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Induction of anthraquinone biosynthesis in Rubia cordifolia cells by heterologous expression of a calcium‐dependent protein kinase gene
Author(s) -
Shkryl Y.N.,
Veremeichik G.N.,
Bulgakov V.P.,
Zhuravlev Y.N.
Publication year - 2011
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.23077
Subject(s) - anthraquinone , activator (genetics) , callus , heterologous , transgene , biology , secondary metabolism , kinase , biochemistry , heterologous expression , transformation (genetics) , gene , biosynthesis , arabidopsis , gene expression , microbiology and biotechnology , chemistry , botany , mutant , recombinant dna , organic chemistry
Calcium‐dependent protein kinases (CDPKs) play an important role in plant cell responses to stress and pathogenic attack. In this study, we investigated the effect of heterologous expression of the Arabidopsis CDPK gene, AtCPK1 , on anthraquinone production in transgenic Rubia cordifolia cells. AtCPK1 variants (a constitutively active, Ca 2+ ‐independent form and a non‐active form used as a negative control) were transferred to callus cells by agrobacterial transformation. Overexpression of the constitutively active, Ca 2+ ‐independent form in R. cordifolia cells caused a 10‐fold increase in anthraquinone content compared with non‐transformed control cells, while the non‐active form of AtCPK1 had no effect on anthraquinone production. Real‐time PCR measurements showed that the activation of anthraquinone biosynthesis in transgenic calli correlated with the activation of isochorismate synthase gene expression. The activator effect of AtCPK1 was stable during prolonged periods of transgenic cell cultivation (more than 3 years) and the transgenic cultures exhibited high growth. Our results provide the first evidence that a CDPK gene can be used for the engineering of secondary metabolism in plant cells. Biotechnol. Bioeng. 2011; 108:1734–1738. © 2011 Wiley Periodicals, Inc.