Involvement of LeMRP, an ATP-binding cassette transporter, in shikonin transport and biosynthesis in Lithospermum erythrorhizon
| Autor: | Cheng-Yi Tang, Xiaoming Wang, Yong Hua Yang, Yanjun Pang, Rong-Wu Yang, Jiangyan Fu, Jinliang Qi, Gui Hua Lu, Yue-Lin Luo, Yin-Ling Zhu, S.‐J. Chu |
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| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine ATP-binding cassette transporter Plant Science Real-Time Polymerase Chain Reaction 01 natural sciences 03 medical and health sciences chemistry.chemical_compound Biosynthesis Gene Expression Regulation Plant Gene expression Cloning Molecular Phylogeny Ecology Evolution Behavior and Systematics Plant Proteins chemistry.chemical_classification biology Lithospermum Membrane Transport Proteins Sequence Analysis DNA General Medicine Membrane transport Plants Genetically Modified Lithospermum erythrorhizon biology.organism_classification Transformation (genetics) 030104 developmental biology Enzyme Biochemistry chemistry Membrane protein ATP-Binding Cassette Transporters Naphthoquinones 010606 plant biology & botany |
| Zdroj: | Plant Biology. 20:365-373 |
| ISSN: | 1435-8603 |
| Popis: | Shikonin and its derivatives are important medicinal secondary metabolites accumulating in roots of Lithospermum erythrorhizon. Although some membrane proteins have been identified as transporters of secondary metabolites, the mechanisms underlying shikonin transport and accumulation in L. erythrorhizon cells still remain largely unknown. In this study, we isolated a cDNA encoding LeMRP, an ATP-binding cassette transporter from L. erythrorhizon, and further investigated its functions in the transport and biosynthesis of shikonin using the yeast transformation and transgenic hairy root methods, respectively. Real-time PCR was applied for expression analyses of LeMRP and shikonin biosynthetic enzyme genes. Functional analysis of LeMRP using the heterologous yeast cell expression system showed that LeMRP could be involved in shikonin transport. Transgenic hairy roots of L. erythrorhizon demonstrated that LeMRP overexpressing hairy roots produced more shikonin than the empty vector (EV) control. Real-time PCR results revealed that the enhanced shikonin biosynthesis in the overexpression lines was mainly caused by highly up-regulated expression of genes coding key enzymes (LePAL, HMGR, Le4CL and LePGT) involved in shikonin biosynthesis. Conversely, LeMRP RNAi decreased the accumulation of shikonin and effectively down-regulated expression level of the above genes. Typical inhibitors of ABC proteins, such as azide and buthionine sulphoximine, dramatically inhibited accumulation of shikonin in hairy roots. Our findings provide evidence for the important direct or indirect role of LeMRP in transmembrane transport and biosynthesis of shikonin. |
| Databáze: | OpenAIRE |
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