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11.
A. Schots J. De Boer A. Schouten J. Roosien J. F. Zil Verentant H. Pomp L. Bouwman-Smits H. Overmars F. J. Gommers B. Visser W. J. Stiekema J. Bakker 《European journal of plant pathology / European Foundation for Plant Pathology》1992,98(2):183-191
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
12.
Bacillary rickettsia-like organisms (BRLO) were found in the tick Dermacentor marginatus. They are gram-negative and differ from common bacteria and reckettsiae both in the cultivation conditions and morphology. BRLO are non-pathogenic for ticks and guinea pigs. In our studies they were isolated on half-engorged females of D. marginatus, on which they are still maintained. 相似文献
13.
J Marcaník J Kocisová K Bod'a T Pauer M Belák R Skarda 《Archives of Animal Nutrition》1976,26(11):757-763
Histological and histochemical studies were carried out on the gastro-intestinal mucosa of three experimental cows (roughages/maize silage) and three control animals. The animals were slaughtered on termination of the long-term trial. Mucosa samples were taken, for further study, from the rumen, the duodenum, the jejunum, the large intestine and the appendix. Histochemical analysis did not reveal any essential differences in the activities of non-specific esterase, alkaline and acid phosphatase and lactic acid dehydrogenase in the mucosa of the rumen, the large and the small intestine and the appendix of both the experimental animals and the controls. The experimental animals were found to exhibit a higher rate of glutamate-dehydrogenase activity in the ruminal mucosa and in the mucosa of the large and small intestine. A higher succinate dehydrogenase activity was observed in the ruminal mucosa of the experimental animals, relative to that of the controls, while the activity in the intestinal mucosa was decreased. Only slight changes were noted in the activity of the enzymatic systems tested. Electron microscopic studies did not reveal any differences in the ultrastructure of the epithelial cells of the ruminal mucosa in both the experimental animal and the controls. 相似文献
14.
The protective effect of a lyophilized vaccine against Marek's disease ("Keramvac"--Pfizer), prepared from a cell-free turkey's herpesvirus (strain FC 126), was compared in field trials with losses in the group of non-vaccinated chickens. Under the indicated conditions, the vaccine had only a 50.86% effectiveness. The possible causes of the reduced vaccination effect are discussed with regard to the pathomorphological and virological findings suggesting, among others, an increased incidence of the symptoms of the classical form of Marek's disease in the population investigated. 相似文献
15.
Bhupender Singh Fernando Saravia Renée B?ge Heriberto Rodríguez-Martínez 《Acta veterinaria Scandinavica》2005,46(1):1-12
Hormonal asynchronies during oestrus, related to the presence of suprabasal plasma-progesterone (P4) concentrations and a delayed ovulation, interfere with the fertility of repeat-breeder heifers (RBH). Since tubal dysfunction can occur in connection with hormonal asynchronies and constrained availability of fertile spermatozoa at the time of ovulation, the present study tested the hypothesis that frequent sperm deposition from onset of oestrus to ovulation may improve pregnancy rates in RBH. Five RBH and five virgin heifers (VH; controls) were repeatedly artificially inseminated (AI) at 6 h intervals from onset of oestrus to spontaneous ovulation. Hormone analyses revealed suprabasal P4 concentrations and a delay in the occurrence of the luteinising hormone (LH) surge, but a normal cortisol profile in RBH. Compared with controls, RBH presented longer interval from onset of oestrus to ovulation, and therefore, received more AIs. Pregnancy rates in RBH reached control levels (60%; NS), indicating that the hypothesis might be correct. Pregnancy rates in VH were below the expected range, presumably attributed to a deleterious influence of the frequent handling. The study suggests that pregnancy rates can be improved in RBH by frequent AI in relation to spontaneous ovulation. However, this practice of repeated manipulations, while seeming not to show adverse effects, lacks practicality for routine use. 相似文献
16.
María Dolores Parra Fernando Tecles Silvia Martínez-Subiela José Joaquín Cerón 《Journal of veterinary diagnostic investigation》2005,17(2):139-144
An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 microg/ml and 0.0091 microg/ml, respectively, and intra- and interassay coefficients of variation ranged between 6.7-9.9% and 8.5-16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental (R2 = 0.76) and clinical studies (R2 = 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments. 相似文献
17.
Shohei MATSUURA Shigeru HOSHINO Hideaki HAYASHI Tetsuyuki KOHGUCHI Kyoji HAGIWARA Toshihiro OMURA 《Journal of General Plant Pathology》2002,68(1):99-102
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
18.
Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR 总被引:2,自引:0,他引:2
Jesús Mercado-Blanco Dolores Rodríguez-Jurado Encarnación Pérez-Artés Rafael M. Jiménez-Díaz 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(1):1-13
Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants. 相似文献
19.
20.
Marlene Cordero Pedro L. Ramos Lester Hernández Ana I. Fernández Ana L. Echemendía Rudy Peral Gloria González Daniel García Surey Valdés Ana Estévez Keren Hernández 《Phytoparasitica》2003,31(5):478-489
The presence of a begomovirus in potato plants with yellow mottle symptoms was determined for the first time in Cuba. The
incidence of typical begomovirus-like symptoms in potato plants in some regions of Havana province (Güira de Melena, San José
de las Lajas, Güines and Boyeros) during the growing seasons from 1992 to 1998 was in general low. However, in some cultivars
belonging to the National Program for Potato Genetic Improvement, the incidence reached 100%. Yield losses, determined in
1992 and 1994, ranged as high as 19% to 56.33% depending on the cultivar. Characterization of the causal agent was done by
light microscopy, host range (graft and mechanical transmission), DNA hybridizations, polymerase chain reaction, and restriction
fragment length polymorphism analysis. Nucleotide sequence of the amplified fragments revealed the presence ofTomato mottle Taino virus. The virus was transmittedvia tubers and has been detected in mixed infections withPotato virus X and withPotato leaf roll virus.
http://www.phytoparasitica.org posting Oct. 20, 2003. The first two authors contributed equally to this work. 相似文献