Direct Agrobacterium tumefaciens-mediated transformation of Hyoscyamus muticus hairy roots using green fluorescent protein

Colleen D. Merritt, Surabhi Raina, Nina Fedoroff, Wayne R. Curtis

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Hyoscyamus muticus hairy root segments were infected with Agrobacterium tumefaciens ASE containing the binary vector pCGN1548 with a green fluorescent protein (GFP) reporter gene under the control of the CaMV 35S promoter. The roots were incubated on callus-inducing medium to generate transformed cells. Transformants were selected on medium containing 50 and 100 mg/L kanamycin and screened by visual inspection for GFP expression. Highly fluorescent cells were incubated on phytohormone-free medium for regeneration of the hairy root phenotype. This infection technique can be applied directly to existing hairy root cultures which have been previously characterized and selected for desirable physiological traits. These studies also indicate that GFP is not toxic to H. muticus plant tissue and that H. muticus hairy roots have minimal autofluorescence which allows for clear observation of GFP.

Original languageEnglish (US)
Pages (from-to)278-282
Number of pages5
JournalBiotechnology progress
Volume15
Issue number2
DOIs
StatePublished - Mar 1 1999

Fingerprint

Hyoscyamus
Hyoscyamus muticus
Agrobacterium tumefaciens
Green Fluorescent Proteins
Agrobacterium radiobacter
green fluorescent protein
Kanamycin
Plant Growth Regulators
Poisons
Bony Callus
Reporter Genes
Regeneration
kanamycin
reporter genes
plant hormones
Observation
plant tissues
Phenotype
callus
protein synthesis

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

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abstract = "Hyoscyamus muticus hairy root segments were infected with Agrobacterium tumefaciens ASE containing the binary vector pCGN1548 with a green fluorescent protein (GFP) reporter gene under the control of the CaMV 35S promoter. The roots were incubated on callus-inducing medium to generate transformed cells. Transformants were selected on medium containing 50 and 100 mg/L kanamycin and screened by visual inspection for GFP expression. Highly fluorescent cells were incubated on phytohormone-free medium for regeneration of the hairy root phenotype. This infection technique can be applied directly to existing hairy root cultures which have been previously characterized and selected for desirable physiological traits. These studies also indicate that GFP is not toxic to H. muticus plant tissue and that H. muticus hairy roots have minimal autofluorescence which allows for clear observation of GFP.",
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Direct Agrobacterium tumefaciens-mediated transformation of Hyoscyamus muticus hairy roots using green fluorescent protein. / Merritt, Colleen D.; Raina, Surabhi; Fedoroff, Nina; Curtis, Wayne R.

In: Biotechnology progress, Vol. 15, No. 2, 01.03.1999, p. 278-282.

Research output: Contribution to journalArticle

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