
TY  - JOUR
A1  - Upadhyaya, Narayana M. 
A1  - Zhou, Xue-Rong 
A1  - Zhu, Qian-Hao 
A1  - Ramm, Kerrie 
A1  - Wu, Limin 
A1  - Eamens, Andrew 
A1  - Sivakumar, Ramani 
A1  - Kato, Tsuneo 
A1  - Yun, Dow-Won 
A1  - Santhoshkumar, Chellian 
A1  - Narayanan, Kottaram K. 
A1  - Peacock, James W. 
A1  - Dennis, Elizabeth S. 
T1  - An <emph type="2">iAc&#47;Ds</emph> gene and enhancer trapping system for insertional mutagenesis in rice
JF  - Functional Plant Biology
JO  - Functional Plant Biol.
Y1  - 2002
VL  - 29
IS  - 5
SP  - 547
EP  - 559
KW  - 
UR  - https://doi.org/10.1071/PP01205 
N2  - We evaluated a two-component transposon iAc/Ds system for generating a library of insertional mutants in rice. The constructs used have gene or enhancer trapping properties, plasmid rescue and T-DNA/Ds launching pad reporter facilities. Mutagenic iAc/Ds lines were produced by three methods: crossing iAc and Ds containing lines; co-transformation with iAc and Ds constructs; and super-transformation of iAc transgenic calli with Ds constructs. First and second generation screening populations, derived from crosses (F2 and F3) or double transformation (DtT1 and DtT2), were analysed for stable insertion lines containing Ds transposed to locations unlinked to iAc. The average frequencies of putative stable insertion (PSI) lines in the F2, DtT1, F3 and DtT2 populations were 6.61, 5.58, 11.47 and 7.05% respectively, with large variations in these frequencies in screening populations derived from different mutagenic lines. Further analyses indicated that 41, 33, 65 and 64% of the PSI lines, respectively, have Ds transposed to locations unlinked to the original Ds launching pad. Using the plasmid rescue system, sequences flanking Ds from 137 PSI lines were obtained. Sixty-eight of these lines had unique insertions in genomic regions, of which 18 were known sequences. Because the average frequency of proven stable insertion lines in any of our screening populations has been less than 5%, we suggest that additional features should be incorporated in this two-component iAc/Ds system to increase the screening efficiency, and to make it suitable for large-scale insertional mutagenesis and determination of gene function in rice. 
ER  -

