Plant husbandry – Process
Reexamination Certificate
2000-03-30
2002-05-28
Campbell, Bruce R. (Department: 1661)
Plant husbandry
Process
Reexamination Certificate
active
06393763
ABSTRACT:
FIELD OF INVENTION
This invention relates to a method for maximization of artemisinin production by the plant
Artemisia annua
belonging to family: Asteraceae. More particularly, the invention is developed through agro-technology involving method of optimizing the planting time, transplanting scheduling, population density, number of harvests and harvesting schedule leading to enhanced yields of artemisinin and related metabolites which have pharmaceutical value of anti-infectives, particularly as antimalarial drug.
BACKGROUND OF INVENTION
The plant
Artemisia annua
(family: Asteraceae ) is valued for producing a sesquiterpenoid lactone endoperoxide named ‘artemisinin’ which is a promising antimalarial drug and effective against
Plasmodium falciparum, P. vivax
and drug resistant parasites at nanomolar concentration. The compounds &agr; and &bgr; arteethers which are synthesized from dihydroartemisinin by etherification with ethanol, were developed as antimalarial drug in India by Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow and Central Drug Research Institute (CDRI), Lucknow after phase III clinical trial. Artemisinins are active against
Schistosoma mansoni
and
S. japonicum
, in-vitro and in experimental animal models. These compounds are also active against
Leishmania major, Toxoplasma gondii
and
Pneumocystis carinii
in-vitro and against
P.carinii
in-vivo. Artemisinins have immunosuppressive activity and also potential anticancer activity. Considering the importance of artemisinins which is tedious and difficult to synthesize chemically an all out programme was undertaken to develop
Artemisia annua
plant varieties with high artemisinin content, development of agrotechnology for increase yield of these compounds, followed by improved extraction procedure. In this direction the inventors were successful in developing and releasing a variety named “Jeevan Raksha” from an isolated population containing high artemisinin in the foliage (0.5 to 1.0%) (Sushil Kumar et al. 1999. Journal of Medicinal and Aromatic Plant Sciences. 21:47-48.) This plant “Jeevan Raksha” not only produces high artemisinin but also maintains the synchronized conversion to higher level of artemisinin during May to October. As the content of artemisinin fluctuates from zero level at the time of planting to more than 0.4 to 1.00% during May and June with subsequent functions of increase till October, it was necessary to scientifically develop cultivation methodology for the crop to maximize the vigour of the foliage and biosynthesis of artemisinin by systematic scheduling. For this purpose the inventors carried out planned experiments with variation in planting times, population density and number of harvest from the crop to increase the yield from limited area within optimum span of time.
OBJECT OF THE INVENTION
The main object of the invention relates to a method for maximize the artemisinin yield of the plant
Artemisia annua
by scheduling transplanting time and following a multiple harvesting schedule coinciding with the higher artemisinin accumulation in the plant.
In another object of the invention relates to provide a method for maximizing the yield of artemisinin to the tune of 85-95 kg/ha by following the schedule of the invention.
In yet another object of the invention relates to a method wherein the yield of artemisinin is maximized by maintaining the schedule defined in the invention for different transplanting time of the plant.
SUMMARY OF THE INVENTION
To meet the above objects and others, the present invention provides a method for maximizing the artemisinin yield of the plant
Artemisia annua
by scheduling transplanting time and following a multiple harvesting schedule coinciding with the higher artemisinin accumulation in the plant, wherein the method comprises of the following steps;
(a) sowing seeds of
Artemisia annua
plant on raised bed nursery during second and third week of December and maintaining the optimum moisture throughout,
(b) transplanting seedlings thus obtained bearing at least 5-15 leaves into the main field fertilized with NPK @ 80,40,40 kg/ha to achieve a population density of 50,000 to 2,00,000 per ha followed by light irrigation in the second week of March and irrigation every fortnight there after,
c) harvesting the crop four times by cutting the plant tops leaving 75-100 cm part of plant for further regeneration, the said harvests are performed in a manner that the first harvest is done in fourth week of May, second harvest in third week of July, third harvest in second week of September and fourth harvest in third week of October of each year, and at each harvesting time care is taken to care at least one green branch, and
d) extracting artemisinin from the plant tissue immediately after each harvest.
DETAILED DESCRIPTION
It is therefore a intent of the invention to provide the crop of
Artemisia annua
, a process for increasing the foliage harvest coupled with the maximum artemisinin yield.
The phenotype of a plant is the consequence of interaction between the genotype and the environment. The genotype is the genetic make up which directs the plant to express all characters. But proper expression needs the signal from the environment to express at a particular time, stage and tissue. This is called temporal and spatial expression of genetic traits, which includes synthesis of important chemicals and secondary metabolites like artimisinin in case of
Artemisia annua
. Based on this fact, it is correct to predict that a plant is not likely to produce and accumulate secondary metabolite just after germination from the seed. Same is true in case of the plant
Artemisia annua
requiring a certain stage of maturity to start producing artemisinin. Now at the time of production of the compound artemisinin, the plant should have sufficient foliage to accumulate this secondary metabolite. Further, the fluctuating environmental conditions also fluctuate the synthesis of these products. A better genotype responds to these signals in a more vigorous way compared to a weak genotype. At the macro level of commercial crop cultivation in the field environment cannot be manipulated as in the case of cultures in the laboratory. However, critical factors like planting date, number of harvests, harvesting schedules and population density can be manipulated for adjusting to the environmental conditions for maximizing the crop as well as critical chemical production.
With these objective, planned experiments were carried out on the high yielding plant variety “Jeevan Raksha” of
Artemisia annua
, to optimize the agrotechnological process to maximize the yield of herbage and thus artemisinin in a single season so that the method, sequence and optimized scheduling approach can be utilized universally with varying crop seasons in different climatic regions globally by skilled modifications.
REFERENCES:
J.C. Laughlin Effect of Agronomic practices on plant yield and anti-malarial constituents of Artemisia annua L. Acta Horticulturae 331, 1993.*
J.C. Laughlin The influence of distribution of antimalarial constituents in Artemisia annu I. on time and method of harvest. Acta Horticulturae 390 Nov. 1995.*
N. Delabays et al. The evolution of the artemisinin content in Artemisia Annua L. during one growing season. Atti convegno internazionale: Coltivazione e miglioramento di piante officinali, Trento, Italy, 2-3 guigno 1994, (1996) pp 635-638 4 ref.*
Muni Ram et al. Effect of planting Time on the Yield of Essential Oil and Artemisinin in Artemisia annua under Subtropical conditions. J. Essent. Oil Res., 9, 193-197 (Mar/Apr 1997).
Darokar Mahendra Pandurang
Gupta Madan Mohan
Gupta Shiv Kumar
Jain Dharam Chand
Khanuja Suman Preet Singh
Campbell Bruce R.
Council of Scientific and Industrial Research
Merchant & Gould P.C.
Para A
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