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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
5.
A. Costagliola D. Russo A. A. Izzo P. Urbani V.
di
Marzo A. Cecio 《Anatomia, histologia, embryologia》2005,34(Z1):13-13
The endocannabinoid anandamide may regulate intestinal motility through activation of CB1 receptors. Anandamide is then inactivated by fatty acid amide hydrolase (FAAH), a membrane bound enzyme. Under pathological conditions, inactivation of such enzymatic activity may lead to inhibition of the intestinal motility. Here, preliminary reports on the distribution of Fatty Acid Amide Hydrolase (FAAH) immunoreactivity in the mouse gastrointestinal neurons, and the pharmacological effects of N‐arachidonoylserotonin (AA‐5HT), a selective inhibitor of FAAH, are reported. FAAH was revealed by an indirect immunofluorescence. Laminar preparations containing the myenteric or the submucous plexus adhered, were peeled off after the whole gut wall had been stretched out and fixed in 4% paraformaldehyde. They were subsequently incubated with a polyclonal anti‐serum directed against a region near the N‐terminus of the human FAAH and revealed by a FITC‐conjugated goat anti‐rabbit secondary anti‐serum. FAAH‐immunoreactive neurons were observed within the myenteric ganglia throughout the GIT. The positive nerve cells varied in size and density of immunoreactivity. Stomach and large intestine showed the highest neuronal density. AA‐5HT significantly reduced both gastric emptying and gastrointestinal tract transit. Such inhibitory effect was reduced by the C1 receptor antagonist SR141716A. Both morphological and pharmacological results suggest that FAAH may play a critical role in controlling gut anandamide levels. 相似文献
6.
Tsutomu MATSUMOTO Yuichiro NARA Hiromitsu FURUYA Harumi TAKAHASHI Kiichi TAIRAKO Hideki YAMAMOTO 《Journal of General Plant Pathology》2002,68(4):382-384
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
7.
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
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9.
Eiji Tanaka Chihiro Tanaka Atsushi Ishihara Yasumasa Kuwahara Mitsuya Tsuda 《Journal of General Plant Pathology》2003,69(1):1-6
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
10.
Piero Roggero Hervé Lot Sylvie Souche Riccardo Lenzi Robert G. Milne 《European journal of plant pathology / European Foundation for Plant Pathology》2003,109(3):261-267
Big-vein disease (BV) of lettuce has been attributed to infection by Lettuce big-vein virus (LBVV), vectored by the soil fungus Olpidium brassicae. The finding of a second soil-borne virus in lettuce, Mirafiori lettuce virus (MiLV), led to a re-investigation of the role of LBVV in big-vein disease, with evidence emerging that both MiLV and LBVV are vectored by O. brassicae, and that MiLV, not LBVV, is the cause of BV (Lot et al. (2002), Phytopathology 92: 288–293). The two viruses have coat proteins of similar size but have different morphologies and are serologically unrelated. We tested individual lettuce plants in BV-prone fields and protected crops in France and Italy for the presence of the two viruses, using DAS-ELISA and antisera specific for each virus. Both MiLV and LBVV were found at high incidence, often together but sometimes separately. Symptoms were frequently found to be associated with MiLV alone or both viruses, but rarely LBVV alone. However, no absolute correlation emerged, because sometimes MiLV was present in the absence of symptoms, and vice versa. To clarify the situation, individual lettuce plants were examined over a period of time in two further surveys. In surveys of protected crops in France, plants with big-vein were always ELISA-positive for MiLV, or else symptomless plants positive for MiLV were later seen to develop big-vein symptoms. Presence or absence of LBVV appeared to have no effect on symptom development. In surveys of open fields in Italy, all combinations were found: presence of both viruses, apparent absence of both viruses, or presence of each one alone, in plants that developed BV. At the end of the observation period, nearly all plants had BV and contained both viruses. 相似文献