The use of the AFLP technique in combination with BSA for identification of markers linked with dwarfness in rye
Helena Kubicka
sekretariat@ob.pan.plOgród Botaniczny — Centrum Zachowania Różnorodności Biologicznej PAN, Warszawa (Poland)
Renata Lewandowska
Ogród Botaniczny — Centrum Zachowania Różnorodności Biologicznej PAN, Warszawa (Poland)
Abstract
The aim of this work was to search for potential molecular markers linked with the recessive dwarfness ds1 gene in rye, with the aid of the AFLP technique in combination with the Bulk Segregant Analysis (BSA) method. Over 22 thousand DNA fragments were identified — differentiating inbred lines of extreme phenotypes (8/40K — dwarfness and 8/40N — control) as well as the segregating population DWSK-1F2, using 239 selective primer pairs for restrictases EcoRI/Mse1. Next, the presence of common markers for the tested parental forms and F2 offsprings as well as parental forms of the mapping population DS2 and RXL10 was analyzed, which enabled reduction of the number of used starter pairs to 68. Further analysis of a narrowed segregating population (DWSK-1F2), consisting of 21 plants, limited the amount of primer pairs to 51.88 markers linked with the dwarfness gene were obtained, which segregated with theratio of 3:1 for χ2 = 0.50 at p = 0.01. They were tested again for 15 starter pairs on the mapping population. For 13 differentiating bands, the segregation of 3:1 was observed and these DNA fragments were classified as dominant AFLP markers linked with the researched dwarfness gene in rye. In the future the identified AFLP markers will be tested on a wider segregated population with the aim of transfering them to the existing genetic map of rye.
Keywords:
AFLP, BSA, dwarfness, molecular markers, ryeReferences
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Authors
Helena Kubickasekretariat@ob.pan.pl
Ogród Botaniczny — Centrum Zachowania Różnorodności Biologicznej PAN, Warszawa Poland
Authors
Renata LewandowskaOgród Botaniczny — Centrum Zachowania Różnorodności Biologicznej PAN, Warszawa Poland
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