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標題: 共同轉殖蘇力菌殺蟲晶體蛋白、熱休克蛋白、超氧化歧化酵素及過氧化氫酵素基因至結球白菜
Co-transformation of Bt-toxin, Heat Shock Protein, Superoxide Dismutase, and Catalase Genes into Chinese Cabbage (Brassica campestris L. ssp. Pekinensis) via Agrobacterium Mediated Transformation
作者: 蔡元純
Yuan-Chun Yeh
Chia-Chune Wu
Menq-Jiau Tseng
關鍵字: 過氧化氫酵素;Chinese cabbage;農桿菌共同轉移法;蘇力菌殺蟲晶體蛋白;熱休克蛋白;超氧化歧化酵素;Co-transformation;Bt-toxin;Heat Shock Protein;Superoxide Dismutase;Catalase
Project: 興大園藝, Volume 28, Issue 4, Page(s) 29-50.
Horticulture NCHU, Volume 28, Issue 4, Page(s) 29-50.
本研究利用農桿菌共同轉移法,研究同時轉移攜帶有rbcS為啟動子及大豆葉綠磯訊息 transit peptide 之蘇力菌殺蟲晶體蛋白(bt)、結球白菜之起氧化歧化酵素 (sod62)、過氧化氫酵素 (cat78) 及番茄之熱休克蛋白 (hsc70) 等四種基因至同一棵之'濱綠'結球白菜並探討轉運蛋白質到葉綠體之可行性。
轉殖結球白菜再生植株經過高溫處理 (42℃ 4天) 之後,有hsc70基因之轉殖植株明顯可以抵抗高溫。具有轉殖bt基因之結球白菜再生植株葉片經餵食小菜蛾幼蟲,證實具有抗蟲之效果。以800ppb之SO2燻蒸轉殖植株三天,具有sod及cat基因之結球白菜再生植株,對於SO2之危害較未轉殖或只轉殖單一sod或cat基因之轉殖株具有抗性。本研究結果証實利用農桿菌混合感染培殖體可達到多基因同時轉移到同一棵結球白菜,且能正常表現有功能之蛋白質,使轉殖植株具有抗蟲、耐高溫及耐逆境之特性。

Taiwan is located at the sub-tropic area. In summer, the high temperature and humidity cause very serious of plant disease and insect pests. Attempts had been made to co-transfer the Bt-toxin gene (bt), superoxide dismutase gene (sod62) of Chinese cabbage, catalase gene (cat78) of Chinese cabbage, and heat shock protein gene (hsc70) of tomato into the Chinese cabbage (Brassica campestrus L. ssp. Pekinensis cv. Tropical Pride). The objectives of this study are to establish the technology of gene co-transfer and targeting the foreign protein into the chloroplast by chloroplast transit peptide, and to study the possibility for improvement of Chinese cabbage with insect, disease, and stresses resistance by genes co-transformation. The results of PCR, Southern and Northern hybridization indicated that the bt, hsc70, sod62 and cat78 genes had been inserted into the genome of Chinese cabbage, and expressed. The result of isozyme profiles assay revealed that the transformed SOD62 and CAT78 were present in the transformed plants. The SOD and CAT activities in leaves and chloroplast of transformed plants were higher than those of the controls. Increase in amounts of HSC70 protein were found in the transformed plants as analyzing by western blot hybridization. Preliminary results of bioassay indicated that the increases in tolerance to high temperature (42℃, 4 days), SO2 (800 ppb, 3 days), and insect resistance (Plutella xylostella) were found in the transgenic plants. This study demonstrated that two, three and four foreign genes could be co-transferred into a Chinese cabbage plant simultaneously via Agrobacterium mediated transformation.
ISSN: 0255-5921
Appears in Collections:第28卷 第04期

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