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wheatgrass (Elymus caninus L.) in central and northern Europe, using isozymes, RAPD and microsatellite markers. — Theor. Appl. Genet. 103 : 191–196. Bianchi D. S. Search for evidence

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Molecular characterization of proliferating carnation plant tissues revealed association with a pararetroviroid-like agent. Carnation small virod-like RNA termed as CarSV RNA is unique among plant viroidlike RNA s by having a homologous DNA counterpart in the plant genome. Previously, CarSV DNA related sequences had been detected in the plant genome fused to microsatellite-like genomic sequences. Here we present data that tissue proliferation symptoms can be seen only on those plants that are expressing the RNA form of this agent. As CarSV RNA adopts a hammerhead structure in both polarities and the plant genome naturally encodes a functional hammerhead ribozyme, it can be concluded that these ribozyme sequences are cleaved out from the plant genome. This could lead to the evolution of small RNA s that replicate autonomously by the rolling circle mechanism and later could spread horizontally by vegetative propagation of the plant material, or by using a hitchhiked plant pararetrovirus fused to its genome.

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Studies involving morphological description with both dominant (RAPD) and codominant (SSR, isoenzyme) molecular markers were made on 28 maize inbred lines of known genetic background with a final aim of prediction of heterosis. The genetic distance and degree of relationship between the lines was determined using cluster analysis. Only a very limited extent of allele polymorphism could be detected in isoenzyme analyses as the 28 lines formed only 16 gel electrophoretic groups, indicating that certain lines had identical isoenzyme patterns. On the basis of RAPD and gene-specific microsatellite (SSR) markers, however, all the lines could be distinguished from each other. When the lines were grouped according to genetic background it was found that although the individual marker systems only partially reflected the actual relationships between the lines, a joint processing of the data, supplemented with morphological data, revealed a close correlation between the groups formed on the dendrogram and the genetic background.

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A húgyhólyagrák a második leggyakrabban előforduló urológiai jellegű tumoros megbetegedés. A mai gyakorlatban kezelésének módját és a beteg várható életkilátásait szinte kizárólag a szövettani diagnózis határozza meg. A tumor invazivitásának megítélése azonban gyakran nehézségekbe ütközik. Újabb molekuláris vizsgálatok lehetőséget nyújthatnak e betegség korai felfedezésére, kérdéses esetekben a szövettani diagnózis megerősítésére, illetve a tumor várható viselkedésének előrejelzésével segíthetnek a terápia optimalizálásában. Az e disszertációban bemutatott vizsgálatok a hólyagrákok aktuális diagnosztikai és prognosztikai problémáira keresnek válaszokat. Ehhez pedig a molekuláris biológia modern módszereit hívják segítségül. Munkánk során 334 hólyagtumoros beteg és 117 kontroll személy tumor-, vér- és/vagy vizeletmintáit vizsgáltuk. A hólyagtumor genetikai eltéréseit PCR-alapú mikroszatellita-allélvesztés vizsgálattal (12 marker felhasználásával) illetve FISH-technikával (4 DNS-próba alkalmazásával) diagnosztikai céllal kerestük vizeletben. A tumormintákból elvégzett teljes genomi mikroszatellita-allélvesztés vizsgálat – melyhez 400 markert használtunk – az invazivitással korreláló markerek kiválasztása által a szövettani diagnózis megerősítését célozta. Továbbá vizsgáltuk a VEGF, Ang-1, Ang-2 és a Tie2 prognosztikus értékét. Megállapítottuk, hogy a vizeletből történő mikroszatellita-allélvesztés vizsgálat és a FISH-vizsgálat érzékeny és specifikus módszer a hólyagtumor nem invazív kimutatására. A teljes genom mikroszatellita-allélvesztés vizsgálata során összeállítottunk egy olyan négy tagból álló markerkombinációt, mellyel az összes vizsgált tumor azonosítható volt. Kiválasztottuk azokat a mikroszatellita-markereket, melyek a tumor invazivitásával mutatnak összefüggést. Megállapítottuk, hogy a tumor indukálta angiogenezisben részt vevő molekulák expressziójának és szérumkoncentrációjának meghatározása alkalmas eszköz lehet a betegség várható viselkedésének pontosabb előrejelzésére.

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126 410 415 Hill-Ambroz, K.L., Brown-Guedira, G.L., Fellers, J.P. 2002. Modified rapid DNA extraction protocol for high throughput microsatellite

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, M. 2002. Characterisation of polymorphic microsatellite markers from Aegilops tauschii and transferability to the D-genome of bread wheat. Theor. Appl. Genet. 104 :1164–1172. Bernard M

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It is well established that the ingestion of cereal prolamins, such as gluten, causes the characteristic symptoms of celiac disease (CD) in people predisposed to it. DNA-based PCR method provides new ways to detect gluten in processed foodstuffs, such as bread. The aim of this work was to adapt a new primer pair combination and to initiate a carefully elaborated PCR methodology to experiment with DNA-based analysis. At first, the purity of cleaned DNA was verified using B49317 and A49855 chloroplast DNA primer pair. Then TR01/2 wheat specific PCR primer pair was used for checking the origin of the DNA, and P1/2 microsatellite (SSR) adapted primer pair for detecting allergen (gluten) specific residues. Method optimisation was achieved with cereal flour samples, then bread and dry pasta products from wheat were used, which were analysed as heat-treated samples with three primer pairs. The gluten specific primer pair was tested on cross-reactive cereals such as rye, barley, triticale and on some questionable cereals, such as oat, and pseudo-cereals, e.g. buck wheat and amaranth.

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Nearly twenty thousand wheat lines were phenotyped for the presence of leaf tip necrosis (LTN), a phenotypic trait linked to adult plant leaf rust resistance (APR) genes, viz. Lr34, Lr46 and Lr67 having pleiotropic association with multiple disease resistance genes. Thirty-six lines showed varied expression of LTN and moderate level of leaf rust severity at adult plant stage with slow rusting (disease progress at a retarded rate). Seedling resistance test (SRT) revealed susceptible and mixed infection types, a characteristic of adult plant resistance (APR) genes. Further molecular confirmation for the presence of these genes using available microsatellite markers revealed that of the 36 lines, five lines carried Lr46+ alone and five other lines carried Lr67+ alone. Seven lines carried the combination of Lr34+ and Lr67+ while six lines confirmed to carry the combination of Lr46+ and Lr67+. Remarkably three lines carried all the three APR genes, viz. Lr34+, Lr46+ and Lr67+. All these stocks can be a source of APR multiple disease resistance genes. Ten lines were not confirmed to carry any of the genes but still had LTN and SRT results showing an infection type typical of APR genes and these can be the source of identifying newer APR genes. The resistance based on minor APR genes when combined with a few additional minor genes in the background of high yielding cultivars is expected to have high level of race non-specific resistance and to be durable.

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Pre-harvest sprouting (PHS) in bread wheat (Triticum aestivum L.) is one of the major abiotic constraints influencing production of high quality grain. Selection for pre-harvest sprouting (PHS) resistance in bread wheat (Triticum aestivum L.) in early generations is difficult because it is expressed as a quantitatively inherited trait and subject to environmental effects. The objectives of this study were to validate a major quantitative trait locus (QTL) for PHS resistance on chromosome 4A in bread wheat and to isolate near-isogenic lines for this QTL using marker-assisted selection. A total of 60 Canadian wheat cultivars and experimental lines were screened with three SSR markers in a QTL region for PHS resistance. The SSR markers DuPw004, barc170 and wmc650 explained 67%, 75% and 60% of total variation in germination (%), respectively, among different wheat genotypes. Marker assisted back crossing with DuPw004 reduced the population size in BC1F1 and BC2F1 generation by 41% and 59%, respectively. A survey of pedigrees of different genotypes revealed that the parental line RL4137 is a major source of increased PHS resistance in a number of western Canadian wheat cultivars. Microsatellite markers (DuPw004, barc170 and wmc650) will be useful for plant breeders to pyramid QTL from different PHS resistance sources.

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Acta Agronomica Hungarica
Authors: Z. Jurković, K. Dugalić, M. Viljevac, I. Piližota, A. Vokurka, B. Puškar, and I. Pejić

Cipriani, G., Lot, G., Huang, W. G., Marrazzo, M. T., Peterlunger, E., Testolin, R. (1999): AC/GT and AG/CT microsatellite repeats in peach [ Prunus persica (L.) Batsch]: isolation, characterisation and cross-species amplification in Prunus

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