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首页> 外文期刊>Journal of Experimental Botany >Pathway engineering of Brassica napus seeds using multiple key enzyme genes involved in ketocarotenoid formation.
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Pathway engineering of Brassica napus seeds using multiple key enzyme genes involved in ketocarotenoid formation.

机译：利用参与类酮体形成的多个关键酶基因对甘蓝型油菜种子进行途径工程。

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Brassica napus (canola) plants were genetically manipulated to increase the amount and composition of carotenoids in seeds by using seven key enzyme genes involved in ketocarotenoid formation, which originated from a soil bacterium Pantoea ananatis (formerly called Erwinia uredovora 20D3), and marine bacteria Brevundimonas sp. strain SD212 and Paracoccus sp. strain N81106 (formerly called Agrobacterium aurantiacum). The seven key gene cassettes, in which each gene was surrounded by an appropriate promoter and terminator, were connected in a tandem manner, and the resulting constructs (17 kb) were inserted into a binary vector and used for transformation of B. napus. Surprisingly, 73-85% of the regenerated plants retained all seven genes, and formed orange- or pinkish orange-coloured seeds (embryos), while untransformed controls had light yellow-coloured seeds with predominant accumulation of lutein. Three of the transgenic lines were analysed further. The total amount of carotenoids in these seeds was 412-657 og gp# fresh weight, which was a 19- to 30-fold increase compared with that of untransformed controls. The total amount of ketocarotenoids was 60-190 og gp# fresh weight. o-Carotene was the predominant carotenoid, with significant amounts of l-carotene, echinenone, phytoene, lutein, and canthaxanthin also detected in the transgenic seeds. The ratio of hydroxylated carotenoids to overall carotenoids was quite small relative to the ratio of ketocarotenoids to overall carotenoids. Interestingly, expression of many endogenous carotenogenic genes was also altered in the transgenic seeds, suggesting that their expression was affected by an increase in carotenoid biosynthesis.
机译：通过使用七个参与类酮缩酮形成的关键酶基因，对甘蓝型油菜（canola）植物进行了基因操纵，以增加种子中类胡萝卜素的含量和组成，该关键酶基因源自土壤细菌Pantoea ananatis（以前称为Erwinia uredovora 20D3）和海洋细菌Brevundimonas sp。 SD212菌株和Paracoccus sp。菌株N81106（以前称为Aurbacterium aurantiacum）。串联连接七个基因盒，其中每个基因被适当的启动子和终止子包围，并将得到的构建体（17 kb）插入二元载体中，用于转化甘蓝型油菜。令人惊讶的是，73-85％的再生植物保留了所有七个基因，并形成了橙色或粉红色的橙色种子（胚），而未转化的对照则具有浅黄色种子，其中主要是叶黄素的积累。进一步分析了三个转基因品系。这些种子中的类胡萝卜素总量为412-657 og gp＃新鲜重量，与未转化的对照相比增加了19至30倍。酮类胡萝卜素的总量为60-190 ug gp＃鲜重。邻胡萝卜素是主要的类胡萝卜素，在转基因种子中也检测到大量的l-胡萝卜素，海胆烯酮，八氢番茄红素，叶黄素和角黄素。相对于酮类胡萝卜素与总类胡萝卜素的比例，羟基化类胡萝卜素与总类胡萝卜素的比例非常小。有趣的是，转基因种子中许多内源性类胡萝卜素基因的表达也发生了变化，表明它们的表达受到类胡萝卜素生物合成增加的影响。

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