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首页> 外文期刊>Molecular Breeding >The production of marker-free genetically engineered broccoli with sense and antisense ACC synthase 1 and ACC oxidases 1 and 2 to extend shelf-life
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The production of marker-free genetically engineered broccoli with sense and antisense ACC synthase 1 and ACC oxidases 1 and 2 to extend shelf-life

机译：使用无义和反义ACC合酶1和ACC氧化酶1和2生产无标记的基因工程西兰花以延长保质期

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摘要

The production of transgenic broccoli (Brassica oleracea) with increased shelf-life using an Agrobacterium rhizogenes-mediated co-transformation protocol is reported. An Agrobacterium rhizogenes Ri vector, pRi!855:GFP was constructed to allow expression of the green fluorescent protein to identify insertion of Ri T_L-DNA into plant cells. The Brassica oleracea ACC synthase 1 and ACC oxidase 1 and 2 cDNAs in sense and antisense orientations were co-transformed into GDDH33, a doubled haploid calabrese-broccoli cultivar. Transformation efficiency was 3.26%, producing 150 transgenic root lines, of which 18 were regenerated into mature plants. The floral buds from T_0 broccoli heads were assayed for post-harvest production of ethylene and chlorophyll levels. Buds from T_0 lines transformed with ACC oxidase 1 and 2 constructs produced significantly less post-harvest ethylene at 20 deg C than the untransformed plants and chlorophyll loss was significantly reduced over a 96 h post-harvest period. The T_0 plants transformed with sense and antisense ACC synthase I had a significantly reduced 24 h post-harvest ethylene peak and delayed chlorophyll loss. A positive correlation between post-harvest bud ethylene production and chlorophyll loss was described bya regression. This demonstrates that the shelf-life of a very perishable vegetable may be increased up to 2 days at 20 deg C by reducing post-harvest ethylene production.
机译：报道了使用发根农杆菌介导的共转化方案生产具有增加的保质期的转基因西兰花（芸苔）。构建发根农杆菌Ri载体pRi1855：GFP以允许绿色荧光蛋白的表达以鉴定RiT_L-DNA插入植物细胞中。将甘蓝型油菜ACC合酶1和ACC氧化酶1和2 cDNA在有义和反义方向上共转化为双倍单倍体卡拉布兰西兰花品种GDDH33。转化效率为3.26％，产生了150个转基因根系，其中18个已再生为成熟植物。检测了来自T_0西兰花头的花蕾的收获后乙烯和叶绿素水平的产生。在20摄氏度下，用ACC氧化酶1和2构建体转化的T_0品系的芽比未转化的植物产生的乙烯明显减少，并且在收获后的96小时内叶绿素的损失明显减少。用有义和反义ACC合酶I转化的T_0植物在收获后24 h的乙烯峰显着降低，并延迟了叶绿素的损失。回归描述了采后芽乙烯产量与叶绿素损失之间呈正相关。这表明，通过减少收获后的乙烯产量，极易腐烂的蔬菜的货架期可以在20摄氏度下延长至2天。

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