Anisodus luridus hairy root cultures were established to test biological effects of acetylsalicylic acid (ASA) and ultraviolet ray-B (UV-B) on gene expression, tropane alkaloid (TA) biosynthesis and efflux. The TAs-pathway gene expression was ASA dosage dependant. The expression of PMT, TRI and CYP80F1 showed no significant difference in hairy root cultures in treatment of 0.01 and 0.1 mM ASA, compared with those without ASA treatment; while 0.01 or 0.1 mM ASA slightly upregulated H6H expression. All the four genes including PMT, TRI, CYP80F1 and H6H had a dramatic increase in 1 mM ASA-treated hairy root cultures compared with control. The expressing levels of all the four genes were much significantly higher in 1 mM ASA-treated hairy root cultures than those in 0.01 and 0.1 mM ASA-treated ones. As expected, hairy root cultures treated with 1 mM ASA had the highest capacity of TAs biosynthesis, in which the content of scopolamine and hyoscyamine reached respectively 57.2 and 14.7 μg g−1 DW. Surprisingly, it was found that 1 mM ASA dramatically induced the efflux of scopolamine. In the liquid medium with 1 mM ASA, the content of scopolamine was 153.4 μg flask−1, about 6.2 folds compared with that of control. At the same time, hyoscyamine was detected at trace levels in liquid medium. In the UV-B stressed hairy root cultures, all the four genes had a very strong increase of gene expression that led to more accumulation of scopolamine and lower accumulation of hyoscyamine. Only trace amounts of hyoscyamine and scopolamine were detected in the liquid medium when hairy root cultures were stressed under UV-B, and this suggested that UV-B did not affect TAs efflux.
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This work was financially supported by the NSFC Project (31370333), the National 863 Hi-Tech Plans (2011AA100605; 2011AA100607), Chongqing Sciences and Technology Project (CSTC2012GGYYJS80013), the Program for New Century Excellent Talents in University (NECT-12-0930) and the Fundamental Research Funds for the Central Universities (XDJK2013A024).
Baifu Qin and Lili Ma have contributed equally to this work.
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Supplemental Fig. 1 PCR analysis of A. luridus hairy roots. analysis for the presence of rolB gene (423 bp) and rolC gene (626 bp) , respectively, M, DL-2000 Marker (100-2,000bp); P, positive control (strain C58C1 containing A. rhizogenes Ri plasmid pRiA4); N, negative control (wild type root), T1-T4, transformed hairy roots of A. luridus. (JPEG 307 kb)
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Qin, B., Ma, L., Wang, Y. et al. Effects of acetylsalicylic acid and UV-B on gene expression and tropane alkaloid biosynthesis in hairy root cultures of Anisodus luridus . Plant Cell Tiss Organ Cult 117, 483–490 (2014). https://doi.org/10.1007/s11240-014-0454-z
- Anisodus luridus
- Gene expression
- Alkaloid biosynthesis