Rapid induction of Agrobacterium tumefaciens-mediated transgenic roots directly from adventitious roots in Panax ginseng

Abstract

Root segments from seedlings of Panax ginseng produced adventitious roots directly when cultured on 1/2 MS solid medium lacking NH 4 NO 3 and containing 3.0 mg l -1 IBA. Using this adventitious root formation, we developed rapid and efficient transgenic root formation directly from adventitious root segments in P. ginseng. Root segments were co-cultivated with Agrobacterium tumefaciens (GV3101) caring b-glucuronidase (GUS) gene. Putative transgenic adventitious roots were formed directly from root segments on medium with 400 mg l -1 cefotaxime and 50 mg l -1 kanamycin. Kanamycin resistant adventitious roots were selected and proliferated as individual lines by subculturing on medium with 300 mg l -1 cefotaxime and 50 mg l -1 kanamycin at two weeks subculture interval. Frequency of transient and stable expression of GUS gene was enhanced by acetosyringon (50 mg l -1 ) treatment. Integration of transgene into the plants was confirmed by the X-gluc reaction, PCR and Southern analysis. Production of transgenic plants was achieved via somatic embryogenesis from the embryogenic callus derived from independent lines of adventitious roots. The protocol for rapid induction of transgenic adventitious roots directly from adventitious roots can be applied for a new Agrobacterium tumefaciens-mediated genetic transformation protocol in P. ginseng.

The improvement of ginsenoside accumulation in Panax ginseng as a result of γ-irradiation

“…The PNX and DDS genes were the most highly expressive in the MAR 5-9 line. The OSC genes are involved in the biosynthesis of phytosterol synthase, oleanane-type synthase, and dammarenediol synthase [38,39]. The DDS gene was involved in the synthesis of ginsenoside, indicating that the DDS gene was highly expressive in the MAR 5-9 line [37].…”

Establishment of adventitious root culture of Stevia rebaudiana Bertoni in a roller bottle system

“…2b). This may have occurred because of the decrease of nitrate concentration (Sudha and Seeni 2001;Han and Choi 2009). The nitrate sources in the MS edium are potassium nitrate (KNO 3 ) and ammonium nitrate (NH 4 NO 3 ), the efficiency of medium strength reduction to increase biomass was explained by Danesh et al (2006).…”

Ginsenoside Production and Morphological Characterization of Wild Ginseng (Panax ginseng Meyer) Mutant Lines Induced by γ-irradiation (60Co) of Adventitious Roots

“…Due to certain limitations in genus Panax breeding, it is difficult to produce large amounts of roots under field conditions [14]. Hence, some scientists followed different biotechnological methods such as root culture, Agrobacterium - mediated hairy root production, and bioreactor- scale production.…”