Chloroplast transformation in oilseed rape.
The chloroplast transformation vector pNRAB carries two expression cassettes for the spectinomycin resistance gene aadA and the insect resistance gene cry1Aa10. The two cassettes are sited between the rps7 and ndhB targeting fragments. Biolistic delivery of the vector DNA, followed by spectinomycin selection, yielded chloroplast transformants at a frequency of four in 1000 bombarded cotyledon petioles. PCR analysis and Southern blot of PCR products confirmed the site-specific integration of aadA and cry1Aa10 into the chloroplast genomes of transgenic oilseed rape. When transgenic oilseed rape leaves were fed to second instar Plutella xylostera larvae, 47% mortality was observed against this insect and the surviving larvae had significantly lower weight than the control. This is the first report of chloroplast transformation in oilseed rape and the introduction of novel genes between the rps7 and ndhB genes in the chloroplast genome. This offers an opportunity for improvement of oilseed rape by chloroplast genetic engineering.
Hou
Bing-Kai
BK
Institute of Genetics, Chinese Academy of Sciences, Beijing 100101, PR China.
Zhou
Yi-Hua
YH
Wan
Li-Hong
LH
Zhang
Zhong-Lin
ZL
Shen
Gui-Fang
GF
Chen
Zheng-Hua
ZH
Hu
Zan-Min
ZM
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015139:Blotting, Southern; D029688:Brassica napus / Q000235:genetics* / Q000378:metabolism; D002736:Chloroplasts / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D017343:Genes, Plant / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D018087:Plastids; D000198:Spectinomycin / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic; D019076:Transgenes / Q000235:genetics*
D000900:Anti-Bacterial Agents; D018744:DNA, Plant; D010940:Plant Proteins; D000198:Spectinomycin