PulseExploreJournal ClubDebatesTrendingResearchersJournals
Instagram
HomeExploreJournal ClubTrending
Synapse
⌘+K
Synapse
August 1, 1988Proceedings of the National Academy of Sciences1,285 citationsOpen Access

Agrobacterium tumefaciens -mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection

View Full Paper
DVDirk ValvekensMMMarc Van MontaguMLMieke Van Lijsebettens

Key Points

  • To develop an efficient protocol for shoot regeneration and Agrobacterium tumefaciens-mediated genetic transformation of Arabidopsis thaliana root explants.
  • Optimized culture conditions to induce rapid shoot regeneration from Arabidopsis thaliana root cuttings.
  • Inoculated root explants with Agrobacterium tumefaciens harboring Ti plasmids encoding chimeric neo and bar marker genes under kanamycin selection across three ecotypes.
  • Root cuttings regenerated shoots at 100% efficiency, yielding fertile, seed-bearing plants in vitro or in soil.
  • Transgenic seed-producing plants were recovered with 20% to 80% transformation efficiency within 3 months of gene transfer.
  • F1 seedlings exhibited Mendelian segregation of kanamycin resistance, and expression of the herbicide-resistance bar gene was verified via enzymatic assay.

Abstract

Culture conditions were developed that induce Arabidopsis thaliana (L.) Heynh. root cuttings to regenerate shoots rapidly and at 100% efficiency. The shoots produce viable seeds in vitro or after rooting in soil. A transformation procedure for Arabidopsis root explants based on kanamycin selection was established. By using this regeneration procedure and an Agrobacterium tumor-inducing Ti plasmid carrying a chimeric neomycin phosphotransferase II gene ( neo ), transformed seed-producing plants were obtained with an efficiency between 20% and 80% within 3 months after gene transfer. F 1 seedlings of these transformants showed Mendelian segregation of the kanamycin-resistance trait. The transformation method could be applied to three different Arabidopsis ecotypes. In addition to the neo gene, a chimeric bar gene conferring resistance to the herbicide Basta was introduced into Arabidopsis . The expression of the bar gene was shown by enzymatic assay.

Ask AI
Helpful
Bookmark
Share
View Full Paper

Cite This Study

Valvekens et al. (1988) studied this question.

synapsesocial.com/papers/6a08ef4727ceb0c2a2d62000https://doi.org/10.1073/pnas.85.15.5536
Ask AI
Helpful
Bookmark
Share
View Full Paper