Evaluation of the entomopathogenic fungiMetarhizium anisopliae andBeauveria bassiana against the red palm weevilRhynchophorus ferrugineus

The red palm weevil (RPW,Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) is one of the most severe pests of various palm species, including date palms. While examining the susceptibility of RPW to two entomopathogenic fungi,Metarhizium anisopliae andBeauveria bassiana, strains of the former were found to be more virulent than those of the latter, achieving 100% larval mortality within 6–7 days. The most virulent strains ofM. anisopliae were then tested on RPW eggs and adults. Incubation in a substrate treated withM. anisopliae spores increased egg mortality and reduced their hatchability. The total percentage mortality of eggs and hatched larvae was 80–82%, compared with 34% in the controls. RPW adults were challenged with two types of fungal formulation: dry powder and aqueous suspension. Cumulative adult mortality of 100% was achieved in 2–3 weeks for the dry rice-based formulation and in 4–5 weeks for the spore suspension. As a result of decreased longevity, treated females had a shorter oviposition period and three times lower fertility than the controls. Possible strategies for fungus application are discussed in the light of the high susceptibility of eggs and larvae to fungal infection. http://www.phytoparasitica.org posting July 19, 2006.     

白僵菌及其伴生菌发酵液对线虫的毒力研究

采用发酵液浸没法,以4种线虫作为靶标,对6株白僵菌、7株伴生菌菌株进行了杀线虫活性筛选,以期获得新的、高效的杀线虫生防菌资源。其中:处理24 h对南方根结线虫Meloidogyne incognita 2龄幼虫及大豆胞囊线虫Heterodera glycines 2龄幼虫校正死亡率均达80%以上的菌株仅有白僵菌Snef2598及伴生菌Snef5;供试菌株对水稻干尖线虫Aphelenchoides besseyi的毒力均较低,其中仅有1株白僵菌校正死亡率达到40%以上。处理48 h对南方根结线虫2龄幼虫校正死亡率达90%以上的菌株有白僵菌4株和伴生菌5株;对大豆胞囊线虫2龄幼虫校正死亡率超过90%的菌株有白僵菌3株和伴生菌1株;供试菌株对水稻干尖线虫毒力均较低,校正死亡率达60%以上的菌株有白僵菌3株和伴生菌2株;几乎所有供试菌株,对小杆线虫Caenorhabditis sp.的毒力均很高,处理24 h及48 h的校正死亡率均达97%以上,且被杀死的小杆线虫体壁以及虫体内部物质均被不同程度地消解。研究表明,球孢白僵菌菌株Snef2598及其伴生菌菌株Snef5对4种靶标线虫的毒力均较好,具有进一步研究的价值。     

Selection of global Metarhizium isolates for the control of the rice pest Nilaparvata lugens (Homoptera: Delphacidae)

BACKGROUND: This study was initiated to search for fungal candidates for microbial control of brown planthopper (BPH) Nilaparvata lugens Stål, to which little attention has been paid in the past two decades. RESULTS: Thirty‐five isolates of Metarhizium anisopliae (Metschnikoff) Sorokin and M. flavoviride Gams & Rozsypal from different host insects worldwide were bioassayed for their lethal effects against third‐instar BPH nymphs at 25 °C and a 14:10 h light:dark photoperiod at ca 1000 conidia mm^?2. On day 9 post‐treatment, mortality attributable to mycosis ranged from 6.5 to 64.2% and differed significantly among the tested isolates with no apparent relationship to their host origin. Only two BPH‐derived M. anisopliae isolates from the Philippines (ARSEF456) and Indonesia (ARSEF576) killed > 50% of the nymphs. Both isolates were further bioassayed for time–concentration–mortality responses of the nymphs to the sprays of 19–29, 118–164 and 978–1088 conidia mm^?2 in repeated bioassays. The resultant data fitted a time–concentration–mortality model very well. Their LC₅₀ values were estimated as 731 and 1124 conidia mm^?2 on day 7 and fell to 284 and 306 conidia mm^?2, respectively, on day 10. CONCLUSION: The two M. anisopliae isolates are potential biocontrol agents of BPH for further research. This is the first report of the lethal effects of global Metarhizium isolates on the rice pest. Copyright © 2008 Society of Chemical Industry     

微生物杀虫剂联合施用对草地贪夜蛾幼虫的防治效果

为提高微生物杀虫剂对草地贪夜蛾Spodoptera frugiperda的防治效果,利用球孢白僵菌Beauveria bassiana IPPB343、IPPB1237与金龟子绿僵菌Metarhizium anisopliae IPPM330189三种昆虫病原真菌,斜纹夜蛾核型多角体病毒（Spodoptera litura nuclear polyhedrosis viruses,SLNPV）、甘蓝夜蛾核型多角体病毒（Mamestra brassicae nuclear polyhedrosis viruses,MBNPV）2种昆虫病毒和短稳杆菌Empedobacter brevis昆虫病原细菌,设置9种联合施用药剂组合,评价6种微生物杀虫剂单独使用以及不同药剂联合施用组合对草地贪夜蛾幼虫的毒力以及田间防治效果。结果表明,与6种微生物杀虫剂单独使用相比,9种联合施用组合均能提升微生物杀虫剂对草地贪夜蛾的毒力水平与田间防治效果。施药后7 d时,金龟子绿僵菌IPPM330189+MBNPV药剂组合的防治效果最好且增效最明显,对草地贪夜蛾的田间防治效果达到88.75%,协同毒力指数为23.78。此外,金龟子绿僵菌IPPM330189+短稳杆菌悬浮剂药剂组合施药后7 d对草地贪夜蛾的田间防治效果达到87.45%,协同毒力指数也可达到21.72。表明微生物杀虫剂联合施用可显著提升其对草地贪夜蛾的防治效果。     

Evaluation of the biocontrol potential of various Metarhizium isolates against green peach aphid Myzus persicae (Homoptera: Aphididae)

BACKGROUND: Twenty‐three isolates of Metarhizium anisopliae (Metschnikof) Sorokin and M. acridum (Driver & Milner) JF Bischoff, Rehner & Humber from non‐aphid host insects around the globe were evaluated for their aphid biocontrol potential, which is not well known. RESULTS: The apterous adults of green peach aphid Myzus persicae (Sulzer) were exposed to the fungal sprays of 11.5, 99 and 1179 conidia mm^?2 and blank control in three leaf‐dish bioassays. All the tested isolates except one were proven to be infective to the aphid species at 21 ± 1 °C and 14:10 h light:dark photoperiod, causing corrected mortalities of 10.1–95.3% at the high spore concentration. The data from ten isolates causing > 50% mortality at the high concentration were found to fit a time–concentration–mortality model well, yielding parameters for the estimates of their LC₅₀ and LT₅₀ that vary with post‐spray time and spore concentration respectively. Four isolates of M. anisopliae (ARSEF 759, 4132, 2080 and 576) had LC₅₀ values of 44–80 conidia mm^?2 on day 8 and LT₅₀ values of 4.9–6.8 days at 100 conidia mm^?2, with 91–98% of the killed aphids being well mycotised after death. CONCLUSION: The Metarhizium infectivity to M. persicae differs greatly among the tested isolates. The four mentioned isolates with desired virulence and sporulation potential are excellent candidates for microbial control of aphids. Copyright © 2010 Society of Chemical Industry     

Effect of fungicides on the complex of <Emphasis Type="Italic">Fusarium</Emphasis> species and saprophytic fungi colonizing wheat kernels

Fusarium head blight of wheat, often associated with mycotoxin contamination of food and feed is caused by various Fusarium species. The efficacy of fungicide sprays for the control of the disease and mycotoxins varies from being highly effective to even increasing mycotoxin levels. The potential role of antagonistic fungi in this variability was investigated assessing sensitivity of Fusarium species and saprophytic fungi colonizing wheat kernels to fungicides. Saprophytes were tested for their antagonistic activity to the prevalent Fusarium species Fusarium avenaceum, Fusarium culmorum, Fusarium graminearum, and Fusarium poae. Fungal isolates from mature winter wheat kernels were Alternaria alternata, Arthrinium sp., Aspergillus niger, Epicoccum sp., Microdochium spp., Rhizopus oryzae and Trichoderma sp. In dual culture A. niger, R. oryzae and Trichoderma hamatum were more effective in reducing mycelial growth of Fusarium species than Microdochium majus; A. alternata and Epicoccum sp. were ineffective because of slow growth rates. Saprophytic fungi were sensitive to triazoles; however, prothioconazole and tebuconazole had stronger effects on mycelial growth of Fusarium species. ED₅₀ values also indicated significant differences in the sensitivity of Fusarium species to triazoles (range 0.1–1.7 mg l⁻¹). Azoxystrobin and fluoxastrobin were largely ineffective in inhibiting in vitro growth of Fusarium spp.; sensitivity of the other fungi was generally lower, except for M. majus which was highly sensitive. Due to differences in fungicide sensitivity among Fusarium spp. and ear-colonizing fungi antagonistic to Fusarium spp. fungicides are likely to modify the balance within the mycoflora of wheat ears which may also affect the mycotoxin contamination of grain.     

Entomopathogenic fungi and nematodes against larvae of the chestnut weevil,Curculio elephas (Coleoptera: Curculionidae): a laboratory evaluation

Pest control in chestnut orchards is mainly achieved by chemicals, but there is growing focus on the potential use of biological control agents. The larval susceptibility of the chestnut weevil Curculio elephas L. (Coleoptera: Curculionidae) to different strains/species of entomopathogenic fungi and nematodes was evaluated in laboratory by soil cup bioassays. In the experiments with fungi, a wild strain of Metarhizium anisopliae caused the highest larval mortality (about 90%) compared with a commercial and a wild strain of Beauveria bassiana (80% and 77% respectively). Regarding nematodes, the commercial strain of Heterorhabditis bacteriophora was more effective in the control of chestnut larvae (77% insect mortality) than Steinernema carpocapsae (43%).     

Virulence of select entomopathogenic fungi to the brown marmorated stink bug, Halyomorpha halys (Stål) (Heteroptera: Pentatomidae)

The brown marmorated stink bug. Halyomorpha halys, (BMSB) is an exotic pest causing damage to fruits and vegetables. They also transmit diseases. Integrated pest management strategies are lacking for this invasive. For the first time, information is presented on the virulence of several entomopathogenic fungi against BMSB. Three Beauveria bassiana and two Metarhizium anispoliae isolates were bioassayed against adult BMSB. One B. bassiana isolate was the active ingredient in BotaniGard^® which produced 85 and 100% mortality in 9 and 12 days post treatment, respectively. Experimental isolates ERL 1170 and ERL 1540 were also efficacious. Metarhizium anispoliae isolates produced lower mortalities than the B. bassiana isolates. Data presented showed the potential for management of BMSB with entomopathogenic fungi. Copyright © 2012 Society of Chemical Industry     

Field evaluation of entomofungal pathogens against cowpea aphid,Aphis craccivora Koch,and their effect on two coccinellid predators

The present study was carried out to assess the field efficacy of 10 isolates of entomofungal pathogens on management of cowpea aphid, Aphis craccivora. All the tested isolates showed suppressive effect on aphid population. Two-year pooled data revealed lower incidence of 19.37–40.81 aphids/plant in the entomofungal pathogen treated plots compared to higher incidence of 88.07 aphids/plant in untreated control. Among the 10 isolates tested, Vl-8 isolate of Lecanicillium lecanii, Ma-6 isolate of Metarhizium anisopliae and Bb-5a isolate of Beauveria bassiana showed 78.01%, 77.42% and 76.91% reduction of aphid population, respectively, over control and were on par with each other. Significantly higher yields were recorded in the plots treated with Bb-5a, Ma-6 and Vl-8 isolates (1252, 1225 and 1208 kg/ha, respectively) compared to untreated control (916 kg/ha). The predatory coccinellid populations of Cheilomenes sexmaculata and Micromus timidus in entomofungal treated and untreated control plots were almost similar and found statistically non-significant, indicating the safety of fungal pathogens to these coccinellid predators.     

Biological control of the tick fowl Argas persicargas persicus by the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae

The pathogenicity of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae against the tick fowl Argas persicargas persicus was studied. Bioassays revealed significant differences between the two fungi and the A. persicargas persicus stages exposed. The first nymphal instar was more highly susceptible to both fungi than the second nymphal instar. The unfed female of A. persicargas persicus was significantly less susceptible to both fungi than the engorged female. Application of M. anisopliae to sackcloth naturally infested with the tick fowl A. persicargas persicus induced mortality which reached 100% three weeks after treatment under field conditions. The present study suggests that entomopathogenic fungi have good potential for tick fowl, A. persicargas persicus, control.     

球孢白僵菌对松墨天牛的致病力及其与花绒寄甲的相容性

为探明昆虫病原真菌球孢白僵菌Beauveria bassiana与天敌昆虫花绒寄甲Dastarcus helophoroides的相容性,观察4株球孢白僵菌菌株的生长性状、产孢量以及孢子萌发率,并选用Bb3275和Bb202菌株测定其对松墨天牛Monochamus alternatus的致病力以及对花绒寄甲的毒力;同时在模拟环境条件下,分别用Bb3275和Bb202菌株的孢子悬浮液先侵染松墨天牛幼虫,再按1∶1比例将花绒寄甲与受侵染的松墨天牛置于同一环境下饲养,观察花绒寄甲的死亡率。结果显示,孢子悬浮液浓度为1×10⁶孢子/mL至1×10⁸孢子/mL时,Bb3275菌株和Bb202菌株对松墨天牛幼虫和成虫的致病力与对照组相比均有显著差异;经4种浓度Bb3275和Bb202菌株侵染花绒寄甲幼虫和成虫15 d后,仅在高浓度1×10⁸孢子/mL时表现出轻微致死作用,但花绒寄甲的存活率均高于85.00%;模拟条件下,松墨天牛幼虫校正死亡率均在80.00%以上,而同环境下花绒寄甲幼虫和成虫均表现出良好的活力,且侵染率为0。表明球孢白僵菌和花绒寄甲之间具有很好的相容性,可以同时应用于松墨天牛的生物防治。     

Sublethal effect of Beauveria bassiana on feeding and fecundity of the sunn pest,Eurygaster integriceps Puton (Hemiptera: Scutelleridae)

The sunn pest, Eurygaster integriceps Puton, is a major pest of wheat in West and Central Asia. Studies were conducted to investigate the effect of Beauveria bassiana (teleomorph; Cordyceps bassiana) isolates on the sunn pest. Bioassays were done with adult insects collected from overwintering sites at three different times. The effect of sublethal doses of fungi on the insects’ feeding and fecundity was determined. When the sunn pest adults were treated with 2 × 10⁷ conidia mL^?1, all isolates caused significant sunn pest mortality varying between 24 and 100% 12 days after inoculation. There were significant differences between mortality, depending on the date of fungal application. Adult sunn pests treated with B. bassiana at sublethal concentrations caused significantly less damage to wheat leaves than sunn pests in the control. The number of eggs laid by the sunn pest was significantly fewer when wheat plots in plastic greenhouse and field cages were infested with adult sunn pests treated at sublethal concentrations, compared with sunn pests in control plots. Results show that sublethal dosages of B. bassiana reduced sunn pest feeding and fecundity. Applying B. bassiana in sunn pest overwintering sites minimizes the pest populations, which would move to wheat fields in spring.     

Pathogenicity of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> and <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> against <Emphasis Type="Italic">Galleria mellonella</Emphasis>

The greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae) is occasionally found in beehives and is a major pest of stored wax. Entomopathogenic fungi have recently received attention as possible biocontrol elements for certain insect pests. In this study, 90 isolates of Beauveria bassiana and 15 isolates of Metarhizium anisopliae were screened for proteases and lipases production. The results showed significant variations in the enzymatic action between the isolates. In the bioassay, the selected isolates evinced high virulence against the 4th instar of the G. mellonella larvae. The isolates BbaAUMC3076, BbaAUMC3263 and ManAUMC3085 realized 100% mortality at concentrations of 5.5 × 10⁶ conidia ml⁻¹, 5.86 × 10⁵ conidia ml⁻¹, and 4.8 × 10⁶ conidia ml⁻¹, respectively. Strong enzymatic activities in vitro did not necessarily indicate high virulence against the tested insect pest. The cuticle of the infected larvae became dark and black-spotted, indicating direct attack of fungus on the defense system of the insects. The LC₅₀ values were 1.43 × 10³, 1.04 × 10⁵ and 5.06 × 10⁴ for Bba3263AUMC, Bba3076AUMC and Man3085AUMC, respectively, and their slopes were determined by computerized probit analysis program as 0.738 ± 0.008, 0.635 ± 0.007 and 1.120 ± 0.024, respectively.     

In vitro effects of flutriafol and azoxystrobin on Beauvaria bassiana and its efficacy against Tetranychus urticae

BACKGROUND: Testing the compatibility of chemical pesticides and fungal biocontrol agents is necessary if these two agents are to be applied together in the integrated management of plant pests and diseases. In this study, the fungicides azoxystrobin (a strobilurin) and flutriafol (a triazole) were tested in vitro for their effects on germination of conidia and mycelial growth of Beauveria bassiana (Bals.) Vuill. and in bioassay for their effect on fungal activity against Tetranychus urticae Koch. The fungicides were tested at three different concentrations [recommended rate for field use (1 × X) and the dilutions 10^?1× X and 10^?2× X]. RESULTS: Flutriafol inhibited growth of mycelia and germination of the fungal conidia at all concentrations tested in vitro, and also reduced the efficacy of B. bassiana in bioassays against mites. The inhibitive effect of azoxystrobin in vitro varied with the concentration applied. A significant effect was observed at 1 × X and 10^?1× X concentrations on both the germination of conidia and mycelia growth. At 10^?2× X concentration, azoxystrobin showed little effect on B. bassiana. However, when this fungicide was tested in bioassays, none of the concentrations reduced B. bassiana activity against mites. CONCLUSION: Azoxystrobin was most compatible with B. bassiana, while flutriafol was the most harmful. Further studies are required to confirm the negative effect of flutriafol on B. bassiana activity. Copyright © 2010 Society of Chemical Industry     

Aloe ring spot,a new disease of aloe caused by <Emphasis Type="Italic">Haematonectria haematococca</Emphasis> (Berk. &; Broome) Samuels &; Nirenberg (anamorph: <Emphasis Type="Italic">Fusarium</Emphasis> sp.)

A ring spot disease of Aloe vera was found on leaves of potted seedlings of Aloe vera in Hachijojima and Chichijima Islands, Tokyo. From tissue of ring spot lesions, a fungus producing Fusarium-type conidia was consistently isolated. After 1 month, reddish perithecia of nectriaceous fungus had formed on the colonies of this isolate on PDA. These nectriaceous and Fusarium fungi were identified as Haematonectria haematococca and Fusarium sp., respectively. From a single ascospore isolation, the former was confirmed to be the teleomorph of the Fusarium sp. Typical ring spot lesions were reproduced by artificial inoculations using single ascospore and single conidium isolates. Inoculations of five species of genus Aloe revealed that they were highly susceptible except for A. arborescens. This is the first report of a disease on Aloe caused by H. haematococca (anamorph: Fusarium sp.) in Japan, and it was named aloe ring spot.     

Pathogenic fungi of planthoppers associated with rice crops in Argentina

We conducted a survey of pathogenic fungi of planthoppers associated with rice crops in Los Hornos, Buenos Aires, Argentina, and a study of the seasonality and prevalence of these pathogens. Samples were taken in rice for two consecutive years. The plants sampled included rice Oryza sativa L. (Poaceae) and its surrounding weeds. The planthopper Oliarus dimidiatus Berg. (Hemiptera: Fulgoromorpha: Cixiidae) was the most abundant species associated with rice. Two species of entomophthoralean and one species of hypocrealean fungi infected and killed adults of O. dimidiatus in the rice agroecosystem. The fungi were identified as: Pandora sp. (Entomophthorales: Entomophthoraceae), Conidiobolus coronatus (Costantin) Batko (Entomophthorales: Ancylistaceae), and Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Hypocreales: Clavicipitaceae). Pandora sp. was the most predominant pathogen collected. It was recorded from the middle of February to the middle of April 2005, and not found again until the end of March 2006. The report of Pandora sp., C. coronatus and B. bassiana represents the first records of these fungi as pathogens of hoppers associated with rice crops in Argentina.     

绿僵菌与3种杀虫剂混用对沙葱萤叶甲的协同作用

采用点滴法,于室内测定了阿维菌素、茚虫威和鱼藤酮3种杀虫剂单独使用及与绿僵菌混用对沙葱萤叶甲3龄幼虫的协同致死作用,以期为协调使用化学药剂和生防制剂防治沙葱萤叶甲提供参考。3种杀虫剂单独使用时的毒力测定结果表明,阿维菌素对沙葱萤叶甲3龄幼虫的毒力最强,LD50值为4.80 ng/头,其次是茚虫威和鱼藤酮,LD50值分别为10.47和53.21 ng/头。采用不同浓度的3种杀虫剂分别处理绿僵菌24和48 h后,发现药剂对绿僵菌分生孢子萌发均有一定的抑制作用,但抑制率低于20%,且抑制率随着药剂浓度的降低和作用时间的延长而降低,其中茚虫威的平均抑制率最小,分别仅为1.89%±0.57%和0.89%±0.39%。将3种杀虫剂分别与绿僵菌孢子悬液混配使用,对防治沙葱萤叶甲3龄幼虫均具有协同增效作用:其中茚虫威与绿僵菌混用增效作用最强,比单独施用绿僵菌时LT50值缩短了8.12 d;阿维菌素与绿僵菌混用时LT50值缩短了7.60 d;鱼藤酮与绿僵菌混用时LT50值缩短了6.45 d。     

两株虫生真菌对草地贪夜蛾的致病力及高毒力菌株与卵寄生蜂的相容性

为有效防控新入侵我国的重大农业害虫草地贪夜蛾 Spodoptera frugiperda,采用浸叶法测定球孢白僵菌 Beauveria bassiana菌株Bb1237、 Bb201017、 Bb20091317及金龟子绿僵菌 Metarhiziumanisopliae菌株Ma189和莱氏绿僵菌 M. rileyi菌株MrCDTLJ1对草地贪夜蛾3龄幼虫的致病力,筛选对草地贪夜蛾3龄幼虫有较强致病力的菌株,并测定该菌株对草地贪夜蛾3龄幼虫的致病力和对其天敌的致死率。结果显示,球孢白僵菌Bb20091317和莱氏绿僵菌MrCDTLJ1对草地贪夜蛾3龄幼虫的累计校正死亡率显著高于其他菌株,分别为95.59%和97.06%,致死中时LT₅₀分别为3.42 d和5.72 d, LC₅₀分别为7.05×10⁶个/mL和2.25×10⁵个/mL。2种菌在高浓度1×10⁸个/mL下对草地贪夜蛾天敌夜蛾黑卵蜂 Telenomus remus和玉米螟赤眼蜂 Trichogramma ostriniae成蜂的致死率也仅介于15%~20%之间。表明球孢白僵菌Bb20091317和莱氏绿僵菌MrCDTLJ1不仅对草地贪夜蛾有高致病力,同时与夜蛾黑卵蜂和玉米螟赤眼蜂有较好的相容性。     

Induction of Systemic Resistance in Cucumber against Several Diseases by Plant Growth-promoting Fungi: lignification and Superoxide Generation

Five fungal isolates (Trichoderma, Fusarium, Penicillium, Phoma and a sterile fungus) from zoysiagrass rhizosphere that promote plant growth were tested for their ability to induce systemic resistance in cucumber plants against Colletotrichum orbiculare. Roots of cucumber plants were treated with these fungal isolates using barley grain inocula (BGI), mycelial inocula (MI) or culture filtrate (CF). Most isolate/inoculum form combinations significantly reduced the disease except BGI of Trichoderma. These fungal isolates were also evaluated for induction of systemic resistance against bacterial angular leaf spot and Fusarium wilt by treatment with BGI. Penicillium, Phoma and the sterile fungus significantly reduced the disease incidence of bacterial angular leaf spot. Phoma and sterile fungus protected plants significantly against Fusarium wilt. Roots treated with CFs of these fungal isolates induced lignification at Colletotrichum penetration points indicating the presence of an elicitor in the CFs. The elicitor activity of CFs was evaluated by the chemiluminescence assay using tobacco callus and cucumber fruit disks. The CFs of all isolates elicited conspicuous superoxide generation. The chemiluminescence activity of the CF of Penicillium was extremely high, and its intensity was almost 100-fold higher than that of other isolates. The chemiluminescence activity was not lost following treatment with protease or autoclaving or after removal of lipid. The MW 12,000 dialyzed CF fraction was highly effective in eliciting chemiluminescence activity. Chemiluminescence emission from cucumber fruit disks treated with Penicillium was the same as that obtained from tobacco callus, except that the lipid fraction also showed a high activity. Both the MW 12,000 fraction and the lipid fraction induced lignification in the epidermal tissues of cucumber hypocotyls.     

Isolation and selection of Hemileia Vastatrix antagonists

Organic coffee growing is rapidly increasing in Brazil, and many diseases, especially coffee leaf rust (Hemileia vastatrix), are threatening its production. This study is a first step towards a biocontrol program for coffee rust on organically grown plants. In three organic coffee farms in the state of Minas Gerais, 393 microbial strains including 154 bacterial and 239 fungal strains were isolated from leaves, leaf debris, and soil samples, and in 6 month-old coffee plants, 17 of these isolates reduced both the infection frequency (IF) and the number of H. vastatrix urediniospores produced per leaf (UPL) by more than 70 %. The isolates were identified as eight bacteria isolates, seven Bacillus spp. and one Pseudomonas sp., and nine fungal isolates, four Fusarium spp., two Penicillium spp., one Aspergillus sp., one Acremonium sp. and one Cladosporium sp. Each fungal and bacterial isolate was applied 0, 4, 8, 12 or 16 days before and 0, 4, 8, 12 or 16 days after H. vastatrix inoculation, and the efficiency in reducing both IF and UPL was evaluated. The efficiency was higher and lasted longer when the bacterial isolates were applied before H. vastatrix inoculation. Six Bacillus (B10, B25, B143, B157, B171, B175), two Fusarium (F205, F281), and one Pseudomonas (B286) isolates are potentially efficient as biocontrol agents of H. vastatrix and will be tested using field experiments.