In vitro Propagation of Puya berteroniana and Assessment of Genetic Stability Using Molecular Markers
Petra Štréblová1, Iva Viehmannová1, Petra Hlásná Cepková1, Jana Andrlova1, Jan Vitamvas2
1Czech University of Life Sciences Prague, Fac. of Tropical Agrisciences, Dept. of Crop Sciences and Agroforestry, Czech Republic
Puya berteroniana belonging to the family Bromeliaceae is a giant plant with very attractive turquoise flowers, that have a great potential to be used for ornamental purposes. The aim of this study was the development of an optimisation protocol for in vitro propagation and assessment of the genetic fidelity of this plant using inter simple sequence repeat (ISSR) molecular markers. An efficient protocol for in vitro propagation via direct morphogenesis was succesfully established. Plant growth regulators as benzylaminopurine (BAP) and zeatin alone or in combination with α–naphthaleneacetic acid (NAA) were examined for their effects on in vitro offsets induction. Media containing NAA or indole-3-acetic acid (IAA) were used to optimise in vitro rooting of plantlets. For in vitro propagation, the most effective treatment was 1/2 MS medium enriched by 0.1 mg l-1 BAP and for in vitro rooting medium containing 0.3 mg l-1 NAA was optimal. Genetic stability of the in vitro regenerated plants was determined by using ISSR molecular markers. Out of 170 in vitro plantlets, 10 randomly chosen plants from the most effective treatment supplemented by 0.1 mg l-1 BAP were used for DNA screening. 6 ISSR primers produced totally 19 clear and reproducible bands resulting in a total of 253 bands where the banding pattern per each primer was highly uniform and identical to the mother plant from which the tissue cultures had been established. The occurrence of somaclonal variation was not detected trough screening of in vitro regenerants and thus, process of in vitro production can be used for mass production of P. berteroniana.
Keywords: Direct morphogenesis, genetic stability, molecular markers, plant growth regulators, Puya berteroniana
Contact Address: Petra Štréblová, Czech University of Life Sciences Prague, Fac. of Tropical Agrisciences, Dept. of Crop Sciences and Agroforestry, Pod Bendovkou 1776, 26901 Rakovník, Czech Republic, e-mail: petrastreblovagmail.com