Presentation on theme: "by elicitors-induced on greenhouse condition."— Presentation transcript:
1 by elicitors-induced on greenhouse condition. Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition.
2 The experimental purpose Study on the effect of using the elicitors on induce the antioxidant activity and the major isoflavonoids content in White Kwoa Krua in the greenhouse condition.
3 White Kwao Krua Found in northern and north eastern. [Pueraria candollei Grah.var. mirifica (Airy Shaw et. Suvatabandhu) Niyomdham].Found in northern and north eastern.It is considered to be a rejuvenating drug and has a history in folk medicine.Name “White Kwao Krua” was definitively identified as Pueraria candollei in 1952 from which an unusual estrogenic.White Kwao Krua (WKK) has a highconcentration of phytoestrogen.
4 Applications cosmetic industry Pharmaceuticals industry Livestock breast creameye gelskin moisturizer.Pharmaceuticals industryReplacement hormonesLoss bone metabolismLivestockAnimals feed
5 Antioxidant activityAntioxidant compounds play an important role as a health-protecting factor.Antioxidants reduce the risk for chronic diseases including cancer and heart disease.The main characteristic of an antioxidant is its ability to trapfree radicals.superoxide anion radical (O2)hydroxyl radical (OH)peroxyl radical (ROO)Cell metabolism & pollutionsGutteridge (1994)
6 Major Isoflavonoids Genistein Puerarin Igham et al. (1986) PhytonutrientManopause (hormonal replacement)Loss bone (bone metabolism)PuerarinHypertensionantioxidantIgham et al. (1986)
7 Elicitors Molecules that stimulate secondary metabolism Elicitors may form inside or outside plant cells, and aredistinguished as endogenously or exogenously inducers.Depending on their origin, they are classified as biotic or abioticHeike and Dietrich (1995)
8 ElicitorsChitosanSoybean; ppm increased seed isoflavone % and Daidzein 150 %Tawaha et al. (2005)Salicylic acidLupinus ruteus; µM increased Genistein µ g/g DW.Kneer et al.( 1999)Copper (II)chloridePueraria montana; 100 ppm increasedGenistein 5 foldsPueraria lobata; 10 mM for 2 hourincreased Genistein 10 foldsTakashi et al. (1990)Hakanatsuka (1991)
9 Material & Methods Plant Planting WKK in greenhouse at SUT farm Selected 48 plant WKK were age of 1 year oldExperimental planCRD 6 replication with 8 treatmentsElicitor combination (treatments)Water salicylic acid (SA) CuCl2 CuCl2 + SA Chitosan Chitosan + SA Chitosan + CuCl2 Chitosan + CuCl2 +SA
10 Antioxidant determination by DPPH Inhibitory concentration at 50% (IC50)Plot between % radical scavenging VS concentration of each treatmentEquation of nonlinear regression
11 Major isoflavonoids determination by HPLC HPLC conditionHPLC column RP-C18 Agilent® (4.6 mm x 15 cm)Detector Diode array 254 nmFlow rate 1 ml / minVolume injection 10 µlPresser pump 111 barMobile phase (A): 0.1% (v/v) acetic acid in water(B): 0.1% (v/v) acetic acid in acetonitrileTime (min) mobile phase (%)(A) (B)Standard isoflavonoids: puerarin (95 %) and genistein (99.9 %)
17 Conclusion & Discussion The results showed that all combinations had no statistically significant effect on the diameter, fresh weight, dry weight and moisture content of the tuberous roots.it had a statistically significant effect on antioxidant activity and the amount of genistein and puerarin.The combination of chitosan (1000ppm) + copper(II)chloride (300ppm) + salicylic acid (100ppm) gave the highest antioxidant activity (IC50 = 1.99 mg/ml by DPPH).
18 The elicitors did not have the bad effects on root growth of WKK. Conclusion & DiscussionThe combination of chitosan (1000ppm) + copper(II)chloride (300ppm) gave the highest amount of genistein and puerarin were determined by HPLC are 31.9 g/100g in dry weight and g/g in dry weight, respectively.The elicitors can increase antioxidant activity and major isoflavonoids content in tuberous roots of WKK.The elicitors did not have the bad effects on root growth of WKK.