Improved accumulation of ajmalicine and tetrahydroalstonine in Catharanthus cells expressing an ABC transporter

• Published in: Journal of plant physiology Vol. 166; no. 13; pp. 1405 - 1412
• Main Authors: Pomahačová, Barbora, Dušek, Jaroslav, Dušková, Jiřina, Yazaki, Kazufumi, Roytrakul, Sittiruk, Verpoorte, Robert
• Format: Journal Article
• Language: English
• Published: Munich Elsevier GmbH 01.09.2009; Elsevier
• Subjects: ABC transporter; ABC transporters; ajmalicine; Alkaloids; ATP-Binding Cassette Transporters; ATP-Binding Cassette Transporters - genetics; ATP-Binding Cassette Transporters - metabolism; berberine; Berberine - metabolism; Biological and medical sciences; Biological Transport; Catharanthus; Catharanthus - cytology; Catharanthus - genetics; Catharanthus - metabolism; Catharanthus roseus; Coptis japonica; cytology; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Plant; gene overexpression; genetics; indole alkaloids; metabolism; monoterpenoids; Over-expression; physiological transport; Plant physiology and development; Plants, Genetically Modified; Reverse Transcriptase Polymerase Chain Reaction; Secologanin Tryptamine Alkaloids; Secologanin Tryptamine Alkaloids - metabolism; tetrahydroalstonine; Transgenes; transgenic plants; Transport; Vacuoles; Vacuoles - metabolism; Alkaloids; ABC; DW; MIA; MDR; MRP; Catharanthus roseus; Transport; WT; ABC transporter; Over-expression; Raubasine; Accumulation; Medicinal plant; Apocynaceae; Alkaloid; Plant physiology; Dicotyledones; Angiospermae; Spermatophyta; Cell
• ISSN: 0176-1617; 1618-1328; 1618-1328
• DOI: 10.1016/j.jplph.2009.02.015

The biosynthetic pathway of monoterpenoid indole alkaloids in Catharanthus roseus is located throughout various membranes at both the cellular and intercellular levels. ATP-binding cassette (ABC) transporters are known to export vincristine and vinblastine from human cancer cells. It has recently been shown that ABC transporters are also involved in the transport of various monoterpenoid alkaloids in Catharanthus roseus cells. Over-expression of an ABC transporter in this plant might therefore affect the regulation of the alkaloid biosynthetic pathway. CjMDR1, an ABC transporter gene originally isolated from Coptis japonica, was expressed in Catharanthus roseus cell cultures. Cells showing a positive PCR signal of the transgene in both cDNA and genomic DNA samples were subject to transport studies using selected substrates. Unexpectedly, transport of the isoquinoline alkaloid berberine, the main substrate of CjMDR1 transporter in Coptis japonica, was not affected as compared with control and wild-type Catharanthus cells. On the other hand, the endogenous alkaloids ajmalicine and tetrahydroalstonine were accumulated significantly more in Catharanthus roseus cells expressing CjMDR1 in comparison with control lines after feeding these alkaloids.