Artificial chromosome formation in maize (Zea mays L.)

• DOI: 10.1007/s00412-008-0191-3
• Publication Date: 2008-11-17T08:15:25Z
• Authors: Evgueni V. Ananiev; Chengcang Wu; Mark A. Chamberlin; Sergei Svitashev; Chris Schwartz; William Gordon-Kamm; Scott Tingey
• Keywords: Minichromosome; Selectable marker; Retrotransposon; Human artificial chromosome

We report on the construction of maize minichromosomes using shuttle vectors harboring native centromeric segments, origins replication, selectable marker genes, and telomeric repeats. These were introduced into scutellar cells immature embryos by microprojectile bombardment. Several independent transformation events identified containing in addition to normal diploid complement 20 chromosomes. Immunostaining indicated that recruited protein C, which is a specific component centromere/kinetochore complex. Minichromosomes estimated be 15-30 Mb size based cytological measurements. Fluorescent situ hybridization (FISH) showed contain centromeric, telomeric, exogenous unique sequences interspersed with retrotransposons. detected for at least year actively dividing callus cultures, providing evidence their stability through numerous cell cycles. Plants regenerated root tips, confirmation replication transmission during mitosis organogenesis. Assembly artificial chromosomes may provide tool study centromere function foundation developing new high capacity plant functional genomics breeding.