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Construction of a designer chromosome in tobacco

Construction of a designer chromosome in tobacco 122 87 87 7 7 K. G. Campbell E. A. Wernsman W. P. Fitzmurice J. A. Burns Agronomy Department, Ohio Agricultural Research and Development Center The Ohio State University 44691-4096 Wooster OH USA Crop Science Department North Carolina State University 27695-7620 Raleigh NC USA Department of Botany, Crop Science, and Genetics North Carolina State University 27695-7614 Raleigh NC USA Biosource Genetics Corporation, Bowman Gray Technical Center Building 611-13E/011 27102 Winston-Salem NC USA Abstract The tobacco ( Nicotiana tabacum L.) breeding line NC 152 is a doubled haploid that possesses an addition chromosome from N. africana (Merxm. and Buttler). A gene on this chromosome confers potyvirus resistance (Poty R ). Our objective was to use the addition chromosome as a base on which to construct a designer chromosome containing a foreign gene linkage package. A mutant dhfr gene conferring resistance to methotrexate (Mtx) was inserted into NC 152-haploid (n = 25) leaf tissue via Agrobacterium tumefaciens -mediated transformation. After chromosome doubling, 135 NC 152 dhfr transformants (2n = 50) were pollinated with the potyvirus-susceptible (Poty S ) cultivar ‘McNair 944’ (2n = 48). Linkage analysis was performed in the BC 1 generation. Two transformants, NC 152 dhfr -996 and NC 152 dhfr -1517 exhibited complete linkage between Mtx resistance (Mtx R ) and Poty R . Segregants from these two transformants which were Mtx R and Poty R possessed 49 chromosomes, while Mtx sensitive (Mtx S ) and Poty S progeny possessed 48 chromosomes. Eighty percent of the NC 152 dhfr transformants transmitted the dhfr gene as one locus. Other foreign genes can be directed to the addition chromosome through transformation followed by selection for single loci with linkage to Poty R or Mtx R . The integrity of both the foreign-gene linkage package and the rest of the genome will be maintained because recombination between the N. africiana and the N. tabacum chromosomes has not been observed. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png TAG Theoretical and Applied Genetics Springer Journals

Construction of a designer chromosome in tobacco

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References (28)

Publisher
Springer Journals
Copyright
Copyright © 1994 by Springer-Verlag
Subject
Life Sciences; Biotechnology; Agriculture; Biochemistry, general; Plant Biochemistry; Plant Sciences; Plant Genetics & Genomics
ISSN
0040-5752
eISSN
1432-2242
DOI
10.1007/BF00221136
pmid
24190470
Publisher site
See Article on Publisher Site

Abstract

122 87 87 7 7 K. G. Campbell E. A. Wernsman W. P. Fitzmurice J. A. Burns Agronomy Department, Ohio Agricultural Research and Development Center The Ohio State University 44691-4096 Wooster OH USA Crop Science Department North Carolina State University 27695-7620 Raleigh NC USA Department of Botany, Crop Science, and Genetics North Carolina State University 27695-7614 Raleigh NC USA Biosource Genetics Corporation, Bowman Gray Technical Center Building 611-13E/011 27102 Winston-Salem NC USA Abstract The tobacco ( Nicotiana tabacum L.) breeding line NC 152 is a doubled haploid that possesses an addition chromosome from N. africana (Merxm. and Buttler). A gene on this chromosome confers potyvirus resistance (Poty R ). Our objective was to use the addition chromosome as a base on which to construct a designer chromosome containing a foreign gene linkage package. A mutant dhfr gene conferring resistance to methotrexate (Mtx) was inserted into NC 152-haploid (n = 25) leaf tissue via Agrobacterium tumefaciens -mediated transformation. After chromosome doubling, 135 NC 152 dhfr transformants (2n = 50) were pollinated with the potyvirus-susceptible (Poty S ) cultivar ‘McNair 944’ (2n = 48). Linkage analysis was performed in the BC 1 generation. Two transformants, NC 152 dhfr -996 and NC 152 dhfr -1517 exhibited complete linkage between Mtx resistance (Mtx R ) and Poty R . Segregants from these two transformants which were Mtx R and Poty R possessed 49 chromosomes, while Mtx sensitive (Mtx S ) and Poty S progeny possessed 48 chromosomes. Eighty percent of the NC 152 dhfr transformants transmitted the dhfr gene as one locus. Other foreign genes can be directed to the addition chromosome through transformation followed by selection for single loci with linkage to Poty R or Mtx R . The integrity of both the foreign-gene linkage package and the rest of the genome will be maintained because recombination between the N. africiana and the N. tabacum chromosomes has not been observed.

Journal

TAG Theoretical and Applied Genetics – Springer Journals

Published: Feb 1, 1994

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