the plant c. chaerophylla was collected from nepal . dried and powdered roots ( 435 g ) were extracted with methanol in a soxhlet extractor .
the crude base fraction was chromatographed over sio2 gel column eluting with solvents of increasing polarity .
eluants from c6h6-chcl3 ( 15 : 85 ) on crystallisation from methanol furnished colorless granules ( 30 mg ) , rf .
. 180~181 , c20 h23 no4 ( m341 ) , [ ]d-300 ( c , 1.5 , etoh ) .
mass spectrum exhibited molecular ion peak at m / z 341 and retro - diel 's alder cleavage of ring c gave a base peak at m / z 190 .
the hnmr and c nmr spectral data were identical to alkaloid 1-corydalmine ( cava et al . , 1968 ) . it was finally identified as 1-corydalmine by direct comparison with authentic sample ( mixed m.p . , co - tlc and super imposable ir ( fig .
stock solution ( 1000 ppm ) of 1-corydalmine was dissolved in 5 mg of compound initially in a few drops of methanol and chloroform ( 1 : 1 ) in a test tube . required concentrations of the chemical ( 100 , 250 ,
500 , 750 , 1000 and 1500 l ) were made by the dilution of the solution with distilled water .
the chloroform and methanol were evaporated on water bath at 80. a drop ( 30~40 l ) of different concentrations of the chemical was placed separately on grease - free glass slides for studying spore germination .
several fungi , e.g. , alternaria brassicae , a. brassicicola , a. solani , curvularia lunata , curvularia maculance , a curvularia sp .
, h. penniseti and a heterosporium sp . , were isolated from their respective infected plant parts .
small portions of the infected material were incubated in moist petri dishes for 24 h and the fungal growth developed after 24 h on potato dextrose agar ( potato 250 g + dextrose + agar 20 g + distilled water 1,000 ml ) was picked up by an inoculating sterile needle and inoculated on slants which were later purified by single spore isolation technique ( singh et al . , 1990 ) .
approximately 200 - 300 spores of each fungus were picked up from fresh sporulating cultures by an inoculating needle aseptically and mixed in a drop of chemical kept on glass slides .
the spores of erysiphe pisi causing powdery mildew of pea ( pisum sativum ) were directly removed from the infected leaves and mixed similarly with the chemical .
all the slides were kept in petri dishes , which were humidified by fixing moist filter paper on the lower and upper surfaces of the base and lid of the petri dishes .
all the petri dishes were incubated at 25  2 for 24 h. after incubation a drop of cotton blue prepared in lactophenol was put on the drop of the chemical containing spores and finally covered with a cover slip .
the spore germination was observed under nikon binocular research microscope and finally the percent spore germination was calculated .
several fungi , e.g. , alternaria brassicae , a. brassicicola , a. solani , curvularia lunata , curvularia maculance , a curvularia sp . , c. pallscens , fusarium udum , a helminthosporium sp . ,
h. penniseti and a heterosporium sp . , were isolated from their respective infected plant parts .
small portions of the infected material were incubated in moist petri dishes for 24 h and the fungal growth developed after 24 h on potato dextrose agar ( potato 250 g + dextrose + agar 20 g + distilled water 1,000 ml ) was picked up by an inoculating sterile needle and inoculated on slants which were later purified by single spore isolation technique ( singh et al . , 1990 ) .
approximately 200 - 300 spores of each fungus were picked up from fresh sporulating cultures by an inoculating needle aseptically and mixed in a drop of chemical kept on glass slides .
the spores of erysiphe pisi causing powdery mildew of pea ( pisum sativum ) were directly removed from the infected leaves and mixed similarly with the chemical .
all the slides were kept in petri dishes , which were humidified by fixing moist filter paper on the lower and upper surfaces of the base and lid of the petri dishes .
all the petri dishes were incubated at 25  2 for 24 h. after incubation a drop of cotton blue prepared in lactophenol was put on the drop of the chemical containing spores and finally covered with a cover slip .
the spore germination was observed under nikon binocular research microscope and finally the percent spore germination was calculated .
the effect of the chemical was tested at varying concentrations starting from 100 , 250 , 500 , 750 , 1,000 to 1,500 ppm .
the maximum effect of the chemical was seen on the spores of h. penniseti as there was no germination at 1,500 ppm and also only 21% at 1,000 ppm as compared to control ( 87.33% ) .
similarly , among the four curvularia species , c. lunata was the most sensitive as the spore germination was significantly inhibited even at 250 ppm .
significant inhibition of spore germination in a. brassicae was observed at 500 ppm and above and other two species of alternaria showed sensitivity at 750 ppm and above concentrations . while 250 ppm and above doses were significantly inhibitory for the germination of spores of heterosporium species , similar significant effect on erysiphe pisi and f. udum was seen at 1,000 and 1,500 ppm concentrations ( table .
a number of chemical compounds isolated from plants are antifungal ( singh et al . , 1990 ; srivastava et al . , 1994 ; sarma et al . , 1999
; singh et al . , 1999 ; basha et al . , 2002 ; maurya et al . , 2001 , 2002 ;
the fungi included in the present study belong to two different groups . but based on the activity of the chemical , it is at present difficult to conclude as to which group of fungi will be most susceptible to this chemical as the number of fungi from different groups is very low .
hence , only further detailed study on several members of other groups will decide its limit and specificity of efficacy .
nevertheless , as antifungal activity of the present compound has not been reported so far and the efficacy is significantly high at low concentrations , there is enough possibility of using this compound in plant disease control under field conditions .
 1-corydalmine , an alkaloid isolated from roots of corydalis chaerophylla inhibited spore germination of some plant pathogenic as well as saprophytic fungi e.g. alternaria brassicae , a. brassicicola , a. solani , curvularia lunata , c. maculans , c. sp . , c. pallscens , erysiphe pisi , fusarium udum , helminthosporium species , h . penniseti and a heterosporium species . 
 1-corydalmine significantly inhibited spore germination of all the fungi at 100 to 1500 ppm . 
 it was effective against all the fungi at 1500 ppm . 
 c. lunata was highly sensitive to this chemical even at 250 ppm . 