A rapid differential staining technique for Fusarium pseudograminearum in cereal tissues during crown rot infections

A novel fluorescence‐based differential staining procedure has been developed for the rapid visualization of Fusarium pseudograminearum hyphae in cereal tissues. Infected tissues are stained with safranin, which binds to host cell walls, while hyphal tissues are stained with the fluorescent dye solophenyl flavine 7GFE. The method is rapid, does not require clearing of culm cross‐sections, and provides high contrast informative images of fungal and plant structures during pathogenesis.     

A Review of Fine Root Dynamics in Populus Plantations

Production of native and hybridized varieties of Populus has received considerable interest in temperate regions as an alternative to agricultural crops and an additional wood source, while acting as a potential carbon (C) sink to offset emissions of fossil fuel-based greenhouse gases. Research of root system dynamics in Populus species is expanding, however, our understanding of the nature and role of fine roots (FR) is incomplete. The study objective, therefore, was to review the literature regarding FR production, mortality and longevity in Populus, and evaluate the magnitude and significance of the FR fraction to C sequestration. FRs, conventionally defined as less than 2 mm in diameter and responsible for water and nutrient uptake, are an essential component of the tree. Populus FRs are relatively short-lived, with reported lifespans ranging from 30 to 300 days, depending on root diameter, tree species and age, and soil environmental factors. Standing FR biomass fluctuates throughout the growing season. Fine root production generally peaks in mid-summer, and ranges between 1.0 and 5.0 mg ha⁻¹ yr⁻¹, while FR mortality has less seasonal amplitude. Production and mortality dynamics in Populus are highly plastic in response to soil environmental conditions, and although opposing conclusions have been proposed, research suggests soil moisture and nitrogen to be most important. Results from the literature indicate annual FR turnover to the soil C pool may be small (0.2–1.6 mg C ha⁻¹ yr⁻¹), but substantial in maintaining or enhancing C levels in natural and managed stands of Populus.     

Gram-negative identification card for identification of Salmonella, Escherichia coli, and other Enterobacteriaceae isolated from foods: collaborative study

Twelve laboratories evaluated the Gram-Negative Identification (GNI) Card to identify members of the Enterobacteriaceae. Eighty-four isolates, previously isolated from foods, were used in the collaborative study; the isolates represented 12 genera within the Enterobacteriaceae group. Each collaborator streaked each isolate on tryptic soy agar plates for purity. In the method, plates are incubated 18-24 h at 35 degrees C. Isolated colonies are then subcultured to tryptic soy agar slants and incubated 18-24 h at 35 degrees C. An emulsion is made from the growth on the slant in 1.8 mL 0.45% sodium chloride solution. The GNI Card is filled and placed in a reader/incubator. Isolates are identified and an identification is printed. The Vitek System correctly identified 96.7% of Salmonella sp., 97.0% of Escherichia coli, and an average of 93.8% of the other enteric genera. The method using the Vitek System and GNI Card has been approved interim official first action by AOAC as a screening method for the presumptive identification of Salmonella sp., E. coli, and other Enterobacteriaceae isolated from foods.     

Hierarchical scheme for LC-MSn identification of chlorogenic acids

The fragmentation behavior of 18 chlorogenic acids that are not substituted at position 1 has been investigated using LC-MS(4) applied to a methanolic coffee bean extract and commercial cider (hard cider). Using LC-MS(3), it is possible to discriminate between each of the three isomers of p-coumaroylquinic acid, caffeoylquinic acid, feruloylquinic acid, and dicaffeoylquinic acid, and a hierarchical key has been prepared to facilitate this process when standards are not available. MS(4) fragmentations further support these assignments, but were not essential in reaching them. The distinctive behavior of 4-acyl and 3-acyl chlorogenic acids compared with the 5-acyl chlorogenic acids is a key factor permitting these assignments. The fragmentation patterns are dependent upon the particular stereochemical relationships between the individual substituents on the quinic acid moiety. Fragmentation is facilitated by 1,2-acyl participation and proceeds through quinic acid conformers in which the relevant substituents transiently adopt a 1,3-syn-diaxial relationship. Selected ion monitoring at m/z 529 clearly indicated the presence in coffee of six caffeoylferuloylquinic acid isomers, whereas previously only two or three had been demonstrated. The hierarchical key permitted specific structures to be assigned to each of the six isomers. These assignments are internally consistent and consistent with the limited data previously available.     

Discriminating between the six isomers of dicaffeoylquinic acid by LC-MS(n)

The fragmentation behavior of all six dicaffeoylquinic acids (diCQA) has been investigated using LC-MS(4). It is possible to discriminate between each of the isomers including those for which commercial standards are not available. For diCQA, the ease of removal of the caffeoyl residue during fragmentation is 1 approximately = 5 > 3 > 4. The distinctive fragmentation observed for the little-studied 1,4-dicaffeoylquinic acid involves elimination of the C1 caffeoyl residue, repeated dehydrations leading to the aromatization of the quinic acid moiety, and its decarboxylation. It is suggested that this process is initiated by the C1 carboxyl protonating the C5 hydroxyl in the inverted chair conformer, followed by its protonating the C1 caffeoyl residue in the favored chair conformation. The fragmentation of 1-caffeoylquinic acid is indistinguishable from that of the commercially available 5-caffeoylquinic acid, but these two isomers can be distinguished easily by their facile chromatographic resolution on reversed phase packings. The hierarchical key previously developed for characterizing chlorogenic acids has been extended to accommodate 1-caffeoylquinic acid and the 1-acyl dicaffeoylquinic acids.     

Specificity of codling moth (Lepidoptera: Tortricidae) for the host plant kairomone, ethyl (2E,4Z)-2,4-decadienoate: field bioassays with pome fruit volatiles, analogue, and isomeric compounds

Codling moth, Cydia pomonella (L.), is a severe pest of apples, pears, and walnuts worldwide, and new approaches for precise monitoring and management would be beneficial. Ninety-two pome fruit volatiles were formulated in 23 distinct blends, of which a single 4-component blend of 10-carbon esters showed the only significant attraction of moths in field bioassays conducted in both walnut and apple orchards. A single constituent of this blend, ethyl (2E,4Z)-2,4-decadienoate--the "pear ester", was the major contributing attractant. The pear ester attracted both male and female moths in combined numbers that were comparable to the attractiveness of conspecific sex pheromone. Structure-activity tests were conducted in a series of orchard trials to determine the specificity of attraction of codling moths to the pear ester kairomone. No analogue 10-carbon alcohols, aldehydes, acetates, or other esters elicited significant moth capture responses. Tests with various analogue esters with alcohol chain length moiety substitutions of the (2E,4Z)-2,4-decadienoic acid elicited differential capture responses, with the ethyl exceeding the propyl, methyl, butyl, and hexyl analogues. The (E,Z) geometric isomers of this series of (2E,4Z)-2,4-decadienoic acid esters far exceeded the attractiveness of the (E,E) isomers. The pear ester is a potent attractant of both males and females, and codling moths are highly discriminating and specific in their structure-activity-based attraction to this pear-derived kairomone. These specificity attributes should allow this host plant kairomone to contribute to new abilities for female monitoring and the potential of development of novel and highly selective control practices that should decrease the current dependence on the use of broad-spectrum insecticides.     

Variables controlling denitrification from earthworm casts and soil in permanent pastures

Summary Denitrification (using the acetylene block method) was determined in earthworm casts and soils from permanent, drained or undrained pasture plots fertilized with 0 or 200 kg N ha^-1 year^-1 as ammonium nitrate. Rates of N₂O production from soil cores were about three times higher from the fertilized than from the unfertilized plots while drainage had a relatively small effect. Denitrification rates from casts were 3–5 times higher than those from soil irrespective of the drainage treatment. Casts generally had higher NO _inf3 ^sup- , NH _inf4 ^sup+ , and moisture contents, and higher microbial respiration rates than soil. Rates of N₂O production were determined primarily by NO _inf3 ^sup- supply, secondarily by moisture; available C did not appear to limit denitrification in these pastures. Estimates of the potential contribution of casts to denitrification ranges from 10.1% of 29.3 kg ha^-1 year^-1 from the unfertilized, drained plot to 22% of 82.5 kg ha^-1 year^-1 from the fertilized undrained plot.     

Ectoparasitic dermatitis in free-ranging swamp wallabies (Wallabia bicolor) in New South Wales

We report extensive dermatological lesions in three, free-ranging, adult, male swamp wallabies ( Wallabia bicolor ), presented to Taronga Western Plains Zoo wildlife hospital. All three animals were moderately infested with the louse species Heterodoxus ualabati , and two were concurrently infested with a previously undescribed sarcoptiform mite of the genus Diabolicoptes . Histological changes included moderate to marked compact hyperkeratosis, focal deep pyoderma and superficial dermatophytosis. Mild to moderate hyperplastic perivascular and periadnexal dermatitis with marked superficial and follicular hyperkeratosis was evident in the skin of the inguinal region and lateral thigh of the two wallabies infested with the Diabolicoptes species. This is the first report of ectoparasitic dermatitis in free-ranging swamp wallabies and the first report of mites of the genus Diabolicoptes from a macropodid.