Not the one,but the only one: about <Emphasis Type="Italic">Cannabis cryptic virus</Emphasis> in plants showing ‘hemp streak’ disease symptoms

Interveinal chlorosis and leaf margin wrinkling are widespread symptoms of Cannabis sativa. They are traditionally attributed to the so-called hemp streak virus (HSV), but its existence has not been demonstrated yet. To our knowledge, no molecular investigation has so far been performed in order to identify the causal agent of this symptomatology, we therefore decided to use traditional and molecular virology techniques to better characterize symptoms and pursue the etiological agent. No pathogenic virus was found by using targeted PCR reactions and by RNA sequencing, whereas we were able to detect the Cannabis cryptic virus (CanCV) with both techniques. We, therefore, developed an RT-qPCR assay based on a CanCV-specific TaqMan probe and applied it to a wide range of symptomatic and symptomless plants, using a two-step (for quantification), or a one-step (for fast detection) protocol. Both symptoms and the virus were only shown to be transmitted vertically and did not pass via mechanical inoculation or grafting, though we could not find any cause-effect correlation between them. In fact, the virus was found in all the tested hemp samples, and its abundance varied greatly between different accessions and individuals, independently from the presence and severity of symptoms. The suggestion that hemp streak is caused by a virus is therefore questioned. Some abiotic stresses seem to play a role in triggering the symptoms but this aspect needs further investigation. For breeding purposes, a selection of parental plants based on the absence of symptoms proved to be efficient in containment of the disease.     

Soil organic matter dynamics and losses in manured maize-based forage systems

Soil organic matter (SOM) produces positive effects on multiple soil properties. Increasing its level also provides an opportunity to reduce atmospheric concentration of carbon dioxide (CO₂). Recycling animal manure and returning crop residue are among the main practices to enhance organic carbon (C) stock in arable croplands. This study analysed data of soil organic carbon (SOC) stocks from a medium-term field trial (established in 1992) through a Hénin–Dupuis-based equation to determine the proportion of different organic materials retained in the soil as SOM.The treatments included in the experiment simulated different typical management techniques and implied the application to the soil of various organic materials, i.e. cattle slurry, cattle farmyard manure, maize straw, root, maize and ley stubble. Soil organic carbon and total nitrogen (N) (Kjeldhal N) were measured in the 0–30 cm layer during spring 1999 and spring 2003. The relationship between SOC and N stocks measured in 1999 and 2003 and the annual additions of C and N was described by the Hénin–Dupuis-based equation, assuming the presence of two C and N pools (stable and fresh).Our results showed that the application of farmyard manure, slurry and maize straw induced higher C and N content compared to the application of urea without return of crop residue. The different levels of urea application did not produce any significant difference in C and N soil content.We found that approximately 2% of SOC is lost to the atmosphere annually. Furthermore, the amount of C and N retained in the soil each year varied by organic materials: 46% C and 44% N with farmyard manure, 26% C and 11% N with slurry, 28% C and 10% N with root and 6% C with maize straw and ley stubble.These results were used to inform an estimate of C lost or removed to/from atmosphere following manure application and crop residue return for the compilation of greenhouse gas inventories. We found that the most C conservative management is the production and spreading of farmyard manure and that increasing amounts of mineral N fertilizer did not affect the C sequestration extent.     

What is the evidence? The issue of verifying correct needle position during epidural anaesthesia in dogs

Objectives

To review the methods for verifying the needle position while performing epidural anaesthesia in dogs, and to discuss the advantages, disadvantages, usefulness and reliability of each technique in the experimental and clinical research setting.

IgE-Mediated Cross-Reactivity among Leguminous Seed Proteins in Peanut Allergic Children

The immunological cross-reactivity among major protein- and oil-crops, including lupin, lentil, pea, peanut, kidney bean and soybean, has been studied by a combination of in vitro and in vivo experimental approaches: SDS-PAGE separations of legume protein extracts and immuno-blot revelations with 12 peanut-sensitive subjects’ sera, Immuno-CAP and Skin Prick tests on the same subjects. The immuno-blotting data showed a wide range of IgE-binding responses both displayed by one subject towards different plant extracts and among subjects. Differences were both quantitative and qualitative. The prevalent responses of most subjects’ sera were seen with peanut polypeptides, as expected, as well as with various polypeptides of the other legumes, the most recurrent of which were the basic subunits of the 11S globulins. The distribution of in vivo responses generally paralleled those obtained by in vitro approaches with strong responses elicited by peanut, lentil and pea protein extracts, especially by most sensitive subjects, thus providing a consistent overall set of results. In this work, the comparison of various approaches has allowed us to get an overall broad picture of the immunological cross-reactivities among proteins of widely used different seed species and to hypothesize the role of most conserved specific polypeptides.     

Insecticidal activity of scorpion toxin (ButaIT) and snowdrop lectin (GNA) containing fusion proteins towards pest species of different orders

BACKGROUND: The toxicity of a fusion protein, ButalT/GNA, comprising a venom toxin (ButaIT) derived from the red scorpion, Mesobuthus tamulus (F.), and Galanthus nivalis agglutinin (GNA), was evaluated under laboratory conditions against several pest insects. Insecticidal activity was compared with SFI1/GNA, a fusion comprising a venom toxin (SFI1) derived from the European spider Segestria florentina (Rossi) and GNA, which has been previously demonstrated to be effective against lepidopteran and hemipteran pests, and to GNA itself. RESULTS: Injection assays demonstrated that both fusion proteins were toxic to lepidopteran larvae, dipteran adults, coleopteran adults and larvae and dictyopteran nymphs. ButalT/GNA was more toxic than SFI1/GNA in all cases. GNA itself made a minor contribution to toxicity. Oral toxicity of ButalT/GNA towards lepidopteran pests was confirmed against neonate Spodoptera littoralis (Boisd.), where incorporation at 2% dietary protein resulted in 50% mortality and > 85% reduction in growth compared with controls. ButaIT/GNA was orally toxic to Musca domestica L. adults, causing 75% mortality at 1 mg mL^?1 in aqueous diets and, at 2 mg g^?1 it was orally toxic to Tribolium castaneum (Herbst.), causing 60% mortality and a 90% reduction in growth. CONCLUSIONS: Toxicity of the ButaIT/GNA recombinant fusion protein towards a range of insect pests from different orders was demonstrated by injection bioassays. Feeding bioassays demonstrated the potential use of the ButaIT/GNA fusion protein as an orally active insecticide against lepidopteran, dipteran and coleopteran pests. These experiments provide further evidence that the development of fusion protein technology for the generation of new, biorational, anti‐insect molecules holds significant promise. © Crown Copyright 2009. Reproduced with permission of Her Majesty's Stationery Office. Published by John Wiley & Sons, Ltd.     

Inventory of related wild species of priority crops in Venezuela

A prerequisite in any conservation programme of Plant Genetic Resources is estimation of diversity. The inventory of wild and naturalized relatives of priority crops in Venezuela (CWR) is based on the main Catalogues of Flora in the country, selecting taxa closely related to crops, according to the concepts of “gene pool” and “taxonomic group”. We included 47 genera, 217 species and 228 taxa belonging to 28 plant families. Among them, those with higher richness are: Fabaceae, Solanaceae, Araceae, Lauraceae, Dioscoreaceae, Poaceae, Rosaceae and Myrtaceae. Genera with a higher number of species are Xanthosoma, Persea, Dioscorea, Prunus, Psidium, Phaseolus, Solanum, Vigna, Capsicum, Manihot, Theobroma, Ipomoea and Oryza. A total of 26 endemic species are found, which belong to genera Xanthosoma, Persea, Dioscorea, Prunus and Manihot. The primary gene pool of crops include native species from genera such as Manihot, Solanum (Section Petota), Lycopersicon, Ananas, Capsicum, Dioscorea, Xanthosoma, Phaseolus, Theobroma, Ipomoea, Gossypium, Arracacia and Psidium. Genera with native species weakly related to crops are Saccharum, Persea, Ipomoea, Prunus, Vigna, Solanum (Section Melongena) and Daucus. Crop genera without native species in Venezuela are Allium, Musa, Brassica, Spinacia, Helianthus, Pisum, Lactuca, Citrus, Elaeis, Beta, Glycine and Triticum. Only a few taxa have already been evaluated according to the IUCN criteria, and Venezuelan accessions of crop wild relatives in national and international genebanks are very scarce.     

Biochemical and immunochemical characterization of different varieties of amaranth (Amaranthus L. ssp.) as a safe ingredient for gluten-free products

Celiac disease is a food intolerance triggered by the ingestion of gluten-containing cereals; the only therapy is a strict gluten-free diet for life. In recent years, amaranth flour has received considerable attention as an interesting source for the formulation of gluten-free products due to its high nutritional value and low content of prolamins, the toxic proteins for celiacs. The aim of this study was to characterize 40 amaranth varieties using both SDS-PAGE/immunoblotting and ELISA to assess their possible tolerance by celiac subjects. All of the amaranth samples studied showed similar binding affinities for both specific anti-gliadin antibodies and human IgAs. In most amaranth grains, the content of gluten-like proteins measured by ELISA was <20 ppm. The molecular characterization of amaranth proteins suggests that amaranth is safe for celiacs to consume. It is recommended that the most suitable amaranth varieties are those having the lowest content of proteins cross-reacting with anti-gliadin antibodies.