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Evaluation of a cotyledonary node regeneration system for Agrobacterium-mediated transformation of pea (Pisum sativum L.)

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1007/BF02632256
AuthorSearch for: 1; Search for: 1
Affiliation
  1. National Research Council of Canada. NRC Plant Biotechnology Institute
FormatText, Article
Subjecttransformation; pea; Pisum sativum; Agrobacterium tumefaciens; regeneration; progeny testing
Abstract
A rapid regeneration system was used for studies of Agrobacterium-mediated transformation in Pisum sativum L. Cotyledonary node explants were inoculated with Agrobacterium tumefaciens strains containing binary vectors carrying genes for nopaline synthase (NOS),β-glucuronidase (GUS), and neomycin phosphotransferase (NPTII) and placed on selection medium containing either 75 or 150 mg/liter kanamycin. A GUS encoding gene (uidA) containing an intron was used to monitor gene expression from 6 to 21 days postinoculation. GUS activity could be observed 6 days after inoculation in the area of the explant in which regeneration-occurred. Regenerating tissue containing transformed cells was observed in explants on selection medium 21 days postinoculation. Using this system, a single transgenic plant was obtained. Progeny of this plant, which contained two T-DNA inserts, demonstrated segregation for the inserts and for expression of the NOS gene in the selfed R₁ progeny. NPTII activity was observed in the R2 generation, indicating inheritance and expression of the foreign DNA over at least two generations. Attempts to repeat this procedure were unsuccessful.
Publication date
PublisherSpringer Nature
In
  • In Vitro Cellular & Developmental Biology - Plant 29, no. 2 (April 1993): 77–82.
LanguageEnglish
Peer reviewedYes
IdentifierBF02632256
NRCC-35897
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Record created2023-11-06
Record modified2023-11-06
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