ISSN:
1432-203X
Keywords:
Key words Embryogenic suspension cultures
;
β-Glucuronidase
;
Microparticle bombardment
;
Image analysis
;
Ferricyanide
;
ferrocyanide
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Tissue derived from embryogenic suspension cultures of cassava was bombarded with microparticles coated with a plasmid containing the uidA gene, which codes for β-glucuronidase (GUS). After 3 days, the effect of different bombardment parameters was evaluated by comparing the numbers of blue spots that resulted from histological GUS assays. Counting of blue spots was performed using a system comprised of a black and white video camera, a stereoscope and a personal computer. A reproducible counting method was established by optimizing GUS assay conditions, preparation of tissue samples and acquisition of video images in view of attaining the highest possible contrast between the blue spots and the surrounding tissue. The effects of bombardment pressure, microparticle size, number of bombardments, and osmotic pretreatment on GUS expression were investigated. Optimal transient expression of the uidA gene was observed after bombardment at 1100 psi, with a particle size of 1 µm, an osmotic pretreatment and two bombardments per sample. The highest number of blue spots observed was 2400 per square centimeter of bombarded tissue.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01142317
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