The world prevalence of diabetes in 2010 among adults aged 20-79 years is estimated to 6.

4%, affecting 285 millions adults. Between 2010 and 2030, there is an expected 70% increase in numbers of adults with diabetes in developing countries and a 20% increase in developed countries. According to the IDF Diabetes Atlas 4th edition, India has the second highest number of people with Diabetes at 5 crore (50 Million) adults. One in 8 Indian adults either has or is at high-risk of Diabetes. Each year more than 3,96 million people worldwide die from diabetes and its complications . This number is expected to increase by more than 50 percent over next decade. Estimated global healthcare expenditures to treat and prevent diabetes and its complications is at least 376 billion US Dollar (USD) in 2010. By 2030, this number is projected to exceed some 490 billion USD. Diabetes is part of the growing epidemic of non communicable diseases, with a high burden for the society on developing countries in future. Primary prevention is the realistic way to curb the rise in the diabetes pandemic. Life style changes aimed to prevention of diabetes by means of diet and weight control by physical activities and exercises and dietary modification. High calorie intake is found to be the main culprit of diabetes and obesity. Though use of sugar needn't be outlawed for people who are diabetic. However, keeping sugar intake low is however certainly recommended. Thus, replacing sugar with low calorie sweeteners can be beneficial. There are several varieties of artificial low calorie sweeteners available but research has shown that these artificial sweeteners may have several side effects. To overcome this problem one can go for natural sweeteners. These can be as good as the artificial ones in terms of taste and potency. They are however healthier and have less potential to cause long term health problems. One such potent natural sweetener, which has been used from decades is Stevia (Stevia rebaudiana Bertoni). It belongs to Asteraceae family. Its leaves contain diterpene glycosides including well characterized steviosides (5-10%) and rebaudioside A (2-4%), which are extensively used as a natural calorie free alternative to artificial sweeteners. Stevia is considered to have high therapeutic values such as anti-hyperglycemic, anticancerous and anti-hypertensive. Because of these immense medicinal properties, the demand for this plant is increasing day by day. But the problem with this variety is that its leaves have characterised liquorice like aftertaste. And it has been reported that it is Rebaudioside A which is the sweetest and the purest compound while the stevioside is responsible for aftertaste bitterness. So, the proposed work is designed to develop a novel variety of sweet herb, Stevia rebaudiana Bertoni with higher contents of stevioside and rebaudioside A without liquorice like aftertaste. Though a novel plant belonging to Stevia rebaudiana Bertoni variety containing four parts by weight or more of rebaudioside-A with respect to one part by weight of stevioside, named as morita has already been developed in Japan using plant breeding approach, which is too laborious and time taking. Therefore, in the present study metabolic engineering approach will be used to alter the biosynthetic pathway in stevia plant leading to the synthesis of

stevioside and rebaudioside A in the ratio of 1:4. And the main focus of the research will be on the over-expression of the genes ent-kaurene 19-oxidase (KO) and UDP-glycosyl transferase 76 G1 leading to an increase not only in stevioside content as a whole, but the rebaudioside A as well.

Title: Cloning and overexpression of ent-kaurene 19-oxidase and UDP-glycosyl transferase 76 G1 genes in Stevia rebaudiana Bertoni and their influence on stevioside and rebaudioside-A contents.