Stable transformation of Picea glauca by particle acceleration

By defining the somatic embryo developmental stage which expressed beta -glucuronidase (GUS) at a high level yet was also competent to form embryogenic callus at a high frequency under selection, we obtained transformed Picea glauca (white spruce) embryogenic callus, embryos and seedlings expressing...

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Bibliographic Details
Published in:Bio/technology (New York, N.Y. 1983) N.Y. 1983), 1993-01, Vol.11 (1), p.84-89
Main Authors: Ellis, D.D. (University of Wisconsin-Madison, Madison, WI), McCabe, D.E, McInnis, S, Ramachandran, R, Russell, D.R, Wallace, K.M, Martinell, B.J, Roberts, D.R, Raffa, K.F, McCown, B.H
Format: Article
Language:English
Subjects:
BACILLUS THURINGIENSIS
BACTERIAL TOXINS
BETA-GLUCURONIDASE
Biological and medical sciences
Biotechnology
CAL
CALLO
CALLUS
CHORISTONEURA FUMIFERANA
DESARROLLO EMBRIONARIO
DEVELOPPEMENT EMBRYONNAIRE
DIRECT DNA UPTAKE
DISEASE RESISTANCE
EMBRIONES VEGETALES
EMBRYON VEGETAL
EMBRYONIC DEVELOPMENT
ENDOTOXINAS
ENDOTOXINE
ENDOTOXINS
FOSFOTRASFERASA
Fundamental and applied biological sciences. Psychology
GENE
GENES
Genetic engineering
Genetic technics
GENETIC TRANSFORMATION
GENETICA
GENETICS
GENETIQUE
GLICOSIDASAS
GLYCOSIDASE
GLYCOSIDASES
HISTOENZYMOLOGY
Methods. Procedures. Technologies
neomycin phosphotransferase ii
PHOSPHOTRANSFERASE
PHOSPHOTRANSFERASES
PICEA GLAUCA
PLANT EMBRYOS
PLANTAS TRANSGENICAS
PLANTE TRANSGENIQUE
REPORTER GENES
RESISTANCE AUX MALADIES
RESISTENCIA A LA ENFERMEDAD
somatic embryos
TOXINAS
TOXINE
TOXINS
TRANSFORMACION GENETICA
TRANSFORMATION GENETIQUE
Transgenic animals and transgenic plants
TRANSGENIC PLANTS
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Description
Summary:By defining the somatic embryo developmental stage which expressed beta -glucuronidase (GUS) at a high level yet was also competent to form embryogenic callus at a high frequency under selection, we obtained transformed Picea glauca (white spruce) embryogenic callus, embryos and seedlings expressing GUS in all cells. Plasmid DNA, containing three chimeric constructs (enhanced cauliflower mosaic virus (CaMV) 35s-GUS, nopaline synthase-neomycin phosphotransferase (NPTII), and CaMV 35s-Bacillus thuringiensis (B.t. ) cryIA endotoxin) was introduced into four developmental stages of white spruce somatic embryos by particle acceleration. Transient expression was observed in all of the stages of somatic embryos tested, but transformed embryogenic callus was induced only from the two most advanced. Embryogenic callus was identified by histochemical staining for GUS as early as 6 weeks following particle acceleration. All GUS positive embryogenic callus lines also showed NPTII activity. Incorporation of the introduced genes into the genome was confirmed by PCR and Southern blot analysis of embryogenic callus and regenerated transformed plants. Plants derived from several transformed embryogenic callus lines are currently undergoing acclimatization in the greenhouse. Sprue budworm (Chorisoneura fumiferana ) feeding trials with embryogenic callus and transformed seedlings indicate a low, sublethal level of B.t. expression.
ISSN:0733-222X
1087-0156
2331-3684
1546-1696
DOI:10.1038/nbt0193-84