ISHS Acta Horticulturae 589: X International Asparagus Symposium
TRANSIENT GENE EXPRESSION IN MAIZE AND ASPARAGUS POLLEN USING MAGNETIC PARTICLES BY PARTICLE GUN |
| Authors: | H. Kakuta, T. Maeda, M. Akiyama, R. Hashimoto, Y. Horikawa |
| Keywords: | particle gun, magnetic-particles, Zea mays L., Asparagus officinalis L., pollen |
| Abstract:
Successful delivery of foreign genes to the pollen of maize and asparagus was achieved using an improved particle gun and magnetic selection of the pollen after bombardment with magnetic particles. The particle gun described by Kakuta et al. and either magnetite particles (Fe3O4) 0.3μ or 0.6μ diameter, or nickel and gold particles 1μm diameter were used. Plasmid pBI221, containing the GUS-gene under the control of the CaMV 35S promotor and NOS polyadenylation signal, was used to investigate transient gene expression in maize pollen. Initially the bombardment with magnetite resulted in a low frequency of expression. However, magnetic separation and concentration of the bombarded pollen increased the frequency compared with that obtained using gold particles. Following the delivery of the sGFP (S65T) plasmid, green fluorescent protein expression was observed in asparagus pollen after magnetic separation, although the frequency of pollen grains expressing sGFP was lower than on maize. Magnetic-microprojectiles could be useful to increasing the efficiency of pollen-mediated transformation for asparagus and other monocotyledons. | |
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