Articles
HERBICIDE RESISTANCE IN ASPARAGUS: IN VITRO CELL SELECTION AND CHARACTERIZATION
Article number
479_14
Pages
115 – 120
Language
Abstract
In vitro cell selection is a feasible method for identifying novel mutants in plants.
Unlike other crops, this approach has not been well established for asparagus.
We report the development of in vitro cell selection techniques to identify mutant asparagus cells with resistance to a sulfonylurea herbicide.
To achieve this we have established cell culture systems for cell selection; defined the toxic concentration for complete growth inhibition of wild-type asparagus cells; isolated and characterized rare, resistant cell colonies; and regenerated plants.
We have used asparagus genotype CRD 168 (a female clone from Limbras I), previously identified as being highly amenable to cell culture.
Friable callus induced from etiolated shoots in darkness was used to initiate cell suspension cultures.
Two ml of single cell suspension was plated onto a cell culture medium supplemented with 8nM (toxic concentration) chlorsulfuron, resulting in 4×105 cells per Petri dish.
Of the 124 Petri dishes established in this manner, 165 growing cell colonies were identified.
Immediate characterization of these selected cell colonies yielded 24 escapes, 98 unstable variants, and 43 resistant variants.
Callus cultures from 34 of these stable variants have retained resistance following 11 months growth in the absence of the selection agent.
Plants have been regenerated from 36 of these stable herbicide-resistant variants.
Unlike other crops, this approach has not been well established for asparagus.
We report the development of in vitro cell selection techniques to identify mutant asparagus cells with resistance to a sulfonylurea herbicide.
To achieve this we have established cell culture systems for cell selection; defined the toxic concentration for complete growth inhibition of wild-type asparagus cells; isolated and characterized rare, resistant cell colonies; and regenerated plants.
We have used asparagus genotype CRD 168 (a female clone from Limbras I), previously identified as being highly amenable to cell culture.
Friable callus induced from etiolated shoots in darkness was used to initiate cell suspension cultures.
Two ml of single cell suspension was plated onto a cell culture medium supplemented with 8nM (toxic concentration) chlorsulfuron, resulting in 4×105 cells per Petri dish.
Of the 124 Petri dishes established in this manner, 165 growing cell colonies were identified.
Immediate characterization of these selected cell colonies yielded 24 escapes, 98 unstable variants, and 43 resistant variants.
Callus cultures from 34 of these stable variants have retained resistance following 11 months growth in the absence of the selection agent.
Plants have been regenerated from 36 of these stable herbicide-resistant variants.
Publication
Authors
D. Ganeshan, A.J. Conner, A.G. Fautrier
Keywords
Asparagus officinalis, chlorsulfuron, sulfonylurea, tissue culture
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