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Genetic transformation of the endangered Tibetan medicinal plant Przewalskia tangutica Maxim and alkaloid production profiling revealed by HPLC

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Abstract

To achieve a high yield of tropane alkaloids (TA) and exploit the alpine plant sustainably, an optimized protocol for induction and establishment of hairy roots culture of Prezwalskia tangutica Maxim was developed through selection of appropriate Agrobacterium strain and the explant type. The hypocotyl is more readily facile to induce the HR than the cotyledon is when infected with the three different agrobacterium strains. MUS440 has an efficiency (of up to 20%), whereas the ATCC10060 (A4) can induce HR on both types of explants with the highest frequency (33.33%), root length (21.17 ± 2.84 cm), and root number (10.83 ± 1.43) per explant than the other strains. The highest HR production resulted from using hypocotyl as explants. Independent transformed HR was able to grow vigorously and to propagate on a no-hormone 1/2MS liquid medium. The presence of pRi rolB gene in transformation of HR was confirmed by PCR amplification. In the liquid medium, the HR growth curve appeared to be “S” shaped, and ADB had increased to 4.633 g/l. Moreover, HPLC analysis showed that HR lines have an extraordinary ability to produce atropine (229.88 mg/100 g), anisodine (4.09 mg/100 g), anisodamine (12.85 mg/100 g), and scopolamine (10.69 mg/100 g), which were all more significant than the control roots. In conclusion, our study optimized the culture condition and established a feasible genetics reactor for P. tangutica green exploration and biological study in the alpine region.

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Acknowledgements

We thank Mr Lawrence Lyon for reading and revising this manuscript. This work was supported by Grants from the National Foundation of Science (31070208), the Training Qualified People Plan ‘‘Hope of West China’’(Y229151211), the Basic Research Programs of Science and Technology in Qinghai Province (2017-ZJ-702), and the Construction Project for Innovation Platform of Qinghai Province (2017-ZJ-Y14).

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Author notes

  1. Tianxiang Lei, Huan Wang, and Songling Li have contributed equally to the work.

Authors and Affiliations

  1. Key Laboratory of Adaptation and Evolution of Plateau Biota (AEPB), Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining, 810008, Qinghai, People’s Republic of China

    Tianxiang Lei, Shilong Chen & Dangwei Zhou

  2. Tibetan Medicine Center, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining, 810008, Qinghai, People’s Republic of China

    Huan Wang & Jianwei Shen

  3. Key Laboratory of Qinghai Plant Molecular Breeding, Chinese Academy of Sciences, Xining, 810008, Qinghai, People’s Republic of China

    Shilong Chen & Dangwei Zhou

  4. University of Chinese Academy of Sciences, Beijing, 100093, People’s Republic of China

    Tianxiang Lei & Dangwei Zhou

  5. Institute of Soil and Fertilizer, Qinghai University, Xining, 810008, Qinghai, People’s Republic of China

    Songling Li & Xiaojian Cai

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  1. Tianxiang Lei
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  2. Huan Wang
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Correspondence to Dangwei Zhou.

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Lei, T., Wang, H., Li, S. et al. Genetic transformation of the endangered Tibetan medicinal plant Przewalskia tangutica Maxim and alkaloid production profiling revealed by HPLC. 3 Biotech 8, 179 (2018). https://doi.org/10.1007/s13205-018-1203-5

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