Host status and reaction of <Emphasis Type="Italic">Medicago truncatula</Emphasis> accessions to infection by three major pathogens of pea (<Emphasis Type="Italic">Pisum sativum</Emphasis>) and alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis>)

Ditylenchus dipsaci, the stem nematode of alfalfa (Medicago sativa), Mycosphaerella pinodes, cause of Ascochyta blight in pea (Pisum sativum) and Aphanomyces euteiches, cause of pea root rot, result in major yield losses in French alfalfa and pea crops. These diseases are difficult to control and the partial resistances currently available are not effective enough. Medicago truncatula, the barrel medic, is the legume model for genetic studies, which should lead to the identification and characterization of new resistance genes for pathogens. We evaluated a collection of 34 accessions of M. truncatula and nine accessions from three other species (two from M. italica, six from M. littoralis and one from M. polymorpha) for resistance to these three major diseases. We developed screening tests, including standard host references, for each pathogen. Most of the accessions tested were resistant to D. dipsaci, with only three accessions classified as susceptible. A very high level of resistance to M. pinodes was observed among the accessions, none of which was susceptible to this pathogen. Conversely, a high level of variation, from resistant to susceptible accessions, was identified in response to infection by A. euteiches.     

Optimisation of larval culture of the mussel <Emphasis Type="Italic">Mytilus</Emphasis><Emphasis Type="Italic">edulis</Emphasis> (L.)

The blue mussel Mytilus edulis is a commercially important species whose fishery and culture generally relies on natural spat collection. Hatchery-production could provide an alternative source of seed, enabling reliable expansion of the industry. Mussel spawning and larval rearing trials were carried out to optimise elements of hatchery production. Culturing fertilised eggs at low density (20–200 eggs cm⁻²) rather than high density (400–720 eggs cm⁻²) significantly improved the quality of first veliger larvae and differences in this improvement were evident between the eggs from different females (maternal effects). Veliger larval growth at 17 or 21°C was significantly faster than growth at 14°C. Feeding veliger larvae an identical total ration either daily or at 2–3 day intervals did not significantly affect their growth. Different microalgal diets (1: Isochrysis sp. (clone T-ISO), 2: Chaetoceros calcitrans forma pumilus, 3: C. muelleri, 4: mixed Isochrysis sp. (clone T-ISO) and C. calcitrans f. pumilus, and 5: mixed Isochrysis sp. (clone T-ISO) and C. muelleri) were tested on veliger larval growth and mixed diets outperformed single-species diets.     

Analysis of rice blast resistance gene <Emphasis Type="Italic">Pi</Emphasis>-<Emphasis Type="Italic">z</Emphasis> in rice germplasm using pathogenicity assays and DNA markers

The Pi-z gene in rice confers resistance to a wide range of races of the rice blast fungus, Magnaporthe oryzae. The objective of this study was to characterize Pi-z in 111 rice germplasm accessions using DNA markers and pathogenicity assays. The existence of Pi-z in rice germplasm was detected by using four simple sequence repeat (SSR) markers (RM527, AP4791, AP5659-1, AP5659-5) closely linked to Pi-z, and was verified using pathogenicity assays with an avirulent strain (IE1k) and two virulent races (IB33 and IB49). Among 111 germplasm accessions evaluated, 73 were found to contain the Pi-z gene using both SSR markers and pathogenicity assays. The remaining 38 germplasm accessions were found to be inconsistent in their responses to the blast races IB33, IEIk and IB49 with expected SSR marker alleles, suggesting the presence of unexpected SSR alleles and additional R gene(s). These characterized germplasm can be used for genetic studies and marker-assisted breeding for improving blast resistance in rice.     

Functional and structural responses of bacterial and fungal communities from paddy fields following long-term rice cultivation

Purpose

Rice paddy soils undergo pedogenesis driven by periodic flooding and drainage cycles that lead to accumulation of organic matter and the stratification of nutrients and oxygen in the soil profile. Here, we examined the effects of continuous rice cultivation on microbial community structures, enzyme activities, and chemical properties for paddy soils along a chronosequence representing 0–700 years of rice cropping in China.

Materials and methods

Changes in the abundance and composition of bacterial and fungal communities were characterized at three depths (0–5, 5–10, and 10–20 cm) in relation to organic carbon, total nitrogen, dissolved organic carbon, microbial biomass carbon/nitrogen, and activities of acid phosphatase, invertase, and urease.

Results and discussion

Both soil organic carbon and total nitrogen increased over time at all three depths, while pH generally decreased. Microbial abundance (bacteria and fungi) and invertase and urease activity significantly increased with the duration of rice cultivation, especially in the surface layer. Fungal abundance and acid phosphatase activity declined with depth, whereas bacterial abundance was highest at the 5–10-cm soil depth. Profiles of the microbial community based on PCR-DGGE of 16S rRNA indicated that the composition of fungal communities was strongly influenced by soil depth, whereas soil bacterial community structures were similar throughout the profile.

Conclusions

Soil bioactivity (microbial abundance and soil enzymes) gradually increased with organic carbon and total nitrogen accumulation under prolonged rice cultivation. Microbial activity decreased with depth, and soil microbial communities were stratified with soil depth. The fungal community was more sensitive than the bacterial community to cultivation age and soil depth. However, the mechanism of fungal community succession with rice cultivation needs further research.

     

Assessing the Impacts of Time-to-Detection Distribution Assumptions on Detection Probability Estimation

Abundance estimates from animal point-count surveys require accurate estimates of detection probabilities. The standard model for estimating detection from removal-sampled point-count surveys assumes that organisms at a survey site are detected at a constant rate; however, this assumption can often lead to biased estimates. We consider a class of N-mixture models that allows for detection heterogeneity over time through a flexibly defined time-to-detection distribution (TTDD) and allows for fixed and random effects for both abundance and detection. Our model is thus a combination of survival time-to-event analysis with unknown-N, unknown-p abundance estimation. We specifically explore two-parameter families of TTDDs, e.g., gamma, that can additionally include a mixture component to model increased probability of detection in the initial observation period. Based on simulation analyses, we find that modeling a TTDD by using a two-parameter family is necessary when data have a chance of arising from a distribution of this nature. In addition, models with a mixture component can outperform non-mixture models even when the truth is non-mixture. Finally, we analyze an Ovenbird data set from the Chippewa National Forest using mixed effect models for both abundance and detection. We demonstrate that the effects of explanatory variables on abundance and detection are consistent across mixture TTDDs but that flexible TTDDs result in lower estimated probabilities of detection and therefore higher estimates of abundance.Supplementary materials accompanying this paper appear on-line.     

Effects of suspended solids in the survival and haematological parameters of pacu juveniles (Piaractus mesopotamicus) in a biofloc technology culture system

This study determined the effects of total suspended solids (TSS) on pacu (Piaractus mesopotamicus) culture in a biofloc technology (BFT) system by performing two experiments. Experiment 1 evaluated the LC50-96 h of pacu at seven TSS levels (0, 1500, 3000, 4000, 5000, 6000 and 7000 mg L^?1). Experiment 2 evaluated haematological variables (glucose, pH, haematocrit, haemoglobin, erythrocytes and haematimetric indices) in juveniles exposed to five different sublethal TSS concentrations (0, 250, 500, 750 and 1000 mg L^?1) for 5 days. Treatments below 5000 mg L^?1 did not cause mortality, and the LC50-96 h of TSS was estimated at 5477 mg L^?1. The TSS concentrations that caused mortality in pacu juveniles were extremely high and rarely maintained in culture systems. Exposure to high TSS concentrations (mainly 750 and 1000 mg L^?1) increased blood glucose, pH, heamatocrit, erythrocytes, haemoglobin values and haematimetric indices on the first day. On the fifth day, most of the evaluated parameters stabilized at different TSS concentrations. In conclusion, pacu exposed to high TSS concentrations for short periods may undergo physiological changes, and TSS concentrations below 250 mg L^?1 are recommended for its culture in a BFT system.     

Characterization of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">melongenae</Emphasis> isolates from Turkey with ISSR markers and DNA sequence analyses

Fusarium oxysporum f. sp. melongenae (Fomg), causal agent of Fusarium wilt of eggplant, is a serious pathogen in open fields and greenhouses. Inter-simple sequence repeat (ISSR) banding profiles, sequence analyses of inter-transcribed-spacer (ITS), translation elongation factor 1-alpha (TEF-1α), and actin (actA) DNA regions were employed in this study to determine genetic diversity and population structure of Fomg isolates obtained from Turkey. For ISSR study, (ACTG)₅, (GACAC)₃, (GACA)₄, (GATA)₄, HVH(TG)₇ and (CA)₈RG primers were selected from a set of 16. Discriminative ability of the primers revealed with various indices including polymorphic information content (PIC), and mean PIC value was calculated as 0.26. The ISSR data revealed 31 loci belonging to 202 Fomg isolates and 14 of them were found to be polymorphic. The isolates on neighbor joining ISSR tree were grouped into two major clusters which separated Fomg and outgroup isolates. Population structure was investigated based on bayesian modeling and results indicated five subpopulations (K = 5, ?K = 205.42). Mean genetic and geographical distances among sampling locations revealed only a weak and insignificant correlation (r = 0.583, P = 0.06). Phylogenetic analyses were carried out with ITS, TEF-1α and actA DNA regions with a selected subset of 30 Fomg, along with one non-host and one outgroup isolates. Since ITS region were not able to provide a meaningful separation, TEF-1α and actA sequences of each organism were concatenated individually to build a dendrogram. The clustering tree successfully separated the Fomg, non-host and outgroup isolates in which all Fomg were located on the same branch, forming a monophyletic group in the dendrogram.     

Plant diversity of secondary forests in response to anthropogenic disturbance levels in montane regions of northeastern China

The effects of anthropogenic disturbances on forest structure and plant diversity of secondary forest ecosystems were evaluated based on the classification by site factors in the montane regions of northeastern China. Forty-five sample plots containing 720 sub-plots of overstory species (8 m × 8 m), 1,440 quadrats of understory species (2 m × 2 m), and 1,440 quadrats of herbaceous layer species (1 m × 1 m) were clustered into five groups (G1, G2, G3, G4 and G5) by site variables with Hierarchical Cluster Analysis. Meanwhile, the disturbance levels corresponding to the five groups were determined according to the factors influencing human disturbances (D0-G2, D1-G1, D2-G3, D3-G5, and D4-G4). Species diversities of overstory, understory, herbaceous layer and overall species were evaluated using species number, Margalef index, Pielou index, Shannon–Wiener index, and Simpson index; and β-diversities (Whittaker index, Cody index). Basal area of stands exhibited a decreasing trend along the disturbance level due to a gradual increase in the extraction of timber by the human disturbances. The indices of species diversity suggested that overstory and understory species were distributed evenly among the groups or disturbance levels. There were no absolutely dominant tree species in the secondary forest ecosystems. The differences in site factors and the current disturbance intensities were not intense enough to lead to loose changes in overstory and understory species. The species diversity indices exhibited the maximum values at D3 (G5) for herbaceous layer species; this may suggest that the current disturbance intensities were intense enough to lead the changes of herbaceous layer species. Three rare and endangered species (Juglans mandshurica, Fraxinus mandshurica and Acanthopanax senticosus) were found within the secondary forests. These rare and endangered species appeared in each clustered group or disturbance level, which suggested that the current disturbance intensity in the study area was not strong enough to influence the distribution of rare and endangered species. The current disturbances in the secondary forests may not lead to a decrease in stability and complexity of the overstory and understory species, but the higher disturbance level may be intense enough to change the habitat fitness for the herbaceous species. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized authors.     

Soil evolution over the Quaternary period in a semiarid climate (Segre river terraces,northeast Spain)

A macro- and micromorphologic study was done on the soils from a stepped sequence of seven dated fluvial terraces in the lower Segre river valley (Lleida, northeast Spain) under a present-day semiarid Mediterranean climate. The soils have evolved from the Holocene through the early Pleistocene, providing an excellent morphostratigraphic framework for evaluating time-dependent factors influencing soil formation in a dry and calcareous environment. Throughout the chronosequence, some properties are regularly age-related specially carbonatation in subsurface horizons. The carbonates occur mainly as micrite, and although microsparite and sparite also appear in the oldest soils, they are replaced by fine-grained calcite by dissolution–reprecipitation processes (micritization process), which is active at present. Some pedological paleofeatures as the presence of sparite and recarbonated argillans in oldest terrace can be interpreted as the reflex of climatic changes during the Quaternary. In spite of this climate variability, the soils display progressive and systematic patterns of carbonate accumulation: on the lowest terraces, the soils do not yet have secondary carbonates but in the beginning Late Pleistocene calcic horizons, with carbonate pendents, are developed; these pendents increase its thickness with age although pendent growth rates decreases from Late Pleistocene to Middle Pleistocene. In the middle of the Late Pleistocene, calcic horizons evolved to petrocalcic horizons, which increase its thickness in the Middle and specially in Early Pleistocene. The presence of calcic and petrocalcic horizons is the primary basis for soil classification. This criterion is applicable not only to the soils of the lower Segre river basin, but also to many soils throughout the semiarid Mediterranean region.     

Lack of biocontrol capacity in a non-pathogenic mutant of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">melonis</Emphasis>

The aim of this study was to assess the biocontrol capacity of rev157, a non-pathogenic mutant of a pathogenic strain of Fusarium oxysporum f. sp. melonis (Fom24). Inoculated in association with the virulent parental strain, the mutant rev157 did not protect the host plant (muskmelon) against infection by Fom24. Applied on flax, a non-host plant, the mutant rev157 was not able to protect it against its specific pathogen F. oxysporum f. sp. lini. On the contrary the parental strain Fom24 did protect flax as well as a soil-borne biocontrol strain (Fo47). Since the mutant rev157 was affected neither in its growth in vitro nor in its capacity to penetrate into the roots, it can be speculated that the mutation has affected traits responsible for interactions within the plant. In F. oxysporum the pair of strains Fom24/rev157 is a good candidate to identify genes involved in the biocontrol capacity of F. oxysporum and to test the hypothesis of a link between capacity to induce the disease and capacity to induce resistance in the plant.