Quan Hong, Xia Ke, Zeng Jun-lan, Chen Min, Lan Xiao-zhong, Liao Zhi-hua
Yao Xue Xue Bao. 2016 Dec;51(12):1913-9.
Atropa belladonna L. is the commercial plant material for production of tropane alkaloids, including hyoscyamine and scopolamine. The wild-type Atropa belladonna is characterized by the hyoscyamine-rich chemotype, in which the hyoscyamine content is much higher than the scopolamine content. It is the common goal for the pharmaceutical industry to increase the content of scopolamine in A. belladonna. Based on the T0 progeny of transgenic A. belladonna with NtPMT and HnH6H overexpression, T1 progeny of transgenic A. belladonna were obtained through self-pollination and used in a field trial. The 461 bp fragment of NtPMT and the 1 077 bpHnH6 H were simultaneously expressed from T1 progeny of transgenic A. belladonna, but were not obtained from the wild-type A. belladonna. At the transcription level, the expression of NtPMT and HnH6H were detected in T1 progeny of transgenic A. belladonna, but were not detected in the wild-type plants. Further, the alkaloids were analyzed by HPLC. In the stems and leaves of T1 progeny of transgenic A. belladonna, hyoscyamine was not detected and scopolamine was detected at very high levels; in the stems and leaves of wild-type A. belladonna, hyoscyamine was detected at much higher levels. In the leaves of T1 progeny of transgenic A. belladonna, the content of scopolamine was 15-36 folds higher than that of wild- type leaves; in the stems of T1 progeny of transgenic A. belladonna, the scopolamine content was 37-108 folds higher than that of wild-type stems. In conclusion, overexpression of NtPMT and HnH6H greatly enhanced conversion of hyoscyamine into high-value scopolamine and improved the commercial value of A. belladonna.
颠茄是生产托烷生物碱（包括莨菪碱和东莨菪碱）的商业植物原料。野生型颠茄的特征是富含莨菪碱的化学型，其中莨菪碱含量远高于东莨菪碱含量。提高颠茄中东莨菪碱的含量是制药行业的共同目标。基于过表达NtPMT和HnH6H的转基因颠茄T0代子代，通过自花授粉获得转基因颠茄T1代子代并用于田间试验。转基因颠茄T1代子代同时表达了461 bp的NtPMT片段和1077 bp的HnH6H片段，但野生型颠茄中未获得。在转录水平上，在转基因颠茄T1代子代中检测到NtPMT和HnH6H的表达，但在野生型植株中未检测到。此外，通过高效液相色谱法分析生物碱。在转基因颠茄T1代子代的茎和叶中，未检测到莨菪碱，检测到高水平的东莨菪碱；在野生型颠茄的茎和叶中，检测到的莨菪碱水平要高得多。在转基因颠茄T1代子代的叶中，东莨菪碱含量比野生型叶高15 - 36倍；在转基因颠茄T1代子代的茎中，东莨菪碱含量比野生型茎高37 - 108倍。总之，NtPMT和HnH6H的过表达极大地增强了莨菪碱向高价值东莨菪碱的转化，提高了颠茄的商业价值。
