<Emphasis Type="Italic">Trichoderma</Emphasis> improves the growth of <Emphasis Type="Italic">Leymus chinensis</Emphasis>

Bio-fertilizer application has been proposed as a strategy for enhancing soil fertility, regulating soil microflora composition, and improving crop yields, and it has been widely applied in the agricultural yields. However, the application of bio-fertilizer in grassland has been poorly studied. We conducted in situ and pot experiments to investigate the practical effects of different fertilization regimes on Leymus chinensis growth, with a focus on the potential microecological mechanisms underlying the responses of soil microbial composition. L. chinensis biomass was significantly (P?<?0.05) increased by treatment with 6000 kg ha^?1 of Trichoderma bio-fertilizer compared with other treatments. We found a positive (R² =?0.6274, P <?0.001) correlation between bacterial alpha diversity and L. chinensis biomass. Hierarchical cluster analysis and nonmetric multidimensional scaling (NMDS) revealed that soil bacterial and fungal community compositions were all separated according to the fertilization regime used. The relative abundance of the most beneficial genera in bio-fertilizer (BOF) (6000 kg ha^?1Trichoderma bio-fertilizer) was significantly higher than in organic fertilizer (OF) (6000 kg ha^?1 organic fertilizer) or in CK (non-amend fertilizer), there the potential pathogenic genera were reduced. There were significant negative (P?<?0.05) correlations between L. chinensis biomass and the relative abundance of several potential pathogenic genera. However, the relative abundance of most beneficial genera were significantly (P?<?0.05) positively correlated with L. chinensis biomass. Soil properties had different effects on these beneficial and on these pathogenic genera, further influencing L. chinensis biomass.     

Dominant Characteristics Between <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> and <Emphasis Type="Italic">Cyclotella</Emphasis> Sp. Accompanying Dilution Process in Eutrophic Lake

Although dilution of lake water has been used for improvement of water quality and algal blooms control, it has not necessarily succeeded to suppress the blooms. We hypothesized that the disappearance of algal blooms by dilution could be explained by flow regime, nutrient concentrations, and their interaction. This study investigated the effects of daily renewal rate (d), nitrogen (N) and phosphorus (P) concentration, and their interaction on the domination between Microcystis aeruginosa and Cyclotella sp. through a monoxenic culture experiment. The simulation model as functions of the N:P mass ratio and dilution rate (D) (calculated from d) was constructed, and the dominant characteristics of both species were predicted based on the model using parameters obtained in a monoculture experiment and our previous study. Results of monoxenic culture experiment revealed that M. aeruginosa dominated in all conditions (d = 5 or 15%; N = 1.0 or 2.5 or 5.0 mg-N L^?1; P = 0.1 or 0.5 mg-P L^?1) and the predicted cell densities were substantially correspondent to experimental data. Under various N:P ratios and D values, characteristics of domination for each species were predicted, indicating that Cyclotella sp. tended to be dominant under high P concentrations (P ≥ 0.36 mg-P L^?1) when the N:P ratio was less than 7.0, and M. aeruginosa could not form algal blooms at the N:P ratio ≤ 7.0 (N ≤ 0.7 mg-N L^?1). It was also suggested that the dilution rate leading to the Cyclotella sp. domination required 0.20 day^?1 or higher regardless of the N:P ratios.

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Graphical Abstract ? M. aeruginosa and Cyclotella sp. could be a superior competitor in nutrient-limited and nutrient-rich conditions, respectively. ? The simulation model in this study indicated that the predicted cell density and nutrient concentration were substantially correspondent to experimental data. ? The model predicted that Cyclotella sp. tended to be dominant at the P ≥ 0.36 mg-P L^?1 when the N:P ratio was less than 7.0, and M. aeruginosa could not form algal blooms at the N:P ratio ≤ 7.0 (N ≤ 0.7 mg-N L^?1).

     

Intracellular chromium accumulation by <Emphasis Type="Italic">Streptomyces</Emphasis> sp. MC1

Streptomyces sp. MC1, previously isolated from sugar cane, has shown ability to reduce Cr(VI) in liquid minimal medium and soil samples. The objective of this work was to demonstrate the intracellular chromium accumulation by Streptomyces sp. MC1 under different culture conditions. This strain was able to accumulate up to 3.54 mg of Cr(III) per gram of wet biomass, reducing the 98% of Cr(VI) and removing 13.9% of chromium from the culture medium supernatants. Streptomyces sp. MC1 chromium bioaccumulation ability was corroborated by using Timm’s reagent technique, a low-cost method, which has been used by first time to detect chromium deposits in bacteria. The results of atomic absorption spectrometry, scanning electron microscopy, and energy dispersive X-ray analysis suggest that the mechanism of Cr(VI) resistance observed in Streptomyces sp. MC1 includes adsorption coupled with reduction to Cr(III), and finally, Cr(III) bioaccumulation. This mechanism have special relevance to remediation of Cr(VI) contaminated environments by Streptomyces sp. MC1.     

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.     

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.     

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.     

Cross-transferability of SSR markers in <Emphasis Type="Italic">Osmanthus</Emphasis>

Developing a molecular tool kit for hybrid breeding of Osmanthus species and related genera is an important step in creating a systematic breeding program for this species. To date, molecular resources have been aimed solely at Osmanthus fragrans with little work to develop markers for other species and cultivars. The objectives of this study were to (1) determine cross-transferability of O. fragrans and Chionanthus retusus derived SSRs in diverse Osmanthus taxa, (2) quantify the influence of locus-specific factors on cross-transferability, and (3) determine the genetic relationships between accessions. We tested 70 SSR markers derived from O. fragrans and C. retusus in 24 accessions of Osmanthus. Sixty-seven markers showed transfer to at least one other Osmanthus species with an overall transfer rate of 84% of loci across taxa. Genotyping with 42 microsatellite markers yielded a total of 367 loci. Number of alleles per locus ranged from 2 to 17 with a mean of 8.7 ± 4.8. Mean observed and expected heterozygosities were 0.560 ± 0.225 and 0.688 ± 0.230, respectively. Percent of polymorphic loci ranged from 40% in Osmanthus delavayi to 100% in O. fragrans. Osmanthus fragrans had the highest mean number of alleles per locus (4.2) while O. delavayi had the lowest (1.1). A reduced suite of eight-markers can distinguish between accessions with non-exclusion probabilities of identity from 3.91E?⁰⁴ to 2.90E?⁰⁷. The SSR markers described herein will be immediately useful to characterize germplasm, identify hybrids, and aid in understanding the level of genetic diversity and relationships within the cultivated germplasm.     

Biodegradation Mechanism of Phenanthrene by Halophilic <Emphasis Type="Italic">Hortaea</Emphasis> sp. B15

This aim of the study is to investigate a halophilic bacterium Hortaea sp. B15, isolated from petroleum-contaminated soil for biodegradation of phenanthrene. Hortaea sp. B15 has the ability to completely degrade phenanthrene (100 mg/L) under salinity 10% within 1-week incubation. The metabolitic product of phenanthrene was identified and assayed by using ultraviolet-visible spectrophotometer and mass spectral analysis. Result revealed that Hortaea sp. B15 metabolized phenanthrene to form 9,10-phenanthrene quinone, salicylic acid, and gentisic acid. Hortaea sp. B15 has an efficient utilization of phenanthrene in high-saline liquid medium. All the results indicated that the fungus has a promising application for the study of high-molecular-weight PAH biodegradation and contaminated saline-alkali soil bioremediation.     

Polymorphism of <Emphasis Type="Italic">Got2</Emphasis> DNA sequences sheds light on <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. intraspecies divergence and origin of <Emphasis Type="Italic">Triticum aestivum</Emphasis> L.

DNA sequences of nuclear gene Got2 was studied in 60 accessions of Aegilops tauschii, 29 of subsp. tauschii and 31 of subsp. strangulata. It was found that Got2 allozyme polymorphism in Ae. tauschii is due to a single, unique, mutation which led to replacement of glutamic acid by isoleucine in residue 256 of the enzyme molecule, encoded by Got2. As revealed by Got2 DNA sequences variation, initially in its history Ae. tauschii was presented by subsp. strangulata, and among phylogenetic lineages of subsp. strangulata, the lineage “t-9¹s” (TauL3) is the most ancient, a relict one. Subspecies tauschii is relatively “young”. Initially it was presented by the lineage marked by combination of allozyme alleles Got2 ¹⁰⁵ and Acph1 ¹⁰⁰. In the past it inhabited the Continental area from Caucasia to Pakistan, but later on it was forced out by newly originated, now—a major lineage of subsp. tauschii, marked by Got2 ¹⁰⁰. This lineage extended the Continental area of the species up to Kirgizstan, but actually failed to penetrate into pre-Caspian area, occupied by subsp. strangulata. These results essentially differ from those obtained previously, using chloroplast DNA (cpDNA) sequences polymorphism. As revealed by cpDNA, the major, “usual”, subsp. strangulata (TauL2) is “younger” than subsp. tauschii, which resided on phylogenetic tree between relict lineage “t-9¹s”of subsp. strangulata—and major subsp. strangulata. But both cpDNA and Got2 DNA sequences indicate that the level of genetic variation in subsp. tauschii is much lower than in subsp. strangulata. According to Got2 DNA sequences variation, it was Ae. tauschii subsp. strangulata lineage “k-109″ which donated genome D to Triticum aestivum L. This lineage includes accessions: k-109 from South-Eastern Precaspian Azerbaijan; KU-2105, KU-2159 from Western Precaspian Iran; KU-2080 from Eastern Precaspian Iran.     

Optimization of <Emphasis Type="Italic">o</Emphasis>-Chlorophenol Biodegradation by Combined Mycelial Pellets Using Response Surface Methodology

In the present study, the immobilizing fermentation characteristics and o-chlorophenol biodegradation of Rhodopseudomonas palustris using mycelial pellets as a biomass carrier were investigated. To improve the o-chlorophenol degradation efficiency of the combined mycelial pellets, eight cultivation variables including glucose concentration, yeast extract concentration, spore inoculum size, pH, and agitation speed were optimized with an integrated strategy involving a combination of statistical designs. First, Plackett-Burman experiments identified glucose, yeast extract, and spore inoculum size as three statistically significant factors important for o-chlorophenol removal. Then, the steepest ascent method was used to access the optimal region of these significant factors. Finally, response surface methodology by Box-Behnken optimization was used to examine the mutual interactions among these three variables to determine their optimal levels. The ideal culture conditions for maximum o-chlorophenol removal according to a second-order polynomial model were as follows: 15.60 g/L glucose, 3.09 g/L yeast extract, and 9% (v/v) spore inoculum size, resulting in an expected o-chlorophenol removal rate of 92.60% with an o-chlorophenol initial concentration of 50 mg/L and 96-h culture time. The correlation coefficient (R ² = 0.9933) indicated excellent agreement between the experimental and predicted values, whereas a fair association was observed between the predicted model values and those obtained from subsequent experimentation at the optimized conditions.     

Phytoremediation Potential of <Emphasis Type="Italic">Helianthus annuus</Emphasis> and <Emphasis Type="Italic">Hydrangea paniculata</Emphasis> in Copper and Lead-Contaminated Soil

This study was conducted to assess the hyperaccumulation and phytoremediation potential of copper (Cu) and lead (Pb) in Hardy ‘Limelight’ Hydrangea (Hydrangea paniculata) and the common sunflower (Helianthus annuus). The study also investigated the capacity of these two plants to transpire the metals in a temperature-controlled greenhouse. Plants were grown for 4 weeks and periodically watered with known elemental concentrations of copper oxide nanoparticles, copper sulfate, and lead nitrate. Both H. annuus and H. paniculata accumulated significant amounts of Cu and Pb to be classified as hyperaccumulator species. H. annuus took up significant amounts of Cu in the shoots, specifically the leaves (Cu max.?=?1368 ppm), and easily translocated it from stem to leaf (translocation factor (TF) ranged from 2.7 to 81.0). Pb was not as easily taken up and translocated (TF?=?0.6) as Cu was by this species. H. paniculata took up Cu and Pb in high concentrations but preferentially stored more metals in the stems (Cu max.?=?1757 ppm; Pb max.?=?780 ppm) than in the leaves (Cu max.?=?126 ppm; Pb max.?=?35 ppm). The translocation ability of H. paniculata was much lower for both metals compared to H. annuus. Both Cu and Pb transpired from H. annuus at concentrations of 0.04 and 0.005 ppm, respectively.     

Geographical distribution of wild <Emphasis Type="Italic">Daucus</Emphasis> species in the natural grasslands of the Argentinian pampas

Three wild carrot species have been reported in the argentinian flora: Daucus montanus Humb. et Bonpl. ex Schult., D. montevidensis Link ex Sprengel and D. pusillus Michx. There is a discrepancy among authors about the distinctive morphological traits of the last two species; thus, it is difficult to ascertain if they are truly two distinct taxa. Based on the available literature and in the search of a paradigmatic site, four collection trips were carried out in 2004 and 2005 in Buenos Aires and Southern Entre Ríos provinces. Populations were sampled at 30 sites, and local environmental parameters and associated plant species were recorded. Morphological observations and chromosome counts were carried out on 10 plants/population. Three morphological phenotypes were distinguished: one in 18 populations, all with 2n = 2x = 18, and two in the remaining 12, with 2n = 2x = 22 or aneusomaty (2n = 2x = 20, 22). Populations of the first phenotype were assigned to wild D. carota and the rest, tentatively, to D. pusillus (D. montevidensis?), till further evaluations are carried out in test sites to verify this tentative conclusion.     

Bioaccumulation and Biosorption of Mercury by <Emphasis Type="Italic">Salvinia biloba</Emphasis> Raddi (Salviniaceae)

Aquatic macrophytes play a key role in nutrient cycling and in the dynamics of aquatic ecosystems. Many species have been evaluated in terms of their potential in phytoremediation processes in environments contaminated by metals. Considering this kind of application, we evaluated the potential for the bioaccumulation and biosorption of mercury by Salvinia biloba as a function of (i) different concentrations of mercury ions in the solution, (ii) the exposure time of live plants and dry biomass to the contaminant, and (iii) different pH’s, besides (iv) analyzing the effects of this metal on morphological and anatomical parameters. Bioaccumulation was evaluated by subjecting live plants to treatments with concentrations of 0.05, 0.1, and 0.2 μg.mL^?1 of mercury in the solution and the control (0 μg.mL^?1), at intervals of 3 days (0, 3, 6, 9, 12, and 15 days) at pH values of 5.5, 6.0, and 6.5. For biosorption, we used the dry biomass applying the same design, only changing the time intervals (0, 4, 8, 12, and 24 h). The bioaccumulated and biosorbed mercury was determined by atomic absorption spectroscopy. High values of mercury were bioaccumulated and biosorbed, and accordingly, as the concentration of mercury ions increases in the solution, the higher the value accumulated by both living plant and dry biomass. The time of exposure and the different pH values presented variation when associated with different concentrations in the bioaccumulation of mercury. Finally, few symptoms of toxicity in living plants were observed, evidencing the resistance of S. biloba to mercury and its potential use as a phytoremediation in water bodies contaminated by this metal.     

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.     

Domestication and saponins contents in a gradient of management intensity of agaves: <Emphasis Type="Italic">Agave cupreata</Emphasis>, <Emphasis Type="Italic">A. inaequidens</Emphasis> and <Emphasis Type="Italic">A. hookeri</Emphasis> in central Mexico

Saponins occur in numerous plants, including agaves, determining benefic and harmful properties to humans; their presence may favor using plants as soap and other products, but also they may cause caustic effects producing contact dermatitis. In domestication, favorable and unfavorable properties of saponins may cause an increase or decrease of their content, respectively. This study quantified and identified saponins among wild and managed populations of three agave species: A. cupreata Trel. et Berger, A. inaequidens Koch with wild and cultivated populations used for mescal production, and A. hookeri Jacobi, existing exclusively cultivated, used for production of the fermented beverage pulque. We studied 272 plants from 19 populations, quantifying contents of crude saponins through spectrometry. In 12 populations, the saponins types were identified by High Performance Liquid Chromatography–Mass-Spectrography-Time-of-Flight HPLC-MS-TOF. The highest crude saponins content was recorded in A. hookeri (26.09 mg/g), followed by A. cupreata (19.85 and 15.17 mg/g in wild and cultivated populations, respectively). For A. inaequidens, we recorded 14.21, 12.95, and 10.48 mg/g in wild, silvicultural managed and cultivated populations, respectively. We identified 18 saponins types, A. inaequidens showing all of them. A hecogenin glycoside (HG1) is found in high amounts in A. hookeri but in low quantities in A. inaequidens and A. cupreata. A. inaequidens had the greatest diversity of saponins. The contents of crude saponins in A. inaequidens and A. cupreata decrease with management intensity, but contrarily to what we expected, it was the highest in A. hookeri. We hypothesize that such high amount could be due to some saponins, probably HG1, may be precursors of sugars.     

Combined bioremediation of soil co-contaminated with cadmium and endosulfan by <Emphasis Type="Italic">Pleurotus eryngii</Emphasis> and <Emphasis Type="Italic">Coprinus comatus</Emphasis>

Purpose

The subjects of this study were to investigate the remediating potential of the co-cultivation of Pleurotus eryngii and Coprinus comatus on soil that is co-contaminated with heavy metal (cadmium (Cd)) and organic pollutant (endosulfan), and the effects of the co-cultivated mushrooms on soil biochemical indicators, such as laccase enzyme activity and bacterial counts.

Materials and methods

A pot experiment was conducted to investigate the combined bioremediation effects on co-contaminated soil. After the mature fruiting bodies were harvested from each pot, the biomass of mushrooms was recorded. In addition, bacterial counts and laccase enzyme activity in soil were determined. The content of Cd in mushrooms and soil was detected by the flame atomic absorption spectrometry (FAAS), and the variations of Cd fractions in soil were determined following the modified BCR sequential extraction procedure. Besides, the residual endosulfan in soil was detected by gas chromatography-mass spectrometry (GC-MS).

Results and discussion

The results indicated that co-cultivation of P. eryngii and C. comatus exerted the best remediation effect on the co-contaminated soil. The biomass of mushroom in the co-cultivated group (T group) was 1.57–13.20 and 19.75–56.64% higher than the group individually cultivated with P. eryngii (P group) or C. comatus (C group), respectively. The concentrations of Cd in the fruiting bodies of mushrooms were 1.83–3.06, 1.04–2.28, and 0.67–2.60 mg/kg in T, P, and C groups, respectively. Besides, the removal rates of endosulfan in all treatments exceeded 87%. The best bioremediation effect in T group might be caused by the mutual promotion of these two kinds of mushrooms.

Conclusions

The biomass of mushroom, laccase activity, bacterial counts, and Cd content in mushrooms were significantly enhanced, and the dissipation effect of endosulfan was slightly higher in the co-cultivated group than in the individually cultivated groups. In this study, the effect of co-cultivated macro fungi P. eryngii and C. comatus on the remediation of Cd and endosulfan co-contaminated soil was firstly reported, and the results are important for a better understanding of the co-remediation for co-contaminated soil.

     

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.     

Copper Binding by Activated Biochar Fibres Derived from <Emphasis Type="Italic">Luffa cylindrica</Emphasis>

The adsorption of copper (Cu(II)) from aqueous solutions by activated Luffa cylindrica biochar fibres has been investigated by means of batch equilibrium experiments and FTIR spectroscopy. The effect of various physicochemical parameters, such as pH, initial metal concentration, ionic strength, mass of the adsorbent, contact time and temperature, has been evaluated by means of batch type adsorption experiments. FTIR spectroscopy, as well as acid-base titrations, was used for the characterization of the material and the surface species formed. According to the experimental results even at pH 3, the relative sorption is above 85% and the adsorption capacity of the activated biochar fibres for Cu(II) is q _max = 248 g kg^?1. Moreover, the interaction between the surface carboxylic moieties and Cu(II) results in the formation of very stable inner-sphere complexes (?G ^o = ?11.2 kJ mol^?1 at pH 3 and ?22.4 kJ mol^?1 at pH 5.5).     

Remarks on genetic diversity and relationship of <Emphasis Type="Italic">Punica protopunica</Emphasis> and <Emphasis Type="Italic">P. granatum</Emphasis> assessed by molecular analyses

Here, two Punica species, viz., P. protopunica Balf. fil., reported as native to Socotra, and P. granatum L., were compared for the first time. Analysis of one P. protopunica and eleven P. granatum accessions was performed using three molecular markers, i.e., sequence related amplified polymorphism (SRAP), target region amplification polymorphism (TRAP), and intron targeted amplified polymorphism (ITAP), along with analysis of pgWD40 sequences, a gene involved in anthocyanin biosynthesis. All markers revealed the relationship between the two species and placed them at 33% similarity. SRAP, TRAP, and ITAP generated a total of 299, 260, and 160 bands, respectively. Of these, 78, 74, and 41 bands were specific for P. protopunica, and 92, 85, and 57 bands, respectively, were shared between both species. Sequence analysis of pgWD40~870 bp amplicons exhibited 100% identity among P. granatum accessions and 98% identity to that of P. protopunica. Phylogenetic analysis of WD40 sequences from monocot and dicot species, including both Punica species confirmed the relation between P. protopunica and P. granatum, supporting earlier reports that P. protopunica could be an ancestral species of P. granatum. Furthermore, the genetic diversity among and within P. granatum accessions from Egypt (3), Mexico (5), and Yemen (3) was assessed. Molecular marker-based relationships among region-bulked accessions was approximately the same (~90% similarity), whereas the degree of genetic variation was altered within each region. Specific bands (alleles) for accessions of each region along with those shared among them were identified. Thus, these bands could be used for pomegranate genotyping and breeding programs.     

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.