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An efficient particle bombardment system for the genetic transformation of asparagus (Asparagus officinalis L.)

Author:
Cabrera-Ponce, J.L., Lopez, L., Assad-Garcia, N., Medina-Arevalo, C., Bailey, A.M., Herrera-Estrella, L.
Source:
Plant cell reports 1997 v.16 no.5 pp. 255-260
ISSN:
0721-7714
Subject:
Asparagus officinalis, herbicide resistance, plant breeding, embryo (plant), callus, genetic transformation, DNA, gene transfer, methodology, transgenic plants, cultivars, genetic variation, plasmids, genetic code, phosphotransferases (phosphomutases), glufosinate, acyltransferases, beta-glucuronidase, culture media, gene expression, enzyme activity, histochemistry
Abstract:
The microprojectile bombardment method was used to transfer DNA into embryogenic callus of asparagus (Asparagus officinalis L.) and to produce stably transformed asparagus plants. Embryogenic callus, derived from UC157 and UC72 asparagus cultivars, was bombarded with tungsten particles coated with plasmid DNA that contained genes encoding hygromycin phosphotransferase, phosphinothricin acetyl transferase and beta-glucuronidase. Putatively transformed calli were identified from the bombarded tissue after 4 months selection on 25 mg/L hygromycin B plus 4 mg/L phosphinothricin (PPT). By selecting embryogenic callus on hygromycin plus PPT the overall transformation and selection efficiencies were substantially improved over selection with hygromycin or PPT alone, where no transgenic clones were recovered. The transgenic nature of the selected material was demonstrated by GUS histochemical assays and Southern blot hybridization analysis. Transgenic asparagus plants were found to withstand the prescribed levels of the PPT-based herbicide BASTA for weed control.
Agid:
1436761