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Downregulation of Cinnamoyl CoA Reductase Affects Lignin and Phenolic Acids Biosynthesis in Salvia miltiorrhiza Bunge

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Abstract

The biosynthesis of salvianolic acid B shares the phenylpropanoid pathway with lignin, and cinnamoyl CoA reductase (CCR; EC 1.2.1.44) is a specific enzyme in the lignin pathway. In this study, a CCR gene (SmCCR1) from Salvia miltiorrhiza Bunge was cloned using DNA walking technology (GenBank ID: JF798634). The full-length SmCCR1 is 2,489 bp long and consists of four introns and five exons encoding a polypeptide of 324 amino acid residues. Sequence alignment revealed that SmCCR1 shares 83 % identity with CCR sequences reported in Camellia oleifera and other plant species. Expression pattern analysis indicated that expression of SmCCR1 can be induced by exposure to Xanthomonas campestris pv. Campestris or methyl jasmonate. To demonstrate its functioning, we selected a 296-bp fragment and established an RNA interference construct that was introduced into S. miltiorrhiza by Agrobacterium tumefaciens-mediated gene transfer. Transgenic plants exhibited dwarfing phenotypes, and both syringyl and guaiacyl lignin monomers were decreased more than 60 %. In contrast, biosynthesis of phenolic acids—danshensu, rosmarinic acid, and salvianolic acid B—was strongly induced by 2.03-, 1.41-, and 1.45-fold, respectively, in the roots of transgenic plants from line CCR-10. Consistent with these phytochemical changes, downregulation of SmCCR1 also affected the expression of related genes in the phenolics and lignin biosynthetic pathways. Our results also provide potential opportunities for engineering danshensu and salvianolic acid B production in S. miltiorrhiza.

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Acknowledgments

This work was supported by the “Fundamental Research Funds for the Central Universities” (Program No. GK200901014) and “Innovation Funds of Graduate Programs, Shaanxi Normal University” (2011CXS034).

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Authors and Affiliations

  1. Key Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi’an, 710062, People’s Republic of China

    Zhengjun Wang, Langjun Cui, Chen Chen, Xiaojing Liu, Yaping Yan & Zhezhi Wang

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  1. Zhengjun Wang
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  2. Langjun Cui
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  3. Chen Chen
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  4. Xiaojing Liu
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Correspondence to Zhezhi Wang.

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Table 1

Primers used in experimental procedures (DOC 39 kb)

Fig. 1

GC–MS analysis of lignin monomer content from stem extract. IS internal standard; G1, G2 G lignin; S1, S2 S lignin (JPEG 4 kb)

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Wang, Z., Cui, L., Chen, C. et al. Downregulation of Cinnamoyl CoA Reductase Affects Lignin and Phenolic Acids Biosynthesis in Salvia miltiorrhiza Bunge. Plant Mol Biol Rep 30, 1229–1236 (2012). https://doi.org/10.1007/s11105-012-0444-4

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