Summary
The ability to recover male gametophyte derived plants, which is necessary to get transformed haploid plants, was verified for a hybrid of maize. Using the isolated microspore culture technique, a 9 × 10−5 plant regeneration frequency was obtained. Maize microspores were bombarded with tungsten particles using a PDS He/1000 apparatus. GUS expression in the microspores was maximum with 1.1 μm diameter tungsten microprojectiles for 1100 and 1350 psi helium pressures at a 6 cm distance between the launch point and the target cells. Increasing the amount of DNA coated on the microparticles from 1.66 to 4 μg DNA/mg of particles allowed a two-fold and four-fold increase of the GUS-expressing microspore frequency for 1100 and 1350 psi helium pressure bombardment, respectively. Optimal concentration of solidifying agent in the bombardment support culture medium was found to be 1%. Cell density ranging from 25000 microspores/bombardment to 100000 microspores/bombardment did not affect the frequency of GUS-expressing microspores. Using these optimal conditions, the maximum frequency of GUS-expressing microspores was found to be about 9 × 10−4, while maintaining an embryo formation frequency about 5 × 10−4.
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Abbreviations
- GUS:
-
β-glucuronidase
- PEG:
-
polyethylene glycol
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Jardinaud, M.F., Souvré, A., Alibert, G. et al. uidA gene transfer and expression in maize microspores using the biolistic method. Protoplasma 187, 138–143 (1995). https://doi.org/10.1007/BF01280242
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DOI: https://doi.org/10.1007/BF01280242