Shoot organogenesis induction from genetically verified individuals of endangered bog pine (Pinus uncinata subsp. uliginosa)

Abstract

The objective of this work was to apply selected genetic markers to verify the genetic purity of bog pine (Pinus uncinata subsp. uliginosa) individuals from the Borkovicka blata population in order to initiate in vitro cultures from them. Key factors of shoot organogenesis induction from seedling shoot apices were examined. The plastid DNA analysis was used as a molecular-genetic tool. The trnL-trnF region of chloroplast DNA was tested by PCR-RFLP method; furthermore, the chloroplast microsatellite region Pt41093 was also analysed. All the tested individuals from the studied population matched the Pinus uncinata subsp. uliginosa haplotype. In bog pine, regeneration and development of axillary shoots were significantly enhanced in the presence of 5 g・l-1 activated charcoal on the WV5 and WPM (Duchefa) media. The highest proliferation (28-30%) of short shoots (brachyblasts) was observed on the WV5 medium in the absence of benzyladenine (BA) or at its low concentration only (0.1 mg・l-1).

Keywords

• dna extraction
• in vitro regeneration
• leaves
• plastid dna
• seedling shoot apices