Effects of the rol C gene on hairy root:: Induction development and tropane alkaloid production by Atropa belladonna
Effects of the rol C gene on hairy root:: Induction development and tropane alkaloid production by Atropa belladonna
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DOI：
10.1021/np990614l
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发表时间：
2000-09-01
影响因子：
5.1
通讯作者：
Fliniaux, MA
中科院分区：
文献类型：
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作者：
Bonhomme, V;Laurain-Mattar, D;Fliniaux, MA
获得了两个系列的颠茄毛状根系，第一个系由含有rol C和npt II基因的根癌农杆菌转化，另一个系由rol ABC和nptⅡ基因转化。获得了13个毛状根，并在无激素培养基上进行了筛选。经PCR分析证实转化成功，并对这些根系的生长速率进行了初步检测。在培养3周和4周后，分别测定东莨菪碱和三七叶皂苷的产量，以评估rol C基因在托烷生物碱形成中的可能作用。仅rol C基因对毛状根的生长速度(增加17倍)起着重要作用。然而，这一效应远低于rol-ABC基因共同诱导的效应(增加75倍)。相反，在颠茄毛状根培养中，rol C基因单独和rol ABC基因结合在一起(总生物碱的平均值：0.36%干重，即是未转化根的12倍)对胡萝卜素生物碱的生物合成具有同等的促进作用。
Two series of Atropa belladonna hairy root lines were obtained, the first one transformed via Agrobacterium tumefaciens harboring rol C and npt II genes, and the other transformed with rol ABC and npt II genes. Thirteen hairy root; lines were obtained and selected on hormone-free medium. The transformation was confirmed by PCR analysis, and these root lines were first examinated for their growth rate. Then hyoscyamine and scopolamine production was measured after 3 and 4 weeks of culture to evaluate the possible role of rol C gene in tropane alkaloid formation. The rol C gene alone played a significant role in the hairy root growth rate (17-fold increase). However this effect was much lower than that induced by the rol ABC genes together (75-fold increase). In contrast, the rol C gene alone was as efficient as the rol ABC genes together (mean value of total alkaloids: 0.36% dry weight, i.e., 12-fold times more than in untransformed roots) to stimulate the biosynthesis of tropane alkaloids in A. belladonna hairy root cultures.