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

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 decontamin...

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Bibliographic Details
Published in:Czech Journal of Genetics and Plant Breeding 2013-01, Vol.49 (3), p.123-129
Main Authors: Tian, N, Liu, S, Ting, H, Huang, J, Krol, S. van der, Bouwmeester, H, Liu, Z
Format: Article
Language:English
Subjects:
absorbance
Acetosyringone
Agrobacterium
Agrobacterium radiobacter
Agrobacterium tumefaciens
anti-malaria
Artemisia annua
Artemisinin
combination
Decontamination
Dimethyl sulfoxide
dna
drug
Explants
Genetic transformation
internodes
Kanamycin
leaves
Malaria
Optical density
Optimization
plants
Regeneration
Rooting
Selective media
Shoots
stem internode
Stems
traditional chinese medicine
Transformations
transgene
Vector-borne diseases
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Summary: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.
ISSN:1212-1975
1212-1975
1805-9325
DOI:10.17221/105/2012-cjgpb