Studies on the Effects of fpf1 Gene on Artemisia annua Flowering Time and on the Linkage between Flowering and Artemisinin Biosynthesis

Abstract

The flowering promoting factor1 (fpf1) from Arabidopsis thaliana was transferred into Artemisia annua L. via Agrobacterium tumefaciens. The fpf1 gene was firstly inserted in the binary vector pBI121 under the control of CaMV 35S promoter to construct the plant expression vector pBIfpf1, then leaf explants of A. annua were infected with A. tumefaciens LBA4404 containing pBIfpf1, and induced shoots. Transgenic plants were obtained through the selection with kanamycin. PCR, PCR-Southern and Southern blot analyses confirmed that the foreign fpf1 gene had been integrated into the A. annua genome. RT-PCR and RT-PCR-Southern analyses suggested that the foreign fpf1 gene had expressed at the transcriptional level. Under short-day conditions, the flowering time of fpf1 transgenic plants was about 20 days earlier than the non-transformed plants; however, no significant differences were detected in artemisinin content between the flowering transgenic plants and the non-flowering non-transgenic plants. These results showed that flowering is not a necessary factor for increasing the artemisinin content, furthermore, there may be no direct linkage between flowering and artemisinin biosynthesis.

Key words

Artemisia annua L. - Asteraceae - Artemisinin - Flower - flowering promoting factor1 (fpf1)

References

• 1 Qinghaosu Antimalarial Coordinating Research Group. Antimalarial studies on Qinghaosu.  Chin Med J. 1979;  92 811-6
• 2 Klayman D L. Qinghaosu (artemisinin): an antimalarial drug from China.  Science. 1985;  228 1049-55
• 3 Ghingra V, Vishweshwar Rao K, Lakshmi Narasu M. Current status of artemisinin and its derivatives as antimalarial drugs.  Life Sci. 2000;  66 279-300
• 4 Schmid G, Hofheinz W. Total synthesis of qinghaosu.  J Am Chem Soc. 1983;  105 624-5
• 5 Abdin M Z, Israr M, Rehman R U, Jain S K. Artemisinin, a novel antimalarial drug: biochemical and molecular approaches for enhanced production.  Plant Med. 2003;  69 289-99
• 6 Paniego N B, Maligne A E, Giulietti A M. V Artemisia annua (Quing-Hao): In vitro culture and the production of artemisinin. In: Bajaj YEP(ed) Medicinal and Aromatic Plants. Biotechnology in Agriculture and Forestry. Vol. 24 Berlin Heidelberg; Springer-Verlag 1993: pp 70-8
• 7 Liersch R, Soicke H, Stehr C, Tuller H V. Formation of artemisinin in Artemisia annua during one vegetation period.  Planta Med. 1986;  52 387-8
• 8 Singh A, Vishwakarma R A, Husain A. Evaluation of Artemisia annua strain for higher artemisinin production.  Planta Med. 1988;  54 475-7
• 9 Chan K L, Chris Tea K H, Jinadasa S, Yuen K H. Selection of high artemisinin yielding Artemisia annua .  Planta Med. 1995;  61 285-7
• 10 Woerdenbag H J, Pras N, Chan N G, Bang B T, Bos R. et al . Artemisinin, related sesquiterpenes, and essential oil in Artemisia annua during a vegetative period in Vietnam.  Planta Med. 1994;  60 272-5
• 11 Ferreira J FS, Simon J E, Janick J. Developmental studies of Artemisia annua: flowering and artemisinin production under greenhouse and field conditions.  Planta Med. 1995;  61 167-70
• 12 Kania T, Russenberger D, Peng S, Apel S K, Melzer S. FPF1 promotes flowering in Arabidopsis .  Plant Cell. 1997;  9 1327-38
• 13 Murashige T, Shook F. A revised medium for rapid growth and bioassay with tobacco tissue culture.  Physiol Plant. 1962;  15 473-97
• 14 Chen D H, Ye H C, Li G F. Expression of a chimeric farnesyl diphosphate synthase gene in Artemisia annua L. transgenic plants via Agrobacterium tumefaciens-mediated transformation.  Plant Sci. 2000;  155 179-85
• 15 Sambrook J, Fritsch E F, Mariatis T. Molecular Cloning: A Laboratory Manual. 2nd ed New York; Cold Spring Harbor Laboratory Press 1989: pp 483-90
• 16 Zhao S S, Zeng M Y. Determination of Qinghaosu in Artemisia annua L. by high performance liquid chromatography.  Chin J Pharm Anal. 1986;  6 3-5
• 17 Vergauwe A, Cammaert R, Vandenberghe D, Genetello C, Inze D, Van Montagu M. et al . Agrobacterium tumefaciens-mediated transformation of Artemisia annua L. and regeneration of transgenic plants.  Plant Cell Rep. 1996;  15 929-33
• 18 Vergauwe A, Van Geldre E, Inze D, Van Montagu M, Van den E eckhout. Factors influencing Agrobacterium tumefaciens-mediated transformation of Artemisia annua L.  Plant Cell Rep. 1998;  18 105-10
• 19 Geng S, Ma M, Ye H C, Liu B Y, Li G F, Chong K. Effects of ipt gene expression on the physiological and chemical characteristics of Artemisia annua L.  Plant Sci. 2001;  160 691-98
• 20 Melzer S, Kampmann G, John C handler, Apel K. FPF1 modulates the competence to flowering in Arabidopsis .  Plant J. 1999;  18 395-405
• 21 Jerreira J FS, Janick J. Floral morphology of Artemisia anuua with special reference to trichomes.  Int J Plant Sci. 1995;  156 807-15