Enhancement of Artemisinin Content through Four Cycles of Recurrent Selection with Relation to Heritability, Correlation and Molecular Marker in Artemisia annua L.

Abstract

Due to the high demand and low yield of the anti-malarial drug artemisinin in natural populations of Artemisia annua (Quinghao), an attempt has been made to enhance the artemisinin content through 4 cycles of recurrent selection (C₀-C₃) using selected genotypic and phenotypic traits. Based on their phenotypic and genotypic characteristics, the top 5 % plants of each cycle were selected, and their seedlings were planted in poly-cross block to produce seeds for the subsequent generation. A significant increase in the artemisinin content (0.15 % in C₀ to 1.16 % in C₃, i.e., about 40 % genetic gain over the generation) was observed. This enhancement was directly correlated with the plant height and branching intensity in all four cycles. Similarly, the PCV (phenotypic coefficient of variation) and GCV (genotypic coefficient of variation) have been observed to have a higher value for artemisinin content. The DNA marker (MAP 12) with relation to artemisinin was also identified for high yielding genotypes in all four cycles of selection. Over the four cycles of recurrent selection, the plant developed an oval appearance (Variety: CIM-Arogya) and a high artemisinin content (1.16 %).

Artemisia annua - Asteraceae - artemisinin - recurrent selection - correlation - molecular marker

References

• 1 McVaugh R. Compositae, Vol. 12. Anderson WR Flora Novo-Galiciana: a descriptive account of vascular plants of western Mexico. Ann Arbor; University of Michigan Press 1984
• 2 Bailey L H, Bailey E Z. Hortus third. New York; Macmillan 1976
• 3 Gray A. Synoptical flora of North America, 1 Part II. Smithsonian Institution, Washington, DC; University Press 1984
• 4 Klayman D L. Weeding out malaria.  Nat Hist. 1989;  10 18-26
• 5 Klayman D L. Artemisia annua: from weed to respectable anti malarial plant. Kinghorn DA, Baladrin MF Human medicinal agents from plants. Washington DC; Am. Chem. Soc. Symp. Ser. 1993: 242-255
• 6 Kumar S, Gupta S K, Gupta M M, Verma R K, Jain D C, Shasany A K, Darokar M P, Khanuja S P S. Method for maximization of artemisinin production by the plant Artemisia annua. US Patent 6393763. 2002
• 7 Sharma A, Bindra R L, Tewari R. Review article: Artemisia annua: cultivation, utilization and chemical studies.  CIMAP Publication. 1991;  46 91
• 8 Klayman D L, Lin A J, Acton N, Scovill N, Hoch J M, Milhous W K, Theorides A D. Isolation of artemisinin (quinghaosu) from Artemisia annua growing in the United States.  J Nat Prod. 1984;  47 715-717
• 9 WHO . Severe Falciparum malaria (severe and complicated malaria, third edition).  Trans R Soc Trop Med Hyg. 2000;  94 (Suppl. 1) SI-1-SI-90
• 10 Kumar S, Gupta S K, Singh P, Bajpai P, Gupta M M, Singh D, Gupta A K, Ram G, Shasany A K, Sharma S. High yield of artemisinin by multiharvest of Artemisia annua crops.  Indian Crops Prod. 2004;  19 71-78
• 11 Khanuja S P S, Paul S, Shasany A K, Gupta A K, Darokar M P, Gupta M M, Verma R K, Ram G, Kumar A, Lal R K, Bansal R P, Singh A K, Bhakuni R S, Tandon S. Genetically tagged improved variety “CIM-Arogya” of Artemisia annua for high artemisinin yield.  J Med Aromatic Plant Sci. 2005;  27 520-524
• 12 Ferreira J F S, Simon J E, Janick J. Relationship of artemisinin content of tissue cultured, greenhouse, grown and field grown plants of Artemisia annua.  Planta Med. 1995;  61 351-355
• 13 Delabays N, Benakis A, Collet G. Selection and breeding for high artemisinin (qinghaosu) yielding strains of Artemisia annua.  Acta Hort. 1993;  330 203-206
• 14 Mathur A K, Kumar S. Micropropagation of Artemisia annua via the inflorencence.  J Herb Spices Med Plants. 1996;  4 61-71
• 15 Gupta M M, Jain D C, Verma R K, Gupta A P. A rapid analytical method for the estimation of artemisinin in Artemisia annua.  J Med Aromatic Plant Sci. 1996;  18 5-6
• 16 Gupta M M, Jain D C, Mathur A K, Singh A K, Verma R K. Isolation of a high artemisinic acid containing plants of Artemisia annua.  Planta Med. 1996;  62 280-282
• 17 Singh R K, Chaudhary B D. Biometrical methods in quantitative genetic analysis. New Delhi; Kalyani Pub 1979
• 18 Khanuja S P S, Shasany A K, Darokar M P, Kumar S. Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils.  Plant Mol Biol Rep. 1999;  17 1-7
• 19 Khanuja S P S, Shasany A K, Srivastava A, Kumar S. Assessment of genetic relationships in Mentha species.  Euphytica. 2000;  111 121-125
• 20 Dudley J W. Seventy six generations of oil and protein percentage in maize. Pollak E, Kempthrone O, Bailey TJ Proceedings of the International Conference on Quantitative Genetics. Ames; Iowa State University Press 1977: 459-474
• 21 Misevic D, Alexander D E. Twenty four cycles of phenotypic recurrent selection for percent oil in maize. Per se and test cross performance.  Crop Sci. 1989;  29 320-324
• 22 Schipper H, Frey K J. Observed gains from three recurrent selection regimes from increased groat oil content of oat.  Crop Sci. 1991;  31 1505-1510
• 23 Frey K J, Holland J B. Nine cycles of recurrent selection for increase groat oil content in oat.  Crop Sci. 1999;  39 1639-1641
• 24 Kumar S, Banerjee S, Dwivedi S, Gupta M M, Verma R K, Jain D C, Khanuja S P S, Mathur A K, Bagchi G D, Zehra M, Mehta V K, Naqvi A A, Paul S, Ram G, Ram M, Saikia D, Sangwan R S, Santha Kumar T R, Shasany A K, Darokar M P, Singh A K, Singh A. Registration of 'Jeevanraksha' and 'Suraksha' varieties of the antimalarial medicinal plant Artemisia annua.  J Med Aromatic Plant Sci. 1999;  21 47-48
• 25 Trigg P I. Quinghaosu (Artemisinin) as an antimalarial drug.  Economics Med Plant Res. 1990;  3 20-55
• 26 Ferreira J F S, Simon J E, Janick J. Developmental studies of Artemisia annua: flower and artemisinin production under greenhouse and field conditions.  Planta Med. 1994;  61 167-170
• 27 Jain D C, Mathur A K, Gupta M M, Singh A K, Verma A K, Gupta A P, Kumar S. Isolation of high artemisinin yielding clones of Artemisia annua.  Phytochemistry. 1996;  43 993-1001
• 28 Wallaart T E, Pars N, Beekman A C, Quax W J. Seasonal variation of artemisinin and its biosynthetic precursor in plants of Artemisia annua of different geographical origin: proof for the existence of chemotypes.  Planta Med. 2000;  66 57-62
• 29 Kulkarni R N, Ramesh S. Development of lemongrass clones with high oil content through population improvement.  J Essent Oil Res. 1992;  4 181-186
• 30 Kulkarni R N. Lemongrass breeding; past progress and future prospects.  J Med Aromatic Plant Sci. 2000;  22 278-284
• 31 Kulkarni R N, Bhaskaran K, Ramesh S. Five cycles of recurrent selection for increased essential oil content in east Indian lemongrass; response to selection and effects on heritability of traits and intertraits correlations.  Plant Breed. 2003;  122 131-135
• 32 Khanuja S P S, Paul S, Shasany A K, Darokar M P, Shukla A K, Gupta M M, Kumar A. Primers and a screening method for identification of artemisinin producing plants. Canadian Patent No: PCT/IN2003/000404 (87). 2005
• 33 Hallauaer A R, Miranda J R. Quantitative genetic in maize breeding, 2nd edition. Ames; Iowa State University Press 1998
• 34 Khanuja S P S, Paul S, Shasany A K, Gupta A K, Darokar M P, Gupta M M, Verma R K, Ram G, Kumar A, Lal R K, Bansal R P, Singh A K, Bhakuni R S, Tandon S. High artemisinin yielding plant genotype “CIM-AROGYA”. US Patent 7375260. 2008