Micropropagation of tulip: production of virus-free stock plants.
We describe here a new tulip micropropagation method based on the cyclic shoot multiplication in presence of the thidiazuron (TDZ), which enables the production of virus-free stock plants, speeds up breeding, and provides new genotypes for the market. In our novel protocol, cyclic shoot multiplication can be performed for 2-3 years by using TDZ instead of other cytokinins, as 6-benzylaminopurine (BAP) and N(6)-(-isopentyl)adenine (2iP). It makes possible to produce 500-2,000 microbulbs from one healthy plant. There are six main stages of tulip micropropagation. Stage 0 is the selection of true-to-type and virus-free plants, confirmed by ELISA. Fragments of flower stems isolated from bulbs are used as initial explants. Shoot multiplication is based on the regeneration of adventitious shoots, which are sub-cultured every 8 weeks. In the Stage 3, the specially prepared shoots are induced by low temperature treatment to form bulbs which finally develop on a sucrose-rich medium at 20 degrees C. Bulbs are then dried for 6 weeks and rooted in vivo. The number of multiplication subcultures should be limited to 5-10 cycles in order to lower the risk of mutation. Virus indexing should be repeated 3-4 times, at the initial stage and then during shoot multiplication. Genetic stability of micropropagated shoots can be confirmed using molecular markers.
Podwyszyńska
Małgorzata
M
Research Institute of Pomology and Floriculture, Skierniewice, Poland.
Sochacki
Dariusz
D
D000064:Acclimatization; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D004797:Enzyme-Linked Immunosorbent Assay; D018506:Gene Expression Regulation, Plant; D005821:Genetic Techniques; D042822:Genomic Instability; D010671:Phenylurea Compounds / Q000494:pharmacology; D010937:Plant Growth Regulators / Q000494:pharmacology; D018517:Plant Roots / Q000254:growth & development; D018520:Plant Shoots / Q000254:growth & development; D012038:Regeneration / Q000187:drug effects; D013830:Thiadiazoles / Q000494:pharmacology; D013997:Time Factors; D031447:Tulipa / Q000187:drug effects / Q000235:genetics / Q000254:growth & development* / Q000821:virology
D010671:Phenylurea Compounds; D010937:Plant Growth Regulators; D013830:Thiadiazoles; C016785:thidiazuron