Pharmaceuticals 2012, 5(2), 133-154; doi:10.3390/ph5020133
Article

Genetic and Phenotypic Analyses of a Papaver somniferum T-DNA Insertional Mutant with Altered Alkaloid Composition

Received: 29 November 2011; in revised form: 15 January 2012 / Accepted: 17 January 2012 / Published: 2 February 2012
(This article belongs to the Special Issue Phytomedicine)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: The in vitro shoot culture of a T-DNA insertional mutant of Papaver somniferum L. established by the infection of Agrobacterium rhizogenes MAFF03-01724 accumulated thebaine instead of morphine as a major opium alkaloid. To develop a non-narcotic opium poppy and to gain insight into its genetic background, we have transplanted this mutant to soil, and analyzed its alkaloid content along with the manner of inheritance of T-DNA insertion loci among its selfed progenies. In the transplanted T0 primary mutant, the opium (latex) was found to be rich in thebaine (16.3% of dried opium) by HPLC analysis. The analyses on T-DNA insertion loci by inverse PCR, adaptor-ligation PCR, and quantitative real-time PCR revealed that as many as 18 copies of T-DNAs were integrated into a poppy genome in a highly complicated manner. The number of copies of T-DNAs was decreased to seven in the selected T3 progenies, in which the average thebaine content was 2.4-fold that of the wild type plant. This may indicate that the high thebaine phenotype was increasingly stabilized as the number of T-DNA copies was decreased. In addition, by reverse transcription PCR analysis on selected morphine biosynthetic genes, the expression of codeine 6-O-demethylase was clearly shown to be diminished in the T0 in vitro shoot culture, which can be considered as one of the key factors of altered alkaloid composition.
Keywords: Papaver somniferum L.; opium alkaloid; Agrobacterium rhizogenes; T-DNA insertional mutant
PDF Full-text Download PDF Full-Text [606 KB, uploaded 2 February 2012 12:02 CET]

Export to BibTeX |
EndNote


MDPI and ACS Style

Kawano, N.; Kiuchi, F.; Kawahara, N.; Yoshimatsu, K. Genetic and Phenotypic Analyses of a Papaver somniferum T-DNA Insertional Mutant with Altered Alkaloid Composition. Pharmaceuticals 2012, 5, 133-154.

AMA Style

Kawano N, Kiuchi F, Kawahara N, Yoshimatsu K. Genetic and Phenotypic Analyses of a Papaver somniferum T-DNA Insertional Mutant with Altered Alkaloid Composition. Pharmaceuticals. 2012; 5(2):133-154.

Chicago/Turabian Style

Kawano, Noriaki; Kiuchi, Fumiyuki; Kawahara, Nobuo; Yoshimatsu, Kayo. 2012. "Genetic and Phenotypic Analyses of a Papaver somniferum T-DNA Insertional Mutant with Altered Alkaloid Composition." Pharmaceuticals 5, no. 2: 133-154.

Pharmaceuticals EISSN 1424-8247 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert