Articles
SOMACLONAL VARIATION IN MICROPROPAGATED TULIPS DETERMINED BY PHENOTYPE AND DNA MARKERS
Article number
714_23
Pages
211 – 220
Language
English
Abstract
The new protocol for tulip micropropagation by cyclic multiplication of adventitious shoots, in the presence of thidiazuron, was developed.
The aim of the study was to verify this method with respect to the genetic stability of the plants produced.
The genetic fidelity of the micropropagated plants was evaluated by phenotypic observation and DNA analysis with RAPD (random amplified polymorphic DNA) and ISSR (inter-simple sequence repeat) techniques.
At first, several genotypes: Blue Parrot, Prominence, their sports and Giewont were used for analysis.
The distantly related cultivars were easily distinguished in both reactions.
But within the family of sports, both RAPD and ISSR analyses did not reveal polymorphism.
Somaclonal variation in micropropagated tulips of Blue Parrot was performed on 22 genotypes, including original cultivar as standard (plant propagated conventionally), micropropagated off-types and randomly selected true-to-types.
Within the flowering plants, no phenotypic off-types were noted in the progeny-line derived from a two-year-old culture and polymorphism on DNA level was not detected.
But in the progeny-lines derived from a four-year-old culture, all plants had a changed color of flowers from purple-violet to red purple.
Within the juvenile plants, derived from 4- to 7-year-old cultures, the off-types with variegated or aberrant leaves occurred.
RAPD and ISSR analyses revealed that this phenotypic variation resulted from genetic changes.
In conclusion: 1) the propagation of tulips with our method can increase the risk of mutation, especially when shoots are micropropagated for more than four years; 2) limiting the period of in vitro propagation to three years and monitoring of the plant material with DNA markers would reduce the risk of production of mutated plants; 3) some somaclones seem to be an interesting material for tulip breeding.
The aim of the study was to verify this method with respect to the genetic stability of the plants produced.
The genetic fidelity of the micropropagated plants was evaluated by phenotypic observation and DNA analysis with RAPD (random amplified polymorphic DNA) and ISSR (inter-simple sequence repeat) techniques.
At first, several genotypes: Blue Parrot, Prominence, their sports and Giewont were used for analysis.
The distantly related cultivars were easily distinguished in both reactions.
But within the family of sports, both RAPD and ISSR analyses did not reveal polymorphism.
Somaclonal variation in micropropagated tulips of Blue Parrot was performed on 22 genotypes, including original cultivar as standard (plant propagated conventionally), micropropagated off-types and randomly selected true-to-types.
Within the flowering plants, no phenotypic off-types were noted in the progeny-line derived from a two-year-old culture and polymorphism on DNA level was not detected.
But in the progeny-lines derived from a four-year-old culture, all plants had a changed color of flowers from purple-violet to red purple.
Within the juvenile plants, derived from 4- to 7-year-old cultures, the off-types with variegated or aberrant leaves occurred.
RAPD and ISSR analyses revealed that this phenotypic variation resulted from genetic changes.
In conclusion: 1) the propagation of tulips with our method can increase the risk of mutation, especially when shoots are micropropagated for more than four years; 2) limiting the period of in vitro propagation to three years and monitoring of the plant material with DNA markers would reduce the risk of production of mutated plants; 3) some somaclones seem to be an interesting material for tulip breeding.
Authors
M. Podwyszynska, K. Niedoba, M. Korbin, A. Marasek
Keywords
Tulipa, genetic stability, in vitro culture, RAPD, ISSR
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