TITLE:
Efficient Somatic Embryogenesis and Organogenesis of Self-Pollination Artemisia annua Progeny and Artemisinin Formation in Regenerated Plants
AUTHORS:
Fatima Alejos-Gonzalez, Kelly Perkins, Malcolm Isaiah Winston, De-Yu Xie
KEYWORDS:
Artemisia annua; Artemisinin; Biosynthesis; Self-Pollination; Somatic Embryogenesis; Organogenesis; HPLC-MS
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.4 No.11,
November
14,
2013
ABSTRACT: To
enhance the understanding of artemisinin biosynthesis, we have successfully
bred self-pollination Artemisia annua plants. Here, we report efficient somatic embryogenesis and organogenesis of
self-pollination plants and artemisinin formation in regenerated plants. The
first through sixth nodal leaves of seedlings are used as explants. On
agar-solidified MS basal medium supplemented with TDZ (0.6 mg/l) and IBA (0.1
mg/l), all explants after inoculation of less than 3 weeks start to form
embryogenic calli, which further produce globular, torpedo, heart and early
cotyledon embryos. In all six positional leaves, explants from the sixth leaf
show the rapidest responses to induction of embryogenic calli and somatic
embryos. On this medium, somatic embryos continuously develop into adventitious
buds, which can form adventitious roots on a rooting medium containing NAA (0.5
mg/l). Meanwhile, on agar-solidified MS basal medium supplemented with BAP (1
mg/l) and NAA (0.05 mg/l), approximately 100% of explants from leaves #3-6 form calli in less than 3 weeks of inoculation and
adventitious buds via organogenesis in 3-4 weeks. In all six positional leaves, explants from the
sixth leaf exhibit the rapidest response to induction of calli and adventitious
buds. Nearly 100% adventitious buds can form adventitious roots on the rooting
medium. Regenerated plants from both somatic embryogenesis and organogenesis
complete self-pollination to produce seeds in 80-90 days of growth in growth chamber. LC-ESI-MS analysis
demonstrates that regenerated plants biosynthesize artemisinin. These results
show the highly efficient regeneration capacity of self-pollination A.
annua plants that can form a new platform to enhance the understanding of
artemisinin biosynthesis and metabolic engineering.