Responses of different geographic populations of two potato tuber moth species to genetic variants of Phthorimaea operculella granulovirus

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.     

The potential of novel African isolates of Phthorimaea operculella granulovirus for the control of Tuta absoluta

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.     

Costa Rican soils contain highly insecticidal granulovirus strains against Phthorimaea operculella and Tecia solanivora

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.     

Biological Control of the Potato Tuber Moth (Phthorimaea operculella Zeller) in the Republic of Yemen Using Granulosis Virus: Biochemical Characterization, Pathogenicity and Stability of the Virus

A granulosis virus infecting the potato tuber moth, Phthorimaea operculella, has been identified, isolated and purified from diseased potato tuber moth larvae collected from potato fields in the highland basin Qa al-Boun of the Republic of Yemen. The granulosis virus was propagated by feeding potato tuber moth larvae with potatoes treated with a concentration of one granulosis virus-infected fourth instar larva to 5l of water (corresponding to an occlusion body (OB) concentration of 2 106 OB ml-1). Restriction enzyme analysis of viral DNA revealed that the isolated virus seemed to correspond to an isolate from the Lima region of Peru. A median LC 50 value of 7.3 104 OB ml-1 was calculated for a purified virus preparation. Different preparations of the granulosis virus were investigated for their persistence in the field on tubers and leaves. A purified virus preparation (PoGV) applied to potato tubers and exposed in the open had a half-life of 1.3 days. On leaves, the activity of granulosis virus spray deposits of an unpurified virus preparation (PoGV I) and of a glycerine-formulated preparation (PoGV II) also declined with exposure time. Mortalities of potato tuber moth larvae of 43% for PoGV I and 49% for PoGV II were recorded when first instar larvae were fed with potato leaves collected 2 days after treatment. After 8 days, 25.8% of the larvae had died from PoGV I treatment and 19.4% from PoGV II. Neither preparation displayed any effect 17 days after application.     

Characterization of a Costa Rican granulovirus strain highly pathogenic against its indigenous hosts,Phthorimaea operculella and Tecia solanivora

A granulovirus isolate collected from diseased Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae) larvae in Costa Rican potato [Solanum tuberosum L. (Solanaceae)] crops was characterized at the molecular and biological level. Restriction endonuclease analysis identified this isolate as a novel P. operculella granulovirus (PhopGV) (Baculoviridae: Betabaculovirus) strain and was designated as PhopGV‐CR1. In addition, PCR amplification of four specific variable genomic regions yielded multiple amplicons for two open reading frames, revealing the presence of different genotypic variants within the virus population. Biologically, PhopGV‐CR1 was highly pathogenic for its two indigenous hosts, although significant differences of up to four‐fold were detected against P. operculella [LD₅₀ = 17.9 occlusion bodies (OBs) mm^?2] and Tecia solanivora (Povolny) (Lepidoptera: Gelechiidae) (LD₅₀ = 69.1 OBs mm^?2). The two P. operculella colonies, from Costa Rica and France, were equally susceptible to PhopGV‐CR1. Serial passage of PhopGV‐CR1 over four generations in T. solanivora increased its pathogenicity by five‐fold in three generations, suggesting an ongoing adaptation to its alternate host.     

Biological Control of the Potato Tuber Moth (Phthorimaea operculella Zeller) in the Republic of Yemen Using Granulosis Virus: Propagation and Effectiveness of the Virus in Field Trials

A granulosis virus (PoGV) infecting the potato tuber moth, Phthorimaea operculella, in the Republic of Yemen was evaluated for its use in an inundative approach to control this pest. In field trials on potatoes, the effectiveness of the virus was examined on-farm and on-station using an unpurified preparation (PoGV I) and a purified glycerine-formulated virus preparation (PoGV II). The virus infection and mortality of potato tuber moth larvae were dependent on the virus concentration applied. After application of 5 1012 occlusion bodies (OB) ha-1 of PoGV I, the proportion of virus-infected larvae increased, fluctuating between 26 and 39% 14-35 days after treatment. At a granulosis virus concentration of 5 1013 OB ha-1, clear symptoms of granulosis virus infection (milky white coloration and reduced vitality) were observed in larvae collected 11 days after treatment. A total of 70% of the fourth instar larvae collected 19 days after treatment with PoGV I were infected. Further observations revealed very high rates of effectiveness in terms of larval mortality. The preparations PoGV I and PoGV II were not found to differ in their degree of effectiveness, which resulted in a mortality of potato tuber moth larvae of 85.3% for PoGV I and 82.5% for PoGV II respectively.     

Copulatory behaviour of the potato tuber moth, Phthorimaea operculella

ABSTRACT. Copulatory behaviour of the potato tuber moth, Phthorimaea operculella , was observed in a small glasshouse. The behavioural chain is started by the female 'calling'. Males responded to the female's sex pheromone by searching in her vicinity. When the males 'recognized' the female, they attempted copulation. This behaviour was released, however, by a wide variety of models, including dried whole male and female bodies, similarly sized rolls of paper with male or female wings attached, rolls of paper covered in wing scales, and even flat surfaces covered in wing scales (provided the surface was much larger than a moth). It is concluded that the mechanical stimulus from the scales is an important stimulus for evoking copulation.     

Tomato fruit size, maturity and alpha-tomatine content influence the performance of larvae of potato tuber moth Phthorimaea operculella (Lepidoptera: Gelechiidae)

Various physical and chemical properties of host plants influence insect larval performance and subsequent adult fitness. Tomato plants are relatively new hosts to the potato tuber moth, Phthorimaea operculella (Zeller), with the fruit being its preferred feeding site. However, it is unclear how the biochemical and physical properties of tomato fruits relate to potato tuber moth performance. Significant amounts of alpha-tomatine were detected in maturing green and ripening fruits of cherry (cv. Ceres) and processing (cv. Serio) types of tomatoes whereas none was detected in a fresh market variety (cv. Marglobe), at comparable stages. alpha-Tomatine is negatively and significantly correlated with development rate (head capsule size) of larvae reared in the fruits of the cherry and processing type tomatoes. Generally, survival, growth and development were significantly superior for larvae reared in the ripening fruits of the fresh market cultivar. At this stage, the fruits of this cultivar are also the largest. Based on these results it is concluded that fruit alpha-tomatine content, as well as fruit size and maturity, all affect performance of P. operculella larvae in the fruits of cultivated tomatoes.     

Potential of a granulovirus isolate to control <Emphasis Type="Italic">Phthorimaea operculella</Emphasis> (Lepidoptera: Gelechiidae)

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.     

马铃薯块茎蛾生物学、生态学与综合治理

马铃薯块茎蛾又称烟草潜叶蛾Phthorimaea operculella,起源于中美洲和南美洲北部地区,现已分布在亚洲、欧洲、北美洲、非洲等100多个国家,是茄科作物的世界性农业害虫,尤其对马铃薯有毁灭性的危害。目前,该虫在我国南方马铃薯产区普遍发生,尤其是在云南、四川、贵州等地区该害虫发生极为严重,且随着气候的变化该虫可能会扩散到其他马铃薯生产区。马铃薯块茎蛾主要进行两性生殖,少数孤雌生殖,其幼虫钻蛀叶片和薯块危害。初孵幼虫无性二态性,4龄幼虫、蛹和成虫均可依据外形特征进行雌雄区分。马铃薯块茎蛾发生世代数取决于当地的农业气候条件,年发生2~12代。马铃薯块茎蛾对温度有广泛的适应性,且在干燥炎热的年份该虫容易大爆发。马铃薯块茎蛾早期防控主要集中在种植抗性品种、深种、灌溉等农业防治措施上,但化学防治依然是马铃薯生产过程中防治马铃薯块茎蛾的主要方式,由于化学农药的广泛使用,该虫对有机磷类、拟除虫菊酯类等杀虫剂均产生了不同程度的抗性;为了减少化学农药的使用,延缓抗药性的发展,发现并筛选到多种对马铃薯块茎蛾具有防治作用的天敌昆虫和昆虫病原微生物。以(E4,Z7) 十三碳二烯基乙酸酯和(E4,Z7,Z10) 十三碳三烯基乙酸酯为主要成分的马铃薯块茎蛾性信息素在马铃薯块茎蛾监测和防治中也取得了较好的效果。桉树、皱叶薄荷等植物源化合物能够抑制马铃薯块茎蛾产卵;转基因抗虫马铃薯、遗传不育技术等绿色防控技术也成为了防控马铃薯块茎蛾的新方法。以往的研究发现使用单一生物防治手段很难达到理想的防控效果,集成与生物防控技术相容的化学物质、自然天敌和病原微生物等技术是有效控制马铃薯块茎蛾种群的重要趋势。本文系统综述了国内外马铃薯块茎蛾发生为害规律及综合防控技术研究进展,以期为马铃薯块茎蛾的持续治理提供参考依据。     

Tethered flight of the potato moth, Phthorimaea operculella

ABSTRACT. The effects of sex, age and mated state on average flight speed, duration and distance were determined for potato moths, Phthorimaea operculella (Zeller) (Lepidoptera, Gelechiidae), tethered to flight mills. Moths were classified as non-fliers (NFs), good-fliers (GFs) and remaining-fliers (RFs) on the basis of their performance over the first two flights. Some moths flew for over 5 h non-stop, while others tethered overnight flew between 20 and 30 km. Speed, duration and distance flown were greater during the first flight. First flight duration and distance flown by females decreased with age, whereas no trend was evident for males. Mated males and females flew slower first flights with increasing age, whereas virgin moths showed no marked trend. The analysis of fliers and NFs revealed that GFs were heavier than both RFs and NFs, GFs were faster than RFs, the percentage of NFs increased with age especially for mated females, and the percentage of GFs decreased with age. Age and mated state are important factors influencing flight performance especially for female moths. The relevance of these results to the field situation and the possible application of tethered flight to tests of potato moth quality are discussed.     

Resistance of wild Lycopersicon species to the potato moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae)

Abstract  Trichome-based host plant resistance of Lycopersicon species to potato moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), was examined in a laboratory bioassay. Neonate P. operculella were placed on the foliage of accessions of L. hirsutum , L. hirsutum f. glabratum , L. pennellii , L. cheesmanii f. minor and L. esculentum for 48 h . At the end of this period, larval mortality, the numbers of larvae emigrating from, and mining into, the leaf surface were recorded as were leaf area and the densities of trichomes. Analysis by categorical logistic regression found accessions differed in levels of resistance to P. operculella ( P  < 0.001). Further analysis using generalised linear models showed mortality was associated with high densities of type VI and low densities of type V trichomes; emigration from the leaf was associated with high densities of type I and type VII trichomes; and the numbers of mines associated with low densities of type I and type VI trichomes and increased leaf area. Although results indicate that certain accessions of L. hirsutum and L. hirsutum f. glabratum may be most appropriate for inclusion in a breeding program aimed at introducing trichome-based host plant resistance of wild Lycopersicon species into the tomato, further research is necessary to fully understand the mechanisms.     

Yield and activity of Autographa californica multicapsid nucleopolyhedrovirus and Phthorimaea operculella granulosis virus in cloned and uncloned cell lines of P. operculella

Three selected uncloned Pop 2, Pop 3, Pop 4 and two cloned cell lines Pop cl1A and Pop cl2B were derived from the original cell line established from Phthorimaea operculella (ORS-Pop-93). Three new non-selected cell lines ORS-Pop-94A, ORS-Pop-94B and ORS-Pop-95 were also established from embryos of the same insect. Differences in morphology, growth rate and polypeptide profile were determined between these cell lines. All the cell lines were susceptible to the Autographa californica nucleopolyhedrovirus (AcMNPV). The cloned cell lines produced higher levels of AcMNPV (TCID-50 and PIB) than the parental cells and at the same rate as the Sf9 reference cell line. Substantial amounts of viral DNA were synthesized in the clone Pop cl 2B after infection with the granulosis virus of the potato tuber moth P. operculella (PTMGV) and a complete multiplication was obtained in the ORS-Pop-95 cell line. The comparison between Pop cell lines which support limited or complete replication of certain baculoviruses can offer insights into some of the molecular barriers which restrict the host range of these viruses. These cell lines with variable susceptibility to baculoviruses could also be used for in vitro recombinations, increasing their virus host range to be used for the control of this pest. This revised version was published online in August 2006 with corrections to the Cover Date.     

Pupation site preference and body orientation of the potato tuber moth, Phthorimaea operculella

ABSTRACT. Pupation site preference and body orientation of potato tuber moth larvae, Phthorimaea operculella Zeller (Geleehiidae), were examined in the laboratory. The larvae preferred a dry, enclosed, dark place as their pupation site. Dryness was the strongest of the three categories of stimulus; the second was stimuli evoking thigmotaxis; the third was darkness. Having selected a pupation site, a larva oriented its head to the open side of the site. If two or more openings were present, final orientation was influenced by light and gravity. These preferences and orientation habits enhance pupal protection, and the ultimate successful exit of the adult pupation crevices.     

马铃薯块茎蛾的交配行为

在温度为(24±2)℃、光周期为L∶D=14∶10、相对湿度为50%～70%环境条件下观察了马铃薯块茎蛾Phthorimaea operculella(Zeller)交配行为。结果表明,1～2日龄、3～4日龄成虫交配高峰期分别发生在进入暗期后的8.0～10.0h、6.5～8.0h内。两性交配率随日龄的增加显著下降。马铃薯块茎蛾雄成虫具有多次交配能力,在连续发生2次以上行为交配时会导致雄蛾有效交配率下降。雄蛾平均有效交配次数可达3.8次/头,最高达6次。增加雄性比例不会显著提高单雌产卵量,但将性比(♀:♂)由1∶1升至2∶1时即可显著降低单雌产卵量。     

Effects of gamma irradiation used to inhibit potato sprouting on potato tuber moth eggs Phthorimaea operculella Zeller (Lep., Gelechiidae)

Abstract: Different age groups of potato tuber moth, Phthorimaea operculella Zeller, eggs were exposed to gamma irradiation at incremental doses up to 150 Gy (the upper dose limit allowed for potato sprout inhibition). Young eggs were more likely to be sensitive to gamma irradiation than older eggs and the sensitivity level declined with age of the eggs. The exposure of 3–3.5-day-old eggs to 150 Gy resulted in a 26% increase in the egg incubation period compared with unirradiated eggs. The minimum dose required to prevent 4–4.5-day-old eggs from hatching was 1 kGy. When eggs were exposed to 75 Gy, 9.7% of larvae survived to the adult stage but emerged as deformed moths. At 100 and 125 Gy, 10.2 and 9.6% of larvae pupated, respectively, although no adult eclosion was recorded, whereas at 150 Gy, the larvae remained in mid-instar stage and eventually died. Gamma irradiation doses applied to inhibit potato sprouting could be also considered as a valuable control tool against potato tuber moth infestations.     

Population biology of the potato tuber moth, Phthorimaea operculella (Lepidoptera: Gelechiidae), in two potato cropping systems in Israel

The life cycle, within-field distribution, crop damage and impact of natural enemies of the potato tuber moth, Phthorimaea operculella (Zeller) were investigated in two potato cropping systems. The two systems differed in soil type (sandy vs. loess) which in turn affected the choice of cultivars, irrigation programme, insecticide application method (ground vs. aerial), and planting and harvest times. From mid-April to the end of May, almost twice as many moths were caught in pheromone traps in sandy than in loess fields. Highest infestation of tubers was found before harvest, and infestation was greater in loess than in sandy fields. Larval densities in foliage and tubers were significantly higher at the margins of the fields than in the centre. A significant positive correlation was found between adult catch and larval infestation on foliage in sandy fields but not in loess. Tuber infestation in sand was positively correlated with foliage infestation. No such correlation was detected in loess. Five parasitic wasps emerged from P. operculella larvae collected from commercial fields and volunteer plants: Diadegma pulchripes (Kokujev) and Temelucha decorata, (Gravenhorst) (Ichneumonidae) and Bracon gelechiae Ashmead and two other unidentified Braconidae. The most abundant predators at the field site were Coccinella septempunctata Linnaeus (Coccinellidae), Chrysoperla carnea Stephens (Chrysopidae), Orius albidipennis (Reuter) (Anthocoridae) and four ant species (Formicidae). Parasitism rate reached 40% and predation was estimated at 79%. Results are discussed with regard to the development of an integrated pest management programme for this important pest.     

Susceptibility of potato plants grown from tubers irradiated with stimulation doses of gamma irradiation to potato tuber moth, Phthorimaea operculella Zeller (Lep., Gelechiidae)

The feeding behaviour of potato tuber moth, Phthorimaea operculella Zeller (Lep., Gelechiidae), reared on leaves and tubers of potato plants, which were irradiated as seed (tubers) with stimulation doses of gamma irradiation (1, 3, 5 and 10 Gy), was studied. Significant differences in the larvae and pupae developmental time, pupal weight, mortality, fecundity and percentage egg hatch, were observed between insects fed on plants which resulted from the irradiated seeds and those of the control. It appears that leaves of potato plants grown from irradiated seeds particularly those of 3 Gy, became more attractive to the larvae, although the resulting tubers, with the exception of those of 10 Gy, became more resistant to potato tuber moth. Storing the tubers at ambient temperature conditions affected the degree of larval sensitivity. The leaves and tubers of 10 Gy-irradiated seeds became more suitable for insect development, indicating that the later dose may inhibit the production of secondary plant metabolities and chemical compounds.     

Histopathology and effect on development of the PhopGV on larvae of the potato tubermoth, Phthorimaea operculella (Lepidoptera: Gelechiidae)

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.     

Effect of gamma irradiation on sperm utilization in twice-mated female Phthorimaea operculella Zeller (Lep., Gelechiidae)

The effects of the second mating on fecundity and fertility of potato tuber moth (PTM) Phthorimaea operculella (Zeller), when females were mated with 450 Gy-irradiated and normal males or vice versa, were studied. The percentage of eggs fertilized by sperm of the second mating (P₂ value) was 0.99, indicating that sperm transferred during the last mating were predominantly utilized in egg fertilization. Females, mated first with irradiated males, remated after 2 days, whereas those mated with normal males, remated after 3.3 days. Fecundity of twice-mated females was higher than those mated only once. Females started to lay their eggs 1.9 days after the first mating, regardless of the type of male. However, virgin females did not lay eggs at all. Duration of copulation varied from 102 to 117 min for normal and irradiated males, respectively. The present study elucidated important aspects of mating behaviour of PTM which could improve the efficiency of its control by the sterile insect technique.