Plant Cell Reports

, Volume 9, Issue 3, pp 121–124 | Cite as

Genetic transformation of foxglove (Digitalis purpurea) by chimeric foreign genes and production of cardioactive glycosides

  • Kazuki Saito
  • Mami Yamazaki
  • Koichiro Shimomura
  • Kayo Yoshimatsu
  • Isamu Murakoshi
Article

Abstract

The chimeric neo and gus genes on a mini Ti vector are efficiently transferred into the genome of fox glove (Digitalis purpurea L.) using a binary vector system based on a rootinducing Ri plasmid, pRi15834. The transgenic state of established transformed roots was confirmed by Southern blot analysis and by detection of agropine and mannopine. The expression of the chimeric genes controlled by the promoters from TR 1′–2′ genes, nos gene and cauliflower mosaic virus 35S RNA was demonstrated by enzymatic and histochemical assays of neomycin phosphotransferase II and ß-glucuronidase. Enzyme-linked immunosorbent assay (ELISA) was carried out using polyclonal antibody reactable against digitoxin to investigate the production of cardenolides. The results of ELISA indicated that the cardioactive glycosides were highly produced in the green transformed hairy roots.

Abbreviations

CaMV

cauliflower mosaic virus

ELISA

enzyme-linked immunosorbent assay

NPT-II

neomycin phosphotransferase II

neo

gene encoding NPT-II from Tn5

GUS

ß-glucuronidase

gus

gene encoding GUS from Escherichia coli

Km

kanamycin

nos

gene encoding nopaline synthase

TR1′–2′

genes 1′ and 2′ of TR-DNA of pTiAch5

Rif

rifampicin

X-gluc

5-bromo-4-chloro-3-indolyl-ß-d-glucuronide

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Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Kazuki Saito
    • 1
  • Mami Yamazaki
    • 1
  • Koichiro Shimomura
    • 2
  • Kayo Yoshimatsu
    • 2
  • Isamu Murakoshi
    • 1
  1. 1.Faculty of Pharmaceutical SciencesChiba UniversityChibaJapan
  2. 2.Tsukuba Medicinal Plant Research StationNational Institute of Hygienic SciencesIbarakiJapan

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