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Current Trends in Biotechnology and PharmacyVol. 5 (2) 1098-1103 April 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1101Fig. 1. Shows crystals of alternaric acid observedunder 10X magnification and crystals observed under40X magnification (inset)Fig. 2(a). Band of crude FCF (from 7 th to 21 st days)seen clearly at 366nm on 18 th day on TLC foil (10X20)showing the presence of alternaric acid HPTLC analysisof purified alternaric acid and figure 2 (b) showsband of alternaric acid on TLC foil (10X10) observedat 366nm wavelengthFig. 3. Shows scanning of HPTLC foil in Scanner 3at 366 nmFig. 4. Shows detached leaf bioassayAbsorption maxima of undiluted sampleAbsorption maxima of undiluted sampleAbsorbance ( o A)Absorbance ( o A)Wavelength (nm)Fig. 5 (a). Shows UV-spectroscopic scan of undilutedalternaric acid from 190-400nmWavelength (nm)Fig. 5 (b). Shows UV-spectroscopic scan of dilutedalternaric acid from 190-400nmPatel et al
Current Trends in Biotechnology and PharmacyVol. 5 (2) 1098-1103 April 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1102can be proved biologically. Spectrophotometricscanning of crystalline alternaric acid showed adistinct peak in diluted samples whereas; inundiluted sample many peaks were seen. Thus,the presence of alternaric acid was confirmed bythis simple analytical technique.Fig. 6. Shows in vivo bioassay Treatedis shown to be 12-(5, 6-dihydro-4-hydroxy-6-methyl-2-oxopyran-3-yl)-4, 5-dihydroxy-3-methyl-9-methylene-12-oxododec-6-ene-5-carboxylicacid. (14).Increase in alternaric acid yield wasmeasured by weighing dried crystals of alternaricacid. The yield obtained was five times higher bythis method than by the previous methodsreported. The presence of colorless alternaric acidin crystalline form could be visualizedmicroscopically under 10X and 40X magnificationsand also through naked eyes. HPTLC is a fastand reliable technique. The advantage of HPTLCis that quantities of substances too small to bedetected by chemical methods could be separatedand thereafter their biological activity can bemeasured by performing bioassay. The presenceof alternaric acid in crude FCF was confirmedby comparing both the Rf values of crude FCFand that of crystalline alternaric acid. Scanningof HPTLC foils showed a distinct peak whichproves the presence and purity of alternaric acid.The detached leaf assay is a simplemethod which allows controlled inoculation withspecific isolates, without destroying the wholeplant. The result of detached leaf bioassay showedsimilar symptoms as shown by the A. solaniinfected leaves in field. The symptoms appearedvery fast and were distinct. Further virulence ofcrystals was checked and confirmed by in vivobioassay. Thus the presence of alternaric acidAcknowledgements We are highly thankful toDr. Subhash Patel, Anand Agricultural University,India for providing the A. solani isolate.References1. FAOSTATdata,2005..http://faostat.fao.org/faostat/form?collection=Production.Crops.Primary & Domain=Production&servlet=1&hasbulk=&version = ext&language=EN. Last updated February 2005.2. Datar, V.V. and Mayee, C.D. (1972).Conidial dispersal of Alternaria solani intomato. Indian Phytopathology 35: 68-70.3. Basu, P.K. (1974). Measuring early blight,its progress and influence on fruit losses innine tomato cultivars. Can Plant Dis Surv.54: 45-51.4. Jones, J.B., Jones, J.P., Stall, R.E. and Zitter,T.A. eds. (1993).Compendium of TomatoDiseases. St Paul, Minnesota, USA:American Phytopathological Society.5. SheRf, A.F. and MacNab, A.A. (1986).Vegetable Diseases and Their Control. JohnWiley & Sons, New York.6. Montemurro, N. and Visconti, A. (1992).Alternaria metabolites – chemical andbiological data. In:Chelkowski J, Visconti A(eds) Alternaria biology, plant disease andmetabolites. Elsevier, Amsterdam, pp 449-558.7. Brian, P.W., Elson, G.W., Hemming, H.G.and Wright, J.M. (1952). The phytotoxicproperties of alternaric acid in relation toIsolation of Alternaric acid
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(December 7-9, 2011)Venue: Karunya
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