Effects of Temperature and Culture Media on Vegetative Growth of an Entomopathogenic Fungus <Emphasis Type="Italic">Isaria</Emphasis> sp. (Hypocreales: Clavicipitaceae) Naturally Affecting the Whitefly, <Emphasis Type="Italic">Bemisia tabaci</Emphasis> in Texas

The effects of temperature and mycological media on mycelial growth and estimates of spore production of an indigenous entomopathogenic fungus, Isaria sp., found during natural epizootics on whiteflies in the Lower Rio Grande Valley of Texas, were investigated. The radial growth (mm/day) of Isaria sp. as a function of temperature fits a linear model; with faster growth on Sabouraud dextrose agar with yeast extract, SDAY slopes (0.23) than on Sabouraud maltose agar, SMA slopes (0.14) from 20 to 30°C, with an optimal temperature of 30°C (SDAY: 4.1 mm, SMA: 3.1 mm). Moderate growth occurred at 25°C (SDAY: 3.4 mm, SMA: 2.7 mm). Growth was lowest at 20°C (SDAY: 1.9 mm, SMA: 1.8 mm). No fungal growth was observed at 35°C and 40°C. However, when Isaria sp. was exposed to 35°C for the first 7 days, it could recover and grow when transferred to 25°C (SDAY: 3.5 mm, SMA: 2.8 mm). No recovery or growth occurred after transfer from 40°C to 25°C. The average conidial production on SDAY after 20 days incubation at 25°C and a photoperiod of 14:10 h light: dark was 1.2 × 10⁸ conidia/cm² with 100% spore viability. When compared on SDAY at 25°C, the radial growth rate of I. javanica ex type CBS 134.22 (5.1 mm/day) was greater than seven Isaria isolates including Isaria sp.; but maximum growth rates were similar among all related Isaria isolates (90–97%). The Isaria sp. fungus tolerates high temperatures (35°C), suggesting that it is naturally selected for the subtropical semi-arid environment, where it could serve as an important natural control agent of the sweet potato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) biotype B, one of the most invasive and economically damaging insects to agriculture. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U. S. Department of Agriculture.     

Variation between laboratory populations in the performance of the parasitoid Encarsia formosa on two host species, Bemisia tabaci and Trialeurodes vaporariorum

We tested the hypothesis that populations of the parthenogenetic parasitic wasp Encarsia formosa Gahan (Hymenoptera: Aphelinidae) differed in their ability to use two different host species, Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) and Trialeurodes vaporariorum Westwood (Homoptera: Aleyrodidae). Of the three wasp populations tested, two populations had been reared for many generations on B. tabaci and one population had been reared for many years on T. vaporariorum. Performance was measured by the number of whitefly nymphs that were successfully parasitized by individual wasps, and performance on either host was measured in separate experiments. There was variation between wasp populations in their performance on the host B. tabaci, with one wasp population reared for many years on this host performing considerably better than the other two populations. There were no significant differences between populations in their use of the preferred host, T. vaporariorum. The experiments were conducted in such a way that we could distinguish heritable differences between populations from environmentally-induced conditioning differences due to the immediate host from which an individual wasp enclosed. In either experiment there were no significant effects of conditioning, although there was a trend within each population for wasps conditioned on T. vaporariorum to have higher performance than those conditioned on B. tabaci. Thirdly, we conducted a selection experiment, initiated with wasps from a single population historically reared on T. vaporariorum, to measure the effect of laboratory rearing on different hosts for 17 generations. We did not see any difference in the performance of wasps on B. tabaci after this period of rearing on either of the two hosts. In summary, populations of E. formosa do differ in their relative performance on B. tabaci. The one population that was tested further did not show any response to selection by rearing, but the ability to respond to selection on performance may not be equal for all populations. The possibility that wasp populations have differential performance on particular hosts may affect the use of this species as a biological control agent.     

Pathogenicity of Aschersonia spp. against whiteflies Bemisia argentifolii and Trialeurodes vaporariorum

Entomopathogenic fungi of the genus Aschersonia are specific for whitefly and scale insects. They can be used as biological control agents against silverleaf whitefly, Bemisia argentifolii and greenhouse whitefly, Trialeurodes vaporariorum. Forty-four isolates of Aschersonia spp. were tested for their ability to sporulate and germinate on semi-artificial media and to infect insect hosts. Seven isolates sporulated poorly (less than 1x10(7) conidia/dry weight) and 10 were not able to infect either of the whitefly species. Several isolates were able to produce capilliconidia. Infection level was not correlated with germination on water agar. After a selection based on spore production and infection, virulence of 31 isolates was evaluated on third instar nymphs of both whitefly species on poinsettia (Euphorbia pulcherrima). Whitefly infection levels varied between 2 and 70%, and infection percentages of B. argentifolii correlated with that of T. vaporariorum. However, mortality was higher for T. vaporariorum than for B. argentifolii, as a result of a higher 'mortality due to unknown causes.' Several isolates, among which unidentified species of Aschersonia originating from Thailand and Malaysia, A. aleyrodis from Colombia, and A. placenta from India showed high spore production on semi-artificial medium and high infection levels of both whitefly species.     

Similarity in flight activity rhythms in coexisting species of Aleyrodidae, Bemisia tabaci and Trialeurodes abutilonea

Experiments were conducted in Arizona, USA to examine the diel patterns of flight activity of two coexisting species of whiteflies, Bemisia tabaci (Gennadius) and Trialeurodes abutilonea (Haldeman) (Aleyrodidae). Aerial populations were monitored during parts of two growing seasons using sticky traps. It was found that on most days when more than 100 whiteflies were captured, the flight activity rhythms of the two species were remarkably similar. During 13 of 14 weeks if aerial populations of one species were used to predict populations of the other, coefficients of determination were highly significant. This is in part a function of the two species having parallel patterns of pupal eclosion and teneral periods of approximately the same duration. The two species both experience 90% eclosion within 1 h of the onset of photophase and both have a teneral period slightly in excess of 4 h at 27°C. As a result populations of the two whiteflies emerge together, become flight ready, and fly at the same time. Arizona populations of these two introduced species appear to have developed certain very similar behavioral and physiological processes under conditions of a rigorous desert climate.

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Zusammenfassung In Arizona (USA) wurden Experimente durchgeführt, um die Muster der täglichen Flugaktivität von zwei co-existierenden Aleurodidenarten, Bemisia tabaci und Trialeurodes abutilonea zu untersuchen. Mit Hilfe von Klebefallen wurden die Adult-Populationen während zweier Wachstumperioden beobachtet. Die Rhythmen der Flugaktivität während der meisten Tage, an denen mehr als 100 Weisse Fliegen gefangen wurden, waren bemerkenswert ähnlich. Die Bestimmungskoeffizienten waren deutlich signifikant in 13 von 14 Wochen, wenn die Populationen der einen Art benutzt wurde, um die Population der anderen Spezies vorherzusagen. Dies rührt zum Teil daher, dass sich beide Arten in ähnlicher Weise entwicklen. Beide schlüpfen zu 90% innerhalb von 3 Stunden nach Beginn der Photophase und beginnen bei 27°C im Mittel nach 4 Stunden und 12 Minuten zu fliegen. Daraus folgt, dass Populationen beider Arten zur selben Zeit schlüpfen, flugbereit werden und zu fliegen beginnen. Es scheint, dass die in Arizona vorhandenen Populationen dieser beiden eingeführten Arten unter dem Einfluss eines rauhen Wüstenklimas einige sehr ähnliche Verhaltensweisen und physiologische Prozesse entwickelt haben.

     

Natural occurrence of entomopathogenic fungi ( Zygomycetes : Entomophthorales ) of aphid ( Hemiptera : Aphididae ) pests of horticultural crops in Argentina

A three-year survey of entomophthoralean pathogenic fungi of aphids from horticultural crops in La Plata, Argentina, was conducted. Nine species of aphids, including Aphis fabae Scopoli, Aphis gossypii Glover, Brevicoryne brassicae (L.), Lipaphis erysimi (Kaltenbach), Macrosiphum euphorbiae (Thomas), Myzus sp., Myzus persicae (Sulzer), Nasonovia ribisnigri (Mosley) and Capitophorus elaeagni (del Guercio) were recorded as hosts of entomopathogenic fungi. Six species of Entomophthorales that infected and killed aphids were found in vegetable crops. The fungal species identified were Conidiobolus obscurus (Hall & Dunn) Remaudière & Keller, Entomophthora planchoniana Cornu, Neozygites fresenii (Nowakowski) Remaudière & Keller, Pandora neoaphidis (Remaudière & Hennebert) Humber, Zoophthora radicans (Brefeld) Batko and Zoophthora sp. Pandora neoaphidis was the most predominant pathogen of aphids and was found throughout the summer (December–March) 2004. The recovery of C. obscurus, N. fresenii and P. neoaphidis represent first records of these fungi for South America.     

Recycling of entomopathogenic nematodes in Delia radicum and in other insects from cruciferous crops

Larvae of Deliaradicum Linnaeus (Diptera: Anthomyiidae) wereexposed to Steinernema feltiae Filipjev(Rhabditida) to study the ability of thenematodes to fulfill their life cycle (recycle)in this host. In addition, the recycling ofSteinernema and Heterorhabditis(Rhabditida) species was studied in larvae ofthe following insects: Meligethes spp.Stephens (Coleoptera: Nitidulidae),Dasyneura brassicae Winnertz (Diptera:Cecidomyidae), Ceutorrhynchus assimilisPaykull, C. pallidactylus Marsham (Coleoptera: Curculionidae) and Mamestrabrassicae Linnaeus (Lepidoptera: Noctuidae).All larval instars of D. radicum werestudied (larvae live and pupate in soil)whereas the final instars of the remaininginsects were studied (larvae only pupate in soil).On average, the following numbers of infectivejuveniles were produced in the different hosts:1,200–1,400 in C. assimilis; 700–1,300 inMeligethes spp. and 47,000 in M.brassicae. In D. radicum, around 400infective juveniles were produced on average inthe smallest larvae and around 3,500 in thelargest larvae. The highest number of nematodesrecorded in a D. radicum larva (thirdinstar) was 9,500 infective juveniles. Thenumber of nematodes produced in D.radicum cadavers was positively related to thesize of the insect host, but large variationwas observed.     

Identification,comparison, and functional analysis of salivary phenol‐oxidizing enzymes in Bemisia tabaci B and Trialeurodes vaporariorum

The differences in the ability of the invading whitefly, Bemisia tabaci (Gennadius) (commonly known as biotype B and hereafter as B) and Trialeurodes vaporariorum (Westwood) (both Hemiptera: Aleyrodidae) to utilize salivary phenol‐oxidizing enzymes – polyphenol oxidase (PPO) and peroxidase (POD) to detoxify plant defensive phenolic compounds were explored. Polyphenol oxidase and POD were found in the saliva of both B and T. vaporariorum. For tomato colonies, the PPO and POD activities in the watery saliva of B were 2.27‐ and 1.34‐fold higher than those of T. vaporariorum. The PPO activities against specific phenolic compounds commonly found in plants were compared. The activities of those from B were significantly greater than those from T. vaporariorum. We also measured PPO activity in both species after they had fed on plants that were undamaged or had been previously damaged with either a plant pathogen [Phytophthora infestans (Mont.) de Bary (Peronosporales)] infection, mechanical damage, B infestation, or exogenous salicylic acid. For B, PPO activities in watery saliva increased 229, 184, 152, and 139% in response to the four treatments, whereas those of T. vaporariorum only increased 133, 119, 113, and 103%, respectively. Biotype B infestation significantly increased the total phenolic content of tomato leaves. Meanwhile, feeding on tomato infestation with B had no significant effect on the survival rate of B, but decreased the survival rate of T. vaporariorum significantly. These results suggest that B has stronger ability utilizing PPO to detoxify high concentrations of phenolics than T. vaporariorum, and this contributes to a significant advantage for B to hold high fitness on plants with induced resistance. Possible roles of salivary PPO in the competition between B and T. vaporariorum are discussed.     

Regulation of cuticle-degrading subtilisin proteases from the entomopathogenic fungi, <Emphasis Type="Italic">Lecanicillium</Emphasis> spp: implications for host specificity

The ability to produce cuticle-degrading proteases to facilitate host penetration does not distinguish per se entomopathogenic fungi from saprophytes. However, adapted pathogens may produce host-protein specific enzymes in response to cues. This possibility prompted an investigation of the regulation of isoforms of the subtilisin Pr1-like proteases from five aphid-pathogenic isolates of Lecanicillium spp. Significant differences were found in substrate specificity and regulation of Pr1-like proteases between isoforms of the same isolate and between different isolates. For example, the pI 8.6 isoform from KV71 was considerably more active against aphid than locust cuticle and was induced specifically by N-acetylglucosamine (NAG). Isoform pI 9.1 from the same isolate was only produced on insect cuticle while most other isoforms were more prominent on chitin containing substrates but not induced by NAG. The ability to regulate isoforms independently may allow production at critical points in host penetration. Appearance of proteases (not subtilisins) with pI 4.2 and 4.4 only on aphid cuticle was a possible link with host specificity of KV71. The absence of C or N metabolite repression in subtilisins from KV42 is unusual for pathogen proteases and may help to account for differences in virulence strategy between aphid-pathogenic isolates of Lecanicillium longisporum (unpublished data).     

Virulence of entomopathogenic hypocrealean fungi infecting <Emphasis Type="Italic">Anoplophora glabripennis</Emphasis>

Twenty isolates of four species of entomopathogenic hypocrealean fungi (Beauveria bassiana, Beauveria brongniartii, Isaria farinosa, and Metarhizium anisopliae) were found to be pathogenic to adults of the Asian longhorned beetle, Anoplophora glabripennis. Survival times for 50% of the beetles tested (ST₅₀) ranged from 5.0 (M. anisopliae ARSEF 7234 and B. brongniartii ARSEF 6827) to 24.5 (I. farinosa ARSEF 8411) days. Screening studies initially included strains of B. brongniartii, which is registered as a microbial control agent in Europe, Asia and South America but not in North America. At that time, we could not confirm that this fungal species is native to North America which added uncertainty regarding future registration of this species for pest control in the USA. Therefore, subsequent bioassays documented median survival times for three M. anisopliae isolates (5–6 days to death) and two of these isolates are suggested for further development because they are already registered for pest control in the USA. An erratum to this article can be found at     

Host plant influences pathogenicity of Beauveria bassiana to Bemisia tabaci and its sporulation on cadavers

Laboratory bioassays were conducted to determine host plant effect on pathogenicity of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuill. (Ascomycota: Hypocreales) to the sweetpotato whitefly, Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae). Fourth instar B. tabaci reared on cucumber, tomato, melon, green pepper, potato, eggplant, marrow, cabbage, bean or cotton, were treated with 1 × 10⁷ conidia/ml B. bassiana EABb 93/14-Tp isolate. Mortality caused by B. bassiana significantly increased with time and it was significantly affected by the host plant on which the nymphs were reared. Mean mortality of nymphs 8 days after inoculation ranged between 52.3±7.3 for nymphs reared on cotton and 91.8±5.8 for nymphs reared on cucumber. Average survival times of nymphs treated with the fungal suspensions were also significantly influenced by the host plant, with a mean of 4.7±0.1 days for nymphs reared on cucumber, 6.6±0.2 days for cotton and 6.9±0.1 days for green pepper. The production of newly formed conidia was also affected by host plant and varied from 111000±8600 conidia/cadaver for nymphs reared on cotton to 597000±28000 conidia/cadaver for those reared on melon.     

Compatibility of the entomopathogenic fungus Lecanicillium muscarium and insecticides for eradication of sweetpotato whitefly, Bemisia tabaci

The compatibility of the entomopathogenic fungus Lecanicillium muscarium and chemical insecticides used to control the second instar stages of the sweetpotato whitefly, Bemisia tabaci, was investigated. The effect on spore germination of direct exposure for 24 h to the insecticides imidacloprid, buprofezin, teflubenzuron and nicotine was determined. Only exposure to buprofezin was followed by acceptable spore germination. However, all chemicals significantly reduced spore germination when compared to a water control. Infectivity of L. muscarium in the presence of dry residues of buprofezin, teflubenzuron and nicotine (imidacloprid is a systemic pesticide) on foliage were also investigated. No significant detrimental effects on the level of control of B. tabaci was recorded when compared with fungi applied to residue free foliage on either tomato or verbena plants. Fungi in combination with imidacloprid gave higher B. tabaci mortality on verbena foliage compared to either teflubenzuron or nicotine and fungi combinations. Use of these chemical insecticides with L. muscarium in integrated control programmes for B. tabaci is discussed.     

The parasitoids of the aleyrodid Bemisia tabaci in Israel: development, host preference and discrimination of the aphelinid wasp Eretmocerus mundus

Developmental history and behavior of Eretmocerus mundus Mercet, a parasitoid of Bemisia tabaci was studied at 25°C. The eggs may be laid under all four nymphal instars but not under the pupa. Yet the second and third instars are preferred. The egg hatches only under the fourth instar or the pupa. Developmental medians at 25°C are: Instar I-2.5, II-4, II-4, prepupa-2 and pupa 8 days. When ovipositing, the female stands at an angle of 90° to the host, with wings raised and inserts the ovipositor under the whitefly nymph. The egg is laid close to the insertion point of the whitefly's proboscis into the leaf. After oviposition, the female apparently marks the host while drumming on it with her hind legs. She distinguishes already parasitized hosts from unparasitized ones and refrains from laying under the former. Discrimination is accomplished after antennal drumming only.

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Les parasitoïdes de Bemisia tabaci (aleyrodidae) en Israel: développement, ponte et sélection des hôtes ches Eretmocerus mundus (aphelinidae)

Résumé Le développment et le comportement de E. mundus, parasitoïde de B. tabaci, ont été étudiés à 25°C. Les oeufs sont pondus sous les quatre stades larvaires (les deuxième et troizième sont préférés) mais pas sous les nymphes. Les oeufs n'éclosent que sous les larves du quatrième stade ou les nymphes. Les temps de développement médiaux sont à 25°, les suivants: stade I: 2,5j; stade II: 4j; stade III: 4j et nymphe 8j. Pendant la ponte, la femelle est à 90° sur son hôte, les ailes dressées, et insère sa tarière sous la larve d'aleurode. L'oeuf est déposé près du point d'insertion de la trompe dans la feuille. Après l'émission, la femelle marque apparemment son hôte pendant qu'elle tambourine avec ses pattes postérieures. Elle distingue les hôtes parasités ou non, et limite sa ponte dans les premiers. La sélection est effectuée seulement après tambourinage antennaire.

     

Endophytic fungi occurring in fennel, lettuce, chicory, and celery — commercial crops in southern Italy

The occurrence of endophytic fungi in fennel, lettuce, chicory, and celery crops was investigated in southern Italy. A total of 186 symptomless plants was randomly collected and sampled at the stage of commercial ripeness. Fungal species of Acremonium, Alternaria, Fusarium, and Plectosporium were detected in all four crops; Plectosporium tabacinum was the most common in all crop species and surveyed sites. The effect of eight endophytic isolates (five belonging to Plectosporium tabacinum and three to three species of Acremonium) inoculated on lettuce plants grown in gnotobiosis was assessed by recording plant height, root length and dry weight, collar diameter, root necrosis, and leaf yellowing. P. tabacinum and three species of Acremonium, inoculated on gnotobiotically grown lettuce plants, showed pathogenic activity that varied with the fungal isolate. Lettuce plants inoculated with the isolates Ak of Acremonium kiliense, Ac of Acremonium cucurbitacearum, and P35 of P. tabacinum showed an increased root growth, compared to the non-inoculated control. The high frequency of P. tabacinum isolation recorded in lettuce plants collected in Bari and Metaponto, and in fennel plants from Foggia agricultural districts, suggests a relationship not only between a crop species and P. tabacinum, but also between the occurrence of the endophyte and the crop rotation history of the soil.     

The extracellular constitutive production of chitin deacetylase in Metarhizium anisopliae: possible edge to entomopathogenic fungi in the biological control of insect pests

The possible contribution of extracellular constitutively produced chitin deacetylase by Metarhizium anisopliae in the process of insect pathogenesis has been evaluated. Chitin deacetylase converts chitin, a beta-1,4-linked N-acetylglucosamine polymer, into its deacetylated form chitosan, a glucosamine polymer. When grown in a yeast extract-peptone medium, M. anisopliae constitutively produced the enzymes protease, lipase, and two chitin-metabolizing enzymes, viz. chitin deacetylase (CDA) and chitosanase. Chitinase activity was induced in chitin-containing medium. Staining of 7.5% native polyacrylamide gels at pH 8.9 revealed CDA activity in three bands. SDS-PAGE showed that the apparent molecular masses of the three isoforms were 70, 37, and 26 kDa, respectively. Solubilized melanin (10microg) inhibited chitinase activity, whereas CDA was unaffected. Following germination of M. anisopliae conidia on isolated Helicoverpa armigera, cuticle revealed the presence of chitosan by staining with 3-methyl-2-benzothiazoline hydrazone. Blue patches of chitosan were observed on cuticle, indicating conversion of chitin to chitosan. Hydrolysis of chitin with constitutively produced enzymes of M. anisopliae suggested that CDA along with chitosanase contributed significantly to chitin hydrolysis. Thus, chitin deacetylase was important in initiating pathogenesis of M. anisopliae softening the insect cuticle to aid mycelial penetration. Evaluation of CDA and chitinase activities in other isolates of Metarhizium showed that those strains had low chitinase activity but high CDA activity. Chemical assays of M. anisopliae cell wall composition revealed the presence of chitosan. CDA may have a dual role in modifying the insect cuticular chitin for easy penetration as well as for altering its own cell walls for defense from insect chitinase.     

Activity of pupal parasitoids of the stable fly Stomoxys calcitrans and prevalence of entomopathogenic fungi in the stable fly and the house fly Musca domestica in Denmark

A survey was conducted on confined dairy cattle farms and a pig farm from May–October in 1999 to determine the activity and relative abundance of pupal parasitoids and the prevalence of entomopathogenic fungi in populations of the haematophagous stable fly, Stomoxys calcitrans (Diptera: Muscidae), in Denmark. Four species of pteromalids were found with Spalangia cameroni as the predominant. The other parasitoids were S. nigripes, S. nigra and Phygadeuon fumator (Ichneumonidae). Peak activity of the parasitoids was observed to be late in the summer and the beginning of autumn (August–September) when approximately 10% of the collected stable fly pupae were parasitised. Adult stable flies were infected with four species of entomopathogenic fungi: Entomophthora muscae, E. schizophorae, Beauveria bassiana and Verticillium lecanii. All fungi occurred in low percentages (max. 4%) and remained at this level throughout the sampling period. Likewise, adult house flies were infected with B. bassiana and V. lecanii,but Metarhizium anisopliae, Paecilomyces fumosoroseus and V. fusisporum were also recorded. The overall hyphomycete prevalence in house flies was 0.3%, and single species rarely exceeded 0.1%. The prevalence remained low in spite of increasing house fly numbers in August–September.     

Factors affecting the occurrence and distribution of entomopathogenic fungi in natural and cultivated soils

Factors affecting the occurrence and distribution of entomopathogenic fungi in 244 soil samples collected from natural and cultivated areas in Spain were studied using an integrated approach based on univariate and multivariate analyses. Entomopathogenic fungi were isolated from 175 of the 244 (71.7 %) soil samples, with only two species found, Beauveria bassiana and Metarhizium anisopliae. Of the 244 soil samples, 104 yielded B. bassiana (42.6 %), 18 yielded M. anisopliae (7.3 %), and 53 soil samples (21.7 %) harboured both fungi. Log-linear models indicated no significant effect of habitat on the occurrence of B. bassiana, but a strong association between M. anisopliae and soils from cultivated habitats, particularly field crops. Also, irrespective of habitat type, B. bassiana predominated over M. anisopliae in soils with a higher clay content, higher pH, and lower organic matter content. Logistic regression analyses showed that pH and clay content were predictive variables for the occurrence of B. bassiana, whereas organic matter content was the predictive variable for M. anisopliae. Also, latitude and longitude predicted the occurrence of these same species, but in opposite directions. Altitude was found to be predictive for the occurrence of B. bassiana. Using principal component analysis, four factors (1 to 4) accounted for 86 % of the total variance; 32.8, 22.9, 19.6 and 10.4 % of the cumulative variance explained, respectively. Factor 1 was associated with high positive weights for soil clay and silt content and high negative weights for soil sand content. Factor 2 was associated with high positive weights for soil organic matter content and high negative weights for soil pH. Factor 3 was associated with high positive weights for latitude and longitude of the sampled localities and factor 4, had high positive weights only for the altitude. Bi-plot displays representing soil samples were developed for different factor combinations and indicated that, irrespective of geographical location, absence of both fungal species was determined by alkaline sandy soils with low organic matter content, whereas heaviness of soil texture, acidity and increasing organic matter content led to progressively higher percentages of samples harbouring entomopathogenic fungi. These results could aid decision-making as to whether or not a particular cultivated or natural soil is suitable for using entomopathogenic fungi as a pest control measure and for selecting the fungal species best suited to a particular soil.     

Isolation and characterization of two entomopathogenic fungi attackingDiaphorina citri (Homoptera, Psylloidea) in Indonesia

In an attempt to suppress the propagation of citrus greening disease in Indonesia, we explored pathogens ofDiaphorina citri which vectors the disease. At two orange orchards, manyD. citri adults were found to be dead and covered with fungal mycelia. Two fungi,Paecilomyces fumosoroseus andHirsutella citriformis, were consistently isolated from the infected insects. Molecular phylogeny of their 18S rDNA sequences showed that they belong to the ascomycetous clade of the Clavicipitales/Hypocreales, which embraces many entomopathogenic fungi. When healthy adults ofD. citri were inoculated with conidia of theP. fumosoroseus, the insects died within 6 d.