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
1. Changes in the concentrations of plasma luteinising hormone (LH), prolactin, androgen and progesterone were measured during the ovulatory cycle of the turkey.
2. Single pre‐ovulatory peaks of plasma LH, androgen and progesterone were observed which took 8, 8 and 12 h respectively, to increase and return to base‐line values. The concentration of plasma prolactin tended to be elevated between 6 h before and 6 h after the LH peak with the maximum values occurring after the peak.
3. The changes in the concentrations of plasma LH and progesterone were 3‐ and 7‐fold respectively while 2‐fold changes were observed in the concentrations of plasma androgen and prolactin.
4. The pre‐ovulatory concentration of plasma progesterone and prolactin began to decrease 4 and 6 h respectively, after the pre‐ovulatory peak of LH.
5. Ovulation and oviposition occurred 6 to 8 h and 36.10+ 0.57 h (SEM) ( n= 11) respectively after the pre‐ovulatory peak of LH.
6. In birds kept on 14 h light/d, pre‐ovulatory peaks of LH were initiated only during a 10 to 11‐h period starting within 2 h after the onset of darkness.
7. A comparison between these data and those from the fowl suggest that the egg is retained in the turkey's oviduct for about 3 to 4 h longer than in the fowl. 相似文献
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
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
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
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
Xiphinema diversicaudatum and X. index are vector nematode species of economic importance in viticulture regions as they can transmit Arabis Mosaic, Grapevine Fanleaf and Strawberry Latent Ringspot viruses to grapevine. Wang et al. (2003) designed species-specific diagnostic primers from ribosomal genes for both these vector species as well as a vector and a non-vector species X. italiae and X. vuittenezi, respectively. Our study aimed to confirm the specificity and determine the sensitivity and reliability of the primers for the two vector species, X. diversicaudatumand X. indexwhen challenged with closely related longidorid species and general nematode communities typical of vineyard soil. With one exception, no PCR product was observed when the primers were tested against six Longidorus, one Paralongidorus and one Xiphinema non-target species. Occasionally (three out of eight replicate PCR reactions) a weak PCR product was noted when primers for X. index were tested with L. elongatus. Furthermore, when challenged with a range of non-target nematode species comprising the nematode community typical of viticulture soil, no PCR product was amplified. An experimental dilution series of extracted DNA rigorously demonstrated that DNA from an equivalent single specimen of the target virus-vector species, X. diversicaudatum and/or X. index, could be detected amongst 1000 equivalent non-targetX. vuittenezi. Also, extracted DNA from an equivalent single target specimen was detected when added to DNA extracted from the overall soil nematode community. The primers were assessed further by using serial mixtures of actual nematodes rather than extracted DNA to simulate field soil. Using this method, a single target nematode could be detected amongst 200 non-target specimens. Given their specificity, sensitivity and reliability, it appears that these diagnostic primers will be of great benefit to phytosanitary/quarantine services related to the viticulture industry. 相似文献