Tailor-made biotechnologies: Possibilities for farmer-centered development

Each technology—and also biotechnology—relates to and is developed within a certain context. Besides a technical dimension (the techniques) biotechnology also contains asocial dimension, such as the social relations that reflect themselves in the technology-development, and the aims for which the technology is used.The Center for Agriculture and Biotechnology (CAB), the research department of the Western Farming and Horticultural Organization, is attempting to relate the socio-economic contents of biotechnological developments more to the interests of farmers and sustainable farming styles.     

Novel application of square-wave adsorptive-stripping voltammetry for the determination of xanthohumol in spent hops

This paper reports the development of a novel electrochemical assay for xanthohumol (XN) by square-wave adsorptive-stripping voltammetry (SWAdSV) with a hanging mercury drop electrode. The method showed good repeatability (CV < 2%) and linearity (between 10 and 250 μg L(-1)), as well as suitable limits of detection (2.6 μg L(-1)) and quantification (8.8 μg L(-1)). The method was applied for the quantification of this compound in spent hops, and the results obtained were compared with the HPLC-UV method. XN contents determined by the SWAdSV method were 16 ± 1 and 100 ± 4 μg L(-1) for aqueous and methanolic extracts, respectively. The developed new methodology considerably reduces the analysis time, approximately from 25 min (HPLC-UV method) to 7 min, enabling a high sample throughput. In addition, the detection and quantification limits were approximately 5-fold lower than those obtained with the chromatographic method.     

Silencing of host basal defense response-related gene expression increases susceptibility of Nicotiana benthamiana to Clavibacter michiganensis subsp. michiganensis

Clavibacter michiganensis subsp. michiganensis is an actinomycete, causing bacterial wilt and canker disease of tomato (Solanum lycopersicum). We used virus-induced gene silencing (VIGS) to identify genes playing a role in host basal defense response to C. michiganensis subsp. michiganensis infection using Nicotiana benthamiana as a model plant. A preliminary VIGS screen comprising 160 genes from tomato known to be involved in defense-related signaling identified a set of 14 genes whose suppression led to altered host-pathogen interactions. Expression of each of these genes and three additional targets was then suppressed in larger-scale VIGS experiments and the effect of silencing on development of wilt disease symptoms and bacterial growth during an N. benthamiana-C. michiganensis subsp. michiganensis compatible interaction was determined. Disease susceptibility and in planta bacterial population size were enhanced by silencing genes encoding N. benthamiana homologs of ubiquitin activating enzyme, snakin-2, extensin-like protein, divinyl ether synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase 2, and Pto-like kinase. The identification of genes having a role in the host basal defense-response to C. michiganensis subsp. michiganensis advances our understanding of the plant responses activated by C. michiganensis subsp. michiganensis and raises possibilities for devising novel and effective molecular strategies to control bacterial canker and wilt in tomato.     

Derivation and testing of a model to predict selection for fungicide resistance

A mathematical model was derived to predict selection for fungicide resistance in foliar pathogens of cereal crops. The model was tested against independent data from four field experiments quantifying selection for the G143A mutation conferring resistance to a quinone outside inhibitor (QoI) fungicide in powdery mildew (Blumeria graminis f.sp. hordei) on spring barley (Hordeum vulgare). Fungicide treatments with azoxystrobin differed in the total applied dose and spray number. For each treatment, we calculated the observed selection ratio as the ratio of the frequency of the resistant strain after the last and before the first spray. The model accurately predicted the variation in observed selection ratios with total applied fungicide dose and number of sprays for three of the four experiments. Underprediction of selection ratios in one experiment was attributed to the particularly late epidemic onset in that experiment. When the equation representing epidemic development was modified to account for the late epidemic, predicted and observed selection ratios at that site were in close agreement. On a scatter plot of observed selection ratios on predicted selection ratios, for all four experiments, the 1:1 line explained 89–92% of the variance in the mean of observed selection ratios. To our knowledge, this is the first fungicide resistance model for plant pathogens to be rigorously tested against field data. The model can be used with some degree of confidence, to identify anti‐resistance treatment strategies which are likely to be effective and would justify the resources required for experimental testing.     

The umbric epipedon in the N Apennines,Italy—an example from the Vallombrosa Forest

The umbric epipedon is a diagnostic surface horizon recognized by both the World Reference Base for Soil Resources and the U.S. Soil Taxonomy. It is mainly characterized by a dark color, a moderate to high content of OM, and a base saturation of less than 50%. In the N Apennines, Central Italy, forest soils over 600–700 m a.s.l. often have this epipedon. This paper stresses the morphological, chemical, and biological properties of the epipedon, specifically in the Vallombrosa Forest, where considerable work has been done in the last decade. Here, the umbric epipedon forms on sandstone, in high forests of Abies alba, Fagus sylvatica, and Castanea sativa. It does not have homogeneous properties throughout its thickness and often is divided into two distinct genetic horizons, A1 and A2. The A1 horizon is thinner, darker, and richer in OM, more base‐saturated, and biologically more active than the underlying A2 horizon. The mean residence time of the bulk OM amounts to about a century in the A1 horizon, versus half a millennium in the A2. In both A1 and A2 horizons, the non‐humic fraction prevails in the OM; this could account for the high susceptibility of the umbric epipedon to degrade when the forest is clear‐cut or undergoes extensive uprooting due to windstorms or heavy snow loads. Significant discrepancies between the two A horizons have been found in regard to the microbial community. Umbric epipedons which developed under different tree species show minor differences, mainly concerning the microbial community.     

Volatiles from leaves,fruits, and virgin oil from Olea europaea Cv. Olivastra Seggianese from Italy

The volatiles produced by leaves and fruits of Olea europaea cv. Olivastra Seggianese have been analyzed in two different phenological stages. Furthermore, the volatiles of the virgin olive oil obtained from ripe fruits has been characterized. The volatiles were sampled by means of two different techniques: hydrodistillation and SPME. Differences were observed between the two different collection times, the different organs, and sampling techniques. The major constituents were often aldehydes, particularly (E)-2-hexenal (9.8-48.0%); however, also many terpenoids have been identified, mainly (E,E)-alpha-farnesene (0.2-27.0%), linalool (0-3.6%), beta-caryophyllene (0-8.1%), and valencene (0-2.5%). This is the first investigation on this cultivar.     

Detection of polycyclic aromatic hydrocarbon levels in milk collected near potential contamination sources

Polycyclic aromatic hydrocarbons (PAHs), mainly formed by incomplete anthropogenic organic matter combustion, are ubiquitous in the environment. To assess milk PAH contamination sources, milk samples were collected from the tank milk at farms located near potential contaminating emission sources such as cementworks, steelworks, and motorways. PAH analyses were carried out by gas chromatography coupled to mass spectrometry. Eight PAHs were identified in milk: naphthalene, acenaphthylene, acenaphthene, fluorene, anthracene, fluoranthene, pyrene, and benzo[a]anthracene. For all potential contaminating sources, these eight PAHs were detected with similar profiles and at low concentrations except for fluorene and naphthalene, for which source-molecule interaction is pointed out.     

Superficial keratectomy and cryosurgery as therapy for limbal neoplasms in 13 horses

Objective To determine the usefulness and clinical outcome of a combined procedure of superficial keratectomy and cryosurgery as a treatment for limbal neoplasms in horses. Study design Retrospective study. Animals Thirteen horses with 14 limbal tumors. Methods Medical records of all patients with limbal tumors, referred to the Department of Equine Sciences of Utrecht University between 1995 and 2002, were retrieved. Patient data were analyzed with respect to signalment, tumor surface area and histologic diagnosis. Surgery, performed under general anesthesia, included surgical debulking of the tumor followed by cryosurgery. Long‐term follow‐up information was obtained by re‐evaluation in the clinic, by the referring veterinarian, and/or by telephone enquiry. Results Therapy was successful in 9 out of 14 eyes after the first attempt (64%); one eye needed retreatment (7%), and four eyes were eventually enucleated (29%). The mean follow‐up period was 4.8 years. The initial surface area of the tumor significantly influenced outcome (P < 0.01). Squamous cell carcinoma was the most predominant tumor type (79%). Haflinger horses accounted for 69% of the cases whereas their occurrence in the overall hospital population is approximately 5%. Conclusions and clinical relevance The described technique of superficial keratectomy and cryosurgery is a simple procedure for the treatment of limbal tumors in equine patients that does not require sophisticated equipment. Nor is it tampered by legal restriction, and appears to be effective in tumors with a small surface area (< 2 cm²). Haflinger horses seem to be predisposed for the development of ocular squamous cell carcinoma.