YU Yong2ting, X IE Yuan2yuan, HUAN G L i2li, KAN G Zhen2sheng
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1 35 2 ( ) V o l. 35 N o Journal of N o rthw est A & F U niversity (N aṫ Sci. Ed. ) Feb Β21, 32 α,,, (, ) [ ] M S,, 7, 14, 21 d (26, 150 rgm in), 3, 52 (DN S) Β21, 32,,, Β21, 32,,,,, 7 d, ;, 21 d,, 7 d [ ] ; ; Β21, 32 ;M S ; ; [ ] S [ ] A [ ] (2007) Effe c ts of d iffe re nt com b ina tions of ca rbon a nd nitroge n source s in MS m e d ium on a c tiv itie s of e xtra ce llula r Β21, 32g luca na se p roduce d by ta ke 2a ll pa thoge n YU Yong2ting, X IE Yuan2yuan, HUAN G L i2li, KAN G Zhen2sheng (College of P lant p rotection,n orthw est A & F U niversity, Yang ling, S haanx i , China) Abstract: T he pathogen ic fungi Gaeum annom y ces g ram in is var. tritici (Ggt) w as cu ltu red in modified M S m edium (26, 150 rgm in) w ith differen t com b ination s of carbon and n itrogen sou rces. T he enzym e activity in the cu ltu re filtrate w as detected by 3, 52din itro salicylic acid (DN S) in 7, 14, 21 d after cu ltu ring the fungu s. T he resu lts show ed the enzym e activities w ere obviou sly differen t in the cu ltu re filtrate w ith w heat stalk s, w heat leaves, m aize stalk s, m aize leaves and m aize roo ts as carbon sou rce com b ined w ith differen t n itrogen sou rces. T he h ighest enzym e activities w ere detected in the cu ltu re filtrate w ith w heat leaves as the carbon sou rce and the low est enzym e activities w ere in the cu ltu re filtrate w ith m aize roo ts as the carbon sou rce. T he enzym e activities w ere no t sign ifican tly differen t in the cu ltu re filtrate w ith w heat tissues as compared w ith that w ith m aize tissues. Comparing 4 n itrogen sou rces, the enzym e activities w ere obviou sly h igher in the cu ltu re filtrate w ith o rgan ic n itrogen than that w ith ino rgan ic n itrogen. T he h ighest enzym e act ivit ies w ere detected in the cu ltu re filt ra te w ith beef ex t ract a s n it rogen sou rce 7 d after cu ltu ring the fungu s. T he enzym e act ivit ies detected in the cu ltu re filt ra te w ith p ep tone w ere low and α [ ] [ ] ( ) ; (PCS IRT ) ; [ ] (1980- ),,,, E2m ail: yuyongting@ 126. com [ ] (1961- ),,,, E2m ail: huanglili1@ho tm ail. com
2 2 : Β21, reached the peak 21 d after cu ltu ring the fungu s. T he enzym e activities in the cu ltu re filtrate w ith beef ex t ract decrea sed g radua lly du ring the cu ltu re stage. T he best ca rbon and n it rogen com b ina t ion in the m edium fo r enzym e activities w as beef exact and w heat leave detected 7 d after cu ltu ring the fungu s. Key words: take2all pathogen of w heat; Gaeum annom y ces g ram in is var. tritici; Β21, 32glucanases; M S m edium ; carbon sou rce; n itrogen sou rce (Gaeum annom y ces g ram in is (Sacc. ) von. A rx and O livier var. tritici W alker),,,,, [128 ], [528 ],, Β21, 32, (lign in2tube) Β21, 32 [9210 ], Β21, 32, Β21, 32,,, Β21, 32 [11 ],,,,,, Β21, 32, Gaeum annom y ces g ram in is var. tritici (Ggt) 9826, PDA ( M S ), V an Hoof [12 ] [11 ] M S, 1% M S, 1% (L am inarin) Sigm a, PDA (26 ) 3 5 d, 5 mm 4, 50 ml 150 ml, 3, ( rgm in, ) , 14, 21 d,, rgm in 5 m in, , 52 (DN S) mo lgl ph % 100 ΛL, 37 5 m in 100 ΛL,, m in DN S [11 ] (U ) : 37, m in 1 Λmo l ,, Β21, 32 1 a,, Β21, 32 7 d, U gml,, 14 d (0. 06 U g
3 112 ( ) 35 ml ),, 21 d U gml, %, Β21, 32, 7 d, U gml, 14 d U g ml, %, 21 d, %, U g ml, Β21, 32, 1 Β21, ; - -. ; - -. ; - -. ; a. ; b. F ig. 1 Extracellular Β21, 32glucanase activity detected in the different m edia of w heat stalk as the single Carbon source and beef extract, pep tone, N H 4NO 4 and N H 4C l as nitrogen source, respectively, in the different incubation tim e - -. Beef extract; - -. Pep tone; - -. ammonium nitrate; - -. Ammonium ch lo rine; a. W heat stalk; b. W heat leaves 1 b,, Β21, 32 ;, Β21, 32 ;, Β21, 32, Β21, 32, 1,, Β21, 32, Β21, ,,, Β21, 32 2 a,, Β2 1, 32, 7 d, U gml, 14 d, 21 d U gml, %, ;, 7 d U gml, 14 d U gml, %, 21 d U g ml, ; ( U gml ),,, Β21, 32 ( 2 b), 21 d 10. 7% ;, 21 d 11. 1% 2 c,, Β21, 32, 14 d 7 d, ;,, 21 d, 35. 8% 42. 7%,, d 7 d 12. 9% 38. 7% ;,,
4 2 : Β21, Β21, ; - -. ; - -. ; - -. ; a. ; b. ; c. F ig. 2 Extracellular Β21, 32glucanase activity detected in the different m edia of m aize stalk as the single Carbon source and beef extract, pep tone, N H 4NO 4 and N H 4C l as nitrogen source, respectively, in the different incubation tim e - -. Beef extract; - -. Pep tone; - -. ammonium nitrate; - -. Ammonium ch lo rine; a. M aize stalk; b. M aize leaves; c. M aize roo ts 2,, Β21, 32 ;, ;, ;,,, Β21, 32,,, [11 ] 2, 3, Β21, 32 [9210 ], Β21, 32 [11 ], Β21, 32,,, Β21, 32 Β21, 32, (T richod erm a reesei) Β2 1, 32, [13 ] ; (Coch liobolus ca rbonum ), [12 ] ; San to s [14 ] (Czapek), Β21, 32,, [11 ], Β21, 32,,,, Β21, 32 [ ] [ 1 ],,,. [J ]., 1997 (1) : [ 2 ],. [J ]. :, 2000, 28 (6) : [ 3 ] Garder P A, A ngus J F, P itson G D, et al. A comparison of six mo theds to contro l take2all in w heat[j ]. A ustralian Journal of A gricultural Research, 1998, 49 (8) : [4 ] Cook R J. P roblem s and p rogress in the bio logical contro l of w heat take2all[j ]. P lan Path, 1994, 43: [ 5 ] Do ri S, H ershenho rn J, So lel Z, et al. Characterization of an endopo lygalacturonase associated w ith take2all disease of w heat[j ]. Physio logical and M o lecular P lant Patho logy, 1992, 40 (3) : [ 6 ] Do ri S, So lel Z, Kashm an Y, et al. Cell w all2degrading enzym es p roduced by Gaeum annomy ces g ram inis vaṙ tritici in v itro and in v ivo [ J ]. Physio logical and M o lecular P lant Patho logy, 1995, 46 (3) :
5 114 ( ) 35 [ 7 ] W este G. Extra2cellular enzym e p roduction by various iso lates of Op h iobolus g ram in is Sacc [J ]. Phytopath Z, 1970a, 52: [ 8 ] W este G. Extra2cellular enzym e p roduction by various iso lates of Op hiobolus g ram inis and O. g ram inis vaṙ avenae g. Enzym es p roduced w ithin the ho st tissue [ J ]. Phytopath Z, 1970b, 67: [ 9 ] Kang Z S, H uang L L, Buchenauer H. Cytochem istry of cell w all component alterations in w heat roo ts infected by Gaeum nnnomy ces g ram in ins vaṙ tritici[j ]. J P lant D is P ro tec, 2000, 107 (4) : [ 10 ] H uang L L, Kang Z S, Buchenauer H. Comparison of infection of w heat roo ts by P hialop hora g ram inicola and Gaeum annomy ces g ram inis var. tritici by ultrastructural and cytochem ical studies [J ]. J P lan D is P ro tec, 2001, 108 ( 6) : [ 11 ],,,. Β21, 32 [J ]., 2003, 22 (4) : [ 12 ] V an Hoof A, L eykam J, Schaeffer H J, et al. A single Β21, 32 glucanase secreted by the m aize pathogen Coch liobolus carbonum acts by an exo lytic m echanism [J ]. Physio l M o lec Path, 1991, 39: [ 13 ] Bam fo rth C W. T he adap tability, purification, and p roperties of exo2β21, 32glucanase from the fungus T richod erm a reesei[j ]. B iochem J, 1980, 191: [ 14 ] Santo s T, V illanuebva J R, N om bela C. P roduction and catabo lite rep ression of P enicillium italicum beta2glucana2 ses[j ]. J Bacterio l, 1977, 129 (1) : ( 109 ) [ ] [ 1 ],,,. [J ]. :, 2003, 23 (3) : [ 2 ]. [M ]. :, [3 ]. [ J ]., 1992 (3) : [ 4 ],. [M ]. :, 2003: 226. [ 5 ],. [ J ]., 2002 (5) : [ 6 ]. [M ]. :, 1999: 217. [ 7 ],,. GA 3 2, 42D [J ]., 2000, 20 (3) : [ 8 ]. [M ]. :, 1980: 25. [ 9 ]. [M ]. :, 1985: [ 10 ]. [D ]. :, 2003: [ 11 ],,,. [ J ]., 1981, 17 (2) : 125. [ 12 ],,. [J ], 2003 (5) : 327.
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