Micropropagation of tulip: production of virus-free stock plants

Methods Mol Biol. 2010:589:243-56. doi: 10.1007/978-1-60327-114-1_23.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acclimatization
  • Cell Culture Techniques*
  • Cell Proliferation
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression Regulation, Plant
  • Genetic Techniques
  • Genomic Instability
  • Phenylurea Compounds / pharmacology
  • Plant Growth Regulators / pharmacology
  • Plant Roots / growth & development
  • Plant Shoots / growth & development
  • Regeneration* / drug effects
  • Thiadiazoles / pharmacology
  • Time Factors
  • Tulipa / drug effects
  • Tulipa / genetics
  • Tulipa / growth & development*
  • Tulipa / virology

Substances

  • Phenylurea Compounds
  • Plant Growth Regulators
  • Thiadiazoles
  • thidiazuron