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.     

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.     

Nuclear and cytoplasmic contributions from <Emphasis Type="Italic">Erianthus arundinaceus</Emphasis> (Retz.) Jeswiet in a sugarcane hybrid clone confirmed through genomic in situ hybridization and cytoplasmic DNA polymorphism

A hybrid between Erianthus arundinaceus (Retz.) Jeswiet and Saccharum spontaneum L. which are wild related species of sugarcane (Saccharum L., Family Poaceae), was repeatedly crossed as female parent with sugarcane commercial varieties to develop near commercial sugarcane clones. The cytoplasm type of the hybrid derivatives were confirmed to be of E. arundinaceus through the mitochondrial and chloroplast DNA polymorphism of nad 4/3-4 intron segment and psbC–trnS segment, respectively. The E. arundinaceus × S. spontaneum hybrid with somatic chromosome number 2n = 62 was confirmed to have 30 chromosomes from E. arundinaceus through genomic in situ hybridization (GISH). The (E. arundinaceus × S. spontaneum) × sugarcane hybrid (2n = 118) had 24 chromosomes from E. arundinaceus whereas its next generation hybrid with sugarcane (2n = 108) had only 12 Erianthus chromosomes. The commercial sugarcane hybrid Co 15015, which is the third generation hybrid with 2n = 106 was confirmed to have two E. arundinaceus chromosomes through GISH. It is the first report of sugarcane with both alien cytoplasm and chromosome contributions from E. arundinaceus.     

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.     

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.     

Diverse morphological and cytogenetic variation and differentiation of the two subspecies in <Emphasis Type="Italic">Aegilops geniculata</Emphasis> Roth,a wild relative of wheat

Aegilops geniculata Roth, a wild relative of wheat (2n = 4x = 28, genome UUMM), is distributed over the Mediterranean basin and nearby areas. The species consists of two subspecies, subsp. geniculata and subsp. gibberosa (Zhuk.) Hammer. The former is distributed over the whole species area and has been genetically analyzed, and the latter is endemic to Spain and North Africa and has not been genetically evaluated. In this study, to clarify the genetic variation and delineation of the two subspecies from a biosystematic viewpoint, morphological variation among 23 accessions of subsp. geniculata and three of subsp. gibberosa and chromosome pairing at meiosis and fertility in their intra- and inter-subspecific F₁ hybrids were examined. A principal component analysis based on the 11 spike characteristics clearly divided the 26 accessions into two groups representing the two subspecies. The inter-subspecific F₁ hybrids showed significantly lower frequencies of chromosome pairing, significantly higher frequencies of multivalents, and significantly lower fertilities relative to those of the intra-subspecific F₁ hybrids. It was concluded that wide-ranging cytogenetic variation is included in subsp. geniculata, that subsp. gibberosa, the intra-subspecific variation of which is small, is morphologically and cytogenetically differentiated from subsp. geniculata beyond the range of the intra-subspecific variation of subsp. geniculata, and that the two subspecies are effectively isolated reproductively by hybrid sterility. The results strongly suggested that western North Africa is one of the important diversity centers of Ae. geniculata, where two subspecies were differentiated in the past and grow together in the present.     

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.     

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.     

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).

     

Population genetics of traditional landraces of <Emphasis Type="Italic">Cucurbita pepo</Emphasis> L., 1753 in the cloud forest in Baja Verapaz,Guatemala

Little is known regarding the effect of fragmentation and human agricultural management on the genetic variation and gene flow of Cucurbita pepo L., 1753 in moderate fragmented areas in central Guatemala. We hypothesize that the genetic variation of C. pepo is affected by forest fragmentation and by traditional agricultural management. Therefore, we aim to determine: (1) the genetic diversity and genetic structure of C. pepo in the Cloud Forest Corridor (CFC) (2) the extent of genetic admixture between commercial variety (CV) and traditional landraces (TL) of C. pepo, (3) the effect of habitat fragmentation in the population genetics of C. pepo with a landscape approach, and (4) the potential relationship between traditional management practices and genetic diversity of C. pepo in the CFC. We detected the existence of high level of genetic diversity (AR = 3.43; He = 0.50), inbreeding (Fis = 0.25) and moderate population structure of C. pepo in the CFC (Fst = 0.16). No correlation between landscape and genetics was found. Also, we found high genetic admixture between CV and TL. It seems that human practices, mainly related with seed exchange patterns, could affect genetic diversity of C. pepo in the CFC. C. pepo populations in the CFC are structured, with inbreeding, and show admixture with the CV, an aspect that could affect its genetic diversity. The agricultural management influenced the population genetics of C. pepo in the CFC, but the landscape did not. We suggest that special efforts should be made to preserve the diversity of this important indigenous food source for Guatemalan people as well as their management practices.     

Differentiation among Maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) Landraces from the Tarasca Mountain Chain,Michoacan, Mexico and the <Emphasis Type="Italic">Chalqueño</Emphasis> Complex

The classification of Mexican maize (Zea mays L.) begun since the early 20th century, it was consolidated during the middle of this century, but recent additions and rearrangements have been performed by several authors employing new methods of analysis and collections from diverse origin; nevertheless, maize from the State of Michoacan, Mexico has received little attention in regard to its systematic classification. Maize populations from the Tarasca Mountain Chain in Michoacan are commonly considered in literature, as belonging to the Chalqueño race; however, closer observations indicate that significant differences do exist, suggesting the necessity of performing an in-depth study on this respect. Thirty nine native maize populations from the Tarasca Mountain Chain region were evaluated along with 19 typical populations of the Chalqueño, Celaya and Conico races coming from the States of Mexico, Puebla, Hidalgo, Querétaro and Oaxaca. Populations were evaluated in Aranza, Michoacan and Montecillo, Mexico State. Seventeen morphological characters were scored and analyzed by one-way analyses of variance and multivariate techniques. Populations were also genetically analyzed through 17 isozyme loci. Native populations had some alleles not found either in the Chalqueño, Celaya or Conico races, and possess larger genetic diversity. Local populations were congregated into a discrete group apart from the typical Chalqueño populations, suggesting that landraces from the Tarasca Mountain Chain region might not be considered as belonging to the Chalqueño race, but they integrate by themselves a different race.     

Phylogenetic relationship of 40 species of genus Aloe L. and the origin of an allodiploid species revealed by nucleotide sequence variation in chloroplast intergenic space and cytogenetic insitu hybridization

Aloe species, which have been used as medicinal plants, belong to the Asphodelaceae family consisting of 530 species. In this study, genetic diversity and phylogenetic relationships among 40 Aloe species including a putative interspecies hybrid were analyzed using PCR band profiles from eight chloroplast intergenic space markers and nucleotide sequence diversity in the psbK–psbI intergenic region. A phylogenetic tree based on psbK–psbI sequences supported the revised classification of the genus Aloe as polyphyletic with several species be re-allocated into three genera Kumara, Aloidendron, and Aloiampelos. Further, the origin of the putative interspecies Aloe hybrid was characterized through molecular cytogenetics. Fluorescence and genomic insitu hybridization illustrated that the hybrid has a bimodal karyotype with a chromosome complement of 2n = 14, of which complementary halves were derived from two parental species, A. vera and A. arborescens. These findings revealed that the hybrid species was allodiploid. The phylogenetic analysis showed that A. arborescens was the maternal genome donor of the hybrid, as both have identical chloroplast genome sequences. We thus conclude that the allodiploid hybrid should be called A. arborescens × A.vera.     

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.     

Molecular characterization of <Emphasis Type="Italic">Triticum militinae</Emphasis> derived introgression lines carrying leaf rust resistance

Triticum militinae Zhuk. et Migusch. belongs to timopheevii [Triticum timopheevii (Zhuk.) Zhuk.] group of wheats with 2n = 4x = 28 chromosomes and genome formula A^tA^tGG. Triticum militinae Zhuk. et Migusch. is known to carry resistance to fungal diseases including rusts and powdery mildew. Genes from timopheevii wheat can be incorporated into cultivated wheat by either direct hybridization or through development of amphiploids. Three T. militinae derived introgression lines (ILs) Triticum Militinae Derivative (TMD) 6-4, TMD7-5 and TMD11-5 were selected for the current study based on cytological stability. All three ILs showed resistance against wide spectrum of Indian pathotypes of leaf rust. More than 1200 SSR markers were used for genotyping of ILs and parental lines. The ILs showed variable and multiple introgressions in different chromosomes of A, B and D genome of wheat. The introgression points were distributed mostly in the distal regions though significant introgressions were also observed in proximal regions of some chromosomes. The extent of introgression in ILs TMD6-4, TMD7-5 and TMD11-5 was 2.8, 8.3 and 8.6% respectively. The set of ‘informative markers’ in the Molecularly Tagged Chromosome Regions (MTCR) of T. militinae origin can also be used in future for tagging of genes associated with traits of economic importance apart from leaf rust resistance. The transferability of Triticum aestivum L. SSR markers to T. militinae was 96.4% for A genome, 95.8% for B genome and 84.3% for D genome. Transferability of wheat SSR markers to T. militinae can be used in preparing genetic maps in timopheevii group of wheats.     

Catalase and Phosphatase Activities During Hydrocarbon Removal from Oil-Contaminated Soil Amended with Agro-Industrial By-products and Macronutrients

Microbiological activities are essential in the bioremediation of polluted soils. The enzymatic activities of microorganisms are usually used as a biological indicator of soil health. The aim of this work was to observe the catalase, acid phosphatase (AcP), and alkaline phosphatase (AlP) activities in soil that was amended with agro-industrial by-products and macronutrients during the process of total petroleum hydrocarbon (TPH) removal. To this end, microcosm tests were performed with soil and agro-industrial by-products ratios of 100:2:2, for soil:sugarcane bagasse pith:filter cake mud (SSF); 100:2, for both soil:sugarcane bagasse pith (SS); and for soil filter cake mud (SF). The macronutrients—carbon, nitrogen, and phosphorus—in the experimental treatments were adjusted to 100:10:1 with a solution of NH₄NO₃ and K₂HPO₄. The best TPH removal (51.4%) was obtained with SSF at 15 days. In addition, a significant correlation was observed between TPH removal and AlP as well as AcP (r = 0.74, p < 0.0001; r = 0.70, p < 0.0107, respectively). Fungi growth was also correlated with both AlP (r = 0.97, p < 0.0001) and AcP (r = 0.95, p < 0.0001) activities. Besides, bacterial and fungi growth showed a correlation with TPH (r = 0.86, p < 0.001; r = 0.77, p < 0.0034, respectively). It could be said that the agro-industrial by-products and macronutrients contributed to pollutant removal from the oil-polluted soil at relatively short amount of time. In addition, the enzymatic activities were increased after the treatment; in this study, the high sensitivity enzyme was AlP, and it could be used as an indirect indicator of oil pollutant removal.     

Earthworm burrowing activity of two non-Lumbricidae earthworm species incubated in soils with contrasting organic carbon content (Vertisol vs. Ultisol)

The aim of this study was to investigate the burrowing activity of two earthworm species: the endogeic Drawida sinica and one undescribed Amynthas species incubated in Vertisol and Ultisol presenting different soil organic C content. Because of their contrasting feeding behaviours, we hypothesised that soil type would have a bigger influence on the burrowing activity of the endogeic than the anecic species. Repacked soil columns inoculated with earthworms for 30 days were scanned using X-ray tomography and the compiled images used to characterise the burrow systems. After scanning, the saturated hydraulic conductivity (K _sat) was also measured. The Amynthas species burrows were less numerous (30 vs. 180), more vertically oriented (57 vs. 37°), more connected from the surface to the bottom of the columns (73 vs. 5 cm³) and had a higher global connectivity index (83 vs. 28%) than those of D. sinica. The K _sat was threefold faster in columns incubated with Amynthas and was linked to the volume of percolating burrows (R ² = 0.81). The soil type did not influence Amynthas burrow characteristics. In contrast, there were 30% more D. sinica burrows in the Vertisol than in the Ultisol while other burrow characteristics were not affected. This result suggests that these burrows were more refilled with casts leading to shorter and discontinuous burrows. The K _sat was negatively related to the number of burrows (R ² = 0.44) but was not statistically different between the Vertisol and the Ultisol, suggesting a constant impact of this species on the K _sat. We found that a decrease in the amount of soil organic C by 50% had only a small influence on earthworm burrowing activity and no effect on the K _sat.     

Investigation and taxonomy of wild <Emphasis Type="Italic">Fragaria</Emphasis> resources in Tibet,China

Tibet is one of the main distribution centers of wild Fragaria species in China. There are abundant, precious and rare wild strawberry resources. This paper focused on the classification, distribution and evaluation of the 65 wild strawberry accessions collected from 17 counties in Tibet. The main taxonomical characters, such as stolon branching, leaflet type, flower sex, fruit shape, status of achene and sepal, and chromosome number were observed for each accession. All accessions were identified and classified into seven Fragaria species and two forms, including F. daltoniana J. Gay (2x), F. nilgerrensis Schlecht. (2x), F. nubicola Lindl. (2x), F. pentaphylla Losinsk. (2x), F. gracilis A. Los. (4x), F. moupinensis (French.) Card. (4x) and F. tibetica Staudt et Dickoré. (4x). The two white-fruited forms were F. nubicola f. alba (2x) and F. moupinensis f. alba (4x). The analytic hierarchy process (AHP) method was used to perform a comprehensive assessment on the value of seven Fragaria species and two forms. The results showed that F. nilgerrensis had better fruit quality, disease resistance and waterlogging tolerance than any other species in Tibet. The collection and identification of wild strawberry resources in Tibet would be helpful to improve the fruit quality and stress resistance of modern strawberry cultivars.     

Allopatric distributions of tetraploid and hexaploid forms of Aegilops neglecta Req. ex Bertol.

The genus Aegilops L. comprises 22 annual wild species that are closely related to wheat (Hammer in Kulturpflanze 28: 33–180, 1980). Aegilops neglecta Req. ex Bertol. is a member of the Aegilops section of this genus and is distributed from Morocco and Spain in the west to Transcaucasia and western Iran in the east. This species includes tetraploid (2n = 28, genome UUMM) and hexaploid forms (2n = 42, UUMMNN). However, the geographical distributions of the two cytological forms remain unclear. Clarifying the distribution of the two cytological forms is essential for a better understanding of the diffusion of Ae. neglecta and its tetraploid and hexaploid forms. In the present study, chromosome numbers were determined for accessions of Ae. neglecta from a total of 137 populations, located in the western area of the species distribution from the Aegean Islands to Morocco. Taken together with previous studies, the present data reveal a difference in the geographical distribution of tetraploid and hexaploid forms: tetraploid form is distributed in the eastern part of the species area and hexaploid form predominantly occurs in the western part with their border on the western margin of the Aegean Sea. Near the border, tetraploid and mixed populations are sporadically found among hexaploid populations in the Balkan and Peloponnesus Peninsulas, while a few hexaploid and mixed populations are found among tetraploid populations in the East Aegean Islands and West Anatolia.     

Screening species of <Emphasis Type="Italic">Pilocarpus</Emphasis> (Rutaceae) as sources of pilocarpine and other imidazole alkaloids

Plants of the Pilocarpus genus (Rutaceae) are popularly known as jaborandi and are the only source of pilocarpine, an imidazole alkaloid used in eye-drops for the treatment of glaucoma as well as for the stimulation of sweat and lachrymal glands. Alkaloid extracts from leaf samples of seven species of Pilocarpus, from the states of São Paulo and Maranhão in Brazil, were analyzed using HPLC–ESI–MS/MS. The samples contained between 0.88 ± 0.04 and 1.00 ± 0.14% of alkaloids in relation to the dry weight of their leaves, with significant differences in results (P ≤ 0.05) found only between Pilocarpus microphyllus planted in the state of Maranhão and Pilocarpus spicatus, Pilocarpus trachyllophus, Pilocarpus pennatifolius and Pilocarpus jaborandi; as well as between Pilocarpus spicatus and Pilocarpus racemosus. Pilocarpine was not found in P. spicatus, whereas in the other species it ranged from 2.6 ± 0.1 to 70.8 ± 1.2% of total alkaloids. P. microphyllus planted in the state of Maranhão for pilocarpine extraction had the highest total alkaloid content, but it had only 35% of pilocarpine in relation to total alkaloids. Three other species contained more pilocarpine in relation to total alkaloids: P. jaborandi (70.8%), P. racemosus (45.6%) and P. trachyllophus (38.7%); and could be candidates for pilocarpine extraction. Differences in alkaloid content were significant for all these samples (P ≤ 0.05). Imidazole alkaloids were observed and partially characterized based on their retention times and high resolution mass. The seven species analyzed had different imidazole alkaloid profiles, but only one did not present quantifiable pilocarpine contents in its leaves. The Pilocarpus genus shows potential for the prospection of novel alkaloids.     

The Pacific yam (<Emphasis Type="Italic">Dioscorea nummularia</Emphasis> Lam.), an under-exploited tuber crop from Melanesia

This study aims to explore the potential of Dioscorea nummularia Lam. and to gather information as a prelude to its improvement by plant breeding. After a taxonomic clarification and a review of the botanical data, its geographical areas of origin and distribution are determined. A germplasm survey conducted on nine islands of Vanuatu enabled the collection of 110 accessions corresponding to 84 cultivars. These were grouped into: (1) wild forms under domestication, (2) annual types, (3) perennial-type cultivars of D. nummularia and (4) natural interspecific hybrids with D. alata L. Within each group, remarkable morphological variation exists for stem spininess, leaf shape, emerging leaf colour, tuber shape and tuber flesh colour. Selected accessions examined for ploidy had chromosome numbers ranging from 2n = 3x = 60 to 2n = 6x = 120. Chloroplast counts conducted on stomatal guard cells confirmed the variation in ploidy levels. Eight different male cultivars were studied for pollen fertility, but <5% of the pollen grains appeared to be fertile when stained with acetocarmine. However, profuse flowering and synchrony favour pollination and result in fruit set. All accessions are resistant to anthracnose. Viruses were detected but are morphologically not discernible, with no incidence on yield. Physicochemical analyses of tubers from 16 accessions revealed that D. nummularia and interspecific hybrids have high percentages of dry matter (mean of 33.11%) and starch (82.81%). The Pacific yam could be improved through conventional hybridisation or used for interspecific crosses with related economically important species. Recommendations are made for further research.