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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 (VL) and heavy chain (VH) 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.  相似文献   
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Summary Sixty dairy buffaloes (second to fourth lactation) from a large buffalo farm were used to compare the effects of single intramuscular injections of 100 µg gondadotrophin releasing hormone (GnRH), 250 µg GnRH or saline given on day 14post partum. The buffaloes had calved at the end of the breeding season (December). Milk samples for progesterone determination were taken at the time of injection and then three times a week either until first insemination or until around day 90post partum. GnRH given at 14 dayspost partum resulted in quicker completion of uterine involution, earlier resumption of ovarian activity, shorter intervals between calving and conception and a better first service conception rate in non-suckled dairy buffaloes. Differences between the results obtained by a GnRH dose level of 100 µg and 250 µg were non-significant. In the post-treatment period cases of prolonged luteal activity were common in all groups of buffaloes. Therefore the sequential administration of GnRH and prostaglandin is suggested for the management of post-partum reproductive activity in problem herds.
Desempeno Reproductivo De Bufalos Nili-Ravi Despues De Una Sola Inyeccion De GnRH Temprano Despues Del Post-Parto
Resumen Sesenta búfalos lecheros (segunda a cuarta lactancia) pertenecientes a un hato grande fueron utilizados para comparar los efectos de inyecciones intramusculares de 100 µg de GnRH, 250 µg y de solución salina, aplicadas 14 días después del parto. Los búfalos habian parido al final de la estación de pariciones (diciembre). Se tomaron muestras de leche para medir progesterona, en el momento de la inyección y después tres veces por semana hasta la primera inseminación, o hasta cerca de los 90 días post-parto. La GnRH administrada 14 días post-parto, dio como resultado una rápida involución uterina, más rápido retorno a la actividad ovárica, más cortos intervalos entre partos y concepción, y un mejor índice servicio-concepción en búfalos lecheros que no estaban amamantando. Los resultados de dosis de 100 µg y 250 µg no fueron significativos. En el período post-tratamiento, los casos de actividad luteal prolongada fueron comunes en todos los grupos de búfalos. Se sugiere entonces, la administración secuencial de GnRH y prostaglandina, parar el manejo de la actividad reproductiva post-parto en hatos problema.

Performances De Reproduction Du Buffle Nili-Ravi Apres Une Injection Unique PrecocePost Partum De Gonadotrophine (GnRH)
Résumé Soixante bufflesses laitières (de la seconde à la quatrième lactation), appartenant à une grande ferme d'élevage de buffles, ont été utilisées pour comparer les effets d'une injection unique intramusculaire de 100 µg de gonadotrophine, ou de 250 µg du même produit ou d'une solution de sérum physiologique. Les bufflesses avaient mis bas à la fin de la saison de reproduction soit en décembre. Des prélèvements de lait ont été effectués pour déterminer le taux de progestérone au moment de l'injection d'une part, puis trois fois par semaine d'autre part jusqu'à la première insémination ou aux alentours d'une période de 90 jours après le part. La gonadotrophine, libérée sous forme d'hormone (GnRH), injectée 14 jours après la mise bas a eu pour effet un retour complet plus rapide de l'involution utérine et des intervalles plus courts entre la mise bas et la conception ainsi qu'un meilleur taux de conception à la première monte chez les femelles laitières non allaitantes. Les différences entre les résultats obtenus par une dose de 100/µg ou de 250/µg de gonadotrophine (GnRH) n'étaient pas significatives. Pendant la période post-thérapeutique, des cas d'activité lutéale prolongée ont été courants dans tous les groupes de bufflesses. En conséquence, l'administration séquentielle de GnRH et de prostaglandine est conseillée pour contrôler l'activité post-partum de reproduction dans les troupeaux qui présentent des problèmes de cet ordre.
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L11A-Fukushima (L11A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L11A-F and L11A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L11A-F appears to possess the properties necessary for practical use. To manage L11A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L11A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002  相似文献   
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DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001  相似文献   
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Aciculosporium take (Ascomycota; Clavicipitaceae) is a causal agent of witches' broom of bamboo plants. The symptoms of this disease are believed to be induced by plant hormones, particularly auxins. Indole-3-acetic acid (IAA) was identified in cultures of this fungus in an l-tryptophan-supplemented liquid medium. IAA production was confirmed on 30 isolates of A. take from various hosts and locations at levels up to 1 mg/l. The biosynthetic pathway of IAA in A. take culture was examined by analyzing intermediate products and by feeding experiments. The results showed that the indole-3-pyruvic acid pathway (l-tryptophan → indole-3-pyruvic acid → indole acetaldehyde → IAA) was the dominant pathway in A. take. Received: June 3, 2002 / Accepted: July 25, 2002  相似文献   
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 Rice seedling growth, estimated by plant height and root development and discoloration, was better in pasteurized soil than in unpasteurized soil obtained from a flooded rice field. Rice seedlings also grew better in sterilized soil modified by adding roots harvested from the pasteurized soil than in soil modified by adding roots harvested from the unpasteurized soil. The results demonstrate that seedling growth in the rice field soil was inhibited by soil microorganisms, even though no typical symptoms such as seedling blight or damping-off appeared. Pythium aristosporum is suggested to be involved in the inhibition. Thus, it appears that inconspicuous restraint of rice seedling growth could occur in soils of rice paddy fields. Received: May 20, 2002 / Accepted: October 16, 2002 Acknowledgments The authors thank Dr. T. Ichitani, former professor at Osaka Prefectural University, for providing an isolate of Pythium aristosporum for comparison, and Mr. Mitsuaki Sato of Akita Prefectural College of Agriculture for technical assistance.  相似文献   
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Large mammalian herbivores are notorious for their propensity towards population irruptions and crashes, yet many herbivore populations remain relatively stable. I explore how resource heterogeneity within landscapes dampens population instability, using a metaphysiological modelling approach considering patch state distributions. Resource heterogeneity is functionally stabilizing through spreading consumption away from preferred resources before these become critically depleted. Lower-quality resources act as a buffer against starvation during critical periods of the seasonal cycle. Enriching resource quality is destabilizing, even if patch diversity is maintained, because food quantity then becomes the limitation. The potential consequences of landscape fragmentation are explored using the Serengeti ecosystem, characterised by broadscale resource gradients, as a hypothetical example. Further insights provided by the model are illustrated with specific examples concerning the effects of patch scales and waterpoint distribution. A metaphysiological modelling approach enables the basic consequences of landscape heterogeneity to be distinguished from further effects that may arise from specific patch scales and configurations, without the distracting detail of spatially explicit models.  相似文献   
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