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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
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Male experimental rats (100 gm liveweight) were distributed into 10 groups of 8 animals each and received balanced diets, with the exception of lysine which was added to the diets in graded amounts in such a way that the lysine content of the diets ranged from 2.44 to 5.92 gm/16 gm N. After a feeding period of 7 days the animals received 3H- and 14C lysine injected intraperitoneally, 4 animals of each group were investigated for the total CO2 excretion and 14CO2 excretion during the first 2 hrs after the injection and for the urinary excretion of radioactivity (48 hours). The remaining animals in each group were used for determining the plasma amino acids and for establishing the specific radioactivity of free lysine in the liver and muscles after an 1-hour incorporation period. Total CO2 excretion was not found to be influenced by the lysine contents while the level of excretion of 14C activity through CO2 and that of specific 14C activity of CO2 increased with increasing lysine concentrations. This produced a broken curve pattern, showing an increased release of 14CO2 (under maintenance conditions) if the diet contained 4 gm lysine/16 gm N and more. Investigations for the specific 14C activity of free lysine in the liver, the main site of lysine oxidation, showed that the increase in 14CO2 release was due to an enhanced rate of lysine catabolism and was not brought about by changes in the pool volume or in specific radioactivity. The levels of urinary 14C excretion were not found to be related to the lysine content of the diets, whereas the curve pattern of 3H excretion observed 5 to 8 hrs after injection was similar to that of 14CO2 excretion. The lysine content of blood plasma and the content of free lysine in the liver increased continuously with increasing levels of dietary lysine. The methodological studies made in the present paper showed that in scientific research a determination of amino acid requirements on the basis of CO2 oxidation data may be a very exact and sensitive method. It will also yield values for maintenance requirements. 相似文献
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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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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
8.
Hiromitsu Furuya Tsutomu Matsumoto Shin-ichi Fuji Hideki Naito 《Journal of General Plant Pathology》2003,69(2):115-119
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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Norman Owen-Smith 《Landscape Ecology》2004,19(7):761-771
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