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Sjefke J. H. M. Allefs Eliza R. De Jong Dion E. A. Florack Coosje Hoogendoorn Willem J. Stiekema 《Molecular breeding : new strategies in plant improvement》1996,2(2):97-105
Tachyplesin I is a 2.3 kDa antimicrobial peptide isolated from Southeast Asian horseshoe crabs. Bacterial suspensions containing 1×106 colony-forming units/ml of six isolates of pectolytic Erwinia spp., the causal pathogens of potato soft rot and blackleg, were killed in vitro by 1.4 to 11.1 g/ml of tachyplesin I. In an attempt to enhance resistance to Erwinia spp., each of the potato cultivars Bintje, Karnico and Kondor were transformed with two gene constructs encoding different precursor tachyplesin I proteins under the control of a cauliflower mosaic virus 35S promotor. Northern and western blot analysis showed that the tachyplesin I gene was expressed in transgenic plants. Small tubers of 17 transgenic clones were screened twice for soft rot resistance to Erwinia carotovora ssp. atroseptica. Under aerobic or anaerobic conditions, transgenic clones showed slightly less rot than control tubers.Abbreviations AP
acidic carboxyl terminal polypeptide
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Eca
Erwinia carotovora ssp. atroseptica
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Ecc
E. carotovora ssp. carotovora
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Ech
E. chrysanthemi
- IF
intercellular fluid
- SP
signal peptide
- TPNI (tpnI)
tachyplesin I 相似文献
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T. M. Golas R. M. C. Feron R. G. van den Berg G. M. van der Weerden C. Mariani J. J. H. M. Allefs 《Plant Systematics and Evolution》2010,285(1-2):103-110
Solanum dulcamara (bittersweet) is one of the few native species of Solanum present in Europe. It is a common weed that occupies a wide range of habitats and is often found in the direct vicinity of cultivated potatoes (Solanum tuberosum), where it could transmit diseases. A broad sampling of European S. dulcamara accessions was carried out to gain insight into the population structure and crossing preferences of this species. Three amplified fragment length polymorphism (AFLP®) primer combinations generating 288 polymorphic fragments were used to analyze 79 bittersweet accessions (245 individuals). Dendrograms revealed a low level of genetic polymorphism in the bittersweet populations, caused partially by the out-crossing nature of this species. 相似文献
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The Rpi-blb2 gene from Solanum bulbocastanum is an Mi-1 gene homolog conferring broad-spectrum late blight resistance in potato 总被引:7,自引:0,他引:7
van der Vossen EA Gros J Sikkema A Muskens M Wouters D Wolters P Pereira A Allefs S 《The Plant journal : for cell and molecular biology》2005,44(2):208-222
The necessity to develop potato and tomato crops that possess durable resistance against the oomycete pathogen Phytophthora infestans is increasing as more virulent, crop-specialized and pesticide resistant strains of the pathogen are rapidly emerging. Here, we describe the positional cloning of the Solanum bulbocastanum-derived Rpi-blb2 gene, which even when present in a potato background confers broad-spectrum late blight resistance. The Rpi-blb2 locus was initially mapped in several tetraploid backcross populations, derived from highly resistant complex interspecific hybrids designated ABPT (an acronym of the four Solanum species involved:S. acaule, S. bulbocastanum, S. phureja and S. tuberosum), to the same region on chromosome 6 as the Mi-1 gene from tomato, which confers resistance to nematodes, aphids and white flies. Due to suppression of recombination in the tetraploid material, fine mapping was carried out in a diploid intraspecific S. bulbocastanum F1 population. Bacterial artificial chromosome (BAC) libraries, generated from a diploid ABPT-derived clone and from the resistant S. bulbocastanum parent clone, were screened with markers linked to resistance in order to generate a physical map of the Rpi-blb2 locus. Molecular analyses of both ABPT- and S. bulbocastanum-derived BAC clones spanning the Rpi-blb2 locus showed it to harbor at least 15 Mi-1 gene homologs (MiGHs). Of these, five were genetically determined to be candidates for Rpi-blb2. Complementation analyses showed that one ABPT- and one S. bulbocastanum-derived MiGH were able to complement the susceptible phenotype in both S. tuberosum and tomato. Sequence analyses of both genes showed them to be identical. The Rpi-blb2 protein shares 82% sequence identity to the Mi-1 protein. Significant expansion of the Rpi-blb2 locus compared to the Mi-1 locus indicates that intrachromosomal recombination or unequal crossing over has played an important role in the evolution of the Rpi-blb2 locus. The contrasting evolutionary dynamics of the Rpi-blb2/Mi-1 loci in the two related genomes may reflect the opposite evolutionary potentials of the interacting pathogens. 相似文献
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Expression of giant silkmoth cecropin B genes in tobacco 总被引:10,自引:0,他引:10
Dion Florack Sjefke Allefs Rik Bollen Dirk Bosch Bert Visser Willem Stiekema 《Transgenic research》1995,4(2):132-141
Cecropin B is a small antibacterial peptide from the giant silkmothHyalophora cecropia. To reveal the potential of this peptide for engineering bacterial disease resistance into crops, several cecropin B gene constructs were made either for expression in the cytosol or for secretion. All constructs were cloned in a plant expression vector and introduced in tobacco viaAgrobacterium tumefaciens. A cDNA-derived cecropin B gene construct lacking the amino-terminal signal peptide was poorly expressed in transgenic plants at the mRNA level, whereas plants harbouring a full-length cDNA-derived construct containing the insect signal peptide, showed increased cecropin B-mRNA levels. Highest expression was found in plants harbouring a construct with a plant-gene-derived signal peptide. In none of the transgenic plants could the cecropin B peptide be detected. This is most likely caused by breakdown of the peptide by plant endogenous proteases, since a chemically synthesized cecropin B peptide was degraded within seconds in various plant cell extracts. This degradation could be prevented by the addition of specific protease inhibitors and by boiling the extract prior to adding the peptide. In addition, anionic detergents, in contrast to cationic, zwitter-ionic or non-ionic detergents, could prevent this degradation. Nevertheless, transgenic tobacco plants were evaluated for resistance toPseudomonas solanacearum, the causal agent of bacterial wilt of many crops, andP. syringae pv.tabaci, the causal agent of bacterial wildfire, which are highly susceptible to cecropin Bin vitro. No resistance was found. These experiments indicate that introduction and expression of cecropin B genes in tobacco does not result in detectable cecropin B protein levels and resistance to bacterial infections, most likely due to degradation of the protein by endogenous proteases. 相似文献
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Tomasz Michal Golas Anne Sikkema Jack Gros Richard M. C. Feron Ronald G. van den Berg Gerard M. van der Weerden Celestina Mariani J. J. H. M. Allefs 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2010,120(4):797-808
Initial screening of 14 Solanum dulcamara accessions enabled the identification of individuals resistant and susceptible to Phytophthora infestans. Crosses between contrasting genotypes resulted in three F2–BC1 populations segregating for resistance to late blight in a laboratory assay and under field conditions. Genetic profiling
of one of these populations using 128 AFLP primers generated three markers linked to the resistant phenotype. Blast analysis
of the sequenced markers resulted in a plausible gene position on the distal end of the long arm of chromosome 9 that could
be confirmed by CAPS markers. Thus, we describe a first resistant gene, named Rpi-dlc1, from S. dulcamara, a Solanum species native to Europe. In addition, one population was tested for broadness of resistance responses using a set of seven
additional P. infestans isolates, varying in virulence. This indicated the possible presence of additional Rpi genes. 相似文献
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T. M. Golas H. van de Geest J. Gros A. Sikkema N. D’Agostino J. P. Nap C. Mariani J. J. H. M. Allefs I. Rieu 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2013,126(1):59-68
Phytophthora infestans, the causal agent of late blight, remains the main threat to potato production worldwide. Screening of 19 accessions of Solanum dulcamara with P. infestans isolate Ipo82001 in detached leaf assays revealed strong resistance in an individual belonging to accession A54750069-1. This plant was crossed with a susceptible genotype, and an F1 population consisting of 63 individuals was obtained. This population segregated for resistance in 1:1 ratio, both in detached leaf assays and in an open-field experiment. Presence of the formerly mapped Rpi-dlc1 gene as the cause of the observed segregating resistance could be excluded. Subsequently, AFLP analyses using 128 primer combinations enabled identification of five markers linked to a novel resistance gene named Rpi-dlc2. AFLP markers did not show sequence similarity to the tomato and potato genomes, hampering comparative genetic positioning of the gene. For this reason we used next-generation mapping (NGM), an approach that exploits direct sequencing of DNA (in our case: cDNA) pools from bulked segregants to calculate the genetic distance between SNPs and the locus of interest. Plotting of these genetic distances on the tomato and potato genetic map and subsequent PCR-based marker analysis positioned the gene on chromosome 10, in a region overlapping with the Rpi-ber/ber1 and -ber2 loci from S. berthaultii. Pyramiding of Rpi-dlc2 and Rpi-dlc1 significantly increased resistance to P. infestans, compared with individuals containing only one of the genes, showing the usefulness of this strategy to enhance resistance against Phytophthora. 相似文献
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The late blight resistance locus Rpi-bib3 from Solanum bulbocastanum belongs to a major late blight R gene cluster on chromosome 4 of potato 总被引:1,自引:0,他引:1
Park TH Gros J Sikkema A Vleeshouwers VG Muskens M Allefs S Jacobsen E Visser RG van der Vossen EA 《Molecular plant-microbe interactions : MPMI》2005,18(7):722-729
Late blight, caused by Phytophthora infestans, is one of the most devastating diseases in cultivated potato. Breeding of new potato cultivars with high levels of resistance to P. infestans is considered the most durable strategy for future potato cultivation. In this study, we report the identification of a new late-blight resistance (R) locus from the wild potato species Solanum bulbocastanum. Using several different approaches, a high-resolution genetic map of the new locus was generated, delimiting Rpi-blb3 to a 0.93 cM interval on chromosome 4. One amplification fragment length polymorphism marker was identified that cosegregated in 1,396 progeny plants of an intraspecific mapping population with Rpi-blb3. For comparative genomics purposes, markers linked to Rpi-blb3 were tested in mapping populations used to map the three other late-blight R loci Rpi-abpt, R2, and R2-like also to chromosome 4. Marker order and allelic conservation suggest that Rpi-blb3, Rpi-abpt, R2, and R2-like reside in the same R gene cluster on chromosome 4 and likely belong to the same gene family. Our findings provide novel insights in the evolution of R gene clusters conferring late-blight resistance in Solanum spp. 相似文献
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Herman C. H. Schoenmakers Maarten Koornneef Sjefke J. H. M. Alefs Wendy F. M. Gerrits Dianne van der Kop Isabelle Chérel Michel Caboche 《Molecular & general genetics : MGG》1991,227(3):458-464
Summary Five nitrate reductase-deficient mutants of tomato were isolated from an M2 population after ethyl-methanesulphonate (EMS) seed treatment by means of selection for chlorate resistance. All mutations were monogenic and recessive and complementation analysis revealed that they were non-allelic. Biochemical and molecular characterization of these mutants showed that four of them are cofactor mutants while one is an apoenzyme mutant. 相似文献