Regeneration of transformed shoots from electroporated soybean (Glycine max (L.) Merr.) protoplasts View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1991-06

AUTHORS

Sarwan K. Dhir, Seema Dhir, Ann Pizanis Sturtevant, Jack M. Widholm

ABSTRACT

Stable transformation of soybean (Glycine max (L.) Merr.) protoplasts isolated from immature cotyledons was achieved following electroporation with plasmid DNA carrying chimeric genes encoding ß-glucuronidase (GUS) and hygromycin phosphotransferase (HPT) under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Transformed colonies were stringently selected by growing 15-day-old protoplast-derived cells in the presence of 40 μg/ml of hygromycin-B for 6 weeks. Over 93% of the resistant cells and colonies exhibited GUS activity, indicating that the two marker genes borne on a single plasmid were co-introduced and co-expressed at a very high freguency. This transformation procedure reproducibly yields transformants at frequencies of 2.9-6.8 × 10(-4) (based on the number of protoplasts electroporated) or 23.0% (based on the number of control microcalli formed) counted after 6 weeks of selection. After repeated subculturing on regeneration medium, shoots were induced from 8.0% of the transformed calli. Southern hybridization confirmed the presence of both the GUS and hygromycin genes in the transformed calli and shoots. More... »

PAGES

97-101

Journal

TITLE

Plant Cell Reports

ISSUE

2

VOLUME

10

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf00236466

DOI

http://dx.doi.org/10.1007/bf00236466

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1006510759

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/24221403


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