Metabolic engineering of Lilium × formolongi using multiple genes of the carotenoid biosynthesis pathway
Metabolic engineering of Lilium × formolongi using multiple genes of the carotenoid biosynthesis pathway
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DOI：
10.1007/s11816-010-0147-y
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发表时间：
2010-08
影响因子：
2.4
通讯作者：
P. Azadi;Ntui Valentine Otang;D. Chin;I. Nakamura;M. Fujisawa;H. Harada;N. Misawa;M. Mii
中科院分区：
文献类型：
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作者：
P. Azadi;Ntui Valentine Otang;D. Chin;I. Nakamura;M. Fujisawa;H. Harada;N. Misawa;M. Mii
利用农杆菌介导法将含有7个酶基因的pCrtZW-N8idi-crtEBIY转化到台湾百合，该基因受CaMV 35S启动子调控。在转化子中，愈伤组织和叶片中均检测到酮类胡萝卜素，呈橙色。在转基因愈伤组织中，类胡萝卜素总量[133.3μg/g鲜重(FW)]是野生型愈伤组织的26.1%。转基因植株的叶绿素含量和光合作用效率显著降低，但经过几个月的继代培养，它们已经变成绿叶植株，叶绿素和光合作用效率显著提高。转基因橙色和绿色植株叶片中类胡萝卜素的总含量分别为102.9和135.2μg/gFW，相当于野生型的5.6%和7.4%。在转基因愈伤组织和橙叶中均检测到紫锥菜酮、角黄素、3‘-羟基紫锥酮、3-羟基紫锥酮和虾青素等酮类胡萝卜素。在从橙色转基因植株向绿色植株过渡的过程中，观察到酮类胡萝卜素的种类和组成发生了显著的变化。尽管3‘-羟基紫锥酮、3-羟基紫锥酮、虾青素和腺苷不存在，而紫锥菊酮和角黄素的含量较低，但有趣的是，类胡萝卜素生物合成的上调导致了叶片中类胡萝卜素浓度的增加(+31.4%)。
Lilium×formolongiwas genetically engineered byAgrobacterium-mediated transformation with the plasmid pCrtZW-N8idi-crtEBIY, which contains seven enzyme genes under the regulation of the CaMV 35S promoter. In the transformants, ketocarotenoids were detected in both calli and leaves, which showed a strong orange color. In transgenic calli, the total amount of carotenoids [133.3 μg/g fresh weight (FW)] was 26.1-fold higher than in wild-type calli. The chlorophyll content and photosynthetic efficiency in transgenic orange plantlets were significantly lowered; however, after several months of subculture, they had turned into plantlets with green leaves that showed significant increases in chlorophyll and photosynthetic efficiency. The total carotenoid contents in leaves of transgenic orange and green plantlets were quantified at 102.9 and 135.2 μg/g FW, respectively, corresponding to 5.6- and 7.4-fold increases over the levels in the wild-type. Ketocarotenoids such as echinenone, canthaxanthin, 3′-hydroxyechinenone, 3-hydroxyechinenone, and astaxanthin were detected in both transgenic calli and orange leaves. A significant change in the type and composition of ketocarotenoids was observed during the transition from orange transgenic plantlets to green plantlets. Although 3′-hydroxyechinenone, 3-hydroxyechinenone, astaxanthin, and adonirubin were absent, and echinenone and canthaxanthin were present at lower levels, interestingly, the upregulation of carotenoid biosynthesis led to an increase in the total carotenoid concentration (+31.4%) in leaves of the transgenic green plantlets.