Genetically directed differential subtraction chain products related to in vitro response of immature embryos of rye (Secale cereale L.): isolation, characterization, and expression analysis

作者：Hromada Judycka Aneta; Bolibok Bragoszewska Hanna; Rakoczy Trojanowska Monika^*

来源：Plant Cell, Tissue and Organ Culture, 2010, 100(2): 131-138.

DOI：10.1007/s11240-009-9626-7

• Genetically Directed Differential Subtraction Chain
• Somatic embryogenesis
• Tissue culture response
• Immature embryos

Winter rye (Secale cereale L.) is known to be recalcitrant to tissue culture response (TCR). Moreover, the mechanisms controlling TCR are poorly recognized. In the present study, a Genetically Directed Differential Subtraction Chain (GDDSC) strategy was used to isolate genomic regions associated with TCR. Two pairs of bulks, R-NR and E > 90-E < 25, were prepared, and for each pair, the bulk was used both as a tester and as a driver. After eight rounds of subtraction, 45 unique GDDSC products were obtained. To verify the connection between GDDSCs and TCR two approaches were applied: Real-Time RT-PCR analysis and genetic mapping. The expression profiles of four out of six investigated products agrees with the phenotype and subtraction direction. Two from the developed GDDSC-SCAR markers showed polymorphism in lines L9 and L318, the parental components of a mapping population. The polymorphic SCAR GDDSC markers were mapped on the rye chromosome 4R (SCAR-GDDSC NR 440BamHI) and 6R (SCAR-GDDSC E < 25 340BamHI). The marker E < 25_340B9 was localized in the border region of QTL for embryogenic callus production.

• 出版日期2010-2