Abstract
Background: Artemisia annua is well known for its anti-malarial bio-active compound
artemisinin. Development of elite planting material of A. annua and its agro-technology can fulfill the
requirement of Artemisinin-based Combination Therapy (ACT) dosages worldwide.
Objectives: To develop an efficient in-vitro propagation protocol for A. annua and assess the field performance
of in-vitro propagated plants for their growth and artemisinin yield.
Methods: The in-vitro propagation protocol of A. annua was developed using the nodal segment in
four steps viz: initiation, multiplication, rooting and hardening. In-vitro propagated plants were transplanted
with open-pollinated seed raised plants in an experimental field trial having soil supplementation
of Farm Yard Manure (FYM), vermicompost and NPK.
Result: Maximum 92% shoots were initiated in Murashige and Skoog medium (MS) with 0.44 μM
6-benzyl aminopurine (BA) and highest 281.33 ± 09.75 micro-shoots/inoculum obtained in MS with
15.54 μM BA. The maximum number of roots was found in MS with 100 mg/L activated charcoal
while 78.20% of plants survived in the sand: soil: vermicompost (1:1:1) mixture. The highest dry leaf
yield (6.37 t/ha) was observed in in-vitro propagated plants grown with vermicompost, while highest
artemisinin content (1.11 ± 0.10) and artemisinin yield (65.05 kg/ha) was found in the in-vitro propagated
plants grown with FYM after 120 days of transplantation.
Conclusion: This study reports an efficient, cost-effective and rapid in-vitro propagation protocol for
A. annua as well as enhanced artemisinin yield through the cultivation of in-vitro propagated plants
using organic soil supplement inputs. This would lead to an increase in the production of artemisinin
yield and fulfill the demand of Artemisinin-based Combination Therapy (ACT).
Keywords:
Micropropagation, HPTLC, farm yard manure (FYM), vermicompost, agriculture, medicinal plant.
Graphical Abstract
[1]
Anonymous . Chemical studies on qinghaosu (artemisinin): China cooperative group on Qinghaosu and its derivatives as anti-malarials. J Tradit Complement Med 1982; 2: 3-8.
[4]
WHO World Malaria Report 2018. Geneva: World Health Organization 2018.
[5]
WHO The World Health Report. Geneva: World Health Organization 2001.
[11]
Noumi E. Traditional medicines for HIV/AIDS and Opportunistic infections in North-West Cameroon: Case of skin infections. Public Health 2011; 1: 44.
[19]
Sharma G, Shanker V, Agrawal V. An efficient microprapagation protocol of an elite clone EC-353508 of Artemisia annua L. an important antimalarial plant. Int J Pharma Bio Sci 2011; 2: 205-14.
[21]
Mohammad A, Alam P, Ahmad MM, Ali A, Ahmad J, Abdin MZ. Impact of plant growth regulators (PGRs) on callogenesis and artemisnin content in Artemisia annual. Plants Ind J Biotech 2014; 13: 26-33.
[23]
Georgieva ZE, Atanasov NT, Hristova PD, Konstantinova ZM, Todorova CG. Efficient protocol for mass micropropagation of Artemisia annua L GSC. Biol Pharm Sci 2018; 5(2): 59-68.
[24]
Kumar S, Banerjee S, Dwivedi S, et al. Registration of Jeevanraksha and Suraksha varieties of the anti-malarial medicinal plant Artemisia annua. Curr Res Med Aromat Plants 1999; 21: 47-8.
[26]
Chandravanshi M, Sahu Y, Agrawal A, Raja W. In vitro Micropropagation of important commorcial medicinal plant: Plumbago zeylanica. Adv Biol Res 2014; 8: 139-42.
[27]
Nik SMM, Keshavarzi MHB, Heidari M, Yarahmadi GR. Studies of changes in chlorophyll content in Artemisia annua L. under the effects of biological and chemical fertilizers. Adv Biol Res (Faisalabad) 2011; 2: 196-202.
[29]
Agrawal A. evaluation and development of simple sequence repeats (ssr) molecular marker for Artemisia annua. PhD dissertation 2013.
[31]
Sairkar P, Chandravanshi MK, Shukla NP, Mehrotra NN. Mass production of an economically important medicinal plant Stevia rebaudiana using in vitro propagation techniques. J Med Plants Res 2009; 3: 266-70.
[33]
Damtew Z, Tesfaye B, Bisrat D. Leaf, essential oil and artemisinin yield of Artemisia (Artemisia annua L) as influenced by harvesting age and plant population density. World J Agric Sci 2011; 7: 404-12.
[35]
Sangoil L. Understanding plant density effects on maize growth and development: An important use to maximize grain yield. Sci Rural 2000; 39: 59-68.
[36]
Mensah DB, Dorcas QM, Kwame BR, Kodjo DS. Comparative study on the field performance of fhia-01 (hybrid dessert banana) propagated from tissue culture and conventional sucker in Ghana. J Plant Dev 2012; 19: 41-6.