To investigate the role of inducible linalool in Arabidopsis-insect interactions, the FANES 1 linalool synthase （LIS） cDNA from strawberry with plastid targeting and a synthetic intron （LIS＇） was placed under the...To investigate the role of inducible linalool in Arabidopsis-insect interactions, the FANES 1 linalool synthase （LIS） cDNA from strawberry with plastid targeting and a synthetic intron （LIS＇） was placed under the control of the wound inducible proteinase inhibitor 2 （PI2） promoter from potato. The construct pBin-PPi2-LIS＇ was transformed to Arabidopsis thaliana ecotype Columbia 0. Kanamycin resistant TO seedlings were confirmed for the presence and transcription of the LIS＇ gene by PCR analysis on genomic DNA and by RT-PCR analysis on RNA. Genomic and RT-PCR products were sequenced to confirm correct splicing of the synthetic intron. The expression of active linalool synthase by the PPI2-LIS＇ gene construct in the transgenic lines was assessed by measuring linalool emission using solid phase micro-extraction （SPME） GC-MS measurements after induction with methyl jasmonate. Among 30 tested independent T2 transgenic lines, 10 exhibited linalool production. Linalool expression could be induced by methyl jasmonate treatment, but not by diamondback moth larvae.展开更多
基金partially supported by IAC grant(the Netherlands)partially funded by the Key Laboratory of Vegetable Genetics and Physiology,Ministry of Agriculture(China)the National Natural Science Foundation of China(30370938).
文摘To investigate the role of inducible linalool in Arabidopsis-insect interactions, the FANES 1 linalool synthase （LIS） cDNA from strawberry with plastid targeting and a synthetic intron （LIS＇） was placed under the control of the wound inducible proteinase inhibitor 2 （PI2） promoter from potato. The construct pBin-PPi2-LIS＇ was transformed to Arabidopsis thaliana ecotype Columbia 0. Kanamycin resistant TO seedlings were confirmed for the presence and transcription of the LIS＇ gene by PCR analysis on genomic DNA and by RT-PCR analysis on RNA. Genomic and RT-PCR products were sequenced to confirm correct splicing of the synthetic intron. The expression of active linalool synthase by the PPI2-LIS＇ gene construct in the transgenic lines was assessed by measuring linalool emission using solid phase micro-extraction （SPME） GC-MS measurements after induction with methyl jasmonate. Among 30 tested independent T2 transgenic lines, 10 exhibited linalool production. Linalool expression could be induced by methyl jasmonate treatment, but not by diamondback moth larvae.
