Stable transformation of barley callus using biolistic® particle bombardment and the phosphinothricin acetyltransferase (bar) gene

Carol M. Stiff; Andrzej Kilian; Huaping Zhou; David A. Kudrna; Andris Kleinhofs

doi:10.1007/BF00048130

Stable transformation of barley callus using biolistic^® particle bombardment and the phosphinothricin acetyltransferase (bar) gene

Carol M. Stiff, Andrzej Kilian, Huaping Zhou, David A. Kudrna, Andris Kleinhofs

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

Suspension culture cells of barley (Hordeum vulgare L. cv. Klages) were collectec on filters and bombarded with gold particles carrying the Basta^® resistance (bar) gene, or both bar and the β-glucuronidase (uidA) genes in a Biolistic^® Particle Accelerator. Filters carrying the bombarded tissues were selected on basal medium with increasing concentrations (maximum of 200 mg I^-1) of phosphinothricin. Surviving calli were assayed for the presence of phosphinothricin acetyltransferase activity. All positive calli were shown to have the bar gene integrated into their genome. Nine independent stable transformation events have been analyzed with substantial variation in the integration pattern and transgene copy number. Some albino plantlets were produced, but no mature plants were obtained.

Original language	English (US)
Pages (from-to)	243-248
Number of pages	6
Journal	Plant Cell, Tissue and Organ Culture
Volume	40
Issue number	3
DOIs	https://doi.org/10.1007/BF00048130
State	Published - Mar 1995
Externally published	Yes

Keywords

Hordeum vulgare L.
accelerated particle
integration
methylation
suspension culture

ASJC Scopus subject areas

Horticulture

Access to Document

10.1007/BF00048130

Cite this

@article{dec8e438e9e745548e3640b134f851dd,

title = "Stable transformation of barley callus using biolistic{\textregistered} particle bombardment and the phosphinothricin acetyltransferase (bar) gene",

abstract = "Suspension culture cells of barley (Hordeum vulgare L. cv. Klages) were collectec on filters and bombarded with gold particles carrying the Basta{\textregistered} resistance (bar) gene, or both bar and the β-glucuronidase (uidA) genes in a Biolistic{\textregistered} Particle Accelerator. Filters carrying the bombarded tissues were selected on basal medium with increasing concentrations (maximum of 200 mg I-1) of phosphinothricin. Surviving calli were assayed for the presence of phosphinothricin acetyltransferase activity. All positive calli were shown to have the bar gene integrated into their genome. Nine independent stable transformation events have been analyzed with substantial variation in the integration pattern and transgene copy number. Some albino plantlets were produced, but no mature plants were obtained.",

keywords = "Hordeum vulgare L., accelerated particle, integration, methylation, suspension culture",

author = "Stiff, {Carol M.} and Andrzej Kilian and Huaping Zhou and Kudrna, {David A.} and Andris Kleinhofs",

year = "1995",

month = mar,

doi = "10.1007/BF00048130",

language = "English (US)",

volume = "40",

pages = "243--248",

journal = "Plant Cell, Tissue and Organ Culture",

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publisher = "Springer Science and Business Media B.V.",

number = "3",

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TY - JOUR

T1 - Stable transformation of barley callus using biolistic® particle bombardment and the phosphinothricin acetyltransferase (bar) gene

AU - Stiff, Carol M.

AU - Kilian, Andrzej

AU - Zhou, Huaping

AU - Kudrna, David A.

AU - Kleinhofs, Andris

PY - 1995/3

Y1 - 1995/3

N2 - Suspension culture cells of barley (Hordeum vulgare L. cv. Klages) were collectec on filters and bombarded with gold particles carrying the Basta® resistance (bar) gene, or both bar and the β-glucuronidase (uidA) genes in a Biolistic® Particle Accelerator. Filters carrying the bombarded tissues were selected on basal medium with increasing concentrations (maximum of 200 mg I-1) of phosphinothricin. Surviving calli were assayed for the presence of phosphinothricin acetyltransferase activity. All positive calli were shown to have the bar gene integrated into their genome. Nine independent stable transformation events have been analyzed with substantial variation in the integration pattern and transgene copy number. Some albino plantlets were produced, but no mature plants were obtained.

AB - Suspension culture cells of barley (Hordeum vulgare L. cv. Klages) were collectec on filters and bombarded with gold particles carrying the Basta® resistance (bar) gene, or both bar and the β-glucuronidase (uidA) genes in a Biolistic® Particle Accelerator. Filters carrying the bombarded tissues were selected on basal medium with increasing concentrations (maximum of 200 mg I-1) of phosphinothricin. Surviving calli were assayed for the presence of phosphinothricin acetyltransferase activity. All positive calli were shown to have the bar gene integrated into their genome. Nine independent stable transformation events have been analyzed with substantial variation in the integration pattern and transgene copy number. Some albino plantlets were produced, but no mature plants were obtained.

KW - Hordeum vulgare L.

KW - accelerated particle

KW - integration

KW - methylation

KW - suspension culture

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