Development of biomarkers inAporrectodea caliginosa andLumbricus rubellus for detecting exposure to benzo[a]pyrene

Objectives and Methods. The sensitivity of two biomarkers, the neutral red retention assay (NRRA) and cytochrome P450 were evaluated in the earthwormAporrectodea caliginosa for their potential to detect exposure to benzo[a]pyrene (BaP). Cytochrome P450 was also evaluated in the earthwormLumbricus rubellus, and measured using the substrate ethoxycoumarin. Optimal assay conditions (pH, and temperature) were determined, followed by exposure of earthworms to 20 mg/kg BaP (a typical concentration at contaminated sites in New Zealand). Ethoxycoumarin-O-dealkylase (ECOD) activity was measured after 7, 14, and 21 days exposure. The NRRA was evaluated in earthworms exposed to 0.2, 20, and 100 mg BaP/kg, and biomarker responses were compared with effects on body weight. Results and Discussion Benzo[a]pyrene failed to induce ECOD activity in either earthworm species, and therefore it is not useful as a biomarker of BaP exposure and was not evaluated further. In all cases, the NRRA was significantly affected in the absence of any effects on earthworm body weight, indicating that this assay can detect exposure to BaP at a range of concentrations comparable to those found at contaminated sites. The NRRA should be linked to reproductive endpoints, then it can be used as an early warning indicator of adverse effects. Establishing biomarker stability under environmental conditions is an important step in biomarker development. Therefore, the effects of soil type (sandy soil, silt loam, and a clay soil), moisture content (15–30%), and temperature (5–20°) on the NRRA were determined. In all cases, there was no effect on the NRRA, indicating that this assay is very stable under varying environmental conditions. Conclusion and Outlook In conclusion, cytochrome P450 activity does not appear to be a useful indicator of PAH exposure in eitherA. caliginosa or L.rubellus, and due to the inherently low activity, it is not suitable as a routine biomarker for detecting environmental contamination by these compounds. In contrast, the NRRA in the earthworm A.caliginosa is a promising indicator of PAH exposure at the concentrations likely to be found in contaminated sites in New Zealand, and therefore has potential for evaluating these contaminated sites. If the NRRA can be linked to ecologically important life-cycle endpoints, such as reproduction, then it could be used as an early warning indicator of adverse effects at contaminated sites, i.e., by measuring biomarker responses in earthworms from a ‘contaminated area’ and comparing these with earthworms from a matched control area.     

Turf-banked terrace treads and risers, turf exfoliation and possible relationships with advancing treeline

Fine-scale geomorphic/pedogenic processes at alpine treeline may facilitate the initial stages of conifer invasion of alpine tundra. Turf-banked terraces and turf exfoliation associated with solifluction may provide both the topographic protection and the seedbed necessary for conifer establishment above treeline. The morphometry, stoniness and surface penetrability of turf-banked terraces were recorded. Whereas differences among sites exist due to topographic constraints, differences in soil penetrability depend on turf exfoliation. Exfoliated turf risers are significantly more penetrable than non-exfoliated solifluction risers and adjacent treads. These penetrable microsites provide favorable conditions for seed germination. The process of turf exfoliation may play a role in conifer advance into tundra during climatic change.     

Biochemical diversity in the genus Coffea L.: chlorogenic acids,caffeine and mozambioside contents

Summary The extent and nature of biochemical diversity in green coffee beans was established by a large sampling of genus Coffea, representing nine species from the Guineo-Congolian region, nine species from east Africa and seven species from Madagascar. HPLC analyses were used to determine the contents of caffeine, chlorogenic acids and mozambioside. Data processed by principal component analyses showed the existence of two metabolic pathways. One leads to the synthesis of small quantities of chlorogenic acids (< 2.5% dmb) and caffeine ( < 0.3 % dmb) while the other leads to large concentrations of chlorogenic acids (> 4.5% dmb) and caffeine (> 0.4% dmb). Their distribution in the genus Coffea is discussed in relation to the biogeographic origin of the plant material.     

Genetic diversity among watermelon (Citrullus lanatus and Citrullus colocynthis) accessions

Genetic diversity was estimated among 42 U.S. PlantIntroduction (PI) accessions of the genusCitrullus (of these, 34 PIs are reported tohave disease resistance), and 5 watermelon cultivars, using 30RAPD primers. These primers produced 662 RAPD markers that could berated with high confidence. Based on these markers, geneticsimilarity coefficients were calculated and a dendrogram wasconstructed using the unweighted pair-group method witharithmetic average (UPGMA). The analysis delineated threemajor clusters. The first cluster consisted of a group of fivewatermelon cultivars, a group of C.lanatus var. lanatusaccessions, and a group of C.lanatus var. lanatusaccessions that contained some C.lanatus var. citroidesgenes. The second cluster consisted of the C.lanatus var. citroidesaccessions, while the third cluster consisted of theC. colocynthis accessions.The two C. lanatus clustersdifferentiated from each other and from the C.colocynthis cluster at the level of 58.8%and 38.9% genetic similarity, respectively. Assessment ofgenetic diversity among accessions that have been reported to havedisease resistance indicated that resistance to either anthracnose,downy mildew, powdery mildew, or watermelon mosaic virus is foundamong all major groups of Citrullus PIs.Additionally, resistance to gummy stem blight or Fusarium wilt mayexist among C. lanatus var.citroides PIs. This study demonstrates thatmolecular markers can be useful in assessing genetic diversity, andin sorting Citrullus PIs into phylogeneticgroups prior to their evaluation for disease or pestresistance.     

Involvement of dehydroalanine and dehydrobutyrine in the addition of glutathione to nisin

Nisin variants and fragments were reacted with glutathione, and the products of the reactions were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography/mass spectrometry (LC-MS). Reactions between glutathione and either [Ala5]nisin or [Ala33]nisin resulted in products with two glutathione molecules conjugated to one nisin variant molecule. Only one glutathione molecule was added to [Ala5,Ala33]nisin. Fragmentation of the nisin molecule resulted in nisin 1-12, nisin 1-20, and nisin 1-32 fragments. Each fragment retained two dehydro residues, which subsequently underwent reaction with glutathione. The data indicated that the dehydroalanine residues of nisin are sites of addition for glutathione. Such addition renders the nisin molecule inactive.     

Preparation of tetrahydroagathic acid: a serum metabolite of isocupressic acid, a cattle abortifacient in ponderosa pine

Isocupressic acid (1) was used to synthetically prepare a mixture of (8S,13R,S)-labda-15,19-dioic acid (tetrahydroagathic acid) (5) via a two-step oxidation procedure followed by hydrogenation of the double bonds at C13 and C8. Reduction of the C8,17 double bond was stereospecific producing only the 8S isomer and confirmed by the nOe interaction between the resulting C17 and C20 methyl groups. The 13R and 13S isomers of 5 were separated and analyzed by HPLC/MS, and (13S)-tetrahydroagathic acid was isolated and identified by comparison to a standard prepared by hydrogenation of naturally occurring (13S)-dihydroagathic acid (4). (13R,S)-dihydroagathic acid was prepared by selective sodium metal-catalyzed hydrogenation of the C13,14 allylic double bond of agathic acid (3). The prepared compounds were then used as standards to confirm the presence of 4 and 5 and their respective 13R and 13S isomers in bovine serum samples. Tetrahydroagathic acid was shown to be the only metabolite detected in serum samples taken from a suspected cattle abortion case submitted for diagnosis; and, thus, 5 could be a valuable diagnostic marker for pine needle-induced abortions.     

The Relationship Between Yield and Fructan Exo-Hydrolases Activity in Two Drought Resistant Wheat Cultivars Grown Under Different Fertilizer and Tillage Treatments

Using two drought resistant wheat (Triticum aestivum L.) cultivars, ‘Changwu134’ and ‘Changhan58,’ a field experiment was conducted in ChangWu Agro-ecological Experiment Station on China's Loess Plateau during 2008 and 2009 to compare the effects of different fertilizer and tillage treatments on the fructan contents as well as fructan exo-hydrolase (FEH) activity in relation to wheat yield. We found that ‘Changhan58’ had greater yield and fructan content in the penultimate internode and higher FEH activity than did ‘Changwu134.’ For ‘Changhan58.’ applying 195 kg·ha^?1(120 +75) N plus 45000 kg·ha^?1 of pig manure and 120 kg·ha^?1 phosphate under conservation tillage produced the highest yield (6769 ka/ha), fructan content in penultimate internode, water use efficiency (WUE), as well as FEH activity among the fertilizer and tillage treatments. Therefore, routine soil management for wheat should focus on combined use of manures and inorganic fertilizer to enhance the amount and transportation efficiency of WSC and ultimately ensure greater yield.