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1

Development of an efficient retrotransposon-based fingerprinting method for rapid pea variety identification

100%
Smykal P.
Journal of Applied Genetics
|
2006
|
vol. 47
|
issue 3
221-230
EN
Fast and efficient DNA fingerprinting of crop cultivars and individuals is frequently used in both theoretical population genetics and in practical breeding. Numerous DNA marker technologies exist and the ratio of speed, cost and accuracy are of importance. Therefore even in species where highly accurate and polymorphic marker systems are available, such as microsatellite SSR (simple sequence repeats), also alternative methods may be of interest. Thanks to their high abundance and ubiquity, temporary mobile retrotransposable elements come into recent focus. Their properties, such as genome wide distribution and well-defined origin of individual insertions by descent, predetermine them for use as molecular markers. In this study, several Ty3-gypsy type retrotransposons have been developed and adopted for the inter-retrotransposon amplified polymorphism (IRAP) method, which is suitable for fast and efficient pea cultivar fingerprinting. The method can easily distinguish even between genetically closely related pea cultivars and provide high polymorphic information content (PIC) in a single PCR analysis.
2

Efficiency of transformation of Polish cultivars of pea (Pisum sativum L.) with various regeneration capacity by using hypervirulent Agrobacterium tumefaciens strains

100%
Pniewski T., Kapusta J.
Journal of Applied Genetics
|
2005
|
vol. 46
|
issue 2
139-147
EN
An Agrobacterium-mediated transformation method of pea has been developed for several edible and fodder cultivars of pea (Pisum sativum L.), characterized previously in their potential for regeneration via organogenesis. The most appropriate explant, which was susceptible to Agrobacterium infection and capable of regenerating transgenic plants, turned out to be a slice of an immature embryo, including the embryo axis and the basal part of a cotyledon. Three hypervirulent strains of A. tumefaciens were tested: AgL0, AgL1 and EHA105. Each carried the binary vector pP35SGIB containing the uid gene, with an intron under control of the 35S promoter, and the bar gene conferring resistance to phosphinotricin. Strain AgL0 was found to be efficient for the majority of cultivars, followed by AgL1 and EHA105. Transformation efficiency varied from 0.7 to 4.1%, depending on cultivar and Agrobacterium strain. The transformation efficiency of particular pea cultivars did not clearly correspond to their regeneration capacity, which ? although indispensable ? was not a critical parameter of successful transformation. The presence of integrated genes in pea genomic DNA was detected by the PCR. T-DNA was stably transmitted to the progeny, as it was confirmed by Southern hybridization. The activity of introduced genes was analysed by the histochemical GUS assay and by painting leaves or by spraying transgenic plants with the herbicide Basta.
3

The TDP method of seed yield component analysis in grain legume breeding

75%
Golaszewski J., Idzkowska M., Milewska J.
Journal of Applied Genetics
|
1998
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vol. 39
|
issue 4
299-308
EN
The results of plant breeding trials with populations of fodder pea strains and broad bean hybrids were the basis of consideration on the interrelationship between some traits - the yield structure elements. Developed by Eaton, a relatively new method of yield component analysis called the two-dimensional partitioning method (TDP) was applied to analyse the data. The method, which combines multiple regression and ANOVA, allows for concise tabular presentation and simple interpretation of the distribution of traits in one direction and the sources of variance according to ANOVA model in the other direction. Additionally, the interpretation of the results was supported by such standard statistical techniques as ANOVA, simple and multiple regression and path analysis. The main components of pea yielding were plant height and the number of pods per plant. Among the analysed characters of broad bean the number of nodes with pods on the main stem, which turned out to be the determinant of broad bean yielding, might be strongly affected by environmental conditions. The number of nodes with pods might be considered a selecting character of high potential yielding of broad bean genotypes.
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