Efficient method for Agrobacterium mediated transformation of Artemisia annua L.
Artemisinin, a potent antimalarial natural products isolated from aerial parts of Artemisia annua L. Many patents have been reported that the demand for artemisinin is exponentially increasing year after year due to increased incidences of drug resistant malaria throughout the world. Leaf explants were used frequently as target tissue to generate transgenic of Artemisia. annua L. However, obtaining a large number of transgenic lines through out the year is a laborious and delicate process. To circumvent this, we have developed a highly efficient leaf explant based Agrobacterium mediated transformation of A. annua L. plant. The gus gene was used as screenable marker to assess and optimize the performance of T-DNA delivery. The age of explant, kind of bacterial inoculation, suspension duration, infection times and co-culture conditions were optimized. The co-culture was carried out with Agrobacterium tumefaciens strain EHA105 under desiccation condition in the dark at 25-28 0C for 2-4 days. Complete analysis of transgene insertion demonstrated that the optimized method of transformation from leaf explants of A. annua L. was efficient and highly reproducible.
Pravej
P
Alam
Anis
A
Mohammad
M M
MM
Ahmad
Mather Ali
MA
Khan
Mohd
M
Nadeem
Riyazuddeen
R
Khan
Mohd
M
Akmal
Seema
S
Ahlawat
Centre for Transgenic Plant Development, Department of Biotechnology, Faculty of Science, Jamia Hamdard, New Delhi, India 62. mzabdin@rediffmail.com.
M Z
MZ
Abdin
D060054:Agrobacterium / Q000235:genetics / Q000378:metabolism*; D031172:Artemisia annua / Q000166:cytology / Q000378:metabolism*; D018920:Coculture Techniques; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D018515:Plant Leaves / Q000166:cytology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018520:Plant Shoots / Q000166:cytology; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
D010940:Plant Proteins; D005966:Glucuronidase