| Summary: | 碩士 === 銘傳大學 === 生物科技學系碩士班 === 103 === The plants can’t move around, so often influenced by the environment and climate stress. In Taiwan summer, the typhoon and afternoon thundershower often incursion. The soil water content increased and causing plant stress death. Previously, flood-resistant of eggplant (Solanum melongena L.) and sponge gourd (Luffa cylindrica) enhance ascorbate peroxidase (APX) activity under flooding. SmAPX and LcAPX gene length were 753 and 561 bps.
In this study, SmAPX and LcAPX were under Ubiquitin promoter regulation and via Agrobacterium tumefaciens strain GV3101 transfer to Arabidopsis. The transformation rate of SmAPX and LcAPX in Arabidopsis were 0.23% and 0.47% with antibiotic selection. Further use of PCR, qRT-PCR and western blot analysis shows gene were successfully transformation into plants and can be transcripted to mRNA and translated into protein. The qRT-PCR flood-related gene found LcAPX transgentics sucrose synthase1 (SUS1) level were long-term impact of flooding and SmAPX transgentics were mid -term impact of flooding. The APX activity of SmAPX and LcAPX transgentic lines were 3.6~6.2, 4.8~70.9 times higher than NT plant. The NT plant MDA conents were 3.0~4.2 and 2.6~4.8 times higher than SmAPX and LcAPX transgentic lines. The results of 7 days flooding treatment seeds germination rate were NT (72%) lower than SmAPX transgentic lines (82~92.3%) and LcAPX transgentic lines (82.3~93.8%). The transgentic lines leaves could growth but NT plant leaves were inhibited growth under 5 days flooding. Compared with NT plant, the transgentic lines leaves were lower level of H2O2 accumulation using DAB staining.
Based on the above results that overexpression of APX gene in Arabidopsis could clear H2O2 produced from flooding and enhance transgentic lines under flooding stress tolerance.
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