An improved Agrobacterium tumefaciens mediated transformation of Artemisia annua L. by using stem internodes as explants

Abstract

Tian N., Liu S., Ting H., Huang J., van der Krol S., Bouwmeester H., Liu Z. (2013): An improved Agrobacterium tumefaciens mediated transformation of Artemisia annua L. by using stem internodes as explants. Czech J. Genet. Plant Breed., 49: 123-129.Transformation of Artemisia annua, which produces the sesquiterpenoid endoperoxide artemisinin widely used for the treatment of malaria, has been hampered by the low efficiency of adventitious shoot and root formation on a selective medium containing additional compounds for Agrobacterium decontamination. Here we identified several factors which were all shown to be of importance for optimization of Artemisia annua transformation. Results indicated that stem internodes showed better resistance capacity to Agrobacterium decontaminator than leaves did. Agrobacterium tumefaciens with an optical density (OD) value of 0.2-0.5 plus 100 μmol of acetosyringone per litre of solution gave the best transformation efficiency. Moreover, kanamycin at 30 mg/l in the culture medium was effective in suppressing the growth of non-transformed tissue. Furthermore, transgenic shoots required an early induction of rooting. In addition, dimethyl sulphoxide considerably improved the rooting of shoots. The present work provides rapid and reproducible transformation and regeneration of A. annua.

Evaluation on the effectiveness of 2-deoxyglucose-6-phosphate phosphatase (DOGR1) gene as a selectable marker for oil palm (Elaeis guineensis Jacq.) embryogenic calli transformation mediated by Agrobacterium tumefaciens

“…When oil palm immature embryos were selected on hygromycin, only 4–13% of the plantlets tested showed PCR positive for the hptII gene (Bhore and Shah, 2012). It was also shown in other plants such as garlic, Artemisia annua and sweet potato that around 86–92% of the transgenic plants regenerated were tested positive for the transgene used for selection (Zang et al, 2009; Lagunes-Fortiz et al, 2013; Tian et al, 2013). Therefore, the possibility of escape plants regenerated after selection is common and PCR analysis could be a good early detection approach to remove the unwanted escapes.…”

“…Lignin, flavanoid precursors and acetosyringone are compounds known to induce vir gene expression (Stachel et al, 1985). Pre-treatment by a PM containing acetosyringone has also been reported to improve transformation of other plants such as rice, orchid, garlic, Panax ginseng , gherkin ( Cucumis anguria ), and Artemisia annua and onion (Uzè et al, 1997; Choi et al, 2001; Men et al, 2003a; Kenel et al, 2010; Thiruvengadam et al, 2013; Tian et al, 2013; Xu et al, 2014). Prior to Agrobacterium infection, oil palm EC were bombarded with gold particles to initiate physical injury.…”

Artemisinin-based antimalarial research: application of biotechnology to the production of artemisinin, its mode of action, and the mechanism of resistance of Plasmodium parasites

“…To obtain successful transformants, several parameters for Agrobacterium tumefaciens -mediated transformation, such as the concentration of antibiotics, method and duration of co-cultivation, and phytohormones supplied for plant regeneration, have been optimized [ 66 – 71 ]. Among various explants available for transformation, stem internodes and young inflorescence seem to be the most appropriate [ 70 – 72 ]. Phytohormones α-naphthaleneacetic acid and 6-benzylaminopurine are crucial for GST development in young leaves, and root generation also affects GST size [ 73 ].…”