Characterization of an Ac transposon system based on apt1-m1 (Ac) on the long arm of maize chromosome 9.
Activator/Dissociation (Ac/Ds) transposable elements have been used in maize insertional mutagenesis as a complement to Mutator (Mu). In this study, to further improve the efficiency of the Ac/Ds mutagenesis system, we adopted apt1-m1 (Ac) on the long arm of chromosome 9 (9L) as a donor Ac to create an Ac insertion library. This system is based on the negative selection pressure against the donor Ac, and it was highly efficient for isolating new transposition events. We obtained 9,625 transposition events from 1083 F1 ears with an average transposition rate of 8.66 % (rates ranged from 1.11 to 29.73 %). We also adopted a modified PCR-based genome walking strategy to improve the efficiency of the new method for isolating transposon-flanking sequences. This method is more efficient than the Southern-based method that was used in previous studies. A validation step was developed to distinguish transposon tags derived from newly transposed Ac or Ds elements. Using this PCR-based method, we isolated 67 inheritable flanking sequences from the apt1-m1 (Ac) transposition library; of these, 51 were confirmed as tr-Ac-flanking sequences and 11 were tr-Ds-flanking sequences. Similar to other Ac donors from different loci, the apt1-m1 (Ac) system also exhibited a preference for short distance transposition. In this study, we have further improved the Ac mutagenesis system in maize for gene isolation and functional genomics studies.
School of Life Sciences, Shanghai Key Laboratory of Bio-energy Crop, Shanghai University, 333 Nanchen Road, Shanghai 200444, People's Republic of China.
Fei
F
Wang
Pengfei
P
Li
Yuanping
Y
Tang
Jun
J
Fan
Dabin
D
Xu
Shengming
S
Guo
Zhengkai
Z
Xu
Rentao
R
Song
D001483:Base Sequence; D032461:Chromosomes, Plant / Q000235:genetics*; D004251:DNA Transposable Elements; D018744:DNA, Plant / Q000737:chemistry; D017343:Genes, Plant; D018745:Genome, Plant; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D003313:Zea mays / Q000235:genetics*
D004251:DNA Transposable Elements; D018744:DNA, Plant