ENHANCED PRODUCTION OF TROPANE ALKALOIDS IN SCOPOLIA PARVIFLORA BY INTRODUCING THE PMT (PUTRESCINE N-METHYLTRANSFERASE) GENE

In wild-type Scopolia parviflora (Solanaceae) tissues, only the roots express the enzyme putrescine N-methyltransferase (PMT; EC 2.1.1.53), which is the first specific precursor of the tropane alkaloids. Moreover, the tropane alkaloid levels were the highest in the root (0.9 mg g−1 on a dry weight b...

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Published in:In vitro cellular & developmental biology. Plant 2005-03, Vol.41 (2), p.167-172
Main Authors: LEE, OK-SUN, KANG, YOUNG-MIN, JUNG, HEE-YOUNG, MIN, JI-YUN, KANG, SEUNG-MI, KARIGAR, CHANDRAKANT S., PRASAD, D. THEERTHA, BAHK, JUNG-DONG, CHOI, MYUNG-SUK
Format: Article
Language:English
Subjects:
Alkaloids
atropine
biosynthesis
enzyme activity
gene overexpression
genes
genetic transformation
hairy root
Korean culture
leaves
Messenger RNA
metabolic engineering
Metabolism
Plant roots
Plants
Polymerase chain reaction
Predetermined motion time systems
putrescine N-methyltransferase
Research Reports
Rhizobium rhizogenes
roots
scopolamine
Scopolia
Scopolia parviflora
tissues
tobacco
Transgenic plants
tropane alkaloids
Tropanes
Western blotting
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Summary:In wild-type Scopolia parviflora (Solanaceae) tissues, only the roots express the enzyme putrescine N-methyltransferase (PMT; EC 2.1.1.53), which is the first specific precursor of the tropane alkaloids. Moreover, the tropane alkaloid levels were the highest in the root (0.9 mg g−1 on a dry weight basis), followed by the stem and then the leaves. We metabolically engineered S. parviflora by introducing the tobacco pmt gene into its genome by a binary vector system that employs disarmed Agrobacterium rhizogenes. The kanamycin-resistant hairy root lines were shown to bear the pmt gene and to overexpress its mRNA and protein product by at least two-fold, as determined by polymerase chain reaction (PCR) and Northern and Western blottings, respectively. The transgenic lines also showed higher PMT activity and were morphologically aberrant in terms of slower growth and the production of lateral roots. The overexpression of pmt markedly elevated the scopolamine and hyoscyamine levels in the transgenic lines that showed the highest pmt mRNA and PMT protein levels. Thus, overexpression of the upstream regulator of the tropane alkaloid pathway enhanced the biosynthesis of the final product. These observations may be useful in establishing root culture systems that generate large yields of tropane alkaloids.
ISSN:1054-5476
1475-2689
DOI:10.1079/IVP2004621