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排序方式: 共有127条查询结果,搜索用时 687 毫秒
21.
High level of resistance to potato virus Y by expressing P1 sequence in antisense orientation in transgenic potato 总被引:2,自引:0,他引:2
22.
dsRNA-mediated resistance to Beet Necrotic Yellow Vein Virus infections in sugar beet (Beta vulgaris L. ssp. vulgaris) 总被引:1,自引:0,他引:1
Britt-Louise Lennefors Eugene I. Savenkov Jan Bensefelt Elisabeth Wremerth-Weich Petra van Roggen Stig Tuvesson Jari P. T. Valkonen Jan Gielen 《Molecular breeding : new strategies in plant improvement》2006,18(4):313-325
Rhizomania, one of the most devastating diseases in sugar beet, is caused by Beet Necrotic Yellow Vein Virus (BNYVV) belonging to the genus Benyvirus. Use of sugar beet varieties with resistance to BNYVV is generally considered as the only way to maintain a profitable yield on rhizomania-infested fields. As an alternative to natural resistance, we explored the transgenic expression of viral dsRNA for engineering resistance to rhizomania. Transgenic plants expressing an inverted repeat of a 0.4 kb fragment derived from the BNYVV replicase gene displayed high levels of resistance against different genetic strains of BNYVV when inoculated using the natural vector, Polymyxa betae. The resistance was maintained under high infection pressures and over prolonged growing periods in the greenhouse as well as in the field. Resistant plants accumulated extremely low amounts of transgene mRNA and high amounts of the corresponding siRNA in the roots, illustrative of RNA silencing as the underlying mechanism. The transgenic resistance compared very favourably to natural sources of resistance to rhizomania and thus offers an attractive alternative for breeding resistant sugar beet varieties. 相似文献
23.
24.
Mapping and marker-assisted selection for a gene for extreme resistance to potato virus Y 总被引:7,自引:1,他引:6
J. H. Hämäläinen K. N. Watanabe J. P. T. Valkonen A. Arihara R. L. Plaisted E. Pehu L. Miller S. A. Slack 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,94(2):192-197
The chromosomal location of the major gene Ry
adg
controlling extreme resistance to potato virus Y (PVY) in Solanum tuberosum subsp. andigena was identified by RFLP analysis of a diploid potato population. A total of 64 tomato and potato RFLP markers were screened
with the bulked segregant analysis (BSA) on segregants extremely resistant, hypersensitive or susceptible to PVY. Four markers
TG508, GP125, CD17 and CT168 at the proximal end of chromosome XI showed close linkage with extremely resistant phenotypes.
TG508 was identified as the closest marker linked with the Ry
adg
locus with the maximum map distance estimated as 2.0 cM. The 4 markers linked with the Ry
adg
locus were tested on independent tetraploid and diploid potato clones and were subsequently found useful for marker-assisted
selection for plants containing Ry
adg
.
Received: 5 July 1996 / Accepted: 19 July 1996 相似文献
25.
26.
Resolution and purification of taurine- and GABA-synthesizing decarboxylases from calf brain 总被引:1,自引:0,他引:1
A. A. Heinämäki S. I. Malila K. M. Tolonen K. H. Valkonen R. S. Piha 《Neurochemical research》1983,8(2):207-218
The present work describes a procedure for the co-purification of cysteine sulfinate decarboxylase (CSAD) and glutamate decarboxylase (GAD) from calf brain. A crude enzyme preparation was first made from brain homogenate by acid precipitation and ammonium sulphate fractionation. Subsequent fractionation of the decarboxylase preparation by cation exchange chromatography on CM-Sepharose CL-6B revealed the existence of a specific CSAD enzyme, which has no GAD activity. The GAD activity peak was found to possess CSAD activity. Further fractionation by gel filtration on Sephacryl S-200 separated the specific CSAD activity into two enzyme forms, one of them having a molecular weight of 150,000 and the other of 71,000. GAD activity was eluted from the gel filtration column in a single peak (mol wt 330,000) and showed CSAD activity. The purification of the specific CSAD enzyme was 920-fold and that of GAD activity 850-fold as compared with the starting material, whole calf brain. SDS gel electrophoresis indicated that the purified CSAD and GAD enzymes consisted of two or more subunits. The crude decarboxylase preparation was analysed by isoelectric focusing in ultra-thin polyacrylamide gel in the pH range 3.5-10.0. The most active fraction of CSAD indicated an isoelectric point of 6.5 and that of GAD 6.8. The pH optimum for CSAD activity in the crude preparation was 7.2 and that for GAD activity 7.9. 相似文献
27.
J.P.T. Valkonen K. Wiegmann J.H. Hämäläinen W. Marczewski & K.N. Watanabe 《The Annals of applied biology》2008,152(1):121-130
The tuber‐bearing wild potato species, Solanum stoloniferum, carries a dominant gene, Rysto, which confers extreme resistance (ER) to Potato virus Y (PVY). This gene was introgressed to cultivated potato germplasm (Solanum tuberosum) using accessions of S. stoloniferum maintained in European gene banks. It is mainly used in potato breeding programmes in Europe. Rysto was recently mapped to potato chromosome XII. However, in this study, a different accession of S. stoloniferum (PI275244; Haw1293) was used as a female parent in a cross to obtain a diploid (2n = 2x = 24) potato population of 112 F1 genotypes. From this accession, ER to PVY has been introgressed to the potato breeding programmes at the International Potato Center (Peru). As expected, ER to PVY was inherited in a dominant, monogenic fashion in the F1 population. Marker‐specific choices of DNA polymerase and adjustments of PCR conditions were made to optimise marker detection. The corresponding gene (Rysto) was mapped to the chromosome XII using the previously described and new cleaved amplified polymorphic sequence (CAPS) markers, which are based on the restriction fragment length polymorphism loci GP122 (six markers) and GP269 (one marker), and the simple sequence repeat marker STM0003. Four GP122‐based CAPS markers and STM0003 detected the same genotypes expressing ER to PVY. Because of a few recombinants, that is ER genotypes lacking the markers and the genotypes that react with necrosis but contain the markers, the marker distance from Rysto was estimated as 15.2 cM in this F1 population. However, the distance may be less if necrosis was considered an altered response also controlled by Rysto. The markers also specifically detected independent European potato cultivars that express ER to PVY derived from S. stoloniferum. Phylogenetic analysis of the sequences amplified from the GP122 locus of S. stoloniferum and potato cultivars further confirmed that the Rysto gene from independent accessions of S. stoloniferum can be selected using the same markers and the protocols described in this study. 相似文献
28.
29.
Potato virus A (PVA), a potyvirus with a (+)ssRNA genome translated to a large polyprotein, was engineered and used as a gene vector for expression of heterologous proteins in plants. Foreign genes including jellyfish GFP (Aequorea victoria) encoding the green fluorescent protein (GFP, 27 kDa) and the genes of human origin (Homo sapiens) encoding a soluble resistance-related calcium-binding protein (sorcin, 22 kDa) and the catechol-O-methyltransferase (S-COMT; 25 kDa) were cloned between the cistrons for the viral replicase and coat protein (CP). The inserts caused no adverse effects on viral infectivity and virulence, and the inserted sequences remained intact in progeny viruses in the systemically infected leaves. The heterologous proteins were released from the viral polyprotein following cleavage by the main viral proteinase, NIa, at engineered proteolytic processing sites flanking the insert. Active GFP, as indicated by green fluorescence, and S-COMT with high levels of enzymatic activity were produced. In contrast, no sorcin was detected despite the expected equimolar amounts of the foreign and viral proteins being expressed as a polyprotein. These data reveal inherent differences between heterologous proteins in their suitability for production in plants. 相似文献
30.
Zamyatnin AA Solovyev AG Bozhkov PV Valkonen JP Morozov SY Savenkov EI 《The Plant journal : for cell and molecular biology》2006,46(1):145-154
The bimolecular fluorescence complementation (BiFC) phenomenon has been successfully applied for in vivo protein-protein interaction studies and protein tagging analysis. Here we report a novel BiFC-based technique for investigation of integral membrane protein topology in living plant cells. This technique relies on the formation of a fluorescent complex between a non-fluorescent fragment of the yellow fluorescent protein (YFP) targeted into a specific cellular compartment and a counterpart fragment attached to the integral membrane protein N- or C-terminus or inserted into the internal loop(s). We employed this technique for topological studies of beet yellows virus-encoded p6 membrane-embedded movement protein, a protein with known topology, and the potato mop-top virus-encoded integral membrane TGBp2 protein with predicted topology. The results confirm that p6 is a type III integral transmembrane protein. Using a novel method, the central hydrophilic region of TGBp2 was localized into the ER lumen, whereas the N- and C-termini localized to the cytosol. We conclude that the BiFC-based reporter system for membrane protein topology analysis is a relatively fast and efficient method that can be used for high-throughput analysis of proteins integrated into the endoplasmic reticulum in living plant cells. 相似文献