ABSTRACT
The use of chitosan as an alternative for fungicides has received more attention worldwide. Hence, this study aimed to evaluate in vitro and in vivo antifungal activity of chitosan against Fusarium solani causing root rot in fenugreek. Chitosan treatments ranged from 0.1 to 2gL-1 were tested against F. solani on to potato dextrose agar and in potato dextrose broth. The results revealed that increase in concentrations of chitosan significantly reduced growth, dried biomass, sporulation and spore germination of F. solani. The hyphal swellings and distortion of F. solani mycelia were induced by chitosan. Fenugreek seeds treated with chitosan at 2 gL-1 and 0.5 gL-1 showed reduced F. solani infection and increased seed germination, respectively. In pot and field studies, fenugreek seeds treated with chitosan at 2.0 gL-1 greatly reduced root rot disease severity and also enhanced yield parameters. The activity of defence enzymes, such as chitinase, ß-1, 3-glucanase and total phenol were increased in chitosan treated in fenugreek plants. This increased activity offered protection to fenugreek plants against F. solani to a greater extent. The results showed that chitosan could be used as inducer of defense response and has the potential of controlling fenugreek root rot disease.
ABSTRACT
A selective and rapid multiresidue analysis method is presented for simultaneous estimation of 12 plant growth regulators (PGRs), namely, auxins (indol-3-acetic acid, indol-3-butyric acid, and naphthyl acetic acid), cytokinins (kinetin, zeatin, and 6-benzyladenine), gibberellic acid (GA3), abscisic acid, and synthetic compounds, namely, forchlorfenuron, paclobutrazole, isoprothiolane, and 2,4-dichlorophenoxy acetic acid (2,4-D) in bud sprouts and grape berries at the development stages of 2-3 and 6-8 mm diameters, which are the critical phases when exogenous application of PGRs may be necessary to achieve desired grape quality and yield. The sample preparation method involved extraction of plant material with acidified methanol (50%) by homogenization for 2 min at 15000 rpm. The pH of the extract was enhanced up to 6 by adding ammonium acetate, followed by homogenization and centrifugation. The supernatant extract was cleaned by SPE on an Oasis HLB cartridge (200 mg, 6 cc). The final extract was measured directly by LC/MS/MS with electrospray ionization in positive mode, except for 2,4-D, GA3, and abscisic acid extracts, which required analysis in negative mode. Quantification by multiple reaction monitoring (MRM) was supported with full-scan mass spectrometric confirmation using "information-dependent acquisition" triggered with MRM to "enhanced product ionization" mode of the hybrid quadrupole-ion trap mass analyzer. The LOQ of the test analytes varied between 1 and 10 ng/g with associated recoveries of 80-120% and precision RSD <25% (n = 8). Significant matrix-induced signal suppression was recorded when the responses for pre- and postextraction spikes of analytes were compared; this could be resolved by using matrix-matched calibration standards. The method could successfully be applied in analyzing incurred residue samples and would, therefore, be useful in precisely deciding the necessity and dose of exogenous applications of PGRs on the basis of measured endogenous levels.
