35 2 ( ) V o l. 35 N o. 2 2007 2 Journal of N o rthw est A & F U niversity (N aṫ Sci. Ed. ) Feb. 2007 Β21, 32 α,,, (, 712100) [ ] 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 ; ; [ ] S435. 121 [ ] A [ ] 167129387 (2007) 0220110204 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 712100, 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 [ ] 2006201223 [ ] (30270863) ; (PCS IRT 200558) ; [ ] (1980- ),,,, E2m ail: yuyongting@ 126. com [ ] (1961- ),,,, E2m ail: huanglili1@ho tm ail. com
2 : Β21, 32 111 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, 1 1. 1 1. 1. 1 Gaeum annom y ces g ram in is var. tritici (Ggt) 9826, PDA 1. 1. 2 ( M S ), V an Hoof [12 ] [11 ] 1. 1. 3 1. 1. 2 M S, 1% 1. 1. 4 1. 1. 2 M S, 1% 1. 1. 5 (L am inarin) Sigm a, 1. 2 1. 2. 1 PDA (26 ) 3 5 d, 5 mm 4, 50 ml 150 ml, 3, (26 150 rgm in, ) 1. 2. 2 7, 14, 21 d,, 4 10 000 rgm in 5 m in, 1. 2. 3 3, 52 (DN S) 0. 05 mo lgl ph 5. 5 2 0. 1% 100 ΛL, 37 5 m in 100 ΛL,, 37 30 m in DN S [11 ] (U ) : 37, m in 1 Λmo l 2 2. 1 1,, Β21, 32 1 a,, Β21, 32 7 d, 0. 062 U gml,, 14 d (0. 06 U g
112 ( ) 35 ml ),, 21 d 0. 051 U gml, 7 17. 7%, Β21, 32, 7 d, 0. 027 U gml, 14 d 0. 044 U g ml, 7 63. 0%, 21 d, 7 122. 2%, 0. 06 U g ml, Β21, 32, 1 Β21, 32 - -. ; - -. ; - -. ; - -. ; 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, 32 2. 2 2,,, Β21, 32 2 a,, Β2 1, 32, 7 d, 0. 057 U gml, 14 d, 21 d 0. 03 U gml, 7 47. 4%, ;, 7 d 0. 037 U gml, 14 d 0. 049 U gml, 7 13. 3%, 21 d 0. 075 U g ml, ; (0. 021 0. 027 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%,, 14 21 d 7 d 12. 9% 38. 7% ;,,
2 : Β21, 32 113 2 Β21, 32 - -. ; - -. ; - -. ; - -. ; 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) : 38241. [ 2 ],. [J ]. :, 2000, 28 (6) : 49271. [ 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) : 122521240. [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: 4292437. [ 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) : 2032210. [ 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) : 1892198.
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