Transient expression of the β-glucuronidase gene in embryogenic callus of Picea mariana following microprojection.
A microprojection protocol using the DuPont Biolistic(™) particle delivery system and the β-glucuronidase (GUS) reporter gene fused with the 35S promoter of Cauliflower mosaic virus (CaMV) was developed for Picea mariana callus. Comparison of four tungsten microprojectile sizes showed the highest transient gene expression with 1.11μm diameter particles. Adsorption of DNA on the microcarriers using calcium chloride led to higher GUS gene activity than using polyethylene glycol. GUS gene activity in P. mariana was the highest when cells were treated 5 and 6 days after subculturing to fresh media. The wheat ABA-inducible Em gene promoter yielded 4.5 times higher GUS gene activity than the 35S CaMV promoter. Comparison of transient GUS gene expression among 10 P. mariana embryogenic cell lines from six different open-pollinated families showed comparable gene activity, with the exception of one family showing no GUS gene activity.
Forestry Canada, Petawawa National Forestry Institute, P.O. Box 2000, KOJ 1JO, Chalk River, Ontario, Canada.
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Duchesne
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PJ
Charest