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Establishment of Salvia castanea Diels f. tomentosa Stib. hairy root cultures and the promotion of tanshinone accumulation and gene expression with Ag+, methyl jasmonate, and yeast extract elicitation

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Abstract

Salvia castanea Diels f. tomentosa Stib. is an endemic medicinal plant distributed in China, and the notably high content of tanshinone IIA in the root is proven effective for the therapy of heart diseases. Hairy root induction of this Salvia species was inoculated with Agrobacterium rhizogenes strain ATCC 15834. Transformed hairy root was cultured in 6,7-V liquid medium for growth kinetics assessment and elicitation. An S curve was present in the hairy root cultures based on the fresh and dry weights with an interval of 3 days. An optimum concentration of the applied elicitors (15 μM Ag+, 200 μM methyl jasmonate, and 200 mg l−1 yeast extract elicitor) benefitted both the growth status and tanshinone accumulation in the hairy root cultures. Tanshinone IIA contents were mostly stimulated 1.8-fold and 1.99-fold compared with the control by Ag+ and methyl jasmonate elicitation, respectively. Yeast extract dramatically enhanced dry mass accumulation, while it promoted cryptotanshinone content of 2.84 ± 0.33 mg g−1 dry weight at most in the hairy root cultures. Selected elicitors diversely influenced tanshinone accumulation in the time courses of hairy root cultures within 7 days. Furthermore, transcripts of selected genes in the tanshinone biosynthetic pathway were remarkably upregulated with elicitation. Yeast extract elicitor heightened 13.9-fold of isopentenyl diphosphate isomerase expression level at 12 h, while it increased 16.7-fold of geranylgeranyl diphosphate synthase transcript at 24 h compared with that of the control, which was more effective than Ag+ and methyl jasmonate. This study provided a convenient hairy root culture system of S. castanea Diels f. tomentosa Stib. for tanshinone production for the first time.

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Acknowledgments

The authors are grateful to Mrs. Rong Chen at Science and Technology Bureau of Nyingchi Prefecture, Tibet Autonomous Region, People’s Republic of China, for providing the S. castanea Diels f. tomentosa Stib. seeds and research fellow Yansheng Chen at Northwest A&F University, Yangling, People’s Republic of China, for the plant identification. This work was supported by the National Natural Science Foundation of China (Grant No. 81373908) and the Key Science and Technology Project of Shaanxi Province, People’s Republic of China (Grant No. 2012 KTCL 02-07).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. College of Life Sciences, Northwest A&F University, 712100, Yangling, People’s Republic of China

    Bo Li, Bangqing Wang, Hongyan Li, Liang Peng & Zongsuo Liang

  2. Hanzhong Institute of Botany, 723000, Hanzhong, People’s Republic of China

    Bangqing Wang

  3. College of Pharmacy, Shaanxi University of Chinese Medicine, 712046, Xi’an, People’s Republic of China

    Liang Peng

  4. Institute of Soil and Water Conservation, Chinese Academy of Sciences, 712100, Yangling, People’s Republic of China

    Mei Ru

  5. College of Life Sciences, Zhejiang Sci-Tech University, 310018, Hangzhou, People’s Republic of China

    Zongsuo Liang

  6. Tianjin Tasly Holding Group Co., Ltd., 300410, Tianjin, People’s Republic of China

    Xijun Yan & Yonghong Zhu

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  1. Bo Li
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Li, B., Wang, B., Li, H. et al. Establishment of Salvia castanea Diels f. tomentosa Stib. hairy root cultures and the promotion of tanshinone accumulation and gene expression with Ag+, methyl jasmonate, and yeast extract elicitation. Protoplasma 253, 87–100 (2016). https://doi.org/10.1007/s00709-015-0790-9

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