The relationship between TXS, DBAT, BAPT and DBTNBT gene expression and taxane production during the development of Taxus baccata plantlets

Abstract

Taxol and related taxane accumulation in plants is regulated by the expression of genes involved in their biosynthesis. Although the metabolic pathway leading to taxol has been almost completely elucidated, comparatively little is known about the rate-limiting steps and their regulation. In this paper we report on a study of taxane production in Taxus baccata plantlets grown in vitro for 1 year. The relationship between taxane patterns and the expression of genes encoding the enzymes taxadiene synthase (TXS), 10-deacetylbaccatin III-10β- O-acetyltransferase (DBAT), baccatin III 13- O-(3-amino-3-phenylpropanoyl) transferase (BAPT) and 3′-N-debenzoyl-2′-deoxytaxol-N-benzoyltransferase (DBTNBT), involved in early and late steps of the taxane pathway, has been considered. A far higher content was found in the aerial part of the plantlets than in the roots. The most abundant taxane in the aerial parts was 10-deacetylbaccatin III, which increased as the plantlets grew, indicating a low conversion to baccatin III and taxol. In contrast, the levels of 10-deacetylbaccatin III in the roots remained lower than those of taxol. These results correlated with transcript accumulation of the studied genes, since in the aerial parts the expression of DBAT, which codes for the enzyme that converts 10-deacetylbaccatin III into baccatin III, did not increase with the age of plantlets, unlike that of TXS, BAPT and DBTNBT, suggesting that this gene controls a rate-limiting step in the taxane biosynthetic pathway. The lower taxane levels found in the roots also correlated with gene expression, since only the early pathway gene TXS was induced in this organ during the 1-year growth period.