Publication Date
1993
Description
Dactylis glomerata L., cv, Embryogen-P, has been previously genetically transformed by direct uptake of DNA by suspension culture-derived protoplasts. The objective of the present experiments was 10 use this genotype to obtain transformation by microprojec1ile bombardment. Bmbryogenic suspension cultures and plated basal leaf segments were bombarded with DNA coated tungsten particles propelled by two different helium pressure discharge apparatuses. The DNA plasmids contained genes for chlorosulfuron resistance, hygromycin resistance and !i-glucuronidase (GUS). Transient expiession for GUS was obtained on both suspension cultures and leaf segments. A stable event for GUS and 16 plants which rooted well on 2.5 mg/I chlorosulfuron were obtained from leaf segments. This method of transformation is preferable lo DNA uptake by protoplasls because of an easier and more efficient protocol. II also avoids difficulties in proloplast regeneration and abnormalities often present in protoplast derived plants.
Citation
Conger, B V.; Songstand, D D.; McDaniel, J K.; and Bond, J, "Genetic Transformation of Dactylis glomerata by Microprojectile Bombardment" (2024). IGC Proceedings (1993-2023). 10.
https://uknowledge.uky.edu/igc/1993/session27/10
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Genetic Transformation of Dactylis glomerata by Microprojectile Bombardment
Dactylis glomerata L., cv, Embryogen-P, has been previously genetically transformed by direct uptake of DNA by suspension culture-derived protoplasts. The objective of the present experiments was 10 use this genotype to obtain transformation by microprojec1ile bombardment. Bmbryogenic suspension cultures and plated basal leaf segments were bombarded with DNA coated tungsten particles propelled by two different helium pressure discharge apparatuses. The DNA plasmids contained genes for chlorosulfuron resistance, hygromycin resistance and !i-glucuronidase (GUS). Transient expiession for GUS was obtained on both suspension cultures and leaf segments. A stable event for GUS and 16 plants which rooted well on 2.5 mg/I chlorosulfuron were obtained from leaf segments. This method of transformation is preferable lo DNA uptake by protoplasls because of an easier and more efficient protocol. II also avoids difficulties in proloplast regeneration and abnormalities often present in protoplast derived plants.