Improved efficiency of the walnut somatic embryo gene transfer system View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1990-01

AUTHORS

Gale H. McGranahan, Charles A. Leslie, Sandra L. Uratsu, Abhaya M. Dandekar

ABSTRACT

AnAgrobacterium-mediated gene transfer system which relies on repetitive embryogenesis to regenerate transgenic walnut plants has been made more efficient by using a more virulent strain ofAgrobacterium and vectors containing genes for both kanamycin resistance and beta-glucuronidase (GUS) activity to facilitate early screening and selection. Two plasmids (pCGN7001 and pCGN7314) introduced individually into the disarmedAgrobacterium host strain EHA101 were used as inoculum. Embryos maintained on medium containing 100 mg/l kanamycin after co-cultivation produced more transformed secondary embryos than embryos maintained on kanamycin-free medium. Of the 186 GUS-positive secondary embryo lines identified, 70% were regenerated from 3 out of 16 primary embryos inoculated with EHA101/pCGN7314 and grown on kanamycin- containing medium, 28% from 4 out of 17 primary embryos inoculated with EHA101/ pCGN7001 and grown on kanamycin medium, and 2% from one out of 13 primary embryos inoculated with EHA101/pCGN7001 but not exposed to kanamycin. Because kanamycin inhibits but does not completely block new embryo formation in controls, identification of transformants formerly required repetitive selection on kanamycin for several months. Introduction of the GUS marker gene allowed positive identification of transformant secondary embryos as early as 5-6 weeks after inoculation. DNA analysis of a representative subset of lines (n=13) derived from secondary embryos confirmed transformation and provided evidence for multiple insertion events in single inoculated primary embryos. More... »

PAGES

512-516

References to SciGraph publications

Journal

TITLE

Plant Cell Reports

ISSUE

9

VOLUME

8

Author Affiliations

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf00820198

DOI

http://dx.doi.org/10.1007/bf00820198

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1013012609

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/24226275


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