TY - JOUR
T1 - Constructing defined chromosome segmental duplications in maize
AU - Yu, C.
AU - Danilova, T.
AU - Zhang, J.
AU - Birchler, J.
AU - Peterson, Thomas
PY - 2010/7/1
Y1 - 2010/7/1
N2 - We describe the production of chromosome segmental duplications and deletions of defined length in maize. The method utilizes a collection of previously isolated transposon-induced reciprocal translocations with sequence-defined breakpoints. These balanced translocations involve a first common breakpoint at the site of transposon Ac/fAc insertions on chromosome 1S, and a second variable endpoint elsewhere in the genome. Stocks containing translocations with different endpoints on the same chromosome arms were crossed together to generate heterozygous combinations. Subsequent meiotic segregation generates gametes that contain the original balanced translocations, or pairs of translocation chromosomes that are genetically unbalanced; i.e., they contain a duplication or deletion of the affected segment. Self-pollination produces progeny plants that are genetically balanced, or that contain 0 to 4 copies of the affected segment. The chromosome constitution of progeny plants can be determined using PCR primers flanking the translocation breakpoints. In addition, plants segregating genetically unbalanced gametes exhibit significant pollen grain abnormalities and thus can be easily detected in the field. In this way, we constructed 4 different segmental tetrasomic stocks involving genomic regions of approximately 7.8, 18.7, 21, and 42.6 Mb on chromosomes 5S, 5L, 5L, and 4L, respectively. FISH analysis using specific probes located within the affected segments confirms their aneuploid composition. These and similar lines containing precisely defined segmental duplications and deletions may be useful to study the genetic impact of gene dosage.
AB - We describe the production of chromosome segmental duplications and deletions of defined length in maize. The method utilizes a collection of previously isolated transposon-induced reciprocal translocations with sequence-defined breakpoints. These balanced translocations involve a first common breakpoint at the site of transposon Ac/fAc insertions on chromosome 1S, and a second variable endpoint elsewhere in the genome. Stocks containing translocations with different endpoints on the same chromosome arms were crossed together to generate heterozygous combinations. Subsequent meiotic segregation generates gametes that contain the original balanced translocations, or pairs of translocation chromosomes that are genetically unbalanced; i.e., they contain a duplication or deletion of the affected segment. Self-pollination produces progeny plants that are genetically balanced, or that contain 0 to 4 copies of the affected segment. The chromosome constitution of progeny plants can be determined using PCR primers flanking the translocation breakpoints. In addition, plants segregating genetically unbalanced gametes exhibit significant pollen grain abnormalities and thus can be easily detected in the field. In this way, we constructed 4 different segmental tetrasomic stocks involving genomic regions of approximately 7.8, 18.7, 21, and 42.6 Mb on chromosomes 5S, 5L, 5L, and 4L, respectively. FISH analysis using specific probes located within the affected segments confirms their aneuploid composition. These and similar lines containing precisely defined segmental duplications and deletions may be useful to study the genetic impact of gene dosage.
KW - Chromosome inversion
KW - Chromosome translocation
KW - Maize
KW - Segmental duplication
KW - Transposon
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U2 - 10.1159/000314554
DO - 10.1159/000314554
M3 - Article
C2 - 20551616
AN - SCOPUS:77955292892
SN - 1424-8581
VL - 129
SP - 72
EP - 81
JO - Cytogenetic and Genome Research
JF - Cytogenetic and Genome Research
IS - 1-3
ER -