ISHS Acta Horticulturae 572: XX International Eucarpia Symposium, Section Ornamentals, Strategies for New Ornamentals - Part II
TRANSFORMATION OF CHRYSANTHEMUM (DENDRANTHEMA GRANDI-FLORUM (RAMAT.) KITAMURA) VIA AGROBACTERIUM TUMEFACIENS) |
| Authors: | S. Kudo, N. Shibata, Y. Kanno, M. Suzuki |
| Keywords: | co-cultivation, gene silencing, β-glucuronidase |
| Abstract:
This study attempted transformation of chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) and studied several factors affecting stable transformation of chrysanthemum, including cultivars of chrysanthemum, strains of Agrobacterium tumefaciens, promoters, and conditions of the co-cultivation period. Four days of co-cultivation at 24°C were optimal using A. tumefaciens strain EHA101 and plasmid pIG121Hm. Integration of the transgenes was confirmed by Southern blot analyses about four transformation. The neomycin phosphotransferase gene (nptII) was integrated in all transformants which were selected by kanamycin resistance. One transformant lacked both the  -glucuronidase gene (uidA) and hygromycin B phoshphotransferase gene (hpt), while the other transformants contained all three transgenes.
RNA gel-blot analyses revealed the presence of nptII transcripts in two transformants that were resistant to kanamycin in the rooting test.
Transcripts of the uidA and hpt genes were not detected in any of the transformants, suggesting that differences in promoters or introduced genes might significantly affect expression.
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