CH INESE JOURNAL OF B IOLOGICAL CONTROL 1998, 14 (3) 123 129 (, 310029) THE ROL E OF EXTRACELL ULAR PROTEASES OF ENTOMOPATHOGEN IC FUNGI IN HOST-PATHOGEN IC PROCESS AND THE IR ASSAY TECHN IQUES XU Junhuan FEN G M ingguang XU Q ian (D epartm en t of B io logical Sciences, Zhejiang A gricu ltu ral U n iversity, H angzhou 310029) [ ], ; S476. 12,,, (B idochka, et al., 1992, 1994a, 1994b; Boucias, et al., 1991), (B eauveria bassiana) (B idochka, et al., 1991, 1992; Charn ley, et al., 1991; Khachatou rian s, 1991) DNA,,,,, (tryp sin2like enzym es),, (A scherson ia a ley rod is) (Con id iobolus corona tus) (E ry n ia spp. ) (L ag en id ium g ig an teum ) (N om u rey i riley i) (M eta rh iz ium an isop liae) (V erticillium lecan ii) (Charn ley, et al., 1991; Khachatou rian s, 1991),, (B idochka, et al., 1991, 1992; Gillesp ie, et al., 1992),, (B idochka, et al., 1990;, ;,, 34,, ;,, 40,, 123
1996;, 1998;, 1996), 1,, (B. brong n ia rtii) (P aecilom y ces f a rinosus),,, (Sh im izu, et al., 1993) 18. 5 35. 0KD a, PM SF (phenylmo thylsufonyl fluo ride) (E ry n ia rh iz osp ora) (P and ora d ip terig ena) (P. neoap h id is) (Co le, et al., 1993; Gerlter, 1971) N 2Α2Θ2 2L 2 2 (N 2Α2Θ2to syl2 L 2lysine2ch lo rom ethy ketone ) N 2Α2 2DL 2 2Θ2 2 (N 2Α2benzoyl2DL 2 argin ine2θ2n itro2an ilide) 2,, : (1), ; (2) ; (3) 2. 1 (B idochka, et al., 1987, 1990; Charn ley, et al., 1991; Sṫ L eger, et al., 1987, 1988), (Sṫ L eger, et al., 1986),,, (Sṫ L eger, et al., 1986) ph 7, ph ph 2 4, pk Α, 10mmo lgl,, (M elanop lus sang u in ip es) 7. 9% (,, ) 8. 4% (, ) (B idochka, et al., 1994a),, B idochka ( 1994b ) (esterification) (carboxam idation), (acetylation), 124
(phenylglyoxylation),,,,, (B idochka, et al., 1992),, (H all, et al., 1961),,, (Sṫ L eger, et al., 1987), (Boucias, et al., 1991), 2. 2 100 (A ndersen, 1980; Cox, et al., 1987; W ills, 1987) B idochka (1994b) ( ), 41. 1% ( ) 83. 0% ( ) PM SF,,, (> 31KD a), PM SF, (ph 10), (B idochka, et al., 1988b; Sṫ L eger, et al., 1987) Gerlter (1971), ph 7 9,, (H all, et al., 1961), (> 31KD a),, P r2,, (Sṫ L eger, et al., 1987),,,, 2,, (S accha rom y ces k luy veri) 3, (p ro2pheno l 125
ox idase) (Gillesp ie, et al., 1992), Sh im izu (1993) 4, 988nggm l, 10 8 g m l Yo sh ida (1990) (A n tica rsia g em m a ta lis), (T richop lusia n i) (Boucias, et al., 1987),, 4, ( ) (b ioassay),, (B idochka, et al., 1990;, 1998;, 1996;, 1996),, UV (Cu lex p ip iens) (Robert, et al., 1985), (R hod n ius p rolix us) (Sila, et al., 1986), (Cu rcu lio ca ry ae) (H elioth is z ea) (Charn ley, et al., 1991; Sṫ L eger, et al., 1986), (1998),,, 7 r 2 = 0. 67 ( ), Sṫ L eger (1988) (M and uca sex ta), 60%,, Kucera (1968),,,,,,,,, 5 N 2 (GlcNA c),, (A sp erg illus ory z ae) (K lapper, et al., 1973; D ean, et al., 1983), (B idochka, et al., 1988a) (N eu rosp ora crassa) 126
(Scopes, 1982) 3, 3 (Grove, et al., 1981; H an son, et al., 1975;M arzluf, et al., 1968), 3,, 5mmo lgl (cam P) GlcNA c,, 25% 40%, (B idochka, et al., 1988a),,, GlcNA c,,, N 2 GlcNA c, GlcNA c GlcNA c,, 6 C lark son ( ) P r1 P r2 P r1, P r1 P r1 P r2,, P r1 P r2, (A sp erg illus n id u lans) P r1, P r1 P r1 DNA area crea, PCR DNA area crea,, area crea DNA, 2S2 (glu tath ione2s2tran sferase) P r1, 7 7. 1,,, 2, 2 ς2,,, 2 (B idochka, et al., 1988a): A zoco ll ( 2 ; Calb iochem 2 Beh ring; 50 100 ) 10m g 1. 0m l 2N aoh (1. 0mo lgl, ph 8. 5), 3m l 127
1m l 25m l, 37 30 ( 180rgm in ) W hatm an N o. 2, (Κgnm = 520), (PU ) ς2 (B idochka, et al., 1988b) : ( Ampho line, ph 3 10),, 37 ς2 30 (B idochka, et al., 1990) : 1. 0% 1. 5% (W gv ), 10 8 gm l,, 5Λl, 6 26 3, 15% (W gv ) H gc l2 ( 2mo lgl HC l ), H gc l2, Fo lin (, 1996) :, T ris2hc l (0. 05mo lgl, ph 8. 5) 1% 1. 0m l, 1. 0m l, 37 30, 2m l 0. 4mo lgl Fo lin, 1. 0Λg (PU ) (B idochka, et al., 1992) : 1. 0mmo lgl 2 2 2 2 2 2 (N 2succinyl2A la2a la2p ro2phe2θ2 n itroan ilide) 10m g, 16m l (dim ethy su lfox ide), 10m l T ris2hc l (0. 1mo lgl, ph 8. 5) 1. 8m l 100Λl, 100Λl, 37 10 (180rgm in), 1 3 100Λl, (Κgnm = 410), 3 110Λmo lgm l 21 gm in 21 (PU ), 7. 2 95% (N H 4) 2SO 4,, 5, ( 6KD a ) 24,,, - 20 (B idochka, et al., 1988b). 1998., ( ).,. 1996., 23: 273 278.,. 1996., 23: 279 284. A ndersen S O. 1980. Cuticular Sclero tization. In: M iller T A, ed. Cuticle T echniques in A rth ropods. 185 214. B idochka M J, Khachatourians G G. 1987. A pp l. Environ. M icrobio l., 53: 1679 1684; 1988b, 54: 2699 2704. B idochka M J, Khachatourians G G. 1988a. Exp. M yco l., 12: 161 168. B idochka M J, Khachatourians G G. 1990. J. Invertbṙ Patho l., 56: 362 370; 1991, 58: 106 117; 1992, 59: 165 173; 1994a, 63: 7 13; 1994b, 61: 26 32. Boucias D G, Pendland J C. 1987. Experientia, 43: 336 339. Boucias D G, Pendland J C. 1991. In: Co le G T, Hoch H C, eds. The Fungal Spo re and D isease Initiation in P lants and A nim als. 101 127. Charnley A K, Sṫ L eger R J. 1991. In: Co le G T, Hoch H C, 128 eds. The Fungal Spo re and D isease Initiation in P lants and A nim als. 267 286. Co le S C J, Charnley A K, Cooper R M. 1993. FEM S M icrobio l. L etter, 113: 189 195. Cox D L,W ills J H. 1987. Insect B iochem., 17: 457 468. D ean D D, Dom ans A J. 1983. Exp. M yco l., 7: 31 39. E l2sayed G N, Ignoffo C M, L eathers T D, et al. 1993. M ycopatho logia, 122: 139 142. Gerlter A T. 1971. J. B iochem., 20: 541 546. Gillesp ie J P, Khachatourians G G. 1992. Comp. B iochem. Physio l., 103: 455 463. Grove G, M arzluf G A. 1981. J. B io l. Chem., 256: 463 470. H all D A, Czerkaw sk i J W. 1961. B iochem. J., 80: 128 136. H anson M A, M arzluf G A. 1975. P roc. N atl. A cad. Sci. U SA, 72: 1240 1244. Khachatourians G G. 1991. In: A ro ra D K,A jello L,M ukerji
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