Molecular cytogenetic identification of newly synthetic <Emphasis Type="Italic">Triticum kiharae</Emphasis> with high resistance to stripe rust

Six new amphiploids, Triticum kiharae Dorof. et Migusch. (2n?=?6x?=?42, A^tA^tGGDD), are described in this study. They were developed by the chromosome doubling of F₁ hybrid crosses between Triticum timopheevii Zhuk. (A^tA^tGG) with high resistance to stripe rust and Aegilops tauschii Cosson (DD) by colchicine treatment. These amphiploids showed a high level of fertility of 68–80% and exhibited relatively normal chromosome pairing in meiotic metaphase I. Individual chromosomes of T. kiharae could be identified by multicolor fluorescence in situ hybridization using the combination of oligonucleotides probes Oligo-pSc119.2-1, Oligo-pTa535-1, and Oligo-pTa71-2. T. kiharae exhibited high resistance to predominant stripe rust races CYR34, CYR31, CYR32, CYR33, and SY11-4 both during the seedling and adult stages. However, high molecular weight glutenin subunits from Ae. tauschii parents were only partially expressed in the T. kiharae background. These T. kiharae lines provide novel materials to widen the genetic diversity of the common wheat gene pool.     

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.     

Polymorphism of gliadins in <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. local populations in two primary habitats in Dagestan

Polymorphism of gliadins was investigated in Aegilops tauschii from primary habitats “4”, near Hily, and “6”, near Rukel, in Dagestan, Russia 205 individual plants were analysed (53/50 and 54/48 plants of subsp. tauschii/subsp. strangulata from the habitats “4” and “6”, respectively) and 1/7 and 18/14 different haplotypes were found among the plants of subsp. tauschii/subsp. strangulata from the habitats “4” and “6”, respectively. No direct evidences of cross-pollination were pointed out, although gliadins electrophoretic phenotypes obtained allowed to suggest that it occur in Ae. tauschii with very low frequency. The data obtained revealed that during Ae. tauschii evolutionary history a local habitat could be populated many times by different phylogenetic lineages of the species. It was found that in Dagestan, at the very edge of the species area, several different lineages belonging to different subspecies could for a long time co-exist together in a local habit, and in such case a very high level of genetic variation in Ae. tauschii could be accumulated on a square of less than one hectare. The further studies of genetic variation in Ae. tauschii local populations, based on molecular genetic methods seems to be very prospective for understanding of peculiarities of the species evolution.     

Relationship between spike morphology and habitat of four <Emphasis Type="Italic">Aegilops</Emphasis> species of section Sitopsis

This study examines the relationship between spike morphology and natural habitat for 84 accessions of four Aegilops species, belongs to section Sitopsis, Ae. bicornis, Ae. longissima, Ae. searsii, and Ae. sharonensis in genus Aegilops, section Sitopsis, wild relatives of Triticum aestivum L. These species are considered valuable genetic resources for future cultivation and breeding of domesticated wheat. The goals of the study were to: (1) document variation in spike morphology among these four species; (2) examine the relationship between spike morphology and native habitat; (3) document geographical distribution of distinct spike morphology; and (4) examine the relationship between spike morphology and heading time and value for these four species. The results reveal significant differences in spike morphology among species of section Sitopsis. The most noteworthy variation involved the absence/presence of lateral awn, such that species with lateral awn were restricted in coastal, though species without lateral awn were mainly distributed in inland. This suggests that local climate may be a determinant of variation in lateral awn, and that this trait may be subject to convergent evolution. Differences in heading time in sympatric area were also observed. The differences may enhance species divergence and could represent a lead speciation event. The results of this study will facilitate identification of populations or accessions of wild wheat with favorable traits and/or novel adaptive genes.     

Microbial and Physico-chemical Characteristics Associated with the Incidence of <Emphasis Type="Italic">Legionella</Emphasis> spp. and <Emphasis Type="Italic">Acanthamoeba</Emphasis> spp. in Rainwater Harvested from Different Roofing Materials

The incidence of Legionella and Acanthamoeba spp. was correlated to microbial indicator analysis and physico-chemical characteristics of rainwater harvested from catchment areas constructed from galvanized zinc, Chromadek®, and asbestos, respectively. Quantitative PCR (qPCR) analysis indicated that no significant difference (p?>?0.05) in copy numbers of Legionella spp. and Acanthamoeba spp. was recorded in tank water samples collected from the respective roofing materials. However, significant positive Spearman (ρ) correlations were recorded between the occurrences of Legionella spp. gene copies vs. nitrites and nitrates (p?=?0.05) in all tank water samples. Significant positive correlations were also established between Acanthamoeba spp. vs. barium (p?=?0.03), magnesium (p?=?0.02), sodium (p?=?0.02), silicon (p?=?0.05), arsenic (p?=?0.03), and phosphate (p?=?0.01), respectively. Additionally, while no significant correlations were observed between Legionella spp. vs. the indicator bacteria (p?>?0.05), positive correlations were observed between Acanthamoeba spp. vs. total coliforms (p?=?0.01) and Acanthamoeba spp. vs. Escherichia coli (p?=?0.02), respectively. Results obtained in the current study thus indicate that the incidence of Acanthamoeba and Legionella spp. in harvested rainwater was not influenced by the roofing material utilized. Moreover, it is essential that the microbial quality of rainwater be assessed before this water source is implemented for potable and domestic uses as untreated harvested rainwater may lead to legionellosis and amoebae infections.     

Arbuscular mycorrhizal fungi optimize the acquisition and translocation of Cd and P by cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.) plant cultivated on a Cd-contaminated soil

Purpose

Fruiting vegetables are generally considered to be safer than other vegetables for planting on cadmium (Cd)-contaminated farms. However, the risk of transferring Cd that has accumulated in the stems and leaves of fruiting vegetables is a major issue encountered with the usage of such non-edible parts. The objective of this study was to resolve the contribution of arbuscular mycorrhizal (AM) fungi to the production of low-Cd fruiting vegetables (focusing on the non-edible parts) on Cd-contaminated fields.

Materials and methods

An 8-week pot experiment was conducted to investigate the acquisition and translocation of Cd by cucumber (Cucumis sativus L.) plants on an unsterilized Cd-contaminated (1.6 mg kg^?1) soil in response to inoculation with the AM fungus, Funneliformis caledonium (Fc) or Glomus versiforme (Gv). Mycorrhizal colonization rates of cucumber roots were assessed. Dry biomass and Cd and phosphorus (P) concentrations in the cucumber shoots and roots were all measured. Soil pH, EC, total Cd, phytoavailable (DTPA-extractable) Cd, available P, and acid phosphatase activity were also tested.

Results and discussion

Both Fc and Gv significantly increased (P?<?0.05) root mycorrhizal colonization rates and P acquisition efficiencies, and thus the total P acquisition and biomass of cucumber plants, whereas only Fc significantly increased (P?<?0.05) soil acid phosphatase activity and the available P concentration. Both Fc and Gv significantly increased (P?<?0.05) root to shoot P translocation factors, inducing significantly higher (P?<?0.05) shoot P concentrations and shoot/root biomass ratios. In contrast, both Fc and Gv significantly decreased (P?<?0.05) root and shoot Cd concentrations, resulting in significantly increased (P?<?0.05) P/Cd concentration ratios, whereas only Gv significantly decreased (P?<?0.05) the root Cd acquisition efficiency and increased (P?<?0.05) the root to shoot Cd translocation factor. Additionally, AM fungi also tended to decrease soil total and phytoavailable Cd concentrations by elevating plant total Cd acquisition and soil pH, respectively.

Conclusions

Inoculation with AM fungi increased the P acquisition and biomass of cucumber plants, but decreased plant Cd concentrations by reducing the root Cd acquisition efficiency, and resulted in a tendency toward decreases in soil phytoavailable and total Cd concentrations via increases in soil pH and total Cd acquisition by cucumber plants, respectively. These results demonstrate the potential application of AM fungi for the production of fruiting vegetables with non-edible parts that contain low Cd levels on Cd-contaminated soils.

     

Changes in methane emission and methanogenic and methanotrophic communities in restored wetland with introduction of <Emphasis Type="Italic">Alnus trabeculosa</Emphasis>

Purpose

The dynamics and uncertainties in wetland methane budgets affected by the introduction of Alnus trabeculosa H. necessitate research on production of methane by methanogenic archaea and consumption by methane-oxidizing microorganisms simultaneously.

Materials and methods

This study investigated methane emission in situ by the closed chamber method, and methanogenic and methanotrophic communities using denatured gradient gel electrophoresis (DGGE) and quantitative PCR based on mcrA (methyl coenzyme M reductase), pmoA (particulate methane monooxygenase) genes in the rhizosphere and non-rhizosphere soils in the indigenous pure Phragmites australis T., and A. trabeculosa–P. australis mixed communities in Chongxi wetland.

Results and discussion

Methane flux rate from the pure P. australis community was 2.4 times larger than that of A. trabeculosa–P. australis mixed community in the rhizosphere and 1.7 times larger in the non-rhizosphere, respectively. The abundance of methanogens was lower in the mixed community soils (3.56?×?10³–6.90?×?10³ copies g^?1 dry soil) compared with the P. australis community (1.47?×?10⁴–1.89?×?10⁴ copies g^?1 dry soil), whereas the methanotrophs showed an opposite trend (2.08?×?10⁶–1.39?×?10⁶ copies g^?1 dry soil for P. australis and 6.20?×?10⁶–1.99?×?10⁶ copies g^?1 dry soil for mixed community soil). A liner relationship between methane emission rates against pmoA/mcrA ratios (R ²?=?0.5818, p?<?0.05, n?=?15) was observed. The community structures of the methane-cycling microorganism based on mcrA and pmoA suggested that acetoclastic methanogens belonging to Methanosarcinaceae and a particular type II methanotroph, Methylocystis, were dominant in these two plant communities.

Conclusions

The introduction of A. trabeculosa would promote the proliferation of methanotrophs, especially the dominant Methylocystis, but not methanogens, ultimately diminishing methane emission in the wetland.

     

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).

     

<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.     

Effect of Cadmium Stress on Growth and Electrical Impedance Spectroscopy Parameters of <Emphasis Type="Italic">Cotinus coggygria</Emphasis> Roots

The use of plants for ecological remediation is an important method of controlling heavy metals in polluted land. Cotinus coggygria is a landscape plant that is used extensively in landscaping and afforestation. In this study, the cadmium tolerance level of C. coggygria was evaluated using electrical impedance spectroscopy (EIS) to lay a theoretical foundation for broad applications of this species in Cd-polluted areas and provide theoretical support to broaden the application range of the EIS technique. Two-year-old potted seedlings of C. coggygria were placed in a greenhouse to analyse the changes in the growth, water content and EIS parameters of the roots following treatment with different Cd concentrations (50, 100, 200, 500, 1000 and 1500 mg kg^?1), and soil without added Cd was used as the control. The roots grew well following Cd treatments of 50 and 100 mg kg^?1. The Cd contents increased with the increase in Cd concentration in the soil. However, the lowest root Cd content was found at 4 months of treatment. The extracellular resistance r_e and the intracellular resistance r_i increased first overall and then decreased with the increasing Cd concentration, and both parameters increased with a longer treatment duration. The water content had a significant negative correlation with the Cd content (P?<?0.01) and the r_e (P?<?0.05). C. coggygria could tolerate a soil Cd concentration of 100 mg kg^?1. There was a turning point in the growth, water content and EIS parameters of the C. coggygria roots when the soil Cd concentration reached 200 mg kg^?1. The root water content and r_e could reflect the level of Cd tolerance in C. coggygria.     

Geographic distance and amorphous iron affect the abundance and distribution of <Emphasis Type="Italic">Geobacteraceae</Emphasis> in paddy soils in China

Purpose

Geobacteraceae are important dissimilatory Fe (III)-reducing microorganisms, influencing the cycling of metals, nutrients as well as the degradation of organic contaminants. However, little is known about their distribution, diversity, and abundance of Geobacteraceae and the effects of environment factors and geographic distance on the distribution and diversity of Geobacteraceae in paddy soils remain unclear. Therefore, the objectives of this study were to investigate the distribution, diversity, and abundance of Geobacteraceae in paddy soils and to determine key factors in shaping the Geobacteraceae distribution, environmental factors, geographic distance, or both and to quantify their contribution to Geobacteraceae variation.

Materials and methods

Illumina sequencing and quantitative real-time PCR using a primer set targeting 16S rRNA genes of bacteria affiliated with the family Geobacteraceae were employed to measure the community composition, diversity, and abundance patterns of 16S rRNA genes of Geobacteraceae in 16 samples collected from north to south of China. MRT, Mantel test, and VPA were used to analyze the relationship between communities of Geobacteraceae and environmental factors and geographic distance.

Results and discussion

Quantitative PCR showed that the abundance of 16S rRNA genes of Geobacteraceae ranged from (1.20?±?0.18)?×?10⁸ to 1.13?×?10⁹?±?2.25?×?10⁸ copies per gram of soil (dry weight) across different types of soils. Illumina sequencing results showed Geobacter was the dominant genus within the family of Geobacteraceae. Multivariate regression tree (MRT) analysis showed that soil amorphous iron contributed more (22.46 %) to the variation of dominant species of Geobacteraceae than other examined soil chemical factors such as pH (14.52 %), ammonium (5.12 %), and dissolved organic carbon (4.74 %). Additionally, more geographically distant sites harbored less similar communities. Variance partitioning analysis (VPA) showed that geographic distance contributed more to the variation of Geobacteraceae than any other factor, although the environmental factors explained more variation when combined. So, we detected the uneven distribution of Geobacteraceae in paddy soils of China and demonstrated that Geobacteraceae community composition was strongly associated with geographic distance and soil chemical factors including aFe, pH, Fe, DOC, C:N, and NO₃ ^?-N. These results greatly expand the knowledge of the distribution of Geobacteraceae in environments, particularly in terrestrial ecosystems.

Conclusions

Our results showed that geographic distance and amorphous iron played important roles in shaping Geobacteraceae community composition and revealed that both geographic distance and soil properties governed Geobacteraceae biogeography in paddy soils. Our findings will be critical in facilitating the prediction of element cycling by incorporating information on functional microbial communities into current biogeochemical models.

     

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.     

Soil quality assessment under different <Emphasis Type="Italic">Paulownia fortunei</Emphasis> plantations in mid-subtropical China

Purpose

Paulownia, one of the fastest growing broad-leaved tree species in the world, is widely distributed in the warm temperate regions of China. However, there are few commercial-scale Paulownia plantations, and there is only limited information available about the most suitable soil quality for Paulownia fortunei growth in mid-subtropical, Hunan Province, China.

Materials and methods

To understand the effect of the growth of P. fortunei on soil conditions, 25 soil property parameters under Paulownia plantations were studied in Hunan Province, China. Seventy-two standard plots of eight different stand types were analyzed by three statistical approaches to assess soil quality (SQ) in the different P. fortunei plantations.

Results and discussion

The results revealed that a majority of the soil characteristics when intercropping with oilseed rape and the pure P. fortunei (plantation III) were better than intercropping with Camellia oleifera, orange trees, and Cunninghamia lanceolata (Lamb.). Available calcium, available magnesium, available potassium, available phosphorus, soil thickness, slope, soil organic matter, available sulfur, available copper, dehydrogenase, and available zinc were selected as the minimum data set (MDS). The SQ index (SQI) showed that three classes for soil quality among the eight P. fortunei plantations ranged from 0.48 to 0.88 and these were correlated with standing volume (p?<?0.05).

Conclusions

From the results, we concluded that selected MDS indicators can describe the soil fertility quality of P. fortunei plantations, and that the relationship between SQI and standing volume has a biological significance. P. fortunei plantations intercropped with Camellia oleifera, orange trees, and Cunninghamia lanceolata (Lamb.) caused a deterioration in SQ, but intercropping oilseed rape and pure P. fortunei plantations produced an improvement in SQ.

     

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.     

Diversity of the composition and content of soluble carbohydrates in seeds of the genus <Emphasis Type="Italic">Vicia</Emphasis> (Leguminosae)

Low molecular weight carbohydrates of seeds of 10 species of Vicia, namely: V. angustifolia, V. articulata, V. cordata, V. ervilia, V. johannis, V. macrocarpa, V. monantha, V. narbonensis, V. pannonica and V. sativa were analyzed by the high resolution gas chromatography method. Seeds of the investigated species contain common (glucose, fructose, myo-inositol, sucrose, galactinol, di-galactosyl myo-inositol and raffinose family oligosaccharides—RFOs) and species-specific carbohydrates (d-pinitol and its α-d-galactosides—in V. articulata, V. monantha and V. pannonica or d-ononitol and its galactoside—in V. ervilia). Among the species containing in seeds RFOs as the main α-d-galactosides (V. angustifolia, V. cordata, V. johanensis, V. macrocarpa, V. narbonensis and V. sativa), an additional subgroup can be separated, which contains a set of unknown compounds (found in V. angustifolia, V. cordata and V. macrocarpa). Moreover, several other unidentified carbohydrate-containing compounds were detected exclusively in seeds of V. ervilia. The concentrations of total soluble carbohydrates (TSCs), including sugars, RFOs, cyclitols and galactosyl cyclitols and unknown compounds, in seeds differ significantly (P < 0.05) among the species. RFOs and sucrose are the main fractions in TSCs, with the exception of V. pannonica seeds, containing more galactosyl pinitols, and V. ervilia, in which unknown compounds account for 50% of TSCs. In V. ervilia RFOs occur at the lowest concentration (9.32 mg g^?1 seeds), and the concentration of DGMI (di-galactosyl myo-inositol) is comparable with that of RFOs, which is unusual among all the investigated Vicia species.     

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.     

Diversity and differentiation of <Emphasis Type="Italic">Oryza sativa</Emphasis> and <Emphasis Type="Italic">O. rufipogon</Emphasis> in Indonesia

Analysis of the genetic structure of Indonesian Oryza sativa and O. rufipogon using neighbour-joining trees based on single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) markers revealed that O. sativa in Indonesia is separated from O. rufipogon. Accessions of O. sativa in this study were differentiated into two major groups, indica and tropical japonica, excluding some varieties. SSR and SNP markers revealed the high value of differentiation (F _ST) and genetic distance (D) between indica and tropical japonica and we discovered four loci by SNP markers and one locus by SSR markers that play a role in differentiation between indica and tropical japonica. Interestingly, genetic diversity (H) in O. rufipogon was lower than that in O. sativa, however H in O. rufipogon was the highest and H in tropical japonica was the lowest when O. sativa was divided into two groups. Inbreeding coefficient (Fst) showed evidences that gene flow (Nm) between species and within species might be one of the mechanisms related to the diversification and differentiation of Indonesian rice germplasm by asymmetric pattern between species and within O. sativa as revealed by SSR and SNP markers. In addition, we found evidences on stabilizing selection in Indonesian rice germplasm and they might be the reasons why Indonesian rice germplasm did not differentiate due to source location of landrace. However, we found a weak relation between SSR and SNP markers probably due to highly polymorphic in SSR and the different properties of both markers.     

Predicting soil loss in central and south Italy with a single USLE-MM model

Purpose

The USLE-MM estimates event normalized plot soil loss, A_e,N, by an erosivity term given by the runoff coefficient, Q_R, times the single-storm erosion index, EI₃₀, raised to an exponent b₁?>?1. This modeling scheme is based on an expected power relationship, with an exponent greater than one, between event sediment concentration, C_e, and the EI₃₀/P_e (P_e = rainfall depth) term. In this investigation, carried out at the three experimental sites of Bagnara, Masse, and Sparacia, in Italy; the soundness of the USLE-MM scheme was tested.

Materials and methods

A total of 1192 (A_e,N, Q_REI₃₀) data pairs were used to parameterize the model both locally and considering all sites simultaneously. The performances of the fitted models were established by considering all erosive events and also by distinguishing between events of different severity.

Results and discussion

The b₁ exponent varied widely among the three sites (1.05–1.44) but using a common exponent (1.18) for these sites was possible. The A_e,N prediction accuracy increased in the passage from the smallest erosion events (A_e,N?≤?1 Mg ha^?1, median error =?3.35) to the largest ones (A_e,N?>?10 Mg ha^?1, median error =?1.72). The Q_REI₃₀ term was found to be usable to predict both A_e,N and the expected maximum uncertainty of this prediction. Soil erodibility was found to be mainly controlled by the largest erosion events.

Conclusions

Development of a single USLE-MM model appears possible. Sampling other sites is advisable to develop a single USLE-MM model for a general use.