In vitro propagation and assessment of genetic stability of acclimated plantlets of Cornus alba L. using RAPD and ISSR markers

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• 作者：Agnieszka Ilczuk ; Ewelina Jacygrad
• 关键词：Dogwood ; Micropropagation ; Molecular marker ; Rooting ; Survival rate
• 刊名：In Vitro Cellular & Developmental Biology - Plant
• 出版年：2016
• 出版时间：September 2016
• 年：2016
• 卷：52
• 期：4
• 页码：379-390
• 全文大小：993 KB
• 刊物主题：Plant Sciences; Cell Biology; Developmental Biology; Plant Breeding/Biotechnology; Plant Genetics & Genomics;
• 出版者：Springer US
• ISSN：1475-2689
• 卷排序：52

文摘

Cornus alba L. (white dogwood) is an important ornamental shrub having a wide range of applications such as reforestation programs and soil retention systems. The vegetative propagation of dogwood by cuttings may be slow, difficult, and cultivar dependent; therefore, an improved micropropagation method was developed. Nodal stem segments of C. alba cultivars ‘Aurea’ and ‘Elegantissima’ were cultured on media enriched with six different sources of macronutrients. Media were supplemented with either N6-benzyladenine (BA) or thidiazuron (TDZ) in combination with 1-naphthaleneacetic acid (NAA). Regardless of the cultivar, the best shoot proliferation was observed on Lloyd and McCown medium (woody plant medium (WPM)) at pH 6.2, containing 1.0 mg L−1 BA, 0.1 mg L−1 NAA, and 20–30 g L−1 sucrose. Rooting of regenerated shoots was achieved by an in vitro method when different concentrations of NAA or indole-3-butyric acid (IBA) were tested. Microcuttings were rooted for 8 wk on medium enriched with 0.25 mg L−1 NAA and potted into P9 containers in the greenhouse. The final survival rate of the plants after 20 wk was 80% for ‘Aurea’ and 90% for ‘Elegantissima’. Genetic stability of the micropropagated plants was confirmed by using two DNA-based molecular marker techniques. A total of 30 random amplified polymorphic DNA (RAPD) and 20 inter-simple sequence repeat (ISSR) primers resulted in 197–199 and 184–187 distinct and reproducible band classes, respectively, in ‘Aurea’ and ‘Elegantissima’ plantlets. All of the RAPD and ISSR profiles were monomorphic and comparable with the mother plant.