Starvation induced stress and the susceptibility of the Colorado potato beetle,Leptinotarsa decemlineata,to infection by Beauveria bassiana

Starvation of second instar Colorado potato beetle larvae for 24h immediately after treatment with Beauveria bassiana conidia increased susceptibility to the pathogen and subsequent sporulation of cadavers but decreased time to larval death. In feeding studies, B. bassiana-treatment had no effect on subsequent larval development, and mortality occurred 5-6 days after treatment. Twenty-four hours of starvation alone retarded subsequent larval development but did not affect mortality. Mortality of B. bassiana-treated starvation stressed larvae occurred 4-5 days after treatment. Both B. bassiana treatment and 24h starvation significantly reduced total foliage consumption and daily weight gains. On the day of treatment, B. bassiana had no effect on the efficiency with which food was converted to biomass (ECI). ECI was not affected by B. bassiana or starvation alone on the day following treatment but was significantly affected by a combination of both. When larvae were exposed to a range of limited food quantities, ECI decreased with decreasing food availability but only extreme stress (starvation for 24h) increased susceptibility to B. bassiana. Topical application of Dacryodes excelsa resin (an antifeedant) to potato leaves caused a concentration dependent reduction in foliage consumption and weight gain by second instar larvae but did not affect larval mortality. When larvae were exposed to a fixed concentration of B. bassiana and a range of antifeedant concentrations there were significant linear relationships between 24h larval weight gain and mortality and 24h larval weight gain and sporulation. The interaction between starvation stress and the susceptibility to B. bassiana infection is discussed and its possible implications in pest management considered.     

Comparative efficacy of Beauveria bassiana, Bacillus thuringiensis, and aldicarb for control of Colorado potato beetle in an irrigated desert agroecosystem and their effects on biodiversity

Five weekly applications of Beauveria bassiana (Balsamo) Vuillemin, a genetically engineered isolate of Bacillus thuringiensis Berliner (Raven^®), and aldicarb (Temik^®) were compared for control of Colorado potato beetle, Leptinotarsa decemlineata (Say) in an irrigated desert cropping system. B. thuringiensis was applied using low and high label rates (1.17 and 7.0 l ha^–1). B. bassiana was applied at 5×10¹³ spores ha^–1. Aldicarb (Rh^one-Poulenc), applied at 3.37 kg a.i. ha^–1 provided the greatest beetle control and potato yields (45 metric tons ha^–1), but overall lowest biodiversity in nontarget organisms, particularly predatory Heteroptera. Low and high rates of B. thuringiensis produced fair to excellent beetle control, yielded 33 and 40 metric tons ha^–1 and enabled good survival in predatory Heteroptera and other nontargets. Plots treated with B. bassiana resulted in poor control of beetles prior to row closure after which fair to good control was achieved. Yield in the Beauveria-treated plots was 33 metric tons ha^–1 and effect on biodiversity was comparable to the Bt-treated plots. The lowest number of overwintering adult L. decemlineata was found in the plots treated with bacteria and fungi (0.68–0.84 adults/0.03 m^–3 of soil) and the highest was found in control and aldicarb plots (3.44 and 1.84 adults/0.03 m³ of soil). Aphids and leafhoppers showed higher densities in plots treated with microbial control agents, but were eliminated from plots treated with aldicarb.     

Mortality of Colorado potato beetle (Leptinotarsa decemlineata) after sublethal stress with the CryIIIA delta-endotoxin of Bacillus thuringiensis and subsequent exposure to Beauveria bassiana

Acute or chronic sublethal exposure of Colorado potato beetle larvae to the CryIIIA delta-endotoxin of Bacillus thuringiensis Berliner did not significantly (P > 0.05) alter their subsequent susceptibility to Beauveria bassiana (Balsamo) Vuillemin. During the period of exposure to B. bassiana there was continued mortality from previous exposure to delta-endotoxin, and B. bassiana also caused significant mortality. Acute and chronic exposure to delta-endotoxin significantly prolonged larval development. The weights of prepupae and adults were significantly reduced by exposure to delta-endotoxin, with the greatest effect being from chronic exposure. Separation of the manifestations of stress in time (feeding vs soil stages) and space (toxin damage to the insect gut vs fungal penetration of the cuticle and activity in the hemocoel) may have precluded alteration of insect susceptibility to infection by B. bassiana. Endemic populations of B. bassiana are not expected to influence the development of resistance in the Colorado potato beetle to the delta-endotoxin of B. thuringiensis.     

Influence of fertilization on the Colorado potato beetle, Leptinotarsa decemlineata, in organic potato production

The abundance of the Colorado potato beetle, Leptinotarsa decemlineata (Say), in organically grown potato did not change significantly in response to increasing rates of dehydrated poultry manure. However, peaks of abundance of larvae were shifted forward in time in response to the high rate of organic fertilizer. Tests using excised foliage showed that the shift was not caused by differential larval mortality or longer developmental times. Time allocation to resting, walking, and feeding by adults was similar regardless of fertilizer rate. Adult foliage consumption was unaffected by organic fertilizer rates in no choice tests and significantly affected in few choice tests. A 22% longer larval development time on plants treated with low fertilizer rate than on plants with high rate was the most significant effect. Even though maximum plant height, canopy, biomass, and yield were significantly smaller in the organic than in conventional plots, the suitability of the plants was not affected except for reduced feeding by summer beetles. Summer adults spent less time feeding and consumed two to five times less foliage on organic potato than on inorganically fertilized and conventionally produced plants. The overall influence of fertilizer on Colorado potato beetle populations was limited and therefore can only play a secondary role in management strategies for organic potato. Avoidance of excessive organic fertilizer that promotes short larval development time and extension of the period over which large Colorado potato beetle larvae are present should be recommended.     

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.     

Colorado potato beetle (Coleoptera: Chrysomelidae) feeding, development, and survival to adulthood after continuous exposure to Bacillus thuringiensis subsp. tenebrionis-treated potato foliage from the field

Colorado potato beetle, Leptinotarsa decemlineata (Say), feeding, development, and survival to adulthood were examined after continuously exposing large larvae to Bacillus thuringiensis subsp. tenebrionis-treated potato foliage from the field. In laboratory assays, the overall consumption and the length of period to become prepupae were determined for larvae, which began as 3rd and 4th instars, that were offered potato leaf disks with naturally declining levels of B. thuringiensis residue. In small-cage field experiments, survival to adulthood and the period to adult emergence for beetles confined to potato plants treated with B. thuringiensis beginning as 3rd and 4th instars also were examined. Third instars remaining on plants after a B. thuringiensis application were unlikely to feed and 4th instars consumed only approximately 50% as much foliage as those fed untreated foliage. Many late instars subjected to B. thuringiensis-treated foliage failed to survive to adulthood; 58-83% of these beetles died during the larval stage. Reduced feeding and poor survival of late instars suggest that counts of large larvae after application do not provide a complete picture of the efficacy of the B. thuringiensis treatment. Late instar Colorado potato beetles that were exposed continually to naturally declining levels of B. thuringiensis-treated potato foliage took an average of 1.8-4.5 d longer to become prepupae and 4-8 d longer to emerge as adults compared with those provided with untreated foliage. Delayed emergence of adults that fed on B. thuringiensis-treated potatoes as late instars indicated that development was prolonged in these insects because of ingestion of a sublethal dose of B. thuringiensis.     

Use of the egg parasiteEdovum puttleri [Hym.: Eulophidae] in an IPM system developed for Colorado potato beetle [Col.: Chrysomelidae] control on potatoes,Beltsville, Maryland

Edovum puttleri Grissell in conjunction with the fungal pathogenBeauveria bassiana (Balsamo) Vuillimen, a β-exotoxin ofBacillus thuringiensis varthuringiensis Berliner, and the insect growth regulator triflumuron (Alsystin 4F) were evaluated in a pest management system to control Colorado potato beetle,Leptinotarsa decemlineata (Say). Soil treatments ofB. bassiana had no significant impact on emerging Colorado potato beetle adults. Foliar applications of the β-exotoxin and triflumuron significantly reduced 1 ^st larval generation. Releases ofE. puttleri resulted in 56 % egg mass parasitization, significant reduction of 2 ^nd generation larvae, and significantly higher potato yields.Leptinotarsa texana (Schaeffer) was successfully used as an alternate host for production ofE. puttleri.      

Identification of beta-exotoxin production, plasmids encoding beta-exotoxin, and a new exotoxin in Bacillus thuringiensis by using high-performance liquid chromatography.

An improved high-performance liquid chromatography separation was developed to detect and quantify beta-exotoxin production in Bacillus thuringiensis culture supernatants. Exotoxin production was assigned to a plasmid in five strains, from three subspecies (B. thuringiensis subsp. thuringiensis serotype 1, B. thuringiensis subsp. tolworthi serotype 9, and B. thuringiensis subsp. darmstadiensis serotype 10). A new exotoxin, called type II beta-exotoxin in this report, was discovered in B. thuringiensis subsp. morrisoni serotype 8ab, purified, and partially characterized. This material is more specific than type I beta-exotoxin and is very active against the Colorado potato beetle, Leptinotarsa decemlineata.     

Germination polarity of Beauveria bassiana conidia and its possible correlation with virulence

Three different germination types of conidia; unidirectional, bidirectional and multidirectional, were revealed through microscopic observations for eight Beauveria bassiana isolates germinated on Sabouraud dextrose agar. Canonical correlation analysis indicated that there is a positive correlation between unidirectional germination and virulence against diamondback moth, Plutella xylostella and the Colorado potato beetle, Leptinotarsa decemlineata. Scanning electron microscopy revealed different in vivo behaviors for unipolar- and bipolar-germinated conidia. Unipolar-germinated conidia produced a strong germ tube with mostly appressorium-like structures while bipolar-germinated conidia continued to invasive hyphal growth without any penetration, indicating that germination polarity in one way or another may be an indicator of pathogenic ability.     

Compatibility of a natural enemy, Coleomegilla maculata lengi (Col., Coccinellidae) and four insecticides used against the Colorado potato beetle (Col., Chrysomelidae)

Abstract:   The toxicity of four insecticides used to control the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), imidacloprid (Admire^®), cryolite (Kryocide^®), cyromazine (Trigard^®), and Bacillus thuringiensis var. tenebrionis (Novodor^®), to one of its natural enemies, the 12-spotted lady beetle, Coleomegilla maculata lengi Timberlake (Coleoptera: Coccinellidae) was determined in the laboratory. Toxicity assays against C. maculata adults and larvae consisted of (1) topical applications and (2) exposures to treated foliage and prey, using concentrations up to 10 times the manufacturer's recommendations. Over a 6-day period, cyromazine (insect growth regulator) and B. t . var. tenebrionis (microbial insecticide) had no lethal effects on first and third instars C. maculata . For both larval and adult stages, cryolite (inorganic insecticide) caused very low predator mortality when topically applied and moderate mortality when ingested through contaminated eggs of Colorado potato beetles. Imidacloprid (systemic organic insecticide) was highly toxic to adult and larval C. maculata . Its estimated LD₅₀ at 6 days following treatment, corresponded to 0.02–0.09 times the recommended field concentration, depending on the developmental stage and mode of contamination. These results indicate that integrated pest management programmes for Colorado potato beetles using imidacloprid or, to a lesser degree, cryolite, would be detrimental to C. maculata . Cyromazine and B. t . var. tenebrionis seem to present a better compatibility with the protection of C. maculata populations.     

An ADAM metalloprotease is a Cry3Aa Bacillus thuringiensis toxin receptor

Bacillus thuringiensis insecticidal proteins toxic action relies on the interaction with receptor molecules on insect midgut target cells. Here, we describe an ADAM metalloprotease as a novel type of B. thuringiensis toxin receptor on the basis of the following data: (i) by ligand blot and N-terminal analysis, we detected a Colorado potato beetle Cry3Aa toxin binding molecule that shares homology with an ADAM10 metalloprotease; (ii) Colorado potato beetle brush border membrane vesicles display ADAM activity since it cleaves an ADAM fluorogenic substrate; (iii) Cry3Aa acts as a competitor of the cleavage of the ADAM fluorogenic substrate; (iv) Cry3Aa sequence contains the recognition motif R(345)FQPGYYGND(354) present in ADAM10 substrates. Accordingly, a peptide representative of the recognition motif localized within loop 1 of Cry3Aa domain II (Ac-F(341)HTRFQPGYYGNDSFN(358)-NH(2)) effectively prevented Cry3Aa proteolytic processing and nearly abolished pore formation, evidencing the functional significance of the Cry3Aa-ADAM interaction in relation to this toxin mode of action.     

A novel strain of Bacillus thuringiensis (NCIMB 40152) active against coleopteran insects

Abstract: A novel Bacillus thuringiensis strain, NCIMB 40152, was isolated from dead Tenebrio molitor L. larvae. The isolate is endowed with specific insecticide activity against the Colorado potato beetle ( Leptinotarsa decemlineata (Say), Coleoptera, Crysomelidae), but has no effect against lepidopteran or dipteran insects.
During the sporulation process it produces diamond-shaped crystals and small inclusions. Rare flat, square crystals are alsò produced. The strain can be differentiated from the other reported B. thuringiensis strains active against Coleoptera on the basis of physiological and biochemical tests, flagellar serotype and toxin electrophoretic patterns.
NCIMB 40152 toxin is easily extractable from spore/crystal preparations in neutral distilled water without detergents or reducing agents.     

Citrus limonoid effects on Colorado potato beetle larval survival and development

The effects of citrus limonoids, applied topically to potato (Solanum tuberosum L. var. Katahdin) foliage, on Colorado potato beetle (Leptinotarsa decemlineata) (Say) (Chrysomelidae) larval development, growth, and survival were quantified in laboratory assays and a small-plot field test. In laboratory assays, survival, development rate, and body weight decreased with increasing limonoid concentration, however these measures of larval response did not significantly differ among varying periods of limonoid exposure (three, six, or nine days). Significant limonoid application concentration and frequency effects on survival, development rate, and defoliation were observed in the field test. These results indicate the potential utility of lethal and non-lethal effects of citrus limonoids for management of the Colorado potato beetle.     

Susceptibility of a native and an exotic lady beetle (Coleoptera: Coccinellidae) to Beauveria bassiana

The exotic multicolored Asian lady beetle, Harmonia axyridis, became established and recently spread across much of North America and southern Canada. In a habitat now used by both the invading H. axyridis and a native lady beetle, Olla v-nigrum, we discovered that the native lady beetle was commonly infected by Beauveria bassiana; whereas, the exotic H. axyridis, was not. Laboratory assays revealed that B. bassiana isolates collected from naturally infected O. v-nigrum were pathogenic to adult O. v-nigrum but not to adult H. axyridis. In contrast, the GHA strain of B. bassiana was not significantly pathogenic to O. v-nigrum nor H. axyridis. Late-season field collections revealed significantly higher B. bassiana infection of O. v-nigrum than H. axyridis. Our results lead us to hypothesize that low susceptibility of H. axyridis to B. bassiana (found to infect O. v-nigrum) may provide an intraguild advantage to H. axyridis over O. v-nigrum; this may also occur with other species of native lady beetles and other endemic entomopathogens in different habitats and regions.     

Differential predation by Coleomegilla maculata on Colorado potato beetle strains that vary in growth on tomato

This study tests the hypothesis that the generalist predator Coleomegilla maculata DeGeer causes differential mortality of Colorado potato beetle, Leptinotarsa decemlineata (Say), larvae differing in their degree of genetic adaptation to tomato (Lycopersicon esculentum Mill.) as a host plant. Results of a series of laboratory experiments demonstrate that adult C. maculata can cause higher mortality to nonadapted than adapted Colorado potato beetle larvae. The extent of differential mortality caused by C. maculata depended on age of potato beetle larvae; presence of potato beetle eggs; whether or not the predator had a choice among prey items; and, in choice situations, the ratio of adapted to nonadapted potato beetle larvae. Although adult C. maculata have the potential to prey differentially on tomato-adapted and nonadapted Colorado potato beetle larvae in mixed populations, the magnitude of differential predation in a natural setting could be highly variable.     

基于GIS的马铃薯甲虫扩散与河流关系研究——以新疆沙湾县为例

马铃薯甲虫是一种重要的外来入侵害虫,自1993年传入我国新疆北部地区以来,迅速扩散传播,对我国马铃薯等茄科作物生产造成严重威胁。为明确马铃薯甲虫传播扩散与河流等生态因子之间的关系,结合马铃薯甲虫虫情调查,利用地理信息系统(GIS)的空间分析功能,对新疆沙湾县马铃薯甲虫发生与河流的关系进行研究。结果表明:在沙湾县内,马铃薯甲虫扩散方向与河流流向基本一致,均呈由南向北方向;不同缓冲区内,马铃薯甲虫危害级别差异极显著,缓冲区危害级别从大到小依次:缓冲区2(1-2 km)>缓冲区4(3-4 km)>缓冲区5(4-5 km)>缓冲区1(0-1 km)>缓冲区3(2-3 km);不同河流流域内马铃薯甲虫危害级别也差异极显著,危害级别排序依次是:金沟河>安集海河>玛纳斯河>西岸大渠。     

Response of potato aphid (Homoptera: Aphididae) to synthetic potato-derived Colorado potato beetle (Coleoptera: Chrysomelidae) attractant and natural potato odor

A recently synthesized kairomone blend, based on the volatiles produced by potato (Solanum spp.) plants, has been demonstrated to be attractive to both adult and larval stages of the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). It was subsequently formulated in a viscous inert carrier for field applications and showed potential for aggregating beetles in treated areas of the field. We investigated effects of this kairomone formulation on the potato aphid, Macrosiphum euphorbiae (Thomas) (Homoptera: Aphididae). The response of both winged and wingless adults to natural potato foliage and synthetic kairomone was tested in a Y-tube olfactometer. Aphid response to untreated potato foliage, foliage treated with the kairomone blend, and foliage treated with blank inert carrier also was tested in petri dishes. In addition, aphid densities on field plots treated with kairomone and blank inert carrier were compared with the control plots. The untreated potato foliage was found to be attractive to wingless, but not winged, potato aphids. In the olfactometer, the foliage treated with synthetic Colorado potato beetle kairomone was not attractive to either winged or wingless aphids. In petri dishes, aphids avoided leaflets treated with both kairomone formulation and its blank carrier. There was no statistical difference between any treatments compared in the field.     

Effect of crop rotation distance on populations of Colorado potato beetle (Coleoptera: Chrysomelidae): development of areawide Colorado potato beetle pest management strategies

The Colorado potato beetle, Leptinotarsa decemlineata (Say), overwinters adjacent to field edges and infests nearby fields in the spring, primarily by walking. Crop rotation is known to be an effective cultural control against Colorado potato beetle populations limiting spring infestations. Spatial separation is an important consideration in optimizing the rotational effect because long-distance rotations have been shown to reduce Colorado potato beetle infestations. To determine the effect of long-distance rotations on Colorado potato beetle populations in commercial Wisconsin potato production, a geographic area of >18,200 ha (>45,000) acres in southern Portage County, Wisconsin, was selected as an experimental area for areawide pest management of the Colorado potato beetle. From 1997 to 1999, beetle populations at edges within each potato field in this region were determined by spring and fall field sampling. The rotational distance between current and previous potato fields was measured and analysis was run between the distance and Colorado potato beetle populations. Long-distance rotations of >400 m were an effective cultural control management strategy to limit adult beetle infestations in the spring. This strategy can be optimized when collaborating growers are able to maximize their rotational distances by coordinating their rotational schemes within large areawide, geographic locations. Deploying long-distance rotations within a geographic area over many years would limit Colorado potato beetle populations and could result in a significantly reduced Colorado potato beetle populations entering fields in the spring.     

Genetically modified potatoes expressing Cry 3A protein do not affect aphidophagous coccinellids

Abstract:  Field investigations showed that genetically modified Bacillus thuringiensis (Bt) potato plants ('New Leaf' expressing Cry 3Aa toxic protein) were not damaged by the Colorado potato beetle (CPB) Leptinotarsa decemlineata and contained diverse populations of the aphidophagous coccinellids Coccinella septempunctata , Coccinula quatuordecimpustulata , Hippodamia variegata and Propylea quatuordecimpunctata . The insecticides, alpha-cypermethrin and fipronil, were used for control of CPB in the fields with the non-transgenic standard potatoes 'Santana' and 'Arinda'. Both insecticides caused significant decrease in the abundance of aphidophagous coccinellids. Laboratory experiments revealed that Bt potatoes expressing Cry 3Aa had no effect on the aphid Myzus persicae and that the aphids fed on Bt potatoes had no effect on the larval development and mortality of C. septempunctata .     

Diapausing Colorado potato beetles are devoid of short neuropeptide F I and II

An extract of head ganglia and retrocerebral complexes of nondiapausing and diapausing Leptinotarsa decemlineata was prepared to characterize regulatory neuropeptides involved in adult diapause by using a differential peptidomics approach. To reduce sample complexity, both extracts were roughly separated by means of an identical chromatographic step. MALDI-TOF MS led to the identification of proctolin, an adipokinetic hormone, and short neuropeptide F I and II in the extract of nondiapausing beetles. In combination with nano-ESI-Q-TOF MS(2) evidence was found for the presence of three pyrokinins, the first to be identified in a coleopteran species. Pyrokinins, involved in the induction of embryonic diapause in Bombyx mori, were present in both physiological conditions suggesting that they are of minor importance in the regulation of adult diapause in the Colorado potato beetle. A striking difference, detected by the differential peptidomics approach, between both neuropeptide profiles was the absence of ions corresponding to the short neuropeptide F (sNPF) related peptides, also known as Led-NPF-I and -II, in the extract of diapausing animals. Therefore, we postulate that "short NPFs" are involved in the regulation of adult diapause, displayed by the Colorado potato beetle.