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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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Record number 432048
Title An Agrobacterium-mediated transformation system of pyrethrum (Tanacetum cinerariifolium) based on leaf explants
Author(s) Mao, J.; Cao, L.Y.; Kong, L.F.; Jongsma, M.A.; Wang, C.Y.
Source Scientia Horticulturae 150 (2013). - ISSN 0304-4238 - p. 130 - 134.
DOI https://doi.org/10.1016/j.scienta.2012.10.019
Department(s) PRI BIOS Applied Metabolic Systems
Publication type Refereed Article in a scientific journal
Publication year 2013
Keyword(s) chrysanthemyl diphosphate synthase - plant-growth regulators - tissue-culture - cinerariaefolium - identification - population - selection - protein - stress - yield
Abstract Pyrethrum (Tanacetum cinerariifolium) is well-known for the production of natural pyrethrin insecticides, which are isolated from the dried flower heads. To allow genetic analysis and improvement of pyrethrum in terms of yield and quality by molecular breeding a transformation protocol would be desirable. To achieve this, first an efficient direct shoot regeneration system was set up. Leaf explants formed shoots within 4 weeks on MS medium supplemented with 2.0 mg l-1 6-BA and 0.2 mg l-1 IBA. The best regeneration frequency obtained was 87% with an average number of 2 shoots per explant. Subsequently, a genetic transformation protocol was developed. Kanamycin added to the medium significantly reduced explant regeneration, and no green shoots were formed after 4 weeks incubation at 10 mg l-1 kanamycin. Severe phytotoxic effects of cefotaxime were observed at concentrations >300 mg l-1 to control Agrobacterium. Thus, 10 mg l-1 kanamycin and 200 mg l-1 cefotaxime were selected as optimal for genetic transformation. Agrobacterium-mediated genetic transformation was carried out with a GUS gene under the control of the chrysanthemum RbcS promoter. Two days of pre-culture and 5 min of infection were found to result in the most efficient transformation rates. Ninety-nine percent of leaf explants infected with Agrobacterium showed GUS expression 3–5 days after transfection and 12% of the regenerated shoots had a stable GUS-positive phenotype. The other plants had a transient kanamycin-resistant phenotype and eventually lost the NPT-II and GUS gene expression.
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