‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

The biochemical basis of resistance of potato to soft rot bacteria

Evidence is presented that a number of mechanisms are involved in the resistance of commercial cultivars of potato to soft rottingErwinia spp. These include phytoalexins, phenolics, cell wall modification, and possibly proteins. The role of elicitor-active oligogalacturonides, which are released from plant cell walls by pectic enzymes, is discussed.     

New predators for biocontrol under glass

Several species ofOrius are under investigation in the UK, Europe and North America for the biological control of thrips, especially western flower thrips(Frankliniella occidentalis). The available species are briefly compared and findings on effectiveness and oviposition ofO. laevigatus are reported.     

Interactions among herbivores,microbial insecticides and crop plants

Knowledge of the interactions among lepidopterous insects, microbial insecticides and plants is either lacking or incomplete. In some of the interactions, plant allelochemicals antagonized the insecticidal activity of the microbe by means of feeding reduction. In others, microbe activity was enhanced either by causing insect toxicity or by a potentiation of the crystal protein. Nutritional evaluations of the interactions amongHeliothis virescens. Bacillus thuringiensis crystal protein and tannin showed that microbe toxicity in the larva was antagonized by this phytochemical.     

Nutrition and Health of Dogs and Cats: Evolution of Petfood

Maintaining the health of dogs and cats by feeding wholesome nutritional diets is becoming an important component of responsible pet ownership. Pet owners now seek a long and healthy life for their pet and look to nutrition, as well as to veterinary medicine, to provide such support. Quality of life, measured in terms of reduced incidence of diseases and the ability to maintain an active life, would appear to be able to be enhanced by appropriate nutrition and nutraceutical supplementation. As a consequence numerous improvements in companion animal nutrition have resulted in development of a wide array of foods that provide complete and balanced nutrition. As a result emphasis also has to be placed on product safety and quality parameters, in connection with traceability. The origin of products, including product characteristics and properties, processing conditions and further handling throughout the period chain, is becoming ever increasingly an issue for collective chain management.     

DNA Immunization Against Very Virulent Infectious Bursal Disease Virus with VP2‐4‐3 Gene and Chicken IL‐6 Gene

The present study was to investigate the feasibility and efficiency of the DNA vaccine to protect chickens against very virulent infectious bursal disease virus (vvIBDV) infection. A plasmid DNA carrying VP2‐4‐3 genes of vvIBDV SH95 and a plasmid DNA carrying chicken interleukin‐6 (ChIL‐6) genes were constructed and designated as pALTER‐MAX‐VP2‐4‐3 and pALTER‐MAX‐ChIL‐6 respectively. Several DNA vaccination experiments were performed: 1‐week‐old chickens were intramuscularly injected with only plasmid pcDNA3‐VP2, pALTER‐MAX‐VP2‐4‐3 or mixture with pALTER‐MAX‐ChIL‐6. The chickens at 4 weeks old were orally inoculated with vvIBDV SH95. The results showed that immunization with the mixture of pALTER‐MAX‐VP2‐4‐3 and pALTER‐MAX‐ChIL‐6 three times conferred protection for 90% of chickens. Enzyme‐linked immunosorbent assay (ELISA) antibody titres in chickens immunized together with pALTER‐MAX‐ChIL‐6 were higher than those immunized simply with plasmid pcDNA3‐VP2 or pALTER‐MAX‐VP2‐4‐3. IBDV was not detected in the bursa of the protected chickens at 8 days after challenge by RT‐PCR. The results indicate that protection against vvIBDV can be achieved by using the VP2‐4‐3 gene of vvIBDV as a DNA vaccine. Furthermore, the simultaneous injection of ChIL‐6 plasmid significantly increased the protection after challenge with the very virulent strain.     

Effects of temperature on ascospore germination and penetration of oilseed rape (Brassica napus) leaves by A- or B-group Leptosphaeria maculans (phoma stem canker)

Ascospores of both A-group and B-group Leptosphaeria maculans germinated at temperatures from 5 to 20°C on leaves of oilseed rape. Germination of ascospores of both groups started 2 h after inoculation and percentage germination reached its maximum about 14 h after inoculation at all temperatures. Both the percentage of A-/B-group ascospores that had germinated after 24 h incubation and germ tube length increased with increasing temperature from 5 to 20°C. Germ tubes from B-group ascospores were longer than those from A-group ascospores at all temperatures, with the greatest difference at 20°C. Hyphae from ascospores of both groups penetrated the leaves predominantly through stomata, at temperatures from 5 to 20°C. A-group ascospores produced highly branched hyphae that grew tortuously, whereas B-group ascospores produced long, straight hyphae. The percentage of germinated ascospores that penetrated stomata increased with increasing temperature from 5 to 20°C and was greater for A-group than for B-group L. maculans after 40 h incubation.