Localization of introduced genes on the chromosomes of transgenic barley, wheat and triticale by fluorescence in situ hybridization

C. Pedersen; J. Zimny; D. Becker; A. Jähne-Gärtner; H. Lörz

doi:10.1007/s001220050474

Localization of introduced genes on the chromosomes of transgenic barley, wheat and triticale by fluorescence in situ hybridization

C. Pedersen, J. Zimny, D. Becker, A. Jähne-Gärtner, H. Lörz^*

^*Corresponding author for this work

Section for Plant and Soil Sciences

70 Citations (Scopus)

Abstract

Using fluorescence in situ hybridization (FISH) we localized introduced genes on metaphase chromosomes of barley, wheat, and triticale transformed by microprojectile bombardment of microspores and scutellar tissue with the pDB1 plasmid containing the uid A and bar genes. Thirteen integration sites were detected in the nine lines analysed. Southern analysis showed that three or more copies of the plasmid were present in the lines. In a triticale line containing four copies three different integration sites were identified, indicating that the method described is sensitive enough for the detection of single-copy integrations. There was a slight tendency towards the localization of transgenes in distal chromosome regions. Using the GAA-satellite sequence for chromosome banding, the chromosomes containing the inserted genes were identified in most cases. Two barley lines derived from the same transformant showed a totally different integration pattern. Southern analysis confirmed that the inserted genes were segregating independently, resulting in different integration patterns among the progeny lines. The application of the FISH technique for the analysis of transgenic plants is discussed.

Original language	English
Journal	Theoretical and Applied Genetics
Volume	94
Issue number	6-7
Pages (from-to)	749-757
Number of pages	9
ISSN	0040-5752
DOIs	https://doi.org/10.1007/s001220050474
Publication status	Published - 1 May 1997

Keywords

Chromosomal localization
FISH
Integration pattern
Transgenic barley
Wheat and triticale

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10.1007/s001220050474

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title = "Localization of introduced genes on the chromosomes of transgenic barley, wheat and triticale by fluorescence in situ hybridization",

abstract = "Using fluorescence in situ hybridization (FISH) we localized introduced genes on metaphase chromosomes of barley, wheat, and triticale transformed by microprojectile bombardment of microspores and scutellar tissue with the pDB1 plasmid containing the uid A and bar genes. Thirteen integration sites were detected in the nine lines analysed. Southern analysis showed that three or more copies of the plasmid were present in the lines. In a triticale line containing four copies three different integration sites were identified, indicating that the method described is sensitive enough for the detection of single-copy integrations. There was a slight tendency towards the localization of transgenes in distal chromosome regions. Using the GAA-satellite sequence for chromosome banding, the chromosomes containing the inserted genes were identified in most cases. Two barley lines derived from the same transformant showed a totally different integration pattern. Southern analysis confirmed that the inserted genes were segregating independently, resulting in different integration patterns among the progeny lines. The application of the FISH technique for the analysis of transgenic plants is discussed.",

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T1 - Localization of introduced genes on the chromosomes of transgenic barley, wheat and triticale by fluorescence in situ hybridization

AU - Pedersen, C.

AU - Zimny, J.

AU - Becker, D.

AU - Jähne-Gärtner, A.

AU - Lörz, H.

PY - 1997/5/1

Y1 - 1997/5/1

N2 - Using fluorescence in situ hybridization (FISH) we localized introduced genes on metaphase chromosomes of barley, wheat, and triticale transformed by microprojectile bombardment of microspores and scutellar tissue with the pDB1 plasmid containing the uid A and bar genes. Thirteen integration sites were detected in the nine lines analysed. Southern analysis showed that three or more copies of the plasmid were present in the lines. In a triticale line containing four copies three different integration sites were identified, indicating that the method described is sensitive enough for the detection of single-copy integrations. There was a slight tendency towards the localization of transgenes in distal chromosome regions. Using the GAA-satellite sequence for chromosome banding, the chromosomes containing the inserted genes were identified in most cases. Two barley lines derived from the same transformant showed a totally different integration pattern. Southern analysis confirmed that the inserted genes were segregating independently, resulting in different integration patterns among the progeny lines. The application of the FISH technique for the analysis of transgenic plants is discussed.

AB - Using fluorescence in situ hybridization (FISH) we localized introduced genes on metaphase chromosomes of barley, wheat, and triticale transformed by microprojectile bombardment of microspores and scutellar tissue with the pDB1 plasmid containing the uid A and bar genes. Thirteen integration sites were detected in the nine lines analysed. Southern analysis showed that three or more copies of the plasmid were present in the lines. In a triticale line containing four copies three different integration sites were identified, indicating that the method described is sensitive enough for the detection of single-copy integrations. There was a slight tendency towards the localization of transgenes in distal chromosome regions. Using the GAA-satellite sequence for chromosome banding, the chromosomes containing the inserted genes were identified in most cases. Two barley lines derived from the same transformant showed a totally different integration pattern. Southern analysis confirmed that the inserted genes were segregating independently, resulting in different integration patterns among the progeny lines. The application of the FISH technique for the analysis of transgenic plants is discussed.

KW - Chromosomal localization

KW - FISH

KW - Integration pattern

KW - Transgenic barley

KW - Wheat and triticale

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JO - Theoretical And Applied Genetics

JF - Theoretical And Applied Genetics

IS - 6-7

ER -

Localization of introduced genes on the chromosomes of transgenic barley, wheat and triticale by fluorescence in situ hybridization

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Keywords

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