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.     

Histopathology of infection and colonization of Quercus ilex fine roots by Phytophthora cinnamomi

Quercus ilex is one of the European forest species most susceptible to root rot caused by the oomycete Phytophthora cinnamomi. This disease contributes to holm oak decline, a particularly serious problem in the ‘dehesas’ ecosystem of the southwestern Iberian Peninsula. This work describes the host–pathogen interaction of Q. ilex and P. cinnamomi, using new infection indices at the tissue level. Fine roots of 6‐month‐old saplings inoculated with P. cinnamomi were examined by light microscopy and a random pool of images was analysed in order to calculate different indices based on the measured area of pathogen structures. In the early stages of invasion, P. cinnamomi colonizes the apoplast and penetrates cortical cells with somatic structures. On reaching the parenchymatous tissues of the central cylinder, the pathogen develops different reproductive and survival structures inside the cells and then expands through the vascular system of the root. Some host responses were identified, such as cell wall thickening, accumulation of phenolic compounds in the middle lamella of sclerenchyma tissues, and mucilage secretion blocking vascular cells. New insights into the behaviour of P. cinnamomi inside fine roots are described. Host responses fail due to rapid expansion of the pathogen and a change in its behaviour from biotrophic to necrotrophic.     

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.     

Effects of various densities of Ophiostoma ips inoculations on Pinus sylvestris in north‐western Spain

The aim was to determine the inoculation density above which Scots pine (Pinus sylvestris) is overcome by the blue‐stain fungus Ophiostoma ips that is associated with the bark beetle Ips sexdentatus. In north‐western Spain, stems of 16 Scots pines were inoculated at various densities (0, 400, 800 or 1600 inoculi/m²) along circumferential 100 or 150 cm wide inoculation belts. Each inoculum consisted of a 5 mm diameter cylinder of malt extract agar colonized by the fungus. Three months later, all trees were harvested and trunk resinosis and foliage colour were visually assessed. The percentage of healthy, desiccated, resin soaked, and blue‐stained sapwood, as well as growth productivity indices, were calculated from stem disks cut from within the inoculated zone of each tree. Sapwood‐specific hydraulic conductivity (Ks) of each tree was measured in the middle of the inoculated zone. All parameters of tree vigour changed dramatically to the worse when inoculation densities were above 400 inoculi/m², and foliage changed from green to yellow‐green or yellow when an inoculation density of 800 instead of 400 was used. The percentage loss of sapwood‐specific conductivity (PLC) increased from 30 to 90% and the percentage of healthy, conductive sapwood dropped from 85 to 35% at 800 inoculi/m². No effect of the width of the inoculation belt was observed, and there was no relationship between tree productivity indices and the level of resistance. A non‐linear negative relationship was found between PLC and the percentage of healthy sapwood. It is concluded that tree resistance was overcome and that trees were going to die when the inoculation density was ≥800 inoculi/m².     

An outbreak of Shewanella putrefaciens group in wild eels Anguilla anguilla L. favoured by hypoxic aquatic environments

Microbiological analyses were conducted on wild eels from the L′Albufera Lake (Spain). A total of 174 individuals were collected in two surveys (i.e. year 2008 and autumn–winter 2014) among those caught by local fishermen into the lagoon. The prevalence of Shewanella putrefaciens group was 1.7% in 2008 and rose above 32% in 2014. It was due to an outbreak of shewanellosis that presented a morbidity rate of 64%. S. putrefaciens group strains were isolated as pure cultures from the sick eels that showed white ulcers surrounded by a reddish inflammation, damage of the mouth, extensive skin discoloration, exophthalmia, ascites and bad odour. The S. putrefaciens group was recovered from freshwater samples taken at the L′Albufera system, along autumn–winter 2015. Its counts significantly increased in freshwater parallel to hypoxia and temperature rising. Shewanellae strains were identified as S. putrefaciens and S. xiamenensis by 16S rRNA gene sequencing. These isolates recovered from sick eels or freshwater were virulent for European eel by IP challenge (LD₅₀ 10⁶ CFU g^?1 body weight). They also caused 30–38% cumulative mortality, in European eels challenged by a 2‐h bath (10⁷ CFU mL^?1). These results suggest that shewanellosis could be transmitted through water highlighting the fact that hypoxic conditions increase this bacterium levels in water.     

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.     

Illinois Soil Nitrogen Test Method Optimization for Analysis of Temperate Mineral Grassland Soils

ABSTRACT

A reliable and practical test that can provide timely measurements of the levels of mineralizable nitrogen (MN) in soil is critical for improving the accuracy of N fertilizer applications for grassland and crops. The Illinois soil N test (ISNT) is considered to be a good estimate of MN, once soils are grouped according to soil characteristics such as the drainage type and sampling depth. To date, development and evaluation of the ISNT method has been conducted using arable soils mainly in North America where, in general, soils have lower levels of soil organic matter (SOM) compared to temperate grassland soils. We evaluated the effects of two pre-treatment soil aggregate sizes of <1 mm and <2 mm on the yield and recovery of MN (1) across temperate grassland soil types, and (2) across a 6-h interval diffusion period. No significant difference existed in the concentrations of ISNT-N between the two soil aggregate sizes of each soil type. For both aggregate sample sizes, the recovery of spiked amino sugar-N glucosamine from a temperate grassland soil was generally linear until hour 5, after which the quantities of recovered N diminished. Although N recovery after 6 h of diffusion at 50°C (±1°C) was less than 100% in both aggregate size samples, the response models indicated that the standard ISNT protocol using a 5-h diffusion period is appropriate for temperate grassland soils. The incomplete recovery of N in these mineral soils suggested that the protocol could be further optimized for temperate soils with high organic matter content and additional evaluation of the temperature during diffusion within an enclosed environment may be required using N (spiked glucosamine-N) recovery studies.     

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.