Ectomycorrhizal fungi identification in single and pooled root samples: terminal restriction fragment length polymorphism (TRFLP) and morphotyping compared

We describe here the results of a study conducted to evaluate a terminal restriction fragment length polymorphism (TRFLP) approach targeting rRNA genes for determination of ectomycorrhizal (ECM) communities colonizing the roots of loblolly pine (Pinus taeda L.). Root tips separated from soil cores were classified according to morphological characteristics and DNA was then extracted from each group of morphotyped tips. Labeled primers were used to generate terminal restriction fragments (TRF) for molecular fingerprinting of root colonizing fungi and to determine how well TRFLP could be used to discriminate between ectomycorrhizal types. Morphotypes generally correlated well with specific TRFs and sequence analysis confirmed that TRFs could be used to discriminate among fungal types. Sequence analysis indicated that important ECM fungi including Russulaceae, Thelephorales, and Tricholomataceae could be fingerprinted with TRFLP. In addition, a fixed proportion of the DNA extracted from each morphotype from the same core was used in a pooling experiment used to assess whether previously identified fungal species types could be distinguished from one another within reconstructed communities. Since some morphotypes share TRFs, dual analysis of ITS1 and ITS2 was necessary for accurate fingerprinting of fungal types. Approximately, 5.0±0.3 phylotypes were detected per core with TRFLP-corrected morphotyping as compared to 4.0±0.4 phylotypes using TRFLP on pooled community samples. TRFLP made on experimental sporocarp communities suggested that reduced ECM richness with TRFLP may be partly caused by differences in the amount of DNA available for PCR and primer bias. Nonetheless, TRFLP on pooled morphotypes accounted for more than 93% of colonized root tips. The method can be used to facilitate analysis of mycorrhizal communities using root tips collected from soil cores.     

Antioxidant properties of bran extracts from "Akron" wheat grown at different locations

Bran extracts of Akron wheat grown at four nonirrigated and one irrigated testing locations were examined and compared for their free radical scavenging properties against the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH(*)) and the radical cation ABTS(*)(+), chelating capacities, and total phenolic content (TPC) to determine the potential effects of environmental factors on the antioxidant properties of hard winter wheat. The environmental factors included total solar radiation, average daily solar radiation, and number of hours exceeding 32 degrees C. The results showed that bran samples from different growing locations may significantly differ in their radical scavenging activities against both DPPH(*) and ABTS(*)(+), chelating capacities, and TPC. A significant negative correlation was detected between the chelating activities of the bran samples from the four nonirrigated locations and total solar or daily average solar radiation (r = -0.999 and P = 0.001). These data suggest potential influences of growing conditions on the antioxidant properties of hard winter wheat and the possibility of producing wheat that is strong in a selected antioxidant property by optimizing the growing conditions of a selected wheat variety. More research is required to further investigate the relationship among antioxidant properties and environmental factors using different wheat varieties and larger sample sizes.     

New mosquito larvicidal tetranortriterpenoids from Turraea wakefieldii and Turraea floribunda

The crude methanol extracts of the root barks of Turraea wakefieldii and Turraea floribunda were found to show mosquito larvicidal activity against third-instar larvae of Anopheles gambiae sensu stricto. Four new limonoids comprising a vilasininoid 1 and three havanensinoids 2-4 were isolated from the chloroform fractions of the methanol extracts of T. wakefieldii and T. floribunda, respectively. The structures of the compounds were elucidated by NMR spectroscopy. Compounds 1, 2, and 4 had LD50 values of 7.1, 4.0, and 3.6 ppm, respectively, and were more potent than azadirachtin, which had an LD50 value of 57.1 ppm when tested against larvae of A. gambiae.     

Germination and hyphal growth of Glomus caledonicum on water agar containing benomyl

Concentrations of benomyl greater than 10 ng ml^?1 inhibited germination and hyphal growth of the vesicular-arbuscular mycorrhizal fungus Glomus caledonicum on water agar. This effect was attributable to only the activity of methyl-2-benzimidazole carbamate, one of the two hydrolysis products of benomyl. Hyphal growth but not germination was affected by the pH of water agar buffered with 10 mm 2-(N-morpholino)ethane sulphonic acid.     

Characterisation of agricultural drainage ditch sediments along the phosphorus transfer continuum in two contrasting headwater catchments

Purpose

This study investigated the phosphorus (P) source, mobilisation and transport potential of ditch bed sediments as well as surrounding field and bank soils in two agricultural headwater catchments with contrasting soil drainage capacities. This information is important for discerning the potential for ditches to attenuate or augment transfers of P from upstream sources and thus for developing appropriate management strategies for these features.

Materials and methods

Phosphorus sources were characterised using the Mehlich3-P, water-soluble P and total P tests. Phosphorus mobilisation potential was characterised using the Mehlich3-AL/P, Mehlich3-Ca/P and DESPRAL P tests. Phosphorus transport potential was characterised using data collected on the presence/absence of surface water in ditches during field surveys and downstream turbidity data.

Results and discussion

Ditch sediments had similar P source contents (Mehlich3-P, water-soluble P and total P) to the surrounding field soils and higher P contents than bank soils. However, calcium contents of sediments in the poorly drained catchment reflected the deep sub-soils rather than the surrounding field and bank soils. Mehlich3-Al/P and Mehlich3-Ca/P contents of ditch sediments in the well (non-calcareous) and poorly (calcareous) drained catchments respectively indicated potential for P retention (above thresholds of 11.7 and 74, respectively). However, sediments were less aggregated than field soils and may mobilise more particulate P (PP) during rain events. Nevertheless, the majority of surveyed ditches dried out from March to September 2011; thus, their potential to mobilise PP may be less important than their capacity to attenuate soluble and PP during this time.

Conclusions

In these and similar catchments, soluble P attenuation and particulate P mobilisation should be maximised and minimised, respectively, for example, by cleaning out the sediments before they become saturated with P and encouraging vegetation growth on ditch beds. This study also highlighted the influence of deep sub-soils on soluble P retention in ditches and thus the utility of characterising soils below depths normally included in soil classifications.

     

Hydrologic pathways and chemical composition of runoff during snowmelt in Loch Vale Watershed,Rocky Mountain National Park,Colorado, USA

Intensive sampling of a stream draining an alpine-subalpine basin revealed that depressions in pH and acid neutralizing capacity (ANC) of surface water at the beginning of the spring snowmelt in 1987 and 1988 were not accompanied by increases in strong acid anions, and that surface waters did not become acidic (ANC<0). Samples of meltwater collected at the base of the snowpack in 1987 were acidic and exhibited distinct ‘pulses’ of nitrate and sulfate. Solutions collected with lysimeters in forest soils adjacent to the stream revealed high levels of dissolved organic carbon (DOC) and total Al. Peaks in concentration of DOC, Al, and nutrient species in the stream samples indicate a flush of soil solution into the surface water at the beginning of the melt. Infiltration of meltwater into soils and spatial heterogeneity in the timing of melting across the basin prevented stream and lake waters from becoming acidic.     

Input-Output Budgets of Inorganic Nitrogen for 24 Forest Watersheds in the Northeastern United States: A Review

Input-output budgets for dissolved inorganic nitrogen (DIN) are summarized for 24 small watersheds at 15 locations in the northeasternUnited States. The study watersheds are completely forested, free of recent physical disturbances, and span a geographical region bounded by West Virginia on the south and west, and Maine on the north and east. Total N budgets are not presented; however, fluxes of inorganic N in precipitation and streamwater dominate inputs and outputs of N at these watersheds. The range in inputs of DIN in wet-only precipitation from nearby National Atmospheric Deposition Program (NADP) sites was 2.7 to 8.1 kg N ha^-1 yr^-1 (mean = 6.4 kg N ha^-1 yr^-1; median = 7.0 kg N ha^-1 yr^-1). Outputs of DIN in streamwater ranged from 0.1 to 5.7 kg N ha^-1 yr^-1 (mean = 2.0 kg N ha^-1 yr^-1; median = 1.7 kg N ha^-1 yr^-1). Precipitation inputs of DIN exceeded outputs in streamwater at all watersheds, with net retention of DIN ranging from 1.2 to 7.3 kg N ha^-1 yr^-1 (mean = 4.4 kg N ha^-1 yr^-1; median = 4.6 kg N ha^-1 yr^-1). Outputs of DIN in streamwater were predominantly NO₃-N (mean = 89%; median = 94%). Wet deposition of DIN was not significantly related to DIN outputs in streamwater for these watersheds. Watershed characteristics such as hydrology, vegetation type, and land-use history affect DIN losses and may mask any relationship between inputs and outputs. Consequently, these factors need to be included in the development of indices and simulation models for predicting 'nitrogen saturation' and other ecological processes.     

Autophagy genes are essential for dauer development and life-span extension in C. elegans

Both dauer formation (a stage of developmental arrest) and adult life-span in Caenorhabditis elegans are negatively regulated by insulin-like signaling, but little is known about cellular pathways that mediate these processes. Autophagy, through the sequestration and delivery of cargo to the lysosomes, is the major route for degrading long-lived proteins and cytoplasmic organelles in eukaryotic cells. Using nematodes with a loss-of-function mutation in the insulin-like signaling pathway, we show that bec-1, the C. elegans ortholog of the yeast and mammalian autophagy gene APG6/VPS30/beclin1, is essential for normal dauer morphogenesis and life-span extension. Dauer formation is associated with increased autophagy and also requires C. elegans orthologs of the yeast autophagy genes APG1, APG7, APG8, and AUT10. Thus, autophagy is a cellular pathway essential for dauer development and life-span extension in C. elegans.