Biotransformation of ent-13-epi-manoyl oxides difunctionalized at C-3 and C-12 by filamentous fungi

Biotransformation of ent-3beta,12alpha-dihydroxy-13-epi-manoyl oxide with Fusarium moniliforme gave the regioselective oxidation of the hydroxyl group at C-3 and the ent-7beta-hydroxylation. The action of Gliocladium roseum in the 3,12-diketoderivative originated monohydroxylations at C-1 and C-7, both by the ent-beta face, while Rhizopus nigricans produced hydroxylation at C-7 or C-18, epoxidation of the double bond, reduction of the keto group at C-3, and combined actions as biohydroxylation at C-2/epoxidation of the double bond and hydroxylation at C-7/reduction of the keto group at C-3. In the ent-3-hydroxy-12-keto epimers, G. roseum originated monohydroxylations at C-1 and C-7 and R. nigricans originated the oxidation at C-3 as a major transformation, epoxidation of double bond and hydroxylation at C-2. Finally, in the ent-3beta-hydroxy epimer R. nigricans also originated minor hydroxylations at C-1, C-6, C-7 and C-20 and F. moniliforme produced an hydroxylation at C-7 and a dihydroxylation at C-7/C-11.     

Hydroxylation of the triterpenoid nigranoic acid by the fungus Gliocladium roseum YMF1.00133

The ability of the fungus Gliocladium roseum YMF1.00133 to transform the bioactive nigranoic acid (=(24Z)-9,19-cyclo-3,4-secolanosta-4(28),24-diene-3,26-dioic acid) was investigated. Three new products from the co-cultures of nigranoic acid and G. roseum YMF1.00133 were obtained by employing a combination of Sephadex LH-20 and silica-gel column chromatography. The major metabolite was identified as 15beta-hydroxynigranoic acid, and the minor metabolites as 6alpha,15beta-dihydroxynigranoic acid and 7beta,15beta-dihydroxynigranoic acid by mass spectrometry and NMR spectroscopy. This is the first report of the biotransformation of the A-ring-secocycloartene triterpenoid, nigranoic acid.     

连作大豆根分泌物对根腐病病原菌的化感作用

采用砂培,水培和室内培养等试验方法研究了连作大豆根分泌物对根腐病病原菌的化感作用。结果表明,与对照相比,连作和轮作大豆根分泌物对半镰孢菌,粉红粘帚功和尖镰孢菌尤其是对半裸镰孢菌的生长有明显的化感促进作用,差异达显著或极显著水平,低浓度时,连作大豆根分泌物对半裸镰孢菌和粉红粘帚菌生长的化感促进作用明显大于轮作大豆,差异达显著水平,同一茬口,高浓度根分泌物半裸镰孢菌生长的化感促进作用小于低浓度,而且在连作大豆中差异达显著水平。与对照相比,高浓度的邻苯二甲酸和丙二酸（L5和B5）对半裸镰孢菌,粉红粘帚菌和尖镰孢菌尤其是对半裸镰孢菌的生长有化感抑制作用,差异达显著或极显著水平,而低浓诬的邻苯二甲酸和丙二酸对半裸镰孢菌,粉红粘帚菌和尖镰孢菌的生长有化感促进,部分差异达显著水平。     

两种基因型大豆根分泌物对大豆根腐病菌的化感作用

采用生物模拟试验、化学分析等方法,研究了两种大豆基因型(9536、吉林30)的根分泌物中的糖、氨基酸、有机酸组分对大豆根腐病菌的化感作用.结果表明,两种大豆基因型(9536、吉林30)根分泌物糖组分表现出低浓度显著促进、高浓度显著抑制半裸镰孢菌、尖镰孢菌生长的规律,对粉红粘帚菌的生长影响不明显;氨基酸组分对上述三种病原菌所表现出的促进、抑制规律不同,9536基因型根分泌物氨基酸组分的中、高浓度处理对半裸镰孢菌、粉红粘帚菌、尖镰孢菌的生长表现出显著的抑制作用,而吉林30多表现出显著的促进作用;有机酸组分对半裸镰孢菌、粉红粘帚菌、尖镰孢菌生长都有显著的抑制作用.两种基因型大豆根分泌物(糖、氨基酸、有机酸组分)与根腐病害发生密切相关,大豆基因型不同,根分泌物对根腐病菌的化感促进或抑制作用有差异.     

粉红粘帚霉菌丝体化学成分研究*

用柱层析色谱进行分离,光谱和化学方法进行结构鉴定,从一种明显促进花叶开唇兰Anoectochilus roxburghii (Wall.)Lindl.生长的粘帚霉属真菌——粉红粘帚霉Gliocladium roseum (Link) Bani.的菌丝体分离鉴定了1,3-二棕榈酰基-2-(4,4-二甲基庚二酸单酰基) 甘油酯(I),4,4-二甲基庚二酸(II)等4个化合物。其中,I为新化合物, II 为首次从本属分离得到的天然产物。     

Role of zearalenone lactonase in protection of Gliocladium roseum from fungitoxic effects of the mycotoxin zearalenone

Zearalenone is a mycotoxin with estrogenic effects on mammals that is produced by several species of Fusarium. We found that zearalenone and its derivatives inhibit the growth of filamentous fungi on solid media at concentrations of < or =10 microg/ml. The fungitoxic effect declined in the order zearalenone > alpha-zearalenol > beta-zearalenol. The mycoparasitic fungus Gliocladium roseum produces a zearalenone-specific lactonase which catalyzes the hydrolysis of zearalenone, followed by a spontaneous decarboxylation. The growth of G. roseum was not inhibited by zearalenone, and the lactonase may protect G. roseum from the toxic effects of this mycotoxin. We inactivated zes2, the gene encoding zearalenone lactonase in G. roseum, by inserting a hygromycin resistance cassette into the coding sequence of the gene by means of Agrobacterium tumefaciens-mediated genetic transformation. The zes2 disruption mutants could not hydrolyze the lactone bond of zearalenone and were more sensitive to zearalenone. These data are consistent with a hypothesis that resorcylic acid lactones exemplified by zearalenone act to reduce growth competition by preventing competing fungi from colonizing substrates occupied by zearalenone producers and suggest that they may play a role in fungal defense against mycoparasites.     

粉红粘帚霉菌丝体化学成分研究

用柱层析色谱进行分离,光谱和化学方法进行结构鉴定,从一种明显促进花叶开唇兰Anoectochilus roxburghii (Wall.)Lindl.生长的粘帚霉属真菌——粉红粘帚霉Gliocladium roseum (Link) Bani.的菌丝体分离鉴定了1,3-二棕榈酰基-2-(4,4-二甲基庚二酸单酰基) 甘油酯(I),4,4-二甲基庚二酸(II)等4个化合物。其中,I为新化合物, II 为首次从本属分离得到的天然产物。     

粉红黏帚霉67-1菌株寄生核盘菌菌核的酶学动态研究

对粉红黏帚霉67-1菌株侵染核盘菌菌核过程的多种细胞壁降解酶活性进行了连续测定,以研究几丁质酶等在这一寄生互作体系中的可能作用。结果表明:葡聚糖酶活性变化表现活跃,且随寄生过程呈增加趋势,配对法T检验结果表明,第10d的处理与对照酶活性差异达到最大;几丁质酶、蛋白酶活性变化表现较低,而纤维素酶未检测得到。酶学动态变化与之前石蜡切片显微观察的结果在时间上表现一致;认为葡聚糖酶可能是粉红黏帚霉67-1菌株寄生核盘菌菌核的关键酶。     

Biological Modification of Trichothecene Mycotoxins: Acetylation and Deacetylation of Deoxynivalenols by Fusarium spp

Attempts were made to elucidate the acetyl transformation of novel trichothecene mycotoxins, 3a,7a,15-trihydroxy-12,13-epoxytrichothec-9-en-8-one (deoxynivalenol) and its derivatives, by trichothecene-producing strains of Fusarium nivale, F. roseum, and F. solani. In the peptone-supplemented Czapek-Dox medium, F. roseum converted 3a-acetoxy-7a,15-dihydroxy-12,13-epoxytrichothec-9-en-8-one (3-acetyldeoxynivalenol) to deoxynivalenol. 3-Acetyldeoxynivalenol was also deacetylated by intact mycelia of the three strains in sugar-free Czapek-Dox medium. The growing F. nivale acetylated deoxynivalenol to afford a small amount of 3-acetyldeoxynivalenol. 3a,7a,15-Triacetoxy-12,13-epoxytrichothec-9-en-8-one (7,15-diacetyl-deoxynivalenol), which was then deacetylated to give 7a-acetoxy-3a,15-dihydroxy-12,13-epoxytrichothec-9-en-8-one (7-acetyldeoxynivalenol). It was noted that the ester at C-7 was not hydrolyzed by the fungal mycelium.     

17 beta-estradiol hydroxylation catalyzed by human cytochrome P450 1A1: a comparison of the activities induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in MCF-7 cells with those from heterologous expression of the cDNA.

Exposure of MCF-7 breast cancer cells to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) causes an elevated cytochrome P450 content and a marked increase in the microsomal hydroxylation of 17 beta-estradiol (E2) at the C-2, C-4, C-15 alpha, and C-6 alpha positions. In this study we investigated the involvement of cytochromes P450 of the 1A gene subfamily in this metabolism of E2. Hydroxylation at each of these four positions of E2 was inhibited by P450 1A-subfamily inhibitors, alpha-naphthoflavone, benzo[a]pyrene, and 7-ethoxyresorufin. Northern blots showed that treatment of MCF-7 cells with TCDD resulted in production of the 2.6-kb CYP1A1 mRNA, but not the 3.0-kb CYP1A2 mRNA. Immunoblot analyses with anti-P450 1A antibodies confirmed the production of P450 1A1 protein in TCDD-treated MCF-7 cells. Anti-rat P450 1A IgG inhibited the hydroxylation of E2 at C-2, C-15 alpha, and C-6 alpha, but not hydroxylation at C-4. E2 hydroxylation by human cytochromes P450 1A1 and P450 1A2 was assessed in experiments with microsomes from Saccharomyces cerevisiae after transformation with cDNAs encoding the two cytochromes. The major hydroxylase activities of expressed human P450 1A1 were at the C-2, C-15 alpha, and C-6 alpha positions of E2; expressed human P450 1A2 catalyzed hydroxylation predominately at C-2. While both expressed P450s 1A1 and 1A2 had minor hydroxylase activities at the C-4 position, neither catalyzed a low-Km hydroxylation at C-4 similar to that observed with microsomes from TCDD-treated MCF-7 cells. These results provide strong evidence that P450 1A1 catalyzes the hydroxylations of E2 at the C-2, C-15 alpha, and C-6 alpha in incubations with microsomes from TCDD-treated MCF-7 cells, but suggest TCDD may also induce a cytochrome P450 E2 4-hydroxylase that is distinct from P450 1A1 or P450 1A2.     

The microbiological transformation of 7alpha-hydroxy-ent-kaur-16-ene derivatives by Gibberella fujikuroi

The biotransformation of 7alpha-hydroxy-ent-kaur-16-ene (epi-candol A) by the fungus Gibberella fujikuroi gave 7alpha,16alpha,17-trihydroxy-ent-kaur-16-ene and a seco-ring B derivative, fujenoic acid, whilst the incubation of candicandiol (7alpha,18-dihydroxy-ent-kaur-16-ene) and canditriol (7alpha,15alpha,18-trihydroxy-ent-kaur-16-ene) afforded 7alpha,18,19-trihydroxy-ent-kaur-16-ene and 7alpha,11beta,15alpha,18-tetrahydroxy-ent-kaur-16-ene, respectively. The presence of a 7alpha-hydroxyl group in epi-candol A avoids its biotransformation along the biosynthetic pathway of gibberellins, and directs it to the seco-ring B acids route. The 15alpha-hydroxyl group in canditriol inhibits oxidation at C-19 and direct hydroxylation at C-11(beta). The formation of fujenoic acid, from 7alpha-hydroxy-ent-kaur-16-ene, probably occurs via 7alpha-hydroxykaurenoic acid and 7-oxokaurenoic acid, with subsequent hydroxylation at the C-6(beta) position.     

Competitive ability of the antagonists Ulocladium atrum and Gliocladium roseum at temperatures favourable for Botrytis spp. development

Ulocladium atrum and Gliocladium roseum are fungal antagonists capable of suppressing sporulation of Botrytis spp. on dead plant parts. The effect of temperature (3 to 36 °C) on antagonist conidial germination and mycelial growth was assessed on agar. In addition conidial germination of U. atrum was measured on dead lily leaves. The optimum temperature of both antagonists for both conidial germination and mycelial growth was between 27 and 30 °C. U. atrum was less affected by lower temperatures than G. roseum. At optimum temperature, 50% of conidia of U. atrum and G. roseum germinated within 2.6 and 10.0 hrs, respectively. At low sub-optimal temperatures (6 °C), 50% of conidia germinated within 18 and 96 hours, respectively.In bioassays on dead onion leaves, U. atrum suppressed sporulation of B. cinerea and B. aclada at all temperatures tested (6 to 24 °C) by more than 85%. On dead cyclamen leaves, G. roseum was more efficient than U. atrum at 21 and 24 °C but, in contrast to U. atrum, showed no antagonistic activity at temperatures below 21 °C. On dead hydrangea leaves, U. atrum significantly reduced sporulation of B. cinerea at temperatures as low as 3 and 1 °C. Under Dutch growing conditions, the mean air temperature during leaf wetness periods in onion and lily fields was 15 °C with temperatures only occasionally above 20 °C. In greenhouse crops of cyclamen, the mean temperature during high humidity periods was 17 °C. It is therefore concluded that U. atrum is better adapted than G. roseum to temperatures which occur in the field, in greenhouse crops such as cyclamen, or during cold storage of plant stocks.     

Effect of inoculation time on the survival of spores of Gliocladium roseum on geranium leaves.

Gliocladium roseumis a successful antagonist of Botrytis cinerea and is considered to have the major potential for biocontrol of the pathogen in cropping systems. In order to elucidate the optimal moment of the day to apply the biological control agent, geranium plants were inoculated until run off with a suspension containing 10 e7 conidia of G. roseum + Tritón 100X. The inoculation times were 9 am, 12 am, 3 pm and 6 pm. The number cfu per cm(2) of leaves at inoculation time (time 0) and at 3, 6 y 9 h after inoculation were estimated. Then, the inoculated plants were kept in a Plexiglas, humidity chamber for 24 h. Discs of 8 mm in diameter were cut off from the inoculated leaves and put then into a paraquat medium to examine the development of G. roseum. According to the results dry period length after inoculation time, shows a significant decrease in number of cfu by cm(2) of leaf. No difference was observed between 3, 6 y 9 h after inoculation time.     

Interactions between Glomus mosseae and arbuscular mycorrhizal sporocarp-associated saprophytic fungi

The saprophytic fungi Wardomyces inflatus (Marchal) Hennebert, Paecilomyces farinosus (Holm & Gray) A. H. S. Brown & G. Sm., Gliocladium roseum Bain., sterile dark mycelium (SDM-54), Trichoderma pseudokoningii Rifai and Trichoderma harzianum Rifai were isolated from sporocarps of Glomus mosseae. The effect of saprophytic fungi on G. mosseae spore germination was tested on water agar. Wardomyces inflatus decreased the percent germination of G. mosseae spores; G. roseum, T. pseudokoningii and T. harzianum had no effect on germination; and P. farinosus and SDM-54 increased the percentage of spore germination of G. mosseae after 4 d. Wardomyces inflatus significantly decreased hyphal length of spores which germinated, but no other saprophytic fungi affected hyphal growth. Trichoderma pseudokoningii, T. harzianum, P. farinosus and SDM-54 increased the number of auxiliary cells formed by G. mosseae. The effect of saprophytic fungi on arbuscular mycorrhizal colonization of soybean was studied in a greenhouse trial. The percentage of soybean root length colonized was decreased by W. inflatus, unaffected by SDM-54 and T. harzianum, and increased by P. farinosus. Gliocladium roseum decreased root length colonized when plants were 12 wk old, and T. pseudokoningii increased colonization of roots when plants were 4 wk old. Antagonistic, synergistic and neutral actions of G. mosseae upon the saprophytic fungi were observed. The population of T. harzianum decreased and the populations of T. pseudokoningii and SDM-54 increased in the presence of G. mosseae. Our results indicate a complex interaction between G. mosseae and associated saprophytic fungi.     

Synthesis of four diastereoisomers at carbons 24 and 25 of 3 alpha,7 alpha,12 alpha,24-tetrahydroxy-5 beta-cholestan-26-oic acid, intermediates of bile acid biosynthesis

The synthesis of four stereoisomers at C-24 and C-25 of 3 alpha,7 alpha,12 alpha,24-tetrahydroxy-5 beta-cholestan-26-oic acid is described. Pyridium chlorochromate oxidation of 3 alpha,7 alpha,12 alpha-triacetoxy-5 beta-cholan-24-ol (II) prepared from cholic acid (I) afforded 3 alpha,7 alpha,12 alpha-triacetoxy-5 beta-cholan-24-al (III) which was converted to a mixture of the four stereoisomers (IV-VII) by a Reformatsky reaction with ethyl DL-alpha-bromopropionate followed by alkaline hydrolysis. Separation of these isomers (IV-VII) was achieved by silica gel column chromatography, and subsequent reversed-phase partition column chromatography. The configurations at C-24 were elucidated by conversion of each isomer into (24R)- or (24S)-5 beta-cholestane-3 alpha,7 alpha,12 alpha,24-tetrol (XII or XI) by Kolbe electric coupling, the C-24 configurations of which were determined by modified Horeau's method and 13C-nuclear magnetic resonance spectroscopy. The stereochemistries at C-25 were deduced by comparison of IV-VII with the products of the hydroboration followed by oxidation with alkaline hydrogen peroxide of (24E)-3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholest-24-en-26-oic acid (XIII).     

Substrate specificity for the hydroxylation of polyoxygenated 4(20),11-taxadienes by Ginkgo cell suspension cultures

Three C-14 oxygenated taxanes isolated from callus cultures of Taxus spp., 2alpha,5alpha,10beta,14beta-tetra-acetoxy-4(20),11-taxadiene 3, 2alpha,5alpha,10beta-triacetoxy-14beta-propionyloxy-4(20),11-taxadiene 4, 2alpha,5alpha,10beta-triacetoxy-14beta-(2-methylbutyryl)-oxy-4(20),11-taxadiene 5, and three deacetylated derivatives of 3, 10beta-hydroxy-2alpha,5alpha,14beta-triacetoxy-4(20),11-taxadiene 6, 14beta-hydroxy-2alpha,5alpha,10beta-triacetoxy-4(20),11-taxadiene 7, 10beta,14beta-dihydroxy-2alpha,5alpha-diacetoxy-4(20),11-taxadiene 8, could all be regio- and stereo-selectively hydroxylated at the 9alpha-position by Ginkgo cell suspension cultures to yield a series of new 9alpha,14beta-dihydroxylated taxoids. The effects of functional groups, especially at C-14 of the substrates, on the biotransformation were also investigated. The results revealed that substrates with an acetoxyl group at C-14 could be more efficiently 9alpha-hydroxylated than those with a longer ester chain or a hydroxyl group at C-14. An acetoxyl or hydroxyl group at C-10 had no effect on the conversion rates of the substrates, but substrates with the hydroxyl group (compared with the acetoxyl analogues) could be converted into 9alpha-hydroxylated products more easily.     

Differences in steroid specificity for rat androgen binding protein and the cytoplasmic receptor.

Two proteins in the rat, androgen binding protein (ABP) and the cytoplasmic receptor (CR), have high affinity and limited capacity for binding androgens. To determine the structural requirements for binding with high affinity, each protein was partially purified and the ability of over 100 steroids to compete with [3H]dihydrotestosterone (17 beta-hydroxy-5 alpha-androstan-3-one) for binding sites was assessed. The results indicate marked differences in the steroid specificities of the two proteins. Some alterations of dihydrotestosterone at C-2 or C-2 and C-3 increase binding to ABP two to four-fold. Similarly, the affinity of 17 beta-hydroxy-7 alpha-methyl-4-estren-3-one for ABP increases two-fold when a double bond is created at C-14. Addition of a methyl group in the alpha position at C-7 or C-17, or an ethinyl group at C-17 cause little change in affinity; however, modifications at C-11 and C-17 beta, and deletion of the methyl group at C-10 significantly impair binding to ABP. Binding to the CR is maintained or increased by deletion of the methyl group at C-10. Binding is lessened by modifications at C-3 and C-17 beta. Most alterations at C-2, C-7, C-11, and C-17 alpha have only minor effects on binding to the CR. These studies should provide a molecular basis for predicting the effects of specific structural modifications. When some modifications at C-2 or C-2 and C-3 are combined with changes at C-17 beta, the resulting steroids retain very high affinity for ABP and very limited binding to the CR. Such steroids may provide a means for assessing the function of ABP.     

Evaluation of Gliocladium roseum Against Wood-Degrading Fungi in vitro and on Major Canadian Wood Species

The protection of wood from fungal stain using biological agents has considerable potential for reducing discoloration of freshly sawn logs and lumber, while decreasing fungicide use. A number of biocontrol candidates have been reported worldwide, and Gliocladium roseum is one of such microorganisms. In this study, the bio-activity of G. roseum was investigated against different wood-degrading fungi on agar plates and wafers of 12 major Canadian wood species. Of the four sap-staining fungi tested on agar plates, Ophiostoma piceae and Alternaria alternata showed greater sensitivity than Aureobasidium pullulans or Cladosporium sphaerospermum to G. roseum . On wood wafers, a spore suspension of G. roseum (1 10 6 spores/ ml) provided satisfactory protection of wood from stain on western hemlock ( Tsuga heterophylla ), white spruce ( Picea glauca ), amabilis fir ( Abies amabilis ), balsam fir ( Abies balsamea ) and jack pine ( Pinus banksiana ). The antagonist also restricted the development of moulds and stain on black spruce ( Picea mariana ), lodgepole pine ( Pinus contorta ) and white pine ( Pinus strobus ), but did not protect Douglas fir ( Pseudotsuga menziesii ), red pine ( Pinus resinosa ), white birch ( Betula papyrifera ) and trembling aspen ( Populus tremuloides ). Logs of black spruce and jack pine treated with G. roseum were much less stained than untreated ones after a 4-month period of summer storage in the field. In an anti-decay test, no significant difference was found for weight loss between wood blocks treated with G. roseum and untreated samples. Application of G. roseum with low levels of an anti-sap stain chemical (NP-1) to wood wafers simultaneously did not produce a noticeable improvement in wood protection against stain compared with the chemical treatment alone.     

5 alpha-androstane-3 beta,17 beta-diol hydroxylating enzymes in stroma and epithelium of human benign prostatic hyperplasia (BPH)

As enzymatic hydroxylation of 5 alpha-androstane-3 beta,17 beta-diol (3 beta-diol) may be a factor in controlling the 5 alpha-dihydrotestosterone (DHT) content in the prostate, we were interested in activity and distribution of these enzymes in epithelium and stroma of human benign prostatic hyperplasia (BPH). The enzyme activities were measured after mechanical separation of BPH tissue from 15 patients of various ages into stroma and epithelium, and optimization of the in vitro transformation of 3 beta-diol to hydroxylated products, which were analyzed by HPLC. The main results were: (1) 3 beta-diol was hydroxylated at C-7 alpha, C-7 beta, C-6 alpha, and C-6 beta. (2) The mean Michaelis constant Km (nM +/- SEM) for hydroxylation at C-7 alpha(beta) (168 +/- 21) was significantly lower than at C-6 alpha(beta) (601 +/- 43) without differences between stroma and epithelium. (3) Hydroxylation at alpha position dominated significantly over that at beta. (4) The mean maximal metabolic rate Vmax (pmol . mg protein-1 . h-1) of hydroxylation at C-6 alpha was about 7-fold lower in stroma (3.4 +/- 0.2) than in epithelium (23.8 +/- 4.1), concerning the other hydroxylations, Vmax was about 1.6-fold lower in stroma. (5) With increasing age of the patients there was a significant decrease of the 3 beta-diol hydroxylation in stroma and epithelium. It is discussed that the significantly lower activity of 3 beta-diol hydroxylation in stroma compared to epithelium and the decrease of activity with increasing age might potentiate the DHT accumulation in stroma of BPH.     

Mycoparasites effect on reproductive ability of Sclerotinia sclerotiorum sclerotia.

The ability to parasitise Sclerotinia sclerotiorum and the effect on apothecia production was evaluated for the following antagonists: Trichoderma harzianum; Trichoderma koningii; Gliocladium roseum and Chaetomium globosum. Plastic trays were filled with of steam-sterilized soil. Each one of them was infested with sclerotia of S. sclerotiorum and the culture of the antagonists. The trays were kept in a greenhouse and after 30, 60 and 90 days, evaluations were made. The rates of carpogenic germination, myceliogenic germination, mycoparasitism and destruction were evaluated. To assess carpogenic germination, the sclerotia were put in a growth chamber over moistened filter paper at 20 -/+ 2 degrees C and 12 light hours. The rates of myceliogenic germination and mycoparasitism were evaluated on Petri dishes with 2% APD. Antagonists effect on carpogenic germination was observed one month after the start of the assay. In the evaluation made at 60 and 90 days, T. harzianum; T. koningii and G. roseum kept inhibitory properties. Such inhibition was not observed in the trays containing C. globosum. In the evaluations made at 30 days, mycoparasitism rate was high in the trays with T. harzianum; T. koningii and G. roseum. G. roseum and T. harzianum destroy S. sclerotiorum sclerotia.