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IN VITRO AND IN SITU BIOMASS AND ARTEMISININ PRODUCTION IN WORM WOOD (ARTEMISIA ANNUA L. Var. Chiyong)
ABSTRACT
Artemisia annuahas been identified for its anti- malarial properties and millions of Nigerians are at high risk of being infected by the disease. Its propagation is hindered by shortage of seeds, germination problem and effective protocol forin vitro and in situ propagation. The effect of some plant growth regulators (PGRs) on the in vitro regeneration of Artemisia annua using seed, leaf primordial, node explants and callus initiation were investigated in the Biotechnology Laboratory of the Department of Plant Science Ahmadu Bello University, Zaria. Explants were surface sterilized and inoculated on Murashige&Skoog basal media supplemented with varying concentrations of plant growth regulators. Results of Analysis of Variance for the in vitro experiment indicated significant difference among the treatments (P<0.05).In situ experiment was conducted on the farm of the Institute for Agricultural Research (IAR) ABU, Zaria. during the 2012 rainy season and 2013 dry season to determine the effect of some physical, chemical and hormone treatments as well as levels of organic and inorganic fertilizers on the growth, yield and artemisinin content in A. annua. Results of Analysis of Variance (ANOVA) for the in situ experiment indicated no significant difference between the seasons with respect to germination, shoot and root lengths (P≥0.05). Experiment with GA3 and BAP, concentrations 1.5 mg/l of GA3 and 0.5 mg/l BAP recorded the shortest days to in vitro germination. Highest regeneration percentage was observed at 2.0 mg/l of GA3 and 0.5 mg/l of BAP. A concentration of 2.0 mg/l of GA3 and 0.5 mg/lof BAP had the best vigor. Highest shoot height was observed at 2.0 mg/l concentration of GA3 and 0.5 mg/l of BAP. GA3 produced a significant effect on the seed, leaf and nodal explants with respect to vigour shoot length, days to regeneration and percentage regeneration except on the leaf‘s percentage regeneration. Also, BAP had significant effect on all the parameters studied on all the explants except on seed‘s percentage regeneration. Using BAP and NAA, best vigour was observed with 0.5 mg/l of BAP and 0.5 mg/l of NAA. Highest shoot height was recorded at 0.5 mg/l of BAP and 0.5 mg/l of NAA. However, lowest days to regeneration were observed with equal concentrations of 0.5mg/l of both BAP and NAA. Similarly, highest percentage regeneration was observed at equal concentrations of 0.5 viii mg/l of both BAP and NAA. However, with GA3 and NAA plant growth regulators, concentrations of 1.0 mg/l of GA3 and 0.5 mg/l produced the highest vigour. Also, a concentration of 1.5 mg/l of GA3 and 0.5 mg/l of NAA had the best shoot height. However, 0.5mg/l of GA3 and 0.5 mg/l of NAA had the lowest days to regeneration. Highest germination percent of 96 % was observed during the rainy season trial with 10 % Sulphuric acid treatment. Application of 6 t/ha of poultry manure during the raining season had the highest plant height followed by 4 t/ha of poultry manure during the irrigation trial.Warm water treatments in both rain and irrigation trials at 40 0C for 3 minutes and 600C for 3minutes produced the best vigor. Similarly, rain season trial had the highest number of branches with 50 kg/ha of NPK followed by same quantity of NPK during the irrigation trial. Similarly, rain trial had the best leaf fresh weight after flowering with 6 t/ha of poultry manure followed by irrigation trial with 4 t/ha of poultry manure. The best leaf dry weight before flowering was observed with 25 kg/ha of NPK during the rain trial. It was followed by the same quantity of NPK during the irrigation trial. Similarly, rain had the highest leaf fresh weight before flowering with 2 t/ha of poultry manure followed by the same quantity during the irrigation trial. Also, the rain season had the best result with respect to leaf dry weight after flowering with 8 t/ha of poultry manure followed by the same quantity during the irrigation trial. Treatment 6 t/ha of poultry manure had the highest artemisinin content of 2.1 % followed by treatment 75 kg/ha with 1.51 %. Lowest artemisinin content of 0.4 % was recorded with the control. Plant growth regulators and the organic and inorganic manure had significant effect on thein vitro and in situ regeneration and artemisinin production of A. annua.
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