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1.
The aim of this work was to isolate and characterize novel Phthorimaea operculella granulovirus (PhopGV) strains from Costa Rican soils. Three novel strains, named PhopGV‐CR3, PhopGV‐CR4 and PhopGV‐CR5, were isolated from three locations in Costa Rica, Alvarado, Zarcero and Abangares, respectively, by means of soaking potato tubers with diluted soil samples. An additional strain, PhopGV‐CR2, was identified from diseased larvae from a Tecia solanivora laboratory culture. Restriction fragment length polymorphisms obtained for each isolate with six restriction endonucleases (RENs) allowed their identification as four distinct PhopGV strains. Both REN and Polymerase chain reaction analyses indicated the existence of an array of genotypic variants present in all isolates. Bioassays in P. operculella and T. solanivora showed that PhopGV‐CR3 was well adapted to the two coexisting hosts with high levels of pathogenicity against both pest species. The mean lethal dose values of this strain were 2.8 OBs/mm2 for P. operculella and less than 0.5 OBs/mm2 for T. solanivora. We conclude that PhopGV‐CR3 shows great promise for soil application against these pests in Costa Rican potato crops.  相似文献   

2.
Phthorimaea operculella granulovirus (PhopGV) belongs to the genus Betabaculovirus of the arthropod‐infecting Baculoviridae. PhopGV is able to infect several gelechiid species. Among them are the potato tuber moths Phthorimaea operculella Zeller and Tecia solanivora Povolny (both Lepidoptera: Gelechiidae). In various South American countries, PhopGV‐based biopesticides are used to control either P. operculella or T. solanivora. Many trials have indicated that a particular viral isolate can exhibit very distinct pathogenicity when infecting different host species or different populations of one host species. In this study, we compared host–pathogen interactions using various PhopGV isolates and various populations of P. operculella and T. solanivora. Virus isolates from P. operculella were more pathogenic against their original host species than against T. solanivora. A PhopGV isolated from T. solanivora was less efficient against P. operculella. In addition, virus isolates differed in pathogenicity toward their hosts (i.e., lethal concentrations of isolates ranged from low to high). Unexpectedly, we also found that host populations of one species from distinct geographic origins did not differ significantly in susceptibility to the same PhopGV isolate. This was the case for both host species and for five PhopGV isolates. Comparative restriction fragment length polymorphism (RFLP) analyses of 11 isolates including those used in bio‐assays indicated three main regions of variation in the genome of PhopGV, corresponding to the regions of open reading frame PhopGV046, gene PhopGV129 (egt), and repeat 9 (located between open reading frames PhopGV083 and PhopGV084). Comparison of the nucleotide sequences of the insertions/deletions present in these regions were carried out for the most variable isolate, JLZ9f. The results are discussed in the context of the production and use of PhopGV as a biological agent against these two pest species.  相似文献   

3.
The control efficiency of a Phthorimaea operculella granulovirus isolate from Costa Rica (PhopGV-CR1) against the concurrent insect pests P. operculella (Zeller) and Tecia solanivora (Povolny) (Lepidoptera: Gelechiidae) was evaluated. In warehouses, the best control efficiency was achieved with a powder formulate applied inside bags, which reduced injury over 70 % compared with the untreated controls. In the field, liquid and powder virus formulations significantly reduced injury between 56.2 and 81.7 % compared with the untreated controls, and were as efficient as chemical treatments. The efficiency of formulations stored at ?20 °C for six months (liquid) or at ambient temperature for three months (powder) was maintained, but higher temperatures and/or longer exposure times resulted in loss of pathogenicity. The data presented here favor the inclusion of granulovirus formulations of PhopGV-CR1 in Integrated Pest Management programs against tuberworms in Costa Rica and give clues on storage conditions for the growers in this country.  相似文献   

4.
Phthorimaea operculella granulovirus (PhopGV) is infectious for larvae of different Gelechiidae insect species, including Tuta absoluta and Phthorimaea operculella. As these are major economic pests in North and sub‐Saharan Africa as well as in the Mediterranean area, the development of locally suitable biocontrol agents is essential. We have studied five isolates of PhopGV from Tunisia (Tns16, Tu1.11 and Tu2.11), Kenya (Ken13) and Yemen (Ym14) for their biological activity and the sequence polymorphism of their granulin and ecdysteroid UDP‐glucosyltransferase (egt) genes and allocated the isolates to two different egt types. Infection experiments with neonate larvae of T. absoluta and P. operculella demonstrated their pathogenicity to both host species. The isolate PhopGV Tu1.11 was the most virulent one for T. absoluta but had a relatively low infectivity to two P. operculella populations originating from Italy and Tunisia.  相似文献   

5.
Chemical analysis of pheromone gland extracts followed by behavioural studies in the wind tunnel and by field trapping tests show that the sex pheromone of the Guatemalan potato moth, Tecia (Scrobipalpopsis) solanivora Povolny (Lepidoptera: Gelechiidae), is a blend of (E)‐3‐dodecenyl acetate, (Z)‐3‐dodecenyl acetate, and dodecyl acetate. A 100 : 1 : 20 blend of these compounds, formulated at 1000 µg on rubber septa, captured more males than the main compound alone. This lure was species‐specific and did not capture the potato tubermoth, Phthorimaea operculella. A potato field was treated with a blend of these three compounds at a rate of 28 g ha?1. Male T. solanivora attraction to synthetic pheromone traps was almost completely suppressed for 2 months, demonstrating the potential use of pheromones for control of this economically important insect pest of potato in Central and Southern America.  相似文献   

6.
In this study a Brazilian granulovirus strain, PhopGV, isolated from the potato tuber moth (PTM) Phthorimaea operculella, was investigated regarding its potential for biological control and in vivo production. The relationship between mortality of P. operculella larvae and virus concentration was determined at different temperatures on potato tubers and susceptibility of P. operculella to PhopGV was also determined on potato leaves. Virulence of PhopGV to P. operculella was not affected by temperatures from 18 to 30°C. The median lethal concentration (LC50) of larvae fed on potato foliage treated with PhopGV was not higher than that verified with larvae fed on treated tubers. Optimal conditions for production of virus-infected larvae were obtained by using the virus suspensions of 41 × 105, 6.3 × 105 and 62 × 105 OBs ml−1 at 18, 24 and 30°C, which resulted in 32.0, 31.4 and 34.8% of infected larvae collected, respectively. The maximum percentage of infected larvae recovered from tubers was not affected by temperature. However, time for production of virus-infected larvae was longer at 18°C and shorter at 30°C. Persistence of PhopGV was determined on stored tubers and we observed that the virus remained effective for at least two months, causing up to 84.2% mortality of P. operculella at 1 × 107 OBs ml−1. The pathogen was also highly virulent to tomato pinworm, Tuta absoluta, inflicting high percentage of mortality, delaying larval growth and inhibiting pupation. This Brazilian PhopGV strain has potential to control PTM larvae on potato tubers at a broad range of temperature and can be produced in vivo using virus-treated tubers.  相似文献   

7.
The tomato leafminer Tuta absoluta (Meyrick 1917) (Lepidoptera:Gelechiidae) is a devastating pest, causing losses of up to 100%. An interesting tool for its control is the use of the insect specific viruses of Baculoviridae family. Despite its high potential, its use on T. absoluta larvae has been poorly studied. In this work, Colombian granuloviruses VG013 and VG003 isolated from T. absoluta and Tecia solanivora (Lepidoptera:Gelechiidae) larvae sampled in tomato and potato crops, respectively, were morphologically, biologically and molecularly characterized. Occlusion bodies showed ovoid shape containing one nucleocapsid. Restriction endonuclease analysis revealed a pattern similar to Phthorimaea operculella granulovirus and bioinformatics studies showed that both isolates are variants of that baculovirus specie. Similar mean lethal concentrations (LC50) on T. absoluta larvae were estimated for both viruses, although VG013 exhibited shorter mean time to death than VG003. The latter reached a higher OBs yield in comparison with VG013. These results demonstrated an interesting potential of evaluated betabaculoviruses to control T. absoluta populations and pointed key features to its use under field conditions.  相似文献   

8.

Potato tuber moth, Phthorimaea ( = Gnorimoschema) operculella (Zeller) (Lepidoptera: Gelechiidae), was reared from egg to adult for 3 generations on an artificial diet. At 30±2°c with a 16‐h photoperiod the total life cycle was completed in 23 days, compared with 18.5 days on potato tubers. Average life cycle data from the 3 generations reared on diet (with corresponding data from tubers in parentheses) are: larval period, 13.0(11.8) days; pupal period, 5–6(4–5) days; survival to adult, 76.1(100)%; fecundity, 102(169) eggs/?; pupal wt‐ ? 8.3(11.4) mg, ? 7.7(9.1) mg.  相似文献   

9.
1 Granulovirus PoGV is a strong candidate to substitute for chemical insecticides in integrated pest management (IPM) of the potato tuber moth Phthorimaea operculella Zeller (Lepidoptera: Gelechiidae). Generally, susceptibility to baculoviruses decreases with increasing larval age. For effective field applications, knowledge of the age‐related change in susceptibility is crucial. 2 The relative susceptibility of various instars of P. operculella larvae to PoGV was studied by leaf‐disc bioassays in the laboratory at 26 °C incubation temperature. The LC50 increased from 4 × 106 granules/mL for neonate larvae to 1.2 × 107, 1.1 × 108, 4.4 × 108, and 4.2 × 109 granules/mL, signifying resistance ratios of 3, 29, 110, and 1051, for 5, 6, 7, and 9‐day‐old larvae, respectively. 3 The relationship between log‐LC50 values and log‐larval weight was significantly linear. A logistic function described significantly the relation between larval weights and physiological age measured in degree‐days (DD > 13.4 °C). Both functions allowed prediction of the activity ratio of PoGV for different larval ages by using temperature summation to express physiological age. 4 PoGV was found to be highly active against P. operculella larvae up to a physiological age of approximately 50 DD (>13.4 °C) (i.e. the first third of the total larval development time). Thereafter, the virus rapidly lost its activity against older larvae. Prospects for applying this knowledge in the field are discussed.  相似文献   

10.
Larvae of the potato tubermoth (PTM), Phthorimaea operculella, feed on potato plants and tubers and are a major pest in the tropics and subtropics worldwide, causing up to 100% damage. The PTM granulovirus (PhopGV) provides significant potato protection, but little is known about its effect on larval development or its histopathology. Here we show that only 10% of larvae exited from PhopGV-treated tubers (1.4 × 108 granule/ml), lagging significantly behind controls, and most of these died by 72 h after emergence. Histopathology studies showed the fat body and epidermis were the principal tissues infected. PhopGV morphogenesis was similar to other GVs, the exception being small vesicles between mature granules.  相似文献   

11.
Males of the potato tuberworm moth Phthorimaea operculella (Lepidoptera: Gelechiidae) locate conspecific females by a series of short and straight flights, or ‘hops’. On the floor of a wind tunnel, P. operculella can change their heading angles in response to wind direction shift, suggesting that they detect wind direction and orient upwind when on the ground. It is unlikely that they navigate in wind by vision‐induced optomotor anemotaxis as in many flying moths. To investigate the mechanism of wind direction detection in this species, their orientation behaviour in response to pheromone pulses is observed in a wind tunnel. Intact male moths orient upwind even in complete darkness. After the flagellum of one antenna is amputated, male moths still successfully orient upwind. However, they fail to head upwind when the basal segments of both of their antennae are immobilized with glue. The ability to surge upwind is restored by removing the glue from the antennae. Thus, the results of the present study indicate that males of P. operculella orient upwind in response to mechanoreceptive cues from mechanosensory organs on their antennae. In Lepidoptera, two distinct anemotactic mechanisms of different sensory modalities appear to coexist: optomotor anemotaxis when airborne and the aim‐then‐shoot anemotactic system mediated by antennal mechanoreception when on the ground.  相似文献   

12.
Preliminary screening of extracts of the leaves and stems of four Turraea plant species (Meliaceae) on second instar larvae of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) showed that the methanolic extract of Turraea abyssinica leaves possess good toxic potential with LD50 of 270.7 ppm. Fractionation of this extract led to isolation of a new limonoid derivative, 12α‐diacetoxywalsuranolide (1) and three other known limonoids [1α,7α,12α‐triacetoxy‐4α‐carbomethoxy‐11β‐hydroxy‐14β,15β‐epoxyhavanensin (2), 11‐epi‐21‐hydroxytoonacilide (3), 11β,12α‐diacetoxycedrelone (4)]. The structures of the compounds were established by IR, UV, mass spectrometry and 1D/2D NMR analyses, and in the case of the known compounds, also by comparison with reported data. All the isolates were tested for their larvicidal activities at different dose range against second instar larvae of T. absoluta. They were found to be more active (with LD50 < 7.0 ppm) compared to the reference compound azadirachtin (LD50 value of 7.8 ppm).  相似文献   

13.
Efficient production is essential for developing baculoviruses into marketable bioinsecticides. In this study, we evaluated how juvenile hormone (JH) III and the JH analog pyriproxyfen affected the production efficiency of Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) Nucleopolyhedrovirus (SpltNPV) in SpltNPV‐inoculated early sixth instars. Both the JH III and pyriproxyfen treatments significantly enhanced the body size and weight of S. litura, which resulted in a 1.5‐ to 2‐fold increase in the production of occlusion bodies (OBs). The OBs obtained from JH‐ and pyriproxyfen‐treated larvae were as pathogenic as those produced in untreated larvae. More importantly, JH‐ or pyriproxy‐fen‐treated larvae consumed less than twice the amount of food consumed by two untreated larvae. These results indicate that JH and pyriproxyfen treatments improve the production efficiency of SpltNPV in S. litura. SpltNPV infection affected the development of S. litura not only as larvae, but also during subsequent growth stages and generations.  相似文献   

14.
Abstract

The combined and alone effects of azadirachtin (AZA) and Spodoptera frugiperda multicapsid nucleopolyhedrovirus (SfMNPV) on the mortality of S. frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) were evaluated in the laboratory. For this, diet surface contamination bioassays were performed on S. frugiperda in the third instar. LC50 values for SfMNPV alone were determined to be 430 and 373 viral occlusion bodies (OBs)/mm2 at 192 and 216 h after treatment, respectively. LC50 values for AZA alone were estimated for two periods of continuous exposure (4 or 5 days). In this case, LC50 values were 45.5 and 16.8 mg L?1 at 216 h after treatment (4 or 5 days of larval exposition to insecticide, respectively). We observed that although the interaction of AZA with SfMNPV increased viral pathogenicity, such improvements were of greater magnitude and more consistent at the lower OB concentration used (177 OBs/mm2). Application of SfMNPV (430 OBs/mm2), AZA (26.4 mg L?1) or SfMNPV–AZA mixtures resulted in a significant reduction in the mean weight of larvae treated in the third instar across the experiment, by 23–41%, 17–95% and 26–97%, respectively, compared to control. The duration of larval development during the third and fourth instars increased significantly in larvae exposed to SfMNPV–AZA mixtures and AZA alone compared to SfMNPV alone and control treatments. The yield of OBs/mg weight of larvae treated with SfMNPV alone was 1.8-fold higher than OB yields from insects inoculated with SfMNPV–AZA mixtures. We conclude that AZA + SfMNPV mixtures are unlikely to be useful for the mass production of this virus and laboratory observations on the value of AZA + SfMNPV mixtures as a potentiator of biological insecticides require validation in field studies under commercial growing conditions.  相似文献   

15.
The fall armyworm (FAW), Spodoptera frugiperda (Lepidoptera: Noctuidae), is the most important pest of maize in many countries. Entomopathogenic viruses mainly Baculoviruses family are excellent biological control agents and therefore a viable alternative for managing this pest. The aim of this study was to determine the biological activity of eight native nucleopolyhedrovirus (NPVs) against FAW larvae. Additionally, two of the most virulent isolates (SfCH32 and SfCH15) were characterized biologically by bioassays to estimate their median lethal dose (LD50) and median lethal time (LT50), morphologically by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) and molecularly by restriction enzymes. Three (SfCH15, SfCH18 and SfCH32) of the eight tested native nucleopolyhedrovirus isolates caused mortalities ˃98% at 168-hr post-inoculation (hpi) with a dose of 9.2 × 104 OBs/larva. SfCH15 and SfCH32 isolates showed occlusion bodies (OBs) of irregular shape and size (1.02–2.24 μm). The SfCH15 and SfCH32 isolates showed similar median lethal dose (5.6 × 102–6.4 × 102 OBs/larva). The lowest median lethal time (114.5 hpi) was observed with the SfCH15 isolate at the highest concentration (2.5 × 106 OBs/larva). The DNA restriction profiles for SfCH15 and SfCH32 were different, with their genome size being ~128,000 bp and 132,000 bp, respectively. SfCH15 and SfCH32 isolates showed similar morphological characteristics and the highest virulence against fall armyworm. This study showed that native isolates were highly virulent against S. frugiperda larvae, being similar to other reported strains; however, field studies are required to confirm their insecticidal effect.  相似文献   

16.
Granulovirus (PoGV) is a promising candidate to substitute for chemical insecticides in integrated pest management (IPM) of the potato tuber moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae). A procedure to assess pathogenicity (LC50) and its standardization is crucial for research into the use of pathogens as biocontrol agents. In the present study, an egg‐dip bioassay method was developed and its precision tested in a series of six bioassays over a time period of 18 months. Bioassays were carried out at 25 °C incubation temperature. The probit model as a statistical analysis method for the interpretation of concentration responses proved better than the exponential model. LC50 values ranged from 2.3 × 106 to 107 granules ml?1 with a mean of 5 × 106 granules ml?1 and regression slopes varied between 0.73 and 1.05 with a mean of 0.84. LC50 values and slopes displayed no significant differences according to their 95% confidence limits. The rate of harvested infected larvae showed a linear increase with increasing concentration on a log‐log scale. The mean natural mortality was 15% (SE ± 1.8%) and can be considered as a main factor producing variability among bioassays. The advantages of this bioassay method and its application in PoGV research are discussed.  相似文献   

17.
Heavy infestations of stored potato (Solanum tuberosum L.) tubers by the two potato tuber moth species Symmetrischema tangolias (Gyen) and Tecia solanivora (Povolny) frequently occur in Andean potato‐growing regions of Ecuador. The aim of the study was to develop a biological control strategy for both species using powder formulations made of inert substances, Phthorimaea operculella (Zeller) granulovirus (PhopGV) and Bacillus thuringiensis Berliner subsp. kurstaki (Btk). The LC50 of PhopGV on T. solanivora was 0.33 LE/L, and Btk caused 82.7% mortality at a concentration of 100 g/L in bioassays. The efficacy of talcum, kaolin, calcium carbonate and sand ranged between 76.2% and 98.7%. Calcium carbonate was highly effective to control both species; however, its efficacy was affected by the relative humidity and dropped to 55.4% at relative humidity of 100%. PhopGV at concentrations of five larvae equivalents (LE) per kg kaolin and Btk at a concentration of 60 g Btk/kg talcum caused 95.7% and 88.1% mortality of T. solanivora, respectively. In storage experiments, the efficacy of calcium carbonate alone and in combination with PhopGV (20 LE/kg) and Btk (15 g/kg) caused 95.0–99.8% mortality of T. solanivora in all treatments and reduced infestation on potato tubers by 83.6%–91.0%. In the case of S. tangolias, Btk significantly increased mortality to 96.5% compared with calcium carbonate alone and reduced tuber infestation by 83.4%. Storage of potato tubers in thin layers enhanced the efficacy of the calcium carbonate treatment compared with storage in bags. It was concluded that calcium carbonate alone seems to be appropriate for the control of T. solanivora, and an addition of 15 g Btk/kg would improve the control of S. tangolias. It is suggested to test these new formulations under on‐farm storage conditions.  相似文献   

18.
A strategy for biological control of the fall armyworm, Spodoptera frugiperda, has included the use of baculoviruses principally the nucleopolyhedrovirus SfMNPV, which have been extensively characterised. In contrast, the granulovirus of S. frugiperda (SfGV) has been poorly studied even though it is able to enhance the infectivity and virulence of NPVs. In this work, a Colombian SfGV isolate (VG008) was characterised in comparison with a reference isolate from Brazil (VG014). The viral morphology was characterised by ovoidal-shaped occlusion bodies (OB) that contained one single internal virion. Median lethal concentrations (LC50) and mean times to death (MTD) were 4.5 × 105 OBs/mL and 29 days for VG008 and 1.6 × 105 OBs/mL and 33 days for VG014. Both isolates reduced their insecticidal activity by 94%, after one hour of direct irradiation with ultraviolet light type B. The most prominent protein had an apparent molecular mass of 27 kDa and corresponded with the Granulin. Genomic comparison among isolates from Colombia and Brazil generated by restriction profiles showed differences in the number and size of fragments. Partial sequences of lef-8 and lef-9 genes and complete sequence of gran gene of Colombian SfGV isolate (VG008) showed high similarity values with VG014 and SfGV A12-4 homologous sequences, showing genetic distance lower than 0.015 (Kimura 2-parameter model), which confirmed that the three isolates belong to the same viral species. The characterisation of VG008 isolate demonstrated its high genomic and biological similarity with the Brazilian isolate.  相似文献   

19.
Abstract The potato tuberworm, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), is the most destructive pest of potato, Solanum tuberosum L. (Solanaceae), in tropical and subtropical regions in both field and storeroom situations. The modeling of temperature‐dependent development can be useful in forecasting occurrence and population dynamics of the pests. Published developmental parameters for this pest vary greatly for many reasons. We determined temperature‐dependent development of P. operculella at seven constant temperatures (16, 20, 24, 28, 32, 34 and 36 °C). Developmental period of whole immature stage (egg to the end of the pupal stage) varied from 75.5 days at 16 °C to 17 days at 32 °C. The population failed to survive at 36 °C. The observed data was modeled to determine mathematical functions for simulating P. operculella development in each stage of development and overall. Two linear models, ordinary linear regression and the Ikemoto linear model were used to describe the relationship between temperature and development rate of the different stages of P. operculella and estimating the thermal constant and lower temperature threshold. The lower temperature threshold (t) and thermal constant (k) of whole immature stage were estimated to be 11.6 °C and 338.5 DD by Ikemoto linear model, and the estimated parameters were not substantially different with those estimated by ordinary linear models. Different models provided a better fit to the various developmental stages. Of the eleven nonlinear models fitted, the Beriere‐1, Logan‐6 and Lactin‐1 model was found to be the best for modeling development rate of egg, larva and pupa of P. operculella, respectively. Phenological models based on these findings can be part of a decision‐support tool to improve the efficiency of pest management programs.  相似文献   

20.
There are few insecticidal options for potato tuber moth (PTM), Phthorimaea operculella Zeller (Lepidoptera: Gelechiidae), control late in the growing season. We evaluated the PTM granulovirus (PoGV) and Bacillus thuringiensis Berliner ssp. kurstaki (Btk) for season‐long control of PTM on potato foliage in 2006 and 2007. Compared to untreated controls, 10 weekly applications of PoGV (1013 occlusion bodies/ha) reduced PTM populations in replicated 1‐m3 field cages by 86–96% on pre‐harvest foliage and 90–97% on tubers added to cages shortly before harvest. Infection rates of 82–95% of L4 larvae by PoGV were noted within individual larval cohorts. Equivalently timed Btk treatments (1.12 kg product/ha) were significantly less effective at population suppression, with a 36–76% reduction in larvae recovered from tubers added to cages. A PoGV/Btk alternation was significantly more effective than Btk alone and as effective as PoGV in 2007, but not in 2006. There was some evidence that reduced rate PoGV treatments (10% rate or 50% application frequency) were less effective than the standard program. There were no treatment effects on percentage of tubers growing in the ground that were infested at harvest, which remained comparatively low at ≤8.1%. Bioassays were conducted to evaluate the residual activities of foliar deposits. Early‐season applications were highly effective for the first 24 h (≥93% mortality) with a steady decline in activity over 10 days. A second application, applied later in the season, showed similar patterns, although in this case Btk was less persistent than PoGV, whereas both agents provided significant larval mortality compared with controls over 14 days. Both PoGV and Btk provide alternatives to manage field infestations of PTM prior to harvest, thus reducing the risk of tuber infestations in storage.  相似文献   

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