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发根农杆菌LBA 9402对难以转化的植物白花罂粟的毛状根诱导

Hairy root induction of Papaver somniferum var. album, a difficult-to-transform plant, by A rhizogenes LBA 9402.

作者信息

Le Flem-Bonhomme V, Laurain-Mattar D, Fliniaux M A

机构信息

Laboratoire de Phytotechnologie, Faculté de Pharmacie, Université de Picardie Jules Verne, 1 rue des Louvels, 80037, Amiens Cedex 1, France.

出版信息

Planta. 2004 Mar;218(5):890-3. doi: 10.1007/s00425-003-1196-z. Epub 2004 Jan 23.

Abstract

Two strains of Agrobacterium rhizogenes (15834, LBA 9402) and one Agrobacterium tumefaciens strain [GV 3101 (PMP90RK, p35SGUS-2)] and four culture media were tested and compared for their ability to induce hairy root formation on wounded Papaver somniferum L. hypocotyls. Five weeks after the infection with A. rhizogenes LBA 9402, hairy roots appeared on 80% of the hypocotyls maintained in the hormone-free liquid medium. Six hairy-root cultures were established. Transformation was confirmed by polymerase chain reaction analysis. One clone was analysed for its alkaloid production. The total alkaloid content was higher in the transformed roots (0.46+/-0.06% DW) than in the untransformed roots (0.32+/-0.05% DW). The transformed roots accumulated three times more codeine (0.18+/-0.02% DW) than intact roots (0.05+/-0% DW). Moreover, morphine (0.255+/-0.03% DW) and sanguinarine (0.014+/-0% DW) were found in the liquid culture medium.

摘要

对两株发根农杆菌(15834、LBA 9402)、一株根癌农杆菌[GV 3101(PMP90RK,p35SGUS - 2)]以及四种培养基进行了测试和比较,以研究它们在致伤的罂粟下胚轴上诱导毛状根形成的能力。用发根农杆菌LBA 9402感染五周后,在无激素液体培养基中培养的下胚轴上,80%出现了毛状根。建立了六种毛状根培养物。通过聚合酶链反应分析确认了转化。对一个克隆进行了生物碱产量分析。转化根中的总生物碱含量(0.46±0.06%干重)高于未转化根(0.32±0.05%干重)。转化根积累的可待因(0.18±0.02%干重)是完整根(0.05±0%干重)的三倍。此外,在液体培养基中发现了吗啡(0.255±0.03%干重)和血根碱(0.014±0%干重)。

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