full issue - Association of Biotechnology and Pharmacy
full issue - Association of Biotechnology and Pharmacy
full issue - Association of Biotechnology and Pharmacy
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Current Trends in <strong>Biotechnology</strong> <strong>and</strong> <strong>Pharmacy</strong>Vol. 5 (2) 1098-1103 April 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1098A Simple <strong>and</strong> Rapid Method for Isolation <strong>of</strong> Alternaric Acid fromAlternaria solaniS.J. Patel*, R.B. Subramanian <strong>and</strong> Y.S. JhaB. R. Doshi School <strong>of</strong> Biosciences, Lab. No.103 B, Bakrol Vadtal Road, Satellite Campus,Sardar Patel University, Vallabh Vidhyanagar 388120, India*For correspondence - suchitapatel5@yahoo.comAbstractAlternaric acid is the major toxin <strong>of</strong>Alternaria solani, the causative agent <strong>of</strong> earlyblight <strong>of</strong> tomato. A simple <strong>and</strong> rapid method hasbeen developed for the isolation <strong>of</strong> alternaric acidin crystalline form from the fungal culture filtrate(FCF) with ethanol <strong>and</strong> carbon tetrachloride. Thecrystalline form <strong>of</strong> alternaric acid obtained wassubjected to High Performance Thin LayerChromatography (HPTLC), spectrophotometricanalysis <strong>and</strong> detached leaf assay to confirm thepurity <strong>and</strong> biological activity <strong>of</strong> alternaric acid.This one step protocol is superior to earlierreported methods <strong>and</strong> also gives higher yield.Keywords: Acid, Blight, Tomato, Toxin, TLCIntroductionTomato (Lycopersicon esculentumMill.) belongs to the family Solanaceae.Worldwide, it is the second most consumedvegetable after potato (1). Early blight <strong>of</strong> tomatocaused by the necrotrophic fungus Alternariasolani Sorauer is perhaps the most common foliardisease <strong>of</strong> tomatoes. In USA, Australia, Israel,UK, <strong>and</strong> India, significant reductions in yield (35up to 78%) due to early blight (EB) have beenobserved (2, 3, 4). The symptoms occur on theleaves where circular <strong>and</strong> concentric lesions areproduced. Early blight reduces the photosyntheticarea <strong>and</strong>, in severe cases, can defoliate plants.The fungus survives between crops as myceliaor conidia in soil, plant debris <strong>and</strong> seed (5). Thespores are transported by water, wind, insects,other animals including man, <strong>and</strong> machinery.Eleven toxins have been identified in culturefiltrates <strong>of</strong> A. solani (6). Among these, alternaricacid <strong>and</strong> solanapyrone A, B, <strong>and</strong> C are able toinduce necrotic symptoms similar to EB symptoms(6). Out <strong>of</strong> these, alternaric acid is one <strong>of</strong> themajor toxins found in the filtrates (7) which causeschlorosis <strong>and</strong> necrosis <strong>and</strong> therefore plays a majorrole in early blight symptoms <strong>and</strong> defoliation (8).Alternaric acid does not cause phytotoxicity whensprayed alone on tomato leaves, but it enhancesthe infection process <strong>and</strong> the development <strong>of</strong>necrotic symptoms when added to A. solani sporesuspensions. Alternaric acid was previouslycrystallized by Brian et al. (9). Stoessl <strong>and</strong>Stothers (10) developed a method for extraction<strong>of</strong> alternaric acid from fungal culture filtrate.Previously, Matern et al. (11) tried toisolate alternaric acid from crude FCF. Theprocedure is time consuming <strong>and</strong> the results werenot reliable. Maiero et al. (12) made an attemptto isolate alternaric acid in bulk quantities forscreening <strong>of</strong> resistant varieties, but wereunsuccessful. St<strong>and</strong>ard alternaric acid or alternaricacid in pure form are difficult to obtainIsolation <strong>of</strong> Alternaric acid
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