The use of aggregation pheromone to enhance dissemination of Beauveria bassiana for the control of the banana weevil in Uganda

Candidate strains of Beauveria bassiana were identified for use in integrated pest management of the banana weevil Cosmopolites sordidus. Horizontal field transmission of B. bassiana between banana weevils using different delivery systems, including aggregation pheromones, was investigated. We observed that infected weevils could transmit the fungal pathogen to healthy individuals. Most dead weevils (52%) due to B. bassiana infection were found at the base of banana plants in the leaf sheath or in the soil near banana plants. Significantly more weevils died from the pathogen in plots where B. bassiana was applied in combination with the pheromone than where it was applied alone. Our data demonstrate that C. sordidus aggregation pheromone can be a valuable tool to enhance the dissemination of B. bassiana for the control of C. sordidus.     

The bacterial community associated with adult vine weevil (Otiorhynchus sulcatus) in UK populations growing on strawberry is dominated by Candidatus Nardonella

Otiorhynchus sulcatus (Fabricius) (Coleoptera: Curculionidae), commonly known as black vine weevil or simply vine weevil, is an important pest of soft fruit and ornamental crops. This species is endemic to temperate areas of Europe but has spread to many other areas over the last century, including North America and Australasia. The ability of vine weevils to adapt to such different environments is difficult to reconcile with the parthenogenetic reproduction strategy, which is likely to underpin a low genetic diversity. It is therefore tempting to hypothesize that weevil adaptation to different environments is mediated, at least partly, by the microbial communities inhabiting these insects. As a first step towards testing this hypothesis we characterized the composition of the bacterial microbiota in weevils from populations feeding on strawberry plants across four geographically separate locations in the UK. We performed 16S rRNA gene Illumina amplicon sequencing, generating 2 882 853 high‐quality reads. Ecological indices, namely Chao1 and Shannon, revealed that the populations used for this study harboured a low diversity and an uneven bacterial microbiota. Furthermore, β‐diversity analysis failed to identify a clear association between microbiota composition and location. Notably, a single operational taxonomic unit phylogenetically related to Candidatus Nardonella accounted for 81% of the total sequencing reads for all tested insects. Our results indicate that vine weevil bacterial microbiota resembles that of other insects as it has low diversity and it is dominated by few taxa. A prediction of this observation is that location per se may not be a determinant of the microbiota inhabiting weevil populations. Rather, other or additional selective pressures, such as the plant species used as a food source, ultimately shape the weevil bacterial microbiota. Our results will serve as a reference framework to investigate other or additional hypotheses aimed at elucidating vine weevil adaptation to its environment.     

Treatment of millet crop plant (Sorghum bicolor) with the entomopathogenic fungus (Beauveria bassiana) to combat infestation by the stem borer,Chilo partellus Swinhoe (Lepidoptera: Pyralidae)

Experiments were done to test if Beauveria bassiana can become an endophyte in sorghum and confer protection from stem borer. Four-week-old sorghum seedlings were treated with B. bassiana. The plants were examined for endophytic presence of B. bassiana, 30 and 60 days after treatment. Stem cultures from treated plants showed growth of B. bassiana. PCR amplification using fungal specific primers for a conserved region of β tubulin gene yielded identical 360 bp products from both B. bassiana and treated sorghum plants. In a subsequent experiment, B. bassiana treated and untreated (control) sorghum plants were artificially infested with stem borer (Chilo partellus) larvae 15 days post treatment and the extent of damage was compared. About 40% of the control plants developed dead heart while no plant in the B. bassiana treated plot did. In the surviving control plants, stem tunneling by shoot borer was significantly higher compared to B. bassiana treated sorghum plants.     

Improving biocontrol of black vine weevil (<Emphasis Type="Italic">Otiorhynchus sulcatus</Emphasis>) with entomopathogenic fungi in growing media by incorporating spent mushroom compost

Amending a peat-based growing medium with 10% v/v spent mushroom compost, a source of fungal chitin and other nutrients, prolonged the persistence of entomopathogenic fungi (Metarhizium brunneum Petsch and Beauveria bassiana (Balsamo) Vuillemin; Hypocreales: Clavicipitaceae). This resulted in improved efficacy of M. brunneum against black vine weevil, Otiorhynchus sulcatus F. (Coleoptera: Curculionidae) larvae compared with using inoculum without spent mushroom compost. B. bassiana only controlled larvae when used in combination with spent mushroom compost (75?±?7% reduction in live larvae). Mixing entomopathogenic fungal inoculum with spent mushroom compost and growing medium was as effective in controlling black vine weevil larvae as using spent mushroom compost colonised with M. brunneum or B. bassiana in the growing medium (80?±?12% reduction in live larvae). The former method is preferable since it does not require production and storage of colonised spent mushroom compost, or registration of new substrate formulations of M. brunneum or B. bassiana.     

Persistence of Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato in Vermont (USA) forest soil

Persistence of granular formulations of Beauveria bassiana sensu lato, strains GHA726 and ERL1170, and Metarhizium anisopliae sensu lato, strains ERL824 and ERL701, applied to forest soil was documented. One formulation was prepared by producing fungi on millet and at application adding cooked, dried millet to provide additional nutrients for fungal growth. The other was made by producing fungi on corn starch ‘packing peanuts’, a biodegradable packaging material, adding additional packing peanuts as a nutrient base. The trial was conducted in three sites in a forest predominating in sugar maple (Acer saccharum) in Underhill, VT, USA. In each site, 33 dominant sugar maples were randomly selected. A 1-m² plot located 1 m from the bole of the tree was established. Soil samples were collected before application, and 1 h, 2, 4, 8, 12 and 16 wk post-application. Fungal levels were estimated based on number of colony-forming units (CFUs). A significant increase in CFUs was observed 2 wk post-application in plots treated with isolates GHA726, ERL1170 and ERL824 as a result of fungal reproduction on nutrients in the formulations. Evidence of greater persistence for the B. bassiana than the M. anisopliae isolates was observed, based on CFU levels over time. Isolates formulated with millet proliferated more than those on packing peanuts. Use of entomopathogenic fungi in forest soil has potential for managing soil-dwelling insect pests such as pear thrips, Taeniothrips inconsequens, and the black vine weevil, Otiorhynchus sulcatus, though the type of formulation and the specific isolate used must be selected carefully.     

The endophytic fungal entomopathogens Beauveria bassiana and Purpureocillium lilacinum enhance the growth of cultivated cotton (Gossypium hirsutum) and negatively affect survival of the cotton bollworm (Helicoverpa zea)

The effects of two entomopathogenic fungal endophytes, Beauveria bassiana and Purpureocillium lilacinum, were assessed on the growth of cultivated cotton (Gossypium hirsutum) and development of the cotton bollworm (Helicoverpa zea). In two replicate greenhouse trials, cotton plants were inoculated as seed treatments with two concentrations of B. bassiana or P. lilacinum conidia and evaluated for effects on both plant dry biomass, number of nodes and number of developing flowers (squares). We similarly treated cotton plants and evaluated H. zea performance using no-choice in planta assays starting at the 2nd larval instar. Treatment with both fungal endophytes resulted in a significant increases in plant dry biomass (ANOVA, P = 0.024). Plant developmental stage and number of squares were also significantly enhanced in the endophyte treated plants (ANOVA, P = 0.005 and P = 0.027, respectively). The survivorship of H. zea was significantly different among the endophyte treatment groups (Kaplan–Meier, P = 0.02), where insects feeding on control plants exhibited higher survival than insects on the endophyte treated plants. There were no significant endophyte treatment effects on larval or pupal weights of H. zea individuals. There was no endophyte effect on days to pupation among treatments, but there was a marginal effect on days to eclosion (Kaplan–Meier, P = 0.07). Overall, our results demonstrate (i) the positive plant growth enhancing effects of the target endophyes on cultivated cotton under greenhouse conditions and (ii) the negative effects of endophytic P. lilacinum and B. bassiana on H. zea survivorship and development using whole plant assays.     

Behavioural responses of the vine weevil, Otiorhynchus sulcatus, to semiochemicals from conspecifics, Otiorhynchus salicicola, and host plants

The vine weevil Otiorhynchus sulcatus is a parthenogenetic reproducing species which forages for suitable host plants at night, but is found congregated in dark places during the day. Frass of this weevil species is suspected to contain attractive compounds that are host‐plant related. Using a still‐air olfactometer, we tested adult vine weevils at night for their behavioural response to odours from conspecifics, feeding on a mixture of spindle tree (Euonymus fortunei) and yew (Taxus baccata), and to a sexually reproducing related species (Otiorhynchus salicicola), feeding on a mixture of ivy (Hedera helix) and cherry laurel (Prunus laurocerasus). Their attraction to conspecifics and O. salicicola appeared to be related to frass production. Freshly collected frass from O. sulcatus and from O. salicicola males and females was attractive. Prunus laurocerasus and H. helix have not been observed to be hosts of the vine weevil in the field. However, our tests showed that the vine weevil was attracted to mechanically damaged leaves of both plant species, whereas undamaged leaves were not attractive. Only undamaged young unfolding leaves of H. helix were also attractive. The attraction to odours from mechanically damaged host and non‐host plants suggested the involvement of compounds that are commonly found in many plant species. The involvement of plant compounds and/or aggregation pheromones in attraction to frass of the vine weevil and frass of the related weevil species O. salicicola is discussed.     

On-farm evaluation of Beauveria bassiana for control of Ostrinia nubilalis in Iowa, USA

Ostrinia nubilalis is (Lepidoptera: Crambidae) a severe pest ofcorn in the major corn growing areas of theUnited States. The efficacy of a Beauveria bassiana application, for season-long suppression of O. nubilalis was evaluated in 1996 and 1997 at locationsacross Iowa. Beauveria bassiana,Mycotech 726 (Mycotech Corporation, Butte, MT)formulated on corn grit granules (14–20 mesh)at 2.2 × 10⁹ conidia/g and applied with ahand-held applicator at the rate of 0.4 g/plant(8.8 × 10⁸ conidia/plant). Applicationswere made when plants were in the V7 or R1growth stage. The length of larval tunneling,percentage of plants not infested with O.nubilalis, percentage of plants with anendophyte, and yield from treated and controlplots were determined. Whorl-stage applicationof B. bassiana in 1996 resulted in asignificant reduction in centimeters oftunneling (46–55%) and the percentageplants not infested by O. nubilalis. In1997, B. bassiana caused significantreductions in larval tunneling at all locations(20–53%); however, a significant increase inthe percentage of plants not infested with O. nubilalis occurred at only one location. Treatment of plants with B. bassiana in1997 did not significantly increase thepercentage of plants with an endophyte;however, the trend, with the exception of onesite, was for a greater percentage ofendophytic plants in treated versus untreatedplots. A whorl-stage application of a granularformulation of B. bassiana was mostefficacious in reducing O. nubilalis larval damage.     

Use of a nematode, Heterorhabditis heliothidis, to control black vine weevil, Otiorhynchus sulcatus, in potted plants

Application of aqueous suspensions of infective juvenile Heterorhabditis heliothidis, isolate T327, to the soil resulted in up to 100% parasitisation of larvae of the black vine weevil, Otiorhynchus sulcatus, in potted yew, raspberries and grapes in nurseries, and over 87% parasitisation on potted cyclamens and strawberries. Pupae and newly emerged adults on grapevines were also parasitised. Another isolate, T310, produced 92.5 to 98.5% parasitism of O. sulcatus larvae on potted cyclamens in glasshouse, but was less effective on strawberries. Neoaplectana bibionis was found to be less effective than H. heliothidis T327 strain. The use of these nematodes provides an economical and effective method for controlling O. sulcatus on potted plants in glasshouses and nurseries.     

Grapevine leaf tissue colonization by the fungal entomopathogen <Emphasis Type="Italic">Beauveria bassiana</Emphasis><Emphasis Type="Italic">s.l.</Emphasis> and its effect against downy mildew

A study was conducted to examine whether Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) can colonize grapevine leaf tissues and subsequently confer protection against downy mildew caused by Plasmopara viticola (Berk. and Curt.) Berl. and de Toni. Following the foliar inoculation of plants with conidial suspensions of selected B. bassiana strains, colonization of leaves by the fungus was determined using culture-based and PCR techniques at different time intervals. Seven days following B. bassiana inoculation, grapevine plants were challenged with P. viticola and symptoms were assessed by calculating the disease incidence and severity. Although all tested strains were able to colonize grapevine plants, percent colonization differed significantly among strains. Disease incidence and severity were, on the other hand, significantly reduced in B. bassiana-inoculated plants compared to control plants irrespective of strain. This study is one of very few studies investigating the promising role B. bassiana could play as a plant disease antagonist.     

Compatibility of an entomopathogenic fungus with a predator and a parasitoid in the biological control of greenhouse whitefly

An ongoing debate in biological control consists of whether interference between biological agents can disrupt pest control. This study investigated the outcome of interactions between the entomopathogen Beauveria bassiana with the whitefly predator Dicyphus hesperus and the parasitoid Encarsia formosa, as well as their effect on the control of the greenhouse whitefly Trialeurodes vaporariorum on greenhouse tomato crops. Our objective was to determine whether the generalist B. bassiana would disrupt biological control by interfering with D. hesperus or E. formosa. In experimental greenhouses, whitefly, parasitoid and predator populations were established, and over 27 days, tomato plants were sprayed with three applications of the B. bassiana based product BotaniGard® (5.13×10³ conidia/mm²) or water (control). Populations of greenhouse whitefly and biological control organisms were regularly monitored in control and B. bassiana-treated compartments. Overall, 10.6% of all whiteflies in treated compartments were infected, and 0.98% were both infected and parasitized. There were 31.7 and 22.3% fewer immature and adult whiteflies, respectively, on B. bassiana-treated plants relative to controls. Parasitism by E. formosa and predation by D. hesperus occurred at rates of 7.5 and 2.5%, respectively, in B. bassiana-treated compartments, and 5 and 6%, respectively in control compartments. Our study suggests that applications of B. bassiana for short-term biological control of greenhouse whiteflies are compatible with the concurrent use of E. formosa and D. hesperus on greenhouse tomato crops.     

Establishment of Beauveria bassiana as a fungal endophyte in potato plants and its virulence against potato tuber moth,Phthorimaea operculella (Lepidoptera: Gelechiidae)

The potato tuber moth, Phthorimaea operculella, is the most damaging potato pest in the world and is difficult to control as the larvae are internal feeders in the foliage and tubers. Entomopathogenic fungi that colonize plants as endophytes have lethal and sublethal pathological effects on insect pests. We show that Beauveria bassiana colonizes the aerial parts of potato plants endophytically after inoculation through soil drenching. Endophytic B. bassiana persisted in potato foliage for more than 50 days postinoculation. Bioassays indicated that foliage of B. bassiana-inoculated potato plants were pathogenic against larvae of P. operculella. Sublethal experiments indicated that B. bassiana negatively affected the growth, development, and reproduction of P. operculella. Development experiments showed that the weight of P. operculella pupae reared on B. bassiana-colonized potato plants (4.25 mg) was significantly less than that of those reared on uninoculated control plants (8.89 mg). Compared with newly eclosed larvae fed on control plants, those fed on B. bassiana-inoculated plants had significantly lower survivorship, with only 17.8% developing to the adult stage. Oviposition of P. operculella females reared on B. bassiana endophytically colonized plants was significantly lower (35 eggs/female) than of those reared on uninoculated plants (115 eggs/female). This study demonstrates that endophytic B. bassiana can be a potential biological control agent for the control and management of P. operculella. Comparing pupal weights of P. operculella reared on potato plants inoculated with the B. bassiana strain GZGY-1-3 and on untreated control plants, pupae from the control plants were significantly heavier than those from treated plants.     

The entomopathogenic nematode Steinernema kraussei and Metarhizium anisopliae work synergistically in controlling overwintering larvae of the black vine weevil,Otiorhynchus sulcatus,in strawberry growbags

Previously, the combination of reduced rate of entomopathogenic nematodes (EPN) and fungus caused additive or synergistic mortality to third-instar black vine weevil (BVW), Otiorhynchus sulcatus. In this study, we examined this interaction in unheated glasshouses during winter and compared a combination of commercial formulation of a cold-tolerant EPN, S. kraussei (Nemasys L?) and fungus Metarhizium anisopliae strain V275 against overwintering third-instar BVW. The combination of M. anisopliae with S. kraussei at a rate of 1×10¹⁰ conidia+250,000 nematodes/growbag resulted in additive or synergistic effects, providing 100% control of overwintering larvae.     

Whorl and pollen-shed stage application of Beauveria bassiana for suppression of adult western corn rootworm

The corn rootworm complex is the most damaging insect pest of corn (Zea mays L.). This study was conducted to determine the efficacy of whorl and pollen-shed stage applications of a granular formulation of Beauveria bassiana (Balsamo) Vuillemin for control of adult western corn rootworm (Diabrotica virgifera virgifera Le Conte). The effect of application time (whorl-stage, pollen-shed) and plant surface exposed (leaves and leaf collars; silks; leaves, leaf collars, and silks) on level of beetle fungal infection were investigated. In addition, the number of colony forming units of B. bassiana in the corn leaf collar area was quantified. In the three years (1998–2000) of the study, application of B. bassiana at whorl-stage did not significantly increase beetle fungal infection. Beauveria bassiana applied to plants at pollen-shed in 1998 resulted in a significant increase in beetle infection with 51% of beetles from treated plants infected and 6.0% from control plants. Similar applications at pollen-shed in 1999 and 2000 resulted in very low infection levels. Beauveria bassiana application at pollen-shed stage significantly increased the number of colony forming units per leaf collar during all years of the study. Beetle infection with B. bassiana did not differ consistently among plant surface to which beetles were exposed for either application. Increased fungal load in leaf collars was not correlated with increased levels of adult infection. Increased rates of B. bassiana and application when beetles are present on the plants are likely needed to significantly increase infection rates.     

Systemic insecticides reduce feeding, survival, and fecundity of adult black vine weevils (Coleoptera: Curculionidae) on a variety of ornamental nursery crops

Systemic activity of the neonicotinoids clothianidin, dinotefuran, and thiamethoxam and the anthranilic diamide chlorantraniliprole was tested against adult black vine weevils, Otiorhynchus sulcatus (F.) (Coleoptera: Curculionidae), on Astilbe, Euonymus, Heuchera, Rhododendron, Sedum, and Taxus. Insecticide treatments were applied to the soilless substrate of containerized plants. Bioassays were conducted 12 or 13, 26, and 42 d after treatment (DAT) and ran for 7 d; and feeding, mortality, and weight gain or loss by weevils were evaluated. Foliage was removed from test plants and then placed in arenas with adult black vine weevils. The neonicotinoids reduced feeding and weight gain by adult black vine weevils on most plant species with residual activity 42 DAT on some plant species. At 12 DAT, mortality was caused by the three neonicotinoids on Astilbe and by thiamethoxam on Sedum; and at 26 DAT dinotefuran caused mortality on Astilbe. Chlorantraniliprole reduced feeding on Taxus at 12 DAT, with no activity detected in other bioassays. Another set of bioassays was conducted to examine survival and fecundity of adult black vine weevils during prolonged feeding on Heuchera and Taxus systemically treated with dinotefuran or thiamethoxam. Bioassay procedures were similar to those described above, except they ran continuously for 56 d. Prolonged feeding on dinotefuran and thiamethoxam treated Heuchera and Taxus resulted in high mortality of adult black vine weevils and reduced fecundity. These studies show that the systemic activity of neonicotinoids is influenced by plant species and that systemic neonicotinoids have the potential to suppress black vine weevil populations in containerized nursery crops.     

Efficacy of the entomopathogenic nematodes Steinernema kraussei and Heterorhabditis megidis against the black vine weevil Otiorhynchus sulcatus in open field‐grown strawberry plants

• 1 Entomopathogenic nematodes are commercially available for inundative biological control of many insects, including the black vine weevil Otiorhynchus sulcatus (F.) (Coleoptera: Curculionidae). Currently, there is a lack of commercial application tests in field‐grown crops comparing the efficacy of different species of entomopathogenic nematodes.
• 2 Field trials were carried out under different growing conditions in Ireland and Norway to evaluate the efficacy of two commercially available nematode species on the market for control of the black vine weevil Heterorhabditis megidis and Steinernema kraussei.
• 3 Heterorhabditis megidis was evaluated not only at temperatures ideal for this species (soil temperatures above 10 °C), but also in the low temperature trials with S. kraussei as a ‘positive control’. Steinernema kraussei is sold as a cold active product and was therefore evaluated at low soil temperatures (below 10 °C).
• 4 The overall results indicated that H. megidis was effective as long as temperatures were optimum (not dropping below 10 °C). For S. kraussei, the results obtained were rather disappointing, where control barely reached 50% in the trial with the coldest temperature. Temperature and soil type appeared to be a major limiting factor for the efficacy of both nematode species.
• 5 On the basis of the results and experience obtained in these trials, the future implications for biological control of O. sulcatus with entomopathogenic nematodes in commercial field‐grown strawberry production are discussed.

     

Toxicity of spinosad and Beauveria bassiana to the black cutworm,and the additivity of subletal doses

The main objective of this study was to assess the susceptibility of the black cutworm (Agrotis ipsilon) to the biopesticide spinosad and to a commercial formulation (GHA strain) of the entomopathogenic fungus, Beauveria bassiana. Secondly, we quantified the effects of sublethal doses of spinosad on a number of A. ipsilon fitness parameters, and interactions resulting from simultaneous applications of sub-lethal doses of spinosad and B. bassiana. Under laboratory conditions, A. ipsilon third instar larvae were highly susceptible to spinosad, with an estimated LC₅₀ of 50 ppm. The entomopathogenic fungus, B. bassiana had a lower efficacy with an estimated LC₅₀ of 7×107 spores mL^?1. Topical applications of 5, 7.5 and 10 ppm of spinosad on third instar larvae reduced larval size and increased time to pupation and to emergence. However, pupal and adult weights were not significantly different between treated and control individuals. Additivity was observed from most spinosad–B. bassiana combinations tested, thus indicating compatibility between products. We concluded that spinosad is a promising tool for controlling black cutworm larvae alone or in combination with other products.     

The Entomopathogenic Fungal Endophytes Purpureocillium lilacinum (Formerly Paecilomyces lilacinus) and Beauveria bassiana Negatively Affect Cotton Aphid Reproduction under Both Greenhouse and Field Conditions

The effects of two entomopathogenic fungal endophytes, Beauveria bassiana and Purpureocillium lilacinum (formerly Paecilomyces lilacinus), were assessed on the reproduction of cotton aphid, Aphis gossypii Glover (Homoptera:Aphididae), through in planta feeding trials. In replicate greenhouse and field trials, cotton plants (Gossypium hirsutum) were inoculated as seed treatments with two concentrations of B. bassiana or P. lilacinum conidia. Positive colonization of cotton by the endophytes was confirmed through potato dextrose agar (PDA) media plating and PCR analysis. Inoculation and colonization of cotton by either B. bassiana or P. lilacinum negatively affected aphid reproduction over periods of seven and 14 days in a series of greenhouse trials. Field trials were conducted in the summers of 2012 and 2013 in which cotton plants inoculated as seed treatments with B. bassiana and P. lilacinum were exposed to cotton aphids for 14 days. There was a significant overall effect of endophyte treatment on the number of cotton aphids per plant. Plants inoculated with B. bassiana had significantly lower numbers of aphids across both years. The number of aphids on plants inoculated with P. lilacinum exhibited a similar, but non-significant, reduction in numbers relative to control plants. We also tested the pathogenicity of both P. lilacinum and B. bassiana strains used in the experiments against cotton aphids in a survival experiment where 60% and 57% of treated aphids, respectively, died from infection over seven days versus 10% mortality among control insects. Our results demonstrate (i) the successful establishment of P. lilacinum and B. bassiana as endophytes in cotton via seed inoculation, (ii) subsequent negative effects of the presence of both target endophytes on cotton aphid reproduction using whole plant assays, and (iii) that the P. lilacinum strain used is both endophytic and pathogenic to cotton aphids. Our results illustrate the potential of using these endophytes for the biological control of aphids and other herbivores under greenhouse and field conditions.     

Impact of azadirachtin on vine weevil (Coleoptera: Curculionidae) reproduction

Abstract 1 The dose–response of azadirachtin on vine weevil, Otiorhynchus sulcatus (Fabricius), reproduction is investigated by confining adults to feed on treated Taxus × media leaves, and by counting and evaluating development in the resulting eggs. 2 A dosage‐dependent reduction in oviposition is discovered for foliar surface residues of azadirachtin, with an EC₅₀ of 25–50 parts per million (p.p.m) and 99.2% inhibition of viable egg production with 100 p.p.m. 3 Switching weevils from treated to untreated foliage allows reproductive capability to be restored for weevils that cease egg laying after azadirachtin exposure of 50 p.p.m. Weevils that had already started laying eggs in untreated groups soon cease oviposition once switched to azadirachtin‐treated foliage. 4 A transovarial effect results in a decrease in the percentage of viable eggs as the azadirachtin concentration increases. 5 The amount of feeding on foliage does not appreciably decrease at these hormonally effective concentrations, and adult weevil mortality is only slightly greater in the azadirachtin‐treated groups. Therefore, the overall effect of azadirachtin on weevil populations in the field is difficult to assess, except by collecting weevils to determine whether they are able to lay viable eggs.     

Effect of inoculation method and plant growth medium on endophytic colonization of sorghum by the entomopathogenic fungus <Emphasis Type="Italic">Beauveria bassiana</Emphasis>

A study was conducted to determine the effect of inoculation method and plant growth medium on colonization of sorghum by an endophytic Beauveria bassiana. Colonization of leaves, stems, and roots by B. bassiana was assessed 20-days after application of the fungus. Although B. bassiana established as an endophyte in sorghum leaves, stems, and roots regardless of inoculation method (leaf, seed, or soil inoculation), plant growth medium (sterile soil, non-sterile soil, or vermiculite) apparently influenced colonization rates. Seed inoculation with conidia caused no stem or leaf colonization by the fungus in non-sterile soil but did result in substantial endophytic colonization in vermiculite and sterile soil. Leaf inoculation did not result in root colonization, regardless of plant growth medium. Endophytic colonization was greater in leaves and stems than roots. Endophytic colonization by B. bassiana had no adverse effects on the growth of sorghum plants. Leaf inoculation with a conidial suspension proved to be the best method to introduce B. bassiana into sorghum leaves for plants growing in either sterile or non-sterile soil. Further research should focus on the virulence of endophytic B. bassiana against sorghum stem borers.