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The present study was under taken in order to analyze the chemical mutagenesis on Chilli germplasm. In this regard, K1 variety of chilli was subjected to different mutagenic concentration for inducing mutagenesis. The M3 plants exposed to EMS and DES to produce clear difference from the untreated control, thus indicating that mutagenic treatment produce polymorphic regions in the chilli. For extraction of genomic DNA was adopted an improved protocol of CTAB method with slight modification. A total of ten primers were used to screen the polymorphism among the treated populations line tall, tall with chlorophyll deficient, leaf, flower, GMS and DNA damages in maturity mutants were analyzed with control. Out of ten primers, four primers (PGF02, PGF03, PGF04 AND OP107) were successfully amplified in all the samples used for this study. The successful primers were amplified in to 93 products showing an average of 9.3 bands.
Restocking and stock enhancement programs are now recognized as an important tool for the management of fishery resources. It is important, however, to have an adequate knowledge on the genetic population structure of both the released stock and the wild population before carrying out such programs. In this study, random amplified polymorphic DNA (RAPD) markers were applied to assess genetic diversity and population structure of wild and hatchery populations of the white seabream Diplodus sargus and the common two-banded seabream D. vulgaris (Sparidae). The estimated values for intrapopulation genetic variation, measured using the percentage of polymorphic loci (%P), Shannon index (H’), and Nei’s gene diversity (h), showed high values for all populations. The percentage of genetic variation within D. sargus and D. vulgaris populations, based on coefficient of gene differentiation, reached 82.5% and 90% of the total genetic variation, respectively. An undeniable decrease in genetic variation was found in both hatchery populations, particularly in D. sargus, compared to the wild ones. However, the high values of variation within all populations and the low levels of genetic variation among populations did not indicate inbreeding or depression effects, thus indicating a fairly proper hatchery management. Nevertheless, the results of this study highlight the importance of monitoring the genetic variation of hatchery populations, particularly those to be used in restocking programs. The creation of a genetic baseline database will contribute to a more efficient conservation management and to the design of genetically sustainable restocking programs.
Arnica montana L. is one of the most important herbal plants used in medicine, pharmaceutical and cosmetic industry. The number of studies performed with molecular markers on arnica genotypes is very limited. Because of this fact the aims of presented examination were optimization of protocols DNA isolation from fresh leaves of A. montana and identification of genetic diversity among this plant genotypes. In presented study to obtain pure DNA Plant & Fungi DNA Purification Kit (EURx) were used. To clean obtained DNA long and slow electrophoresis and isolation DNA from gels were used. A. montana genotypes were analyzed using 40 RAPD primers (Operon Technologies), out of which 12 produced high number of polymorphic and repeatable fragments. In total, selected primers produced 120 fragments, among them 111 (92.5%) were polymorphic. The genetic similarity matrices were produced based on RAPD using the Dice’s coefficient. RAPD based genetic similarity was estimated between 0.535 and 0.945. The highest genetic similarity was estimated among GA17 and GA18 genotypes, which are closely located on the obtained dendrogramme.
RAPD and ISSR techniques were used in identification of 26 pear cultivars. As a result of reactions carried out with RAPD 25 primers, 103 polymorphic DNA frag­ments were obtained. The largest number of polymorphic dNa fragments (7-8) was produced in reactions with the following primers: OPT 15, OPG 16 and OPG 19. Identification of cultivars and rootstocks was achievable with the use of OPT 15, OPG 19 and OPU 07 primers. In the reactions performed with 22 ISSR primers, 135 markers of pear cultivars were obtained. The size of fragments varied from 280 to 1790 bp. The greatest number of polymorphic products (9) was obtained in the reac­tions with primers: 830, 840 and 844. Primers 840 and 844 enabled identification of all tested genotypes. Degree of DNA polymorphism was estimated at 56.3% (RAPD) and 71.5% (ISSR). Results of the research confirm the usefulness of both techniques in identifying pear cultivars.
Isozymes were the first widely used molecular markers in plant population analysis. They yielded valuable information on the amount and the structure of genetic variability. DNA technology has provided new types of markers based on DNA sequence, which make it possible to study polymorphisms in a much greater proportion of the genome. This is the reason why the use of isozymes is less popular nowadays. This effect would be justified if all markers provided the same type of information on polymorphism and genetic relationships among populations; otherwise, it would be necessary to use different markers to obtain the complete picture of the genetic structure of populations and species. In this study, we compared data of isozyme and RAPD markers in the populations of two tetraploid species of wild oats: Avena barbata populations collected in Argentina, and Avena murphyi populations collected in Spain and Morocco. The samples were evaluated for 9 isozymatic systems and 10 primers. The structure of genetic variability was studied using Nei’s method, and the relationships between populations were estimated using Hedrick and Jaccard's similarities for isozymes and RAPDs, respectively. As expected, RAPDs were more polymorphic than isozymes, but the information obtained from both markers was weakly correlated. The various reasons for this observation are discussed, but our conclusion is that in order to study the structure of genetic variability, several types of markers should be used.
Application of molecular markers makes the selection process much more effective. Marker assisted selection is an important tool for plant breeders to increase the efficiency of a breeding process, especially for multigenic traits, highly influenced by the environment. Epistasis is the interaction between alleles from two or more loci determining the complex traits, and thus plays an important role in the development of quantitative traits of crops. In this paper, the relationships between RAPD marker-by-marker interactions and 22 quantitative traits of caraway (Carum carvi L.) were analyzed. Significant associations of 116 epistatic markers with at least one trait in 2004 as well as 112 in 2005 were found on the basis of multivariate regression analysis. The proportion of total phenotypic variances of individual trait explained by the marker-by-marker interactions ranged from 25.3% to 96.0%.
The aim of our study was to evaluate the genetic relationship of Rhododendrons from Rhododendron collection at Tomaszkowice in Pogórze Wielickie. Leaves and leaf buds of three evergreen Asiatic species: R. aureum, R. brachycarpum, R. purdomii, six cultivars from Catawbiense Hybridum group (R. catawbiense): Catharine van Tol, Nova Zembla, Album Novum, Boursoult, Old Port, Hachmann’s Charmant and one cultivar from Yakushimanum Hybridum group: Koichiro Wada were investigated. Two plants from each accession were analysed. DNA was isolated by Ziegenhagen and Scholz [1998] protocol, dedicated to the difficult species in terms of high quality DNA extraction. After isolation, DNA was amplified with 19 RAPD primers. We obtained 255 RAPD markers and worked out a dendrogram, which illustrated the level of genetic diversity among the tested plants. Our study confirmed a close relationship between rhododendrons from Catawbiense Hybridum group. Another group was constituted by R. aureum and R. purdomii. The third group consisted of R. brachycarpum and Koichiro Wada cv.
Identification of cultivars is essential both in breeding and to settle cultivar disputes. The purpose of the study has been to examine cultivar identities based on absorption spectra of plant pigments and to confirm a genetic stability with SCoT and RAPD molecular markers in new Polish lines of Chenopodium quinoa Willd. Spectral analysis of pigments extracted from plant inflorescences in quinoa gives an opportunity to confirm the cultivar identity and identification of ‘Faro’ and ‘Titicaca’ cultivars and their new lines. Spectral analysis is an effective method of confirming cultivar identity and it should be used in practice for the identification of cultivars or cultivars lines in Chenopodium quinoa Willd. Analysis of molecular markers indicated by RAPD as well as SCoT technique revealed a high genetic stability of the derivative lines of ‘Faro’ and ‘Titicaca’, while variation was detected in plants representing original cultivars: banding pattern different than predominant was present in three plants of ‘Titicaca’ (genetic distnaces from 7.5% to 55.9%) and in a single plant of ‘Faro’(genetic distance 61.2% as indicated by SCoT technique).
Analizami objęto dwie populacje F2 mieszańców międzyliniowych żyta oraz zestaw 62 rekombinacyjnych linii wsobnych (RIL-F7) żyta wyprowadzonych z mieszańca 541 × Ot1-3. Potwierdzono istnienie sprzężenia między uprzednio wytypowanymi trzema markerami RAPD, a genem kontrolującym męską sterylność w cytoplazmie C. Dodatkowo zidentyfikowano zestaw pięciu nowych markerów RAPD sprzężonych z genem Rfc1, ale wśród nich tylko jeden wykazywał sprzężenie z genem męskiej sterylności wystarczająco silne dla podjęcia prób wykorzystania go przy selekcjonowaniu materiałów hodowlanych.
Klasyczna ocena zmienności zasobów genetycznych uwzględnia cechy fenologiczne, morfologiczne i rozwojowe. Cechy te podlegają wpływom środowiska, stąd też oparte na nich wnioski mogą być mało precyzyjne. Dlatego do oceny zmienności wykorzystuje się również markery biochemiczne i molekularne. Spośród omawianych systemów (izoenzymy, RAPD, ISSR, AFLP, SSR, SNP, SSAP, IRAP, REMAP, RBIP) najczęściej stosowane są izoenzymy, RAPD, ISSR i SSR. Metody molekularne umożliwiają badanie zmienności genetycznej, wnioskowanie na temat filogenezy i pokrewieństwa gatunków oraz śledzenie zmian, jakie zachodzą w puli genowej w zależności od długości i sposobu przechowywania zasobów genetycznych. Markery znajdują powszechne zastosowanie w charakteryzowaniu kolekcji. Mogą być wykorzystywane w identyfikowaniu zduplikowanych obiektów i zanieczyszczeń kolekcyjnych, stanowiąc uzupełnienie klasycznej oceny zmienności. Markery wykorzystuje się także do oceny czystości linii zarówno w zasobach genowych, jak i odmian uprawnych zgłaszanych do badań rejestrowych. Metody molekularne stosowane są do tworzenia „core collection”, która pomimo ograniczonej liczby obiektów w stosunku do kolekcji wyjściowej reprezentuje szeroką zmienność i jest łatwiejsza w ocenie i wykorzystaniu. Selekcja hodowlana z wykorzystaniem markerów (MAS) stanowi cenne uzupełnienie w wyborze materiału kolekcyjnego do krzyżowań oraz w śledzeniu introgresji puli genowej z dzikiego materiału genetycznego do odmian uprawnych. Praca przedstawia przykłady literaturowe powyższych zastosowań markerów molekularnych w ocenie zmienności zasobów genetycznych.
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