Diatomaceous earth increases the efficacy of Beauveria bassiana against Tribolium castaneum larvae and increases conidia attachment

This research tested the suppressive ability of Beauveria bassiana (Balsamo) Vuillemin alone and in combination with diatomaceous earth against the red flour beetle, Tribolium castaneum (Herbst). Adults did not show a dose response to B. bassiana, and the addition of diatomaceous earth (DE) did not result in a significant increase in mortality. Against larvae, however, DE at 190 mg/kg grain enhanced the efficacy of B. bassiana at all concentrations ranging from 33 to 2,700 mg of conidia per kilogram of grain. The presence of DE resulted in 17- and 16-fold decreases in the median lethal concentration of B. bassiana at 56 and 75% RH, respectively. No significant differences in larval mortality in response to B. bassiana and diatomaceous earth alone or in combination were found between 56 and 75% RH. Conidial attachment to larvae was significantly greater with 190 mg/kg DE than without it. The partial analysis of lipids taken up by DE from the larvae revealed the removal of phospholipids and long-chain fatty acids. These results support the hypothesis that diatomaceous earth enhances the efficacy of B. bassiana against larval T. castaneum, at least in part by damaging the insect cuticle, thus increasing conidial attachment and making nutrients more available to conidia for their germination.     

Biocontrol of almond bark beetle (Scolytus amygdali Geurin-Meneville, Coleoptera: Scolytidae) using Beauveria bassiana (Bals.) Vuill. (Deuteromycotina: Hyphomycetes)

AIMS: To formulate the entomopathogenic fungus Beauveria bassiana in invert emulsion, then apply it against adults of almond bark beetle (Scolytus amygdali) under laboratory and field conditions. METHODS AND RESULTS: The effect of formulated B. bassiana in invert emulsion against S. amygdali adults was shown by comparing the mortality percentage of adults exposed to the formulated fungus using a Petri dish treatment method and by field applications to infested peach trees with mortality of adults exposed to the unformulated fungus or the untreated control. Results obtained from both exposure methods have indicated that treatment of S. amygdali adults with the formulated fungus resulted in a significantly higher mean mortality percentage (P < 0.05) when compared with the treatment with the unformulated fungus or the untreated control. This mortality ranged from 81.2 to 100%, 10 days after treatment with the formulated fungus when compared with 6.7 to 49.6% mortality, 10 days after treatment with the control or the unformulated fungus, respectively. Viability of the fungus conidia in invert emulsion was assessed by calculating the germination percentage of the conidia over time. Results indicated a high storage stability shown by a small loss of germination percentage for the formulated conidia of both strains (5.8 to 8.4% over a 12-week period) vs a low storage stability shown by a high loss of germination percentage for the unformulated conidia of the same strains (58.9 to 61.0% over the same period). The presence of B. bassiana in the galleries of beetles following the treatment of infested trees was shown in the present research. CONCLUSIONS: The results obtained have demonstrated a significantly higher level of efficacy of formulated B. bassiana in invert emulsion against S. amygdali adults under laboratory and field conditions. The ingredients of invert emulsion used in the formulation of the fungus had a negligible effect on the viability of formulated conidia when compared with the unformulated. SIGNIFICANCE AND IMPACT OF THE STUDY: Results obtained in the present research are promising and may be exploited commercially to control S. amygdali adults on various species of stone fruit trees, especially peach trees. This type of biocontrol of this insect may be used as an alternative means to chemical control for management of the insect. No adverse environmental impacts of the fungus or its formulation have been observed during application.     

Efficacy of Bauveria bassiana for red flour beetle when applied with plant essential oils or in mineral oil and organosilicone carriers

The carriers mineral oil and Silwet L-77 and the botanical insecticides Neemix 4.5 and Hexacide were evaluated for their impacts on the efficacy of Beauveria bassiana (Balsamo) Vuillemin conidia against red flour beetle, Tribolium castaneum (Herbst), larvae. The dosages of liquid treatments were quantified by both conidia concentration in the spray volume and conidia deposition on the target surface. The latter approach allowed comparison with dry, unformulated conidia. The median lethal concentrations of B. bassiana in 0.05% Silwet L-77 solution or without a carrier were approximately double that for conidia in mineral oil. Carriers had highly significant effects on the efficacy of B. bassiana. The lower efficacy of conidia in aqueous Silwet L-77 may have been the result of conidia loss from the larval surface because of the siloxane's spreading properties. Neemix 4.5 (4.5% azadirachtin) delayed pupation and did not reduce the germination rate of B. bassiana conidia, but it significantly reduced T. castaneum mortality at two of four tested fungus doses. Hexacide (5% rosemary oil) caused significant mortality when applied without B. bassiana, but it did not affect pupation, the germination rate of conidia, or T. castaneum mortality when used in combination with the fungus.     

Recombinant immunosuppressive protein from Pimpla hypochondrica venom (rVPr1) increases the susceptibility of Mamestra brassicae larvae to the fungal biological control agent, Beauveria bassiana

Although fungi are used to control a variety of insect pests, it is accepted that their usage could be increased if their efficacy was greater. The outcome of the interaction of a fungus and a pest insect may be influenced by a number of criteria, including the ability of the insect to mount effective immune responses against the pathogen. In view of this, we aimed to determine if a recombinant immunosuppressive wasp venom protein (rVPr1) can increase the susceptibility of larvae of the lepidopteran pest, Mamestra brassicae, to the fungal biological control agent, Beauveria bassiana. Bioassays indicated that when larvae were injected with 3.5 μl of rVPr1 and 100 B. bassiana conidia (combined injection assays), a significant reduction in survival of larvae occurred compared with each treatment on its own (P=0.006). Similar results were obtained when larvae were dipped in a solution containing 3 × 10(6) B. bassiana conidia per ml and then injected with 3.5 μl of rVPr1 2 days later (topical application assays), (P<0.001). These results indicate that rVPr1 can increase the efficacy of B. bassiana toward a lepidopteran pest, and are discussed within the context of insect immune responses and integrated pest management.     

Arena size, hole density, and capture of Oryzaephilus surinamensis (Coleoptera: Silvanidae) in grain probe traps

The relationship of size of test arena, number of holes in a grain probe trap body and capture of the sawtoothed grain beetle, Oryzaephilus surinamensis (L.), was determined in simulated field tests conducted in an outdoor screen enclosure exposed to natural temperature fluctuations. Polyvinylchloride (PVC) probe bodies were attached to electronic sensor heads, and insect captures were recorded electronically using an electronic grain probe insect counter (EGPIC) system. In comparisons among PVC probe trap bodies with 60, 132, 252, and 492 holes, tested at 18 insects per kilogram in 4.5, 17, and 40 kg of soft wheat in cylindrical arenas (10.2, 20.3, and 30.5 cm in diameter, respectively), number of holes in the probe trap body had no effect on insect capture, but percentage of insects recovered was indirectly related to size of the test arena. Periodicity of insect capture was determined using the time-stamp data that were recorded by the EGPIC system. Circadian rhythm was observed in the periodicity of the capture that corresponded to foraging activity peaks documented for sawtoothed grain beetles, with activity peaks occurring early in the scotophase. There were shifts in times of peak activity among the different test arena sizes that corresponded to differences in temperature in the grain mass. Increases in both temperature and contact between insects and grain probe in the smallest arenas resulted in higher capture of sawtoothed grain beetles. This research documents additional important factors when evaluating capture of sawtoothed grain beetles in grain probe traps.     

利用GFP表达系统检测球孢白僵菌侵染昆虫过程

绿色荧光蛋白(GFP)作为一种重要的报告基因,在病原菌和寄主之间的互作研究中具有良好的应用前景。利用草丁膦抗性基因bar为筛选标记,构建了组成性表达绿色荧光蛋白基因egfp的载体pBG,并转入球孢白僵菌获得成功表达。将转化子接种菜青虫进行生物测定表明,组成性表达egfp不影响球孢白僵菌的毒力。进一步利用冰冻切片和荧光显微观察技术监测球孢白僵菌侵染寄主行为,结果显示,通过荧光观察可清楚的检测到病原菌在昆虫体表的附着、菌丝穿透体壁以及菌丝从寄主体内长出等侵染过程。由此表明,GFP可有效用于球孢白僵菌的遗传标记,进行病原菌的鉴别、侵染致病机理及相关功能基因的表达模式研究。     

利用GFP表达系统检测球孢白僵菌侵染昆虫过程

The green fluorescent protein (GFP) has been expressed in a variety of organisms and has been used extensively as a marker in the study of host-pathogen interaction. We have expressed GFP in the entomopathogenic fungus Beauveria bassiana through transformation with a vector that confers resistance to phosphinothricin. Conidia expressed GFP and were readily detected by fluorescence microscopy. Fluorescent hyphae and conidia were easily distinguished in the tissue of the cabbage worm. GFP-tagged strains of B. bassiana could be used to study the developmental fate of the fungus within its insect hosts. Thus, this method is useful for screening and monitoring the engineered strains even after infection.     

Response of the Wasp Cephalonomia tarsalis (Hymenoptera: Bethylidae) to Beauveria bassiana (Hyphomycetes: Moniliales) as Free Conidia or Infection in Its Host, the Sawtoothed Grain Beetle, Oryzaephilus surinamensis (Coleoptera: Silvanidae)

Cephalonomia tarsalis, an ectoparasitoid, and Beauveria bassiana, an entomopathogenic fungus, are potential biological control agents for the sawtoothed grain beetle, Oryzaephilus surinamensis. Several experiments were conducted to determine whether the two beneficial organisms are compatible. Wasps exhibited little avoidance behavior toward the fungus. Adult wasps oviposited on B. bassiana-infected larvae up to within 1 day of the host's death and the appearance of red fungal pigment. Wasp larvae are susceptible to the fungus and die within 1 day of oviposition on host larvae with mycosis. A 3-h exposure of adult wasps to 100 mg of B. bassiana/kg of wheat resulted in 52.7% mortality. Nevertheless, the wasps entered into grain containing B. bassiana conidia as freely as they entered into conidia-free grain. The mean prevalence of B. bassiana in 46 samples of pooled wheat representing 276 locations was 7.5 colony-forming units/g of wheat. Natural C. tarsalis exposure to B. bassiana in untreated stored wheat is likely to be below lethal quantities, and the introduction of the fungus in insecticidal quantities would have a negative impact on C. tarsalis populations.     

Desiccant dusts synergize the effect of Beauveria bassiana (Hyphomycetes: Moniliales) on stored-grain beetles

Diatomaceous earth (DE) is a desiccant insecticide and most efficacious in low humidity. It acts on insect cuticle by absorbing lipids, and perhaps by cuticular abrasion. Beauveria bassiana (Balsamo) Vuillemin, an entomopathogenic fungus, is most efficacious in high humidity and has a complex interaction with cuticular lipids. Interaction between these materials may enhance insect control performance. Assays with stored-grain beetles were conducted with B. bassiana at rates of 11, 33, 100, and 300 mg of conidia per kilogram of grain with and without single rates of DE that killed 10% or less of the target beetles. The assays revealed synergism in effects on adult Rhyzopertha dominica (F.) and Oryzaephilus surinamensis (L.) at all doses. There was statistically significant synergism for adult Cryptolestes ferrugineus (Stephens) and larval R. dominica but at only one B. bassiana rate for each target. Both amorphous silicon dioxide, a sorptive dust, and diamond dust, an abrasive, showed synergistic interaction with B. bassiana on adult R. dominica. These results may provide a basis for a least-toxic approach to control of stored-product beetles and for efficacy-enhancing formulation of entomopathogenic fungi.     

Evaluation of synergistic interactions between the Colorado potato beetle (Coleoptera: Chrysomelidae) pathogen Beauveria bassiana and the insecticides, imidacloprid, and cyromazine

Laboratory studies investigated the interaction between the fungal entomopathogen Beauveria bassiana (Balsamo) Vuillemin and sublethal doses of the insecticides imidacloprid and cyromazine when applied to larvae of the Colorado potato beetle, Leptinotarsa decemlineata (Say). When second instars were fed potato leaf discs treated with sublethal doses of imidacloprid and a range of doses of B. bassiana, a synergistic action was demonstrated. Similar results were observed when larvae were sprayed directly with B. bassiana conidia and immediately fed leaf discs treated with imidacloprid. No synergistic interaction was detected when larvae were fed leaf discs treated with sublethal doses ofimidacloprid 24 h after application of B. bassiana conidia to larvae. However, a synergistic interaction was detected when larvae were fed leaf discs treated with imidacloprid and sprayed with B. bassiana conidia 24 h later. Although sublethal doses of both imidacloprid and the triazine insect growth regulator (IGR) cyromazine prolonged the duration of the second instar, only imidacloprid interacted with B. bassiana to produce a synergistic response in larval mortality. In leaf consumption studies, the highest dose of B. bassiana tested promoted feeding in inoculated second instars. Feeding was inhibited when larvae were fed foliage treated with sublethal doses of imidacloprid and significantly reduced when fed foliage treated with a sublethal dose of cyromazine. Starvation of larvae for 24 h immediately after B. bassiana treatment produced a similar result to the combined treatment of B. bassiana and imidacloprid and increased the level of mycosis when compared with B. bassiana controls. Imidacloprid treatment affected neither the rate of germination of B. bassiana conidia on the insect cuticle nor the rate at which conidia were removed from the integument after application. The statistical analysis used to detect synergism and the possible role of starvation-induced stress factors underlying the observed synergistic interactions are discussed.     

A Proposed Role for the Cuticular Fatty Amides of Liposcelis bostrychophila (Psocoptera: Liposcelidae) in Preventing Adhesion of Entomopathogenic Fungi with Dry-conidia

Maximum challenge exposure of Liposcelis bostrychophila to Beauveria bassiana, Paecilomyces fumosoroseus, Aspergillus parasiticus or Metarhizium anisopliae resulted in no more than 16% mortality. We investigated several of L. bostrychophila's cuticular lipids for possible contributions to its tolerance for entomopathogenic fungi. Saturated C14 and C16 fatty acids did not reduce the germination rates of B. bassiana or M. anisopliae conidia. Saturated C6 to C12 fatty acids that have not been identified in L. bostrychophila cuticular extracts significantly reduced germination, but the reduction was mitigated by the presence of stearamide. Cis-6-hexadecenal did not affect germination rates. Mycelial growth of either fungal species did not occur in the presence of caprylic acid, was reduced by the presence of lauric acid, and was not significantly affected by palmitic acid. Liposcelis bostrychophila is the only insect for which fatty acid amides have been identified as cuticular components. Stearamide, its major fatty amide, did not reduce germination of B. bassiana or M. anisopliae conidia or growth of their mycelia. Adhesion of conidia to stearamide preparations did not differ significantly from adhesion to the cuticle of L. bostrychophila. Pretreatment of a beetle known to be fungus-susceptible, larval Oryzaephilus surinamensis, with stearamide significantly decreased adhesion of B. bassiana or M. anisopliae conidia to their cuticles. This evidence indicates that cuticular fatty amides may contribute to L. bostrychophila's tolerance for entomopathogenic fungi by decreasing hydrophobicity and static charge, thereby reducing conidial adhesion.     

Field Evaluation of Beauveria bassiana: Its Persistence and Effects on the Pea Aphid and a Non-target Coccinellid in Alfalfa

Several strains of the entomopathogenic fungus Beauveria bassiana have been considered for use as microbial insecticides. Experimental sprays were conducted in an alfalfa field with an aphid-derived strain of B. bassiana to determine its persistence and its effects on pea aphids, Acyrthosiphon pisum (Homoptera: Aphididae) and a non-target aphid predator, Hippodamia convergens (Coleoptera: Coccinellidae). B. bassiana conidia persisted in the field for at least 28 days, when approximately 10% of the original inoculum was still present. In the lower canopy, more conidia were present than on other plant parts and they persisted longer on the leaves in this location. However, conidia were still abundant in the upper canopy, where 97.9% of the aphids and 95.5% of H. convergens larvae were found. Thus, both insect species were exposed to the fungus for at least 1 month. However, pea aphid populations were not affected by the fungus. The predator's incidence was reduced by 75-93% (depending on application rate) early in the season, but was not affected later in the season. Insect life history patterns and weather conditions are likely causes for the differences seen in field effects.     

Development of a novel bioassay for estimation of median lethal concentrations (LC50) and doses (LD50) of the entomopathogenic fungus Beauveria bassiana, against western flower thrips, Frankliniella occidentalis

To conduct laboratory experiments aimed at quantifying secondary acquisition of fungal conidia by western flower thrips (Frankliniella occidentalis), an efficient assay technique using Beauveria bassiana as the model fungus was developed. Various application protocols were tested and it was determined that the percent mortality did not vary among protocols. Peak mortality of second-instar nymphs, under constant exposure to conidia, occurred 5 days post-inoculation. Second-instar thrips that were exposed to conidia within 24 h of the molt to second instar were more susceptible to Beauveria bassiana than thrips exposed after times greater than 24 h post-molt. Conidia efficacy, which was monitored at 24 h intervals, did not differ significantly within 72 h. A test of the final bioassay system was conducted in a series of assays aimed at determining the LD50 of B. bassiana technical powder against second-instar western flower thrips. It was determined that B. bassiana (strain GHA) is highly effective at very low doses (LD50 of 33-66 conidia/insect).     

Hole density and capture of stored-product insect pests in grain probe traps

The relationship between number of holes in a grain probe trap body and capture of stored-grain pests was determined in laboratory tests using adults of the rice weevil, Sitophilus oryzae (L.), the sawtoothed grain beetle, Oryzaephilus surinamensis (L.), and the red flour beetle, Tribolium castaneum (Herbst). Polyvinylchloride (PVC) probe bodies were attached to electronic sensor heads, and insect captures were recorded electronically using an Electronic Grain Probe Insect Counter (EGPIC) system. In comparisons among PVC probe trap bodies with 60-492 holes, tested at 71 insects per kg in 2.8 kg of soft wheat in cylindrical mini-silos, sawtoothed grain beetle and rice weevil captures were directly related to number of holes in the probe trap body, but there was no relationship for red flour beetle capture. Subsequent tests were conducted comparing sawtoothed grain beetle and rice weevil captures in a PVC probe body with 210 holes over a 40-cm long trapping surface with two commercially available probe traps, a polycarbonate (Lexan) probe trap with 180 holes over a 14-cm long trapping surface and a polyethylene (WBII) probe trap with 750 holes over a 34-cm long trapping surface. The highest percentage capture of both species was in the WBII probe trap, but the 210-hole PVC probe body was as effective as the Lexan probe body for rice weevils and sawtoothed grain beetles at 71 and 17 insects per kg of wheat, respectively.     

Dietary stress increases the susceptibility of Tribolium castaneum to Beauveria bassiana

Sanitation being an important component of chemical-free management of stored-products pests, the nutritional stress on insects that results from a clean environment may prove advantageous to the use of microbial controls. Dietary stress by food deprivation or suboptimal diet increased susceptibility of the red flour beetle, Tribolium castaneum (Herbst), larvae to Beauveria bassiana (Balsamo) Vuillemin. Fungus-related mortality increased linearly with the number of days of food deprivation. Rearing of larvae on a rice meal diet resulted in slower development and greater susceptibility to B. bassiana than rearing on whole wheat flour with brewer's yeast. Larvae that were fed for 24 h on flour with B. bassiana conidia consumed significantly less and weighed significantly less than those that were fed fungus-free flour. Thus, the presence of B. bassiana conidia is itself a cause of dietary stress.     

Horizontal transmission of Beauveria bassiana in Anastrepha ludens (Diptera: Tephritidae) under laboratory and field cage conditions

The virulence of two products of the fungus Beauveria bassiana (LCPP and Bassianil) on adult Anastrepha ludens (Loew) (Diptera: Tephritidae) and their effect on the mating performance of infected males was evaluated in laboratory and field cage tests. The horizontal transmission capacity of the fungus during copulation or attempted copulation also was quantified using inoculated males as well as the impact of infection on female fecundity and longevity. Both fungal products were found to be highly virulent (LCPP, mortality = 98.7% at 1 x 10(8) conidia per ml, LT50 = 4.20 d, LC50 = 9.35 x 10(5) conidia per ml; Bassianil, mortality = 99.3% at 1 X 10(8) conidia per ml, LT50 = 4.04 d, LC50 = 2.69 x 10(7) conidia per ml). Mating success of inoculated males was not affected compared with the control group during the 3 d postinoculation. Horizontal transmission to females during the first day was 80.6 and 84.3% through mating and 15.4 and 21.6% through attempts to mate and contact during courtship for the LCPP and Bassianil products, respectively. The fertility of infected females was notably reduced, and longevity did not extend beyond 15 d. Our results suggest the possibility of using sterile flies as fungus vectors in sterile insect technique programs, but the potential benefits and shortcomings of this approach require further investigation.     

Potential of fungi as biocontrol agents of polyphagotarsonemus latus (Acari: Tarsonemidae)

Infection of broad mites, Polyphagotarsonemus latus, by conidia of Beauveria bassiana, Hirsutella thompsonii, and Paecilomyces fumosoroseus conidia was investigated in the laboratory under controlled temperature and moisture conditions. Infection of P. latus by the fungus (LC₅₀) was related positively to the 1.16 × 10⁶ B. bassiana conidia per ml, 2.39 × 10³ H. thompsonii conidia per ml, and 1.29 × 10⁵ P. fumosoroseus conidia per ml. Mortality caused by B. bassiana occurred the fastest among densities fluctuating between 65 and 125 mites per leaf. The pathogens B. bassiana and P. fumosoroseus, adjuvants (oil and molasses), and the acaricide ABG6364, a microbial insecticide containing the Beta-exotoxin of Bacillus thuringiensis were evaluated for efficacy against P. latus in a greenhouse test. The B. bassiana treated plants had the greatest and most persistent effect on percentage mortality of total broad mites present per leaf (88 %) followed by the acaricide ABG6364. Use of adjuvants (oil and molasses) did not increase infection of P. latus.     

Effectiveness of spinosad against seven major stored-grain insects on corn

In January 2005, the United States Environmental Protection Agency registered spinosad as a stored grain protectant. No referenced data on the efficacy of spinosad on corn in suppressing major stored-grain insects have been published. In this paper, we evaluated the efficacy of spinosad against seven major stored-grain insects on shelled corn in the laboratory. Insect species tested were the red flour beetle, Tribolium castaneum （Jacquelin duVal）; rusty grain beetle, Cryptolestesferrugineus （Stephens）; lesser grain borer, Rhyzopertha dominica （F.）; sawtoothed grain beetle, Oryzaephilus surinamensis （L.）; rice weevil, Sitophilus oryzae （L.）; maize weevil, Sitophilus zeamais （Motschulsky）; and Indian meal moth, Plodia interpunctella （Htibner）. Corn kernels were treated with spinosad at 0, 0. 1, 0.5, 1, and 2 active ingredient （a.i.） mg/kg for controlling the seven species. Beetle adults or P. interpunctella eggs were introduced into each container holding 100 g of untreated or insecticide-treated corn. The seven insect species survived well on the control treatment, produced 28 to 336 progeny, and caused significant kernel damage after 49 days. On spinosad-treated corn, adult mortality of C. ferrugineus, R. dominica, 0. surinamensis, S. oryzae, and S. zeamais was 〉 98% at 1 and 2 mg/kg after 12 days. Spinosad at≥ 0.5 mg/kg completely suppressed egg-to-larval survival after 21 days and egg-to-adult emergence of P. interpunctella after 49 days, whereas 16% T. castaneum adults survived at 1 mg/kg after 12 days. Spinosad at 1 or 2 mg/kg provided complete or near complete suppression of progeny production and kernel damage of all species after 49 days. Our results indicate that spinosad at the current labeled rate of 1 mg/kg is effective against the seven stored-grain insect pests on corn.     

Beauveria bassiana -A Potential Mycopesticide for the Efficient Control of Coffee Berry Borer, Hypothenemus hampei (Ferrari) in India

The Coffee Berry Borer (CBB), Hypothenemus hampei (Ferrari) (Coleoptera: Scolytidae) has been a serious insect pest of coffee cultivars C. robusta and C. catimor in India since 1991, causing 40-80% coffee bean loss. To combat this important pest, an indigenous entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin was isolated from dead and moribund coffee berry borers from the wild. The fungus was cultured on yeast extractpeptone supplemented liquid medium. The mycelial mat was harvested from 12-day old cultures and lyophilized. A suspension of the most virulent isolate (Bb2) was prepared in sterile water and used as a mycopesticide. The laboratory studies were conducted on coffee berry borers by applying conidial suspensions at a dosage rate of 1 ×10 6 conidia ml -1 . Pest mortality with the Bb2 isolate increased from 69.3% to 95.3% with an increase in relative humidity (RH) at 25 ±2°C. Field experiments were conducted in a coffee plantation area in the Kodagu district of Karnataka, and the results showed that, under favourable environmental conditions (27- 29°C; 82-91% RH; 10-15 inches rainfall per year), the fungus required only eight days to colonize and kill the target pest. A maximum of 75.6% insect mortality was recorded 24 days after spraying. Large-scale field trials conducted in five plantation plots between September 1995 and September 1998 showed significant insect mortality both in C. robusta and C. catimor cultivars of coffee. The potential use of this indigenous fungal strain of B. bassiana as a mycopesticide for management of CBB in India is discussed.     

N-Acetyl-d-Glucosamine-Mediated Regulation of Extracellular Protease in the Entomopathogenic Fungus Beauveria bassiana

The entomopathogenic fungus Beauveria bassiana GK2016 grown in a liquid medium incorporating gelatin as the sole carbon and nitrogen source produced an extracellular serine protease (molecular weight, 35,000; pI ca. 10). Without gelatin, B. bassiana could utilize N-acetyl-d-glucosamine (GlcNAc; 2-acetamido-2-deoxy-d-glucose) as the sole source of carbon and nitrogen, and GlcNAc availability increased the storage carbohydrate content in mycelia. Synthesis of protease was repressed in gelatin medium containing GlcNAc at levels of >1.07 mumol mg of fungal dry weight. At levels below this, protease synthesis was initiated; subsequently, free amino nitrogen appeared in the medium and diauxic growth was observed. Slow feeding with GlcNAc (35.34 mug ml h) did not repress protease synthesis nor did GlcNAc accumulate in the medium above 0.5 mg ml. Increasing the rate of release of GlcNAc (83.51 mug ml h) resulted in the accumulation of GlcNAc in the medium to 2.0 mg ml, a 45% increase in growth and a decrease in protease synthesis by about 81%. Free amino acids generated from the hydrolysis of gelatin did not repress protease synthesis. These data are interpreted in terms of known interaction of B. bassiana with insect cuticular components. We suggest that the action of extracellular chitinases synthesized by B. bassiana on insect cuticle, and pursuant release of GlcNAc, may have important consequences on the regulation of other extracellular catabolic enzymes such as the protease.