NDLI: Generation of Rye (Secale cereale L.) Plants With Low Transgene Copy Number After Biolistic Gene Transfer and Production of Instantly Marker-Free Transgenic Rye

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Generation of Rye (Secale cereale L.) Plants With Low Transgene Copy Number After Biolistic Gene Transfer and Production of Instantly Marker-Free Transgenic Rye

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Generation of Rye (Secale cereale L.) Plants With Low Transgene Copy Number After Biolistic Gene Transfer and Production of Instantly Marker-Free Transgenic Rye

Content Provider	Springer Nature Link
Author	Popelka, Juan Carlos Xu, Jianping Altpeter, Fredy
Copyright Year	2003
Abstract	Rye is extremely recalcitrant to tissue culture and genetic transformation. We describe the efficient and reproducible production of stably expressing transgenic rye plants after biolistic gene transfer to callus tissue derived from immature embryos. Key factors in the production of transgenic rye plants include the identification of biolistic gene transfer parameters and a selection protocol, which does not affect its regeneration ability. The bar gene was used as a selectable marker and selection was performed by spraying the regenerated shoots with 0.05% Basta solution without any previous selection of tissue cultures. Based on Southern blot analysis, a total of 21 transgenic rye plants with independent transgene integration patterns were produced. A low transgene copy number was observed in most transgenic plants and 40% of the plants had a single transgene copy insert. The high frequency of single transgene copy inserts might be a consequence of the selection system, which is based on the identification of stably expressing transgenic plantlets rather than stably expressing tissue cultures. All transgenic rye lines with single transgene inserts showed stable transgene expression in sexual progenies, but indications of transcriptional and post-transcriptional gene silencing were observed in few transgenic lines with multiple transgene inserts. Tissue culture-based selection was not necessary for the generation of transgenic rye. The identification of 17 transgenic rye plants without using any selectable marker gene by PCR amplification of transgene sequences is also demonstrated. Instant generation of selectable marker-free transgenic rye avoids a negative impact of selective agents on the transgenic tissue cultures, responds to public concerns on the safety of selectable markers and will support multiple transformation cycles for transgene pyramiding.
Starting Page	587
Ending Page	596
Page Count	10
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	12
Issue Number	5
e-ISSN	15739368
Language	English
Publisher	Kluwer Academic Publishers
Publisher Date	2003-01-01
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Human Genetics Plant Sciences Animal Anatomy / Morphology / Histology
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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Generation of Rye (Secale cereale L.) Plants With Low Transgene Copy Number After Biolistic Gene Transfer and Production of Instantly Marker-Free Transgenic Rye