An efficient field screening procedure for identifying transposants for constructing an Ac/Ds-based insertional-mutant library of rice.
An efficient system was developed, and several variables tested, for generating a large-scale insertional-mutagenesis population of rice. The most important feature in this improved Ac/Ds tagging system is that one can conveniently carry out large-scale screening in the field and select transposants at the seedling stage. Rice was transformed with a plasmid that includes a Basta-resistance gene (bar). After the Ds element is excised during transposition, bar becomes adjacent to the ubiquitin promoter, and the rice plant becomes resistant to the herbicide Basta. In principle, one can plant up to one million plants in the field and select those plants that survive after spraying with Basta. To test the utility of this system, 4 Ds starter lines were crossed with 14 different Ac plants, and many transposants were successfully identified after planting 134,285 F2 plants in the field. Over 2,800 of these transposants were randomly chosen for PCR analysis, and the results fully confirmed the reliability of the field screening procedure.
Luan
Wei-Jiang
WJ
China National Rice Research Institute, State Key Laboratory of Rice Biology, Hangzhou, Zhejiang, People's Republic of China.
He
Cheng-Kun
CK
Hu
Guo-Cheng
GC
Dey
Moul
M
Fu
Ya-Ping
YP
Si
Hua-Min
HM
Zhu
Li
L
Liu
Wen-Zhen
WZ
Duan
Faping
F
Zhang
Hong
H
Liu
Wen-Ying
WY
Zhuo
Ren-Ying
RY
Garg
Ajay
A
Wu
Ray
R
Sun
Zong-Xiu
ZX
D000613:Aminobutyrates / Q000494:pharmacology; D003433:Crosses, Genetic; D004351:Drug Resistance; D015723:Gene Library; D006540:Herbicides / Q000494:pharmacology; D020071:Interspersed Repetitive Sequences; D016254:Mutagenesis, Insertional; D012275:Oryza / Q000033:anatomy & histology / Q000187:drug effects / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000033:anatomy & histology / Q000187:drug effects / Q000235:genetics; D016133:Polymerase Chain Reaction; D016324:Sequence Tagged Sites; D014170:Transformation, Genetic; D001616:beta-Galactosidase / Q000032:analysis
D000613:Aminobutyrates; D006540:Herbicides; C003121:phosphinothricin; D001616:beta-Galactosidase