ifferent concentrations, was pipette into the wells of a 96-well microtiter plate containing 140l of test fungal spore suspension in potato dextrose broth, which was placed in an incubator at 28 C. Antifungal activity of each concentration of protein was performed in triplicates. Fungal spore germination was observed microscopically, whereas optical density at 595 nm wavelength was measured to check the spore growth after inoculation for 30 min and 48 hrs. Controls that were devoid of the test protein were tested for comparing the antifungal activity of the rAdTLP. Values of growth inhibition less than 10% were not considered as significant. Growth inhibition is defined as the ratio of the corrected absorbance at 595 nm of 
the control minus the corrected absorbance of the test sample, divided by the corrected absorbance of the control. The corrected absorbance is defined as the absorbance at 48 h minus that at 30 min. IC50 is defined as the protein concentration at which 50% inhibition was reached. For the in vitro plate assay, fungal discs of uniform size were inoculated at the centre of the Potato Dextrose agar media and incubated at 28 C. When the mycelial spread reached 4 cm in 20354118 diameter, four sterile Whatman no.1 filter paper discs of equal size were placed at equal distance from centre. Purified protein was added at various concentrations at the centre of disc on the plate. The elution buffer served as control and the plates were incubated at 28 C. Growth of Expression and purification of AdTLP protein The 726 bp open reading frame was amplified using ORF-F and ORF-R primers with BamHI and XhoI sites, digested and cloned into the corresponding sites of pET32a expression vector. The cloned vector was transformed into cells of E.coli Rosetta gami-2 strain and the transformed cells were grown at 37 C in Luria and Bertani broth with antibiotics chloramphenicol and ampicillin overnight at 200 rpm in an orbital shaker. A 10 mL aliquot from the overnight grown culture was added to 1L of fresh LB medium and grown further at 37 C until the OD600 reached 0.5-0.6. Then, these cells were induced to express MedChemExpress JW 55 recombinant protein by adding 1mM IPTG for 4 h. Following 10381762 the induction, the induced cells were harvested by centrifugation at 5000 rpm for 10 min at 4 C. Recombinant proteins were exclusively found in inclusion bodies, which was confirmed by resuspending 2 mL culture pellet in lysis buffer were selected to analyse the salt and oxidative stress tolerance. T2 transgenic seedlings were grown on half MSH medium containing 125mgL-1 kanamycin and simultaneously, the WT plants were grown on MSH medium without kanamycin. Twenty seedlings each of WT and kanamycin resistant seedling in T2 generation were transferred to different stress treatment plates for each experiment. For salt stress seedlings were transferred to 100 mM, 200 mM and 300 mM NaCl in MSH. For oxidative stress treatment 2% H2O2 in MSH medium was used. Total chlorophyll content was measured spectrophotometrically as described by Arnon. Lipid peroxidation was assessed by measuring thiobarbituric acid reactive substances as described by Heath and Packer. In the case of NaCl treatment, both the chlorophyll and TBARS estimation were done after 12 days of treatment and for H2O2, after 10 days of treatment. -1, 3 Glucanase assay -1, 3 Glucanase assay was done using the method of Looze et al. with some modifications. The activity of rAdTLP proteins was analyzed by mixing 50 g of protein samp