthonia tenuior and Themeda australis) incubated at ambient (400 μL L^?1) and elevated (700 μL L^?1) atmospheric CO₂ concentrations, as well as the effects of elevated CO₂ on mine rehabilitation.

Results and discussion

Soil total N significantly improved following biochar incorporation under all plant species (P < 0.05) except for T. Australis. E. crebra had the highest soil total N (0.197%, 0.198% and 0.212% for B₆₅₀, B₇₅₀ and B₈₅₀, respectively). Different from the negligible influence of elevated CO₂ on soil properties under the grasses and the N-fixing shrubs, elevated CO₂ significantly increased soil water and hot water extractable organic C (WEOC and HWEOC, respectively) and decreased total C under E. crebra, indicating that the nutrient demands were not met.

Conclusions

Biochar addition showed the potential in mine rehabilitation in terms of improving soil N pool, especially with E. crebra. However, it would be more difficulty to rehabilitate mine spoils in future with the rising atmospheric CO₂ concentration.

     

Effects of TiO<Subscript>2</Subscript> Nanoparticles on the Neotropical Cladoceran <Emphasis Type="Italic">Ceriodaphnia silvestrii</Emphasis> by Waterborne and Dietary Routes

The impact of nanoparticles (NPs) in zooplankton is poorly studied, particularly when organisms are exposed through diet. Food, constituted mainly by unicellular algae, can act as an important route of contamination for zooplankton. Since unicellular algae have a high surface area in relation to their volume, NPs can interact with their cell membranes and walls, as well as with exopolysaccharides secreted by them. In the present research, we investigated both the acute effects of waterborne titanium dioxide nanoparticles (TiO₂ NPs), and its chronic effects via dietary exposure on the Neotropical freshwater zooplankton Ceriodaphnia silvestrii Daday, 1902 (Crustacea: Cladocera). The observed acute effects served as support for chronic tests, in which we investigated the effects of TiO₂ NPs on survival and life history parameters (body length, numbers of eggs, and neonates produced) of cladoceran adult females, using the freshwater cosmopolitan chlorophycean Raphidocelis subcapitata as source of contamination of TiO₂ NPs for zooplankton. R. subcapitata cells were exposed to concentrations of 0, 0.01, 1, and 10 mg L^?1 of TiO₂ NPs for 96 h, and then provided as food for females of C. silvestrii until the third brood was released. Significant toxic effects were observed in body length and total number of neonates and eggs produced by females of C. silvestrii at concentrations of 1 and 10 mg L^?1 of TiO₂ NPs. Survival was the most sensitive parameter when exposure was given via food. From the concentration of 0.01 mg L^?1 of TiO₂ NPs, there was a decrease in the survival of C. silvestrii females. The quantification of TiO₂ NPs in algae evidenced that they have retained NPs in their cells, being, therefore, an important route of exposure and toxicity of TiO₂ NPs to the studied microcrustacean.     

Increases in soil CO<Subscript>2</Subscript> and N<Subscript>2</Subscript>O emissions with warming depend on plant species in restored alpine meadows of Wugong Mountain,China

Purpose

Ecosystem restorations can impact carbon dioxide (CO₂) and nitrous oxide (N₂O) emissions which are important greenhouse gasses. Alpine meadows are degraded worldwide, but restorations are increasing. Because their soils represent large carbon (C) and nitrogen (N) pools, they may produce significant amounts of CO₂ and N₂O depending on the plant species used in restorations. In addition, warming and N deposition may impact soil CO₂ and N₂O emissions from restored meadows.

Materials and methods

We collected soils from degraded meadows and plots restored using three different plant species at Wugong Mountain (Jiangxi, China). We measured CO₂ and N₂O emissions when soils were incubated at different temperatures (15, 25 or 35 °C) and levels of N addition (control vs. 4 g m^?2) to understand their responses to warming and N deposition.

Results and discussion

Dissolved organic C was higher in restored plots (especially with Fimbristylis dichotoma) compared to non-restored bare soils, and their soil inorganic N was lower. CO₂ emission rates were increased by vegetation restorations, decreased by N deposition, and increased by warming. CO₂ emission rates were similar for the three grass species at 15 and 25 °C, but they were lower with Miscanthus floridulus at 35 °C. Soils from F. dichotoma and Carex chinensis plots had higher N₂O emissions than degraded or M. floridulus plots, especially at 25 °C.

Conclusions

These results show that the effects of restorations on soil greenhouse gas emissions depended on plant species. In addition, these differences varied with temperature suggesting that future climate should be considered when choosing plant species in restorations to predict soil CO₂ and N₂O emissions and global warming potential.

     

Estimation of natural outcrossing rate and genetic diversity in Lima bean (<Emphasis Type="Italic">Phaseolus lunatus</Emphasis> L. var. <Emphasis Type="Italic">lunatus</Emphasis>) from Brazil using SSR markers: implications for conservation and breeding

Lima bean (Phaseolus lunatus L.) is an important food source in Brazil, especially in the northeast region, where its production and consumption are high. The goals of the present study were to estimate natural outcrossing rates and genetic diversity levels of Lima bean from Brazil, using ten microsatellite loci to obtain information for their conservation and breeding. Fourteen accessions were selected from an experiment in field with open-pollinated and with the presence of pollinating insects. Twelve seeds of each of the 14 selected accessions were grown in screenhouse for tissue harvest and DNA extraction. The multilocus model was used to determine the reproductive system. The outcrossing rate was 38.1 % (t_m = 0.381; t_s = 0.078), and the results indicated a mixed mating system with a predominance of selfing (1 ? t_m = 61.9 %). The biparental inbreeding rate was high (t _m  ? t _s  = 0.303) and the multilocus correlated paternity was quite high (r _p(m) = 0.889), indicating that the progeny was mostly composed of full sibs. The average effective number of pollen donors per maternal plant (N _ep ) was low (1.12), and the fixation index for maternal genotypes (F _m ) was 0.945, indicating that most genitors resulted from inbreeding. The studied families presented considerable genetic variability: A = 6.10;  %P = 30; H _e  = 0.60 and H _o  = 0.077. Total diversity was high (H _T = 0.596), and a portion was distributed within families (H _S = 0.058). In addition, diversity was higher between families (D _ST = 0.538), and genetic differentiation was high (G _ST = 0.902). The results presented here can be used in the implementation of Lima bean conservation and breeding programs in Brazil.     

Elevated CO<Subscript>2</Subscript> alters the abundance but not the structure of diazotrophic community in the rhizosphere of soybean grown in a Mollisol

The effect of elevated CO₂ (eCO₂) on rhizospheric diazotrophic community in cropland has little been studied, although eCO₂ facilitates nodulation and N₂ fixation in legumes. In this study, four soybean cultivars (Xiaohuangjin, Suinong 8, Suinong 14, and Heinong 45) were grown in Mollisols for 65 days under ambient CO₂ (aCO₂) (390 ppm) or eCO₂ (550 ppm). Quantitative PCR and Illumina MiSeq sequencing targeting the nifH gene that reflects the composition of diazotrophic community were determined. Elevated CO₂ significantly increased the abundance of nifH gene copies in the rhizospheres of the Suinong 8 and Heinong 45 cultivars, but not in the Suinong 14 and Xiaohuangjin cultivars. The nifH abundance correlated negatively with nodule density (p?≤?0.01) but positively with nodule size (p?≤?0.01). Elevated CO₂ did not significantly alter the composition of diazotrophic community, nor shift dominant bacterial operational taxonomic units (OTUs). These results indicated that eCO₂ stimulated the growth but did not alter the community composition of diazotrophs in the rhizosphere of soybean, which depended on cultivar and might contribute to nodulation responses to eCO₂.     

Genetic diversity and relationships of wild and cultivated <Emphasis Type="Italic">Zanthoxylum</Emphasis> germplasms based on sequence-related amplified polymorphism (SRAP) markers

The genus Zanthoxylum, belonging to Rutaceae, has a long history of cultivation both for economic and chemical values in China. To effectively conserve and sustainably utilize this genus resource, a study on genetic diversity and relationships of Zanthoxylum germplasms was carried out by employing SRAP markers. We used 16 primer combinations to assess genetic variations and relationships among 175 accessions from eight cultivated provenances, including Shandong, Henan, Shanxi, Shaanxi, Gansu, Sichuan, Guizhou and Yunnan. A total of 145 clear repetitive and intense bands were yielded, and the percentage of polymorphic bands was 100 % for per primer combination, indicating a relatively high diversity among Zanthoxylum germplasms. From a geographic perspective, the highest genetic diversity level was observed within Guizhou provenance (N _a  = 1.97, Ne = 1.52, H = 0.31, I = 0.46) while Henan provenance had the lowest genetic diversity (N _a  = 1.68, Ne = 1.45, H = 0.25, I = 0.37). Based on AMOVA results, the abundant genetic variation was mainly caused by variation of intra-provenances (84.96 %), rather than among provenances (15.038 %). The results indicated low genetic differentiation (G _st  = 0.133) and high gene flow (N _m  = 3.2605) among provenances. The neighbor-joining tree revealed that the 175 accessions could be divided into four groups, and groupings indicated a divergence between the cultivated accessions of Zanthoxylum bungeanum Maxim. and Z. armatum DC. Moreover, three accessions of Z. piperitum DC. var. inerme without prickles introduced from Japan gathered one cluster. Cluster IV is composed of accessions of different geographical origin, including 11 wild species and 10 cultivated accessions of Z. bungeanum. The cluster analysis also reflected a relatively close relationship between the geographical origins and the classification of accessions in cluster I. Structure analysis indicated that collected Zanthoxylum accessions could be divided into two major groups. The information obtained from our research would benefit to make use of Zanthoxylum germplasms and assist the management of a Zanthoxylum germplasms collection.     

Response of CaCl<Subscript>2</Subscript>-extractable heavy metals,polychlorinated biphenyls,and microbial communities to biochar amendment in naturally contaminated soils

Purpose

Biochar can be used to reduce the bioavailability and leachability of heavy metals, as well as organic pollutants in soils through adsorption and other physicochemical reactions. The objective of the study was to determine the response of microbial communities to biochar amendment and its influence on heavy metal mobility and PCBs (PCB52, 44, 101, 149, 118, 153, 138, 180, 170, and 194) concentration in application of biochar as soil amendment.

Materials and methods

A pot (macrocosm) incubation experiment was carried out with different biochar amendment (0, 3, and 6 % w/w) for 112 days. The CaCl₂-extractable concentration of metals, microbial activities, and bacterial community were evaluated during the incubation period.

Results and discussion

The concentrations of 0.01 M CaCl₂-extractable metals decreased (p?>?0.05) by 12.7 and 20.5 % for Cu, 5.0 and 15.6 % for Zn, 0.2 and 0.5 % for Pb, and 1.1 and 8.9 % for Cd, in the presence of 3 and 6 % of biochar, respectively, following 1 day of incubation. Meanwhile, the total PCB concentrations decreased from 1.23 mg kg^?1 at 1 day to 0.24 mg kg^?1 at 112 days after 6 % biochar addition, representing a more than 60 % decrease relative to untreated soil. It was also found out that biochar addition increased the biological activities of catalase, phosphatase, and urease activity as compared with the controls at the same time point. Importantly, the Shannon diversity index of bacteria in control soils was 3.41, whereas it was 3.69 and 3.88 in soils treated with 3 and 6 % biochar soil. In particular, an increase in the number of populations with the putative ability to absorb PCB was noted in the biochar-amended soils.

Conclusions

The application of biochar to contaminated soils decreased the concentrations of heavy metals and PCBs. Application of biochar stimulated Proteobacteria and Bacteroides, which may function to absorb soil PCB and alleviate their toxicity.

     

Genetic diversity of a <Emphasis Type="Italic">Coffea</Emphasis> Germplasm Collection assessed by RAPD markers

Genetic diversity and relationships within and among nine species of Coffea, one species of Psilanthus and the Piatã hybrid from the Coffee Germplasm Collection of Instituto Agronômico de Campinas (IAC), Brazil were assessed using RAPD markers. Genetic diversity and relationships were evaluated by proportion of polymorphic loci (P), Shannon’s genetic index (H′ and G′_ST) and clustering analysis. The overall RAPD variation among all accessions was mostly partitioned between rather than within species. However, C. canephora and C. liberica showed a high genetic diversity within the species (\({\underline{\hbox{H}'}} \) _sp = 0.414 and \({\underline{\hbox{H}'}} \) _sp = 0.380, respectively) and this was highly structured (high \({\underline{\hbox{G}'}} \) _ST). Genetic diversity from C. congensis and C. arabica was also structured, but with lower levels of genetic diversity (\({\underline{\hbox{H}'}} \) _sp = 0.218 and \({\underline{\hbox{H}'}} \) _sp = 0.126, respectively). The results were consistent with agronomic and molecular studies and demonstrated that the IAC Coffea Collection is representative of the phylogenetic structure observed in the genera. This study devises sampling strategies for coffee germplasm collections and provides genetic diversity parameters for future comparisons among them.     

Effect of Metals on Decolorization of Reactive Blue HERD by <Emphasis Type="Italic">Comamonas</Emphasis> sp. UVS

Comamonas sp. UVS was able to decolorize Reactive Blue HERD (RBHERD) dye (50 mg L^?1) within 6 h under static condition. The maximum dye concentration degraded was 1,200 mg L^?1 within 210 h. A numerical simulation with the model gives an optimal value of 35.71?±?0.696 mg dye g^?1 cell h^?1 for maximum rate (V_max) and 112.35?±?0.34 mg L^?1 for the Michaelis constant (K_m). Comamonas sp. UVS has capability of decolorization of RBHERD in the presence of Mg²⁺, Ca²⁺, Cd²⁺, and Zn²⁺, whereas decolorization was completely inhibited by Cu²⁺. Metal ions also affected the levels of biotransformation enzymes during decolorization of RBHERD. Comamonas sp. UVS was also able to decolorize textile effluent with significant reduction in COD. The biodegradation of RBHERD dye was monitored by UV–vis spectroscopy, FTIR spectroscopy, and HPLC.     

Changes in CH<Subscript>4</Subscript> production during different stages of litter decomposition under inundation and N addition

Purpose

Our aim was to examine linkages between mass loss, chemical transformation and CH₄ production during decomposition of leaf litters submerged under water. We hypothesised that (i) labile leaf litters would fuel a rapid, high rate of methane (CH₄) production and that recalcitrant litters would fuel long-lasting but lower emissions, (ii) leaf litters experiencing a greater alteration to chemical properties would stimulate increased CH₄ production and (iii) nitrogen (N) addition would increase CH₄ emissions.

Materials and methods

Litters from six plant species were collected from a riparian ecosystem adjacent to Wyaralong Dam, located in Queensland, Australia, i.e., Lophostemon confertus, Cynodon dactylon, Heteropogon contortus, Chamaecrista rotundifolia, Chrysocephalum apiculatum and Imperata cylindrica. We evaluated the rate of mass loss and CH₄ emissions for 122 days of incubation in inundated microcosms with and without N addition. We quantified the chemical changes in the decomposing litters with ¹³ C-cross polarization and magic angle spinning (CPMAS) nuclear magnetic resonance (NMR) spectrum.

Results and discussion

The inundation treatment of plant litters significantly affected decomposition rates. All litters decomposed in either inundated or aerobic microcosms were quite distinct with regard to the NMR spectra of their initial litters. N addition altered the NMR spectra under both inundation and aerobic conditions. The N treatment only marginally influenced the decomposition rates of I. cylindrica and C. apiculatum litters. The diurnal patterns of CH₄ production in the H. contortus, C. rotundifolia and C. apiculatum litters under inundation incubation could be expressed as one-humped curves, with the peak value dependent on litter species and N treatment. N addition stimulated CH₄ emission by C. rotundifolia and C. apiculatum litters and inhibited CH₄ emission from microcosms containing the litters of the three gramineous species, i.e., I. cylindrica, C. dactylon and H. contortus.

Conclusions

Our results provide evidence that labile leaf litters could fuel a rapid, high rate of CH₄ production and that recalcitrant litters fuelled a lower CH₄ emission. We did not find that leaf litters with altered chemical properties stimulated increased CH₄ production. We also found that N addition was able to increase CH₄ emissions, but this effect was dependent on the litter species.

     

Nitrogen transformation rates and N<Subscript>2</Subscript>O producing pathways in two pasture soils

Purpose

Better understanding of N transformations and the regulation of N₂O-related N transformation processes in pasture soil contributes significantly to N fertilizer management and development of targeted mitigation strategies.

Materials and methods

¹⁵N tracer technique combined with acetylene (C₂H₂) method was used to measure gross N transformation rates and to distinguish pathways of N₂O production in two Australian pasture soils. The soils were collected from Glenormiston (GN) and Terang (TR), Victoria, Australia, and incubated at a soil moisture content of 60% water-filled pore space (WFPS) and at temperature of 20 °C.

Results and discussion

Two tested pasture soils were characterized by high mineralization and immobilization turnover. The average gross N nitrification rate (n_tot) was 7.28 mg N kg^?1 day^?1 in TR soil () and 5.79 mg N kg^?1 day^?1 in GN soil. Heterotrophic nitrification rates (n_h), which accounting for 50.8 and 41.9% of n_tot, and 23.4 and 30.1% of N₂O emissions in GN and TR soils, respectively, played a role similar with autotrophic nitrification in total nitrification and N₂O emission. Denitrification rates in two pasture soils were as low as 0.003–0.004 mg N kg^?1 day^?1 under selected conditions but contributed more than 30% of N₂O emissions.

Conclusions

Results demonstrated that two tested pasture soils were characterized by fast N transformation rates of mineralization, immobilization, and nitrification. Heterotrophic nitrification could be an important NO₃^?–N production transformation process in studied pasture soils. Except for autotrophic nitrification, roles of heterotrophic nitrification and denitrification in N₂O emission in two pasture soils should be considered when developing mitigation strategies.

     

C-banded karyotype of <Emphasis Type="Italic">Thinopyrum bessarabicum</Emphasis> and identification of its chromosomes in wheat background

C-banded pattern in two accessions of Thinopyrum bessarabicum (Save ex Rayss) A. Löve (2n = 2x = 14, E^bE^b) and their idiogram was established. C-banding analysis was further used to identify the chromosomes of Tritipyrum amphiploid (2n = 6x = 42, AABBE^bE^b) and a BC₁F₂ genotype from wheat and Tritipyrum. Two 18S-26S rDNA loci were detected on Th. bessarabicum chromosomes by in situ hybridization using an 18S-26S rDNA probe. E^b chromosomes in Tritipyrum generally were identified by their distinctive C-banding patterns which reflected heterochromatin regions. C-banding procedure resulted in sharp and distinct bands in one or both ends of E^b chromosomes without interval bands. Observed C-bands in E^b genome mainly reflected the telomeric and subtelomeric sequences which also showed more strong signals in genomic in situ hybridization. Results showed the importance of the C-banding technique as a screening tool in identification of addition and substitution lines in the progenies of wheat and Tritipyrum crosses during segregating generations.     

A Comparative Study on Removal Efficiency of Cr(VI) in Aqueous Solution by <Emphasis Type="Italic">Fusarium</Emphasis> sp. and <Emphasis Type="Italic">Myrothecium</Emphasis> sp.

Environmental pollution with chromium is due to residues of several industrial processes. Bioremediation is an alternative actually considered to remove Cr (VI) from the environment, using adapted organisms that grow in contaminated places. Have been conducted studies with fungi mechanisms of interaction with chromium, most of which have focused on processes biosorption, characterized it by passive binding of metal components of the cell surface, and bioaccumulation, wherein the metal entry to cells occurs with energy expenditure. The paper presents the results of studies carried out on sorption of chromium (VI) ions from aqueous solutions by Fusarium sp. and Myrothecium sp. Both biomasses have the ability to take up hexavalent chromium during the stationary phase of growth and as well inactive conditions. Fusarium sp. showed 26% of biosorption with active biomass and 64% in inactive biomass; meanwhile, Myrothecium sp. obtained 97 and 82%, respectively. Both fungi showed adjust to pseudo-second-order model in active (Fusarium sp. R ² = 0.99; Myrothecium sp. R ² = 0.96) and inactive biomass assay (Fusarium sp. R ² = 0.99; Myrothecium sp. R ² = 0.99). The data of the active biomass test also confirmed to the intraparticle diffusion model (Fusarium sp. R ² = 0.98; Myrothecium sp. R ² = 0.93). The results obtained through this investigation indicate the possibility of treating waste effluents containing hexavalent chromium using Fusarium sp. and Myrothecium sp.     

High Zn concentration does not impair biomass,cutting radial growth,and photosynthetic activity traits in <Emphasis Type="Italic">Populus alba</Emphasis> L.

Purpose

Evaluate the efficiency of Populus alba clone Villafranca in the uptake and translocation of Zn from contaminated soils.

Materials and methods

The effects of 48 days of zinc treatment (Zn _t ) on the growth and the photosynthetic activities of P. alba L. clone Villafranca were studied using ZnSO₄ (375 ppm per unit of soil dry weight) added in sand and peat moss substrate at the beginning of the treatment (T ₀) and again after 30 days (T ₁) in order to reach a target Zn concentration of 375 ppm at T ₀ and 750 ppm at T ₁ per unit of soil dry weight.

Results and discussion

Zn uptake in the different organs was analyzed after 30 (T ₁) and 48 days (T ₂) from the beginning of treatment, showing the following order: root ? leaves ≥ woody cutting = stem. The leaf area increased by 12 % in comparison to control plants at the end of second treatment (48 days). Cutting radial growth showed a high synchronicity in the growth rate fluctuation among control and Zn _t plants, but a higher increase in radial diameter of Zn _t cutting was observed starting from day 38 (after 8 days of second Zn _t ) reaching after 48 days 38 % higher than control plants.

Conclusions

Although our data of leaf Zn concentration were in the range usually reported as toxic for plants, Villafranca clone in Zn _t substrate were unaffected in terms of net CO₂ assimilation and stomatal conductance to water vapor.

     

Abundance of transcripts of functional gene reflects the inverse relationship between CH<Subscript>4</Subscript> and N<Subscript>2</Subscript>O emissions during mid-season drainage in acidic paddy soil

Agricultural management significantly affects methane (CH₄) and nitrous oxide (N₂O) emissions from paddy fields. However, little is known about the underlying microbiological mechanism. Field experiment was conducted to investigate the effect of the water regime and straw incorporation on CH₄ and N₂O emissions and soil properties. Quantitative PCR was applied to measure the abundance of soil methanogens, methane-oxidising bacteria, nitrifiers, and denitrifiers according to DNA and mRNA expression levels of microbial genes, including mcrA, pmoA, amoA, and nirK/nirS/nosZ. Field trials showed that the CH₄ and N₂O flux rates were negatively correlated with each other, and N₂O emissions were far lower than CH₄ emissions. Drainage and straw incorporation affected functional gene abundance through altered soil environment. The present (DNA-level) gene abundances of amoA, nosZ, and mcrA were higher with straw incorporation than those without straw incorporation, and they were positively correlated with high concentrations of soil exchangeable NH₄⁺ and dissolved organic carbon. The active (mRNA-level) gene abundance of mcrA was lower in the drainage treatment than in continuous flooding, which was negatively correlated with soil redox potential (Eh). The CH₄ flux rate was significantly and positively correlated with active mcrA abundance but negatively correlated with Eh. The N₂O flux rate was significantly and positively correlated with present and active nirS abundance and positively correlated with soil Eh. Thus, we demonstrated that active gene abundance, such as of mcrA for CH₄ and nirS for N₂O, reflects the contradictory relationship between CH₄ and N₂O emissions regulated by soil Eh in acidic paddy soils.     

Morphological and genetic diversity of shea tree (<Emphasis Type="Italic">Vitellaria paradoxa</Emphasis>) in the savannah regions of Ghana

Vitellaria paradoxa C. F. Gaertn., commonly known as shea tree or Vitellaria, is ranked the most important tree species of the savannah regions in the most African countries due to its ecological and economic importance for livelihoods and national economies. However, the savannah regions are the most vulnerable areas to the global climate change. Moreover, the Vitellaria populations on farmlands are threatened by the dominance of old trees with low or lack of regeneration. In this study both morphological and genetic diversity were assessed using several phenotypic traits and 10 microsatellite markers, respectively, to assess the impact of land use and agro-ecozone types on Vitellaria in Ghana. The land use types were forests and farmlands, and the agro-ecozone types included the Transitional, Guinea, and Sudan savannah zones. The mean values of morphological traits, such as diameter at breast height (DBH) and canopy diameter (CD), were statistically different between forest (DBH = 22.20, CD = 5.37) and farmland (DBH = 39.85 CD = 7.49) populations (P < 0.00001). The Sudan savannah zone with mean petiole length of 4.96 cm showed significant difference from the other zones, likely as a result of adaptation to drier climate conditions. Genetic data analysis was based on 10 microsatellite markers and revealed high genetic diversity of Vitellaria in Ghana: mean expected heterozygosity, H _e was 0.667, and allelic richness, measured as number of effective alleles A _e , was 4.066. Both farmlands and forests were very diverse indicating lack of negative influence of farmer’s selection on genetic diversity. Fixation index was positive for all populations (mean F _IS = 0.136) with farmlands recording relatively higher values than forests in all ecological zone types studied, probably indicating less gene flow in the farmlands. Moderate differentiation (F′ _ST = 0.113) was comparable to other similar tree species. Both land use and ecological zone types influenced genetic differentiation of Vitellaria at varying levels. The species was spatially structured across three ecozones and following climatic gradient. The forest reserves are used in situ conservation for Vitellaria in Ghana. High diversity observed in the most arid zones provides opportunity to find and use appropriate plant materials for breeding climate change resilient trees.     

Contrasting response of two grassland soils to N addition and moisture levels: N<Subscript>2</Subscript>O emission and functional gene abundance

Purpose

Nitrification and denitrification processes dominate nitrous oxide (N₂O) emission in grassland ecosystems, but their relative contribution as well as the abiotic factors are still not well understood.

Materials and methods

Two grassland soils from Duolun in Inner Mongolia, China, and Canterbury in New Zealand were used to quantitatively compare N₂O production and the abundance of bacterial and archaeal amoA, denitrifying nirK and nirS genes in response to N additions (0 and 100 μg NH₄ ⁺–N g^?1 dry soil) and two soil moisture levels (40 and 80 % water holding capacity) using microcosms.

Results and discussion

Soil moisture rather than N availability significantly increased the nitrification rate in the Duolun soil but not in the Canterbury soil. Moreover, N addition promoted denitrification enzyme activities in the Canterbury soil but not in the Duolun soil. The abundance of bacterial and archaeal amoA genes significantly increased as soil moisture increased in the Duolun soil, whereas in the Canterbury soil, only the abundance of bacterial amoA gene increased. The increase in N₂O flux induced by N addition was significantly greater in the Duolun soil than in the Canterbury soil, suggesting that nitrification may have a dominant role in N₂O emission for the Duolun soil, while denitrification for the Canterbury soil.

Conclusions

Microbial processes controlling N₂O emission differed in grassland soils, thus providing important baseline data in terms of global change.

     

Application of Response Surface Methodology and Machine Learning Combined with Data Simulation to Metal Determination of Freshwater Sediment

A comparative study between conventional methods (EPA 3050B and ISO 11466.3) of metal extraction and a simple low-cost method, using aqua regia, was carried out in this work. Six elements (Mn, Cu, Zn, Pb, Ni, and Cd) were determined by flame atomic absorption spectrometry (FAAS) in a certified sample of sediment (CNS 392). Central composite design (CCD) and response surface methodology (RSM), as well as machine learning, were used to find the optimal conditions for metal extraction. The influence of the parameters—volume of nitric acid in aqua regia (v), time of extraction (t), and temperature (T)—on Mn, Cu, Zn, and Pb recoveries was investigated. The best condition for the recovery of all the metals was v = 2.5 mL of HNO₃, t = 2 h, and T = 90 °C. In comparison with the conventional methods, the aqua regia method was found to present better recovery values and lower standard deviations for all the metals studied.     

Comparison of the <Emphasis Type="Italic">Rhodotorula mucilaginosa</Emphasis> Biofilm and Planktonic Culture on Heavy Metal Susceptibility and Removal Potential

This study compares the effect of heavy metals (Hg²⁺, Cu²⁺, and Pb²⁺) on the Rhodotorula mucilaginosa and Saccharomyces boulardii biofilm and planktonic cells. A MBEC^TM-HTP assay was used to test the levels of tolerance to heavy metals. The minimum inhibitory concentration (MICp) and minimum lethal concentration (MLCp) of the R. mucilaginosa and S. boulardii planktonic cells were determined, as well as minimum biofilm eradication concentration (MBEC). Metal removal efficiency was determined by batch biosorption assay. Previous studies had focused on heavy metal tolerance and removal efficiency of planktonic cells from Rhodotorula species only. Hence, our study presents and compares results for metal tolerance and removal efficiency of the R. mucilaginosa planktonic cells and biofilm. Biofilm tolerance was higher than the planktonic cells. The R. mucilaginosa planktonic cells showed the tolerance in the presence of Hg²⁺ (MICp 0.08 mM), Cu²⁺ (MICp 6.40 mM), and Pb²⁺ (MICp 3.51 mM), while the S. boulardii planktonic cells only tolerated Pb²⁺ (MICp 0.43 mM). The R. mucilaginosa biofilm showed the highest tolerance in the presence of Hg²⁺ (MBEC >0.31 mM), Cu²⁺(MBEC >12.81 mM), Pb²⁺ (MBEC >7.12 mM), and obtained results were confirmed by fluorescence microscopy. S. boulardii did not show potential in biofilm formation. The R. mucilaginosa biofilm exhibited better efficiency in removal of all tested metals than the planktonic cells. Metal removal efficiency was in the range from 4.79–10.25% for planktonic cells and 91.71–95.39% for biofilm.