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Transcript
Title: Engineering Brassica with insect-resistance by stacked gene of trypsin inhibitor（sporamin）and chitinase genes M.S. student: Chen Pengjen Advisor: Kai-Wun Yeh Abstract: Cabbage（Brassica oleracea L. Capitata）is the most important leafly vegetable in Taiwan, due to it’s abundant antioxidant compounds and anticancer compounds like glucosinolate. Unfortunately, cabbage can be easily infected with or attacked by insect peats. In order to decrease the risk of cabbage attacked by insects, the goal to create transgenic cabbage by gene stacking methods is inprogress in our lab. Trypsin inhibitor（sporamin）and chitinase genes were in stack constructed using pMSPOA as promoter,. We successfully got ten independent lines through Agrobacterium tumefaciens-mediated transformation,. In trypsin inhibitor or chitinase activity assay, we found that both sporamin and chitinase activity can be increasing 1-2 times at 2 hrs after wounding. In bioassay, we found that transgenic line 4 is more effectivel to defend Diamond-back moth(Plutella xylostella) attack.. In order to alleviate public concerns on the risk of antibiotics resistance gene, recommendations have been made to eliminate all antibiotic marker genes from GM plants. Our results showed that by Agrobacterium tumefaciens-mediated co-transformation with a double T-DNA binary vector in Arabidopsis, several marker-free transformants were successfully obtained. This technology mote will further applied in the transgenic cabbage system.
