Bactrocera dorsalis s.s. is a pestiferous tephritid fruit fly distributed from Pakistan to the Pacific, with the Thai/Malay peninsula its southern limit. Sister pest taxa, B. papayae and B. philippinensis, occur in the southeast Asian archipelago and the Philippines, respectively. The relationship among these species is unclear due to their high molecular and morphological similarity. This study analysed population structure of these three species within a southeast Asian biogeographical context to assess potential dispersal patterns and the validity of their current taxonomic status.
Bactrocera dorsalis s.s. (Hendel) and B. papayae Drew & Hancock, are invasive pests belonging to the B. dorsalis complex. Their species status, based on morphology, is sometimes arguable. Consequently, the existence of cryptic species and/or population isolation may decrease the effectiveness of the sterile insect technique (SIT) due to an unknown degree of sexual isolation between released sterile flies and wild counterparts. To evaluate the genetic relationship and current demography in wild populations for guiding the application of area-wide integrated pest management using SIT, seven microsatellite-derived markers from B. dorsalis s.s. and another five from B. papayae were used for surveying intra- and inter-specific variation, population structure, and recent migration among sympatric and allopatric populations of the two morphological forms across Southern Thailand and West Malaysia.
Four morphologically cryptic species of the Bactrocera dorsalis fruit fly complex (B. dorsalis s.s., B. papayae, B. carambolae and B. philippinensis) are serious agricultural pests. As they are difficult to diagnose using traditional taxonomic techniques, we examined the potential for geometric morphometric analysis of wing size and shape to discriminate between them. Fifteen wing landmarks generated size and shape data for 245 specimens for subsequent comparisons among three geographically distinct samples of each species. Intraspecific wing size was significantly different within samples of B. carambolae and B. dorsalis s.s. but not within samples of B. papayae or B. philippinensis. Although B. papayae had the smallest wings (average centroid size=6.002 mm±0.061 SE) and B. dorsalis s.s. the largest (6.349 mm±0.066 SE), interspecific wing size comparisons were generally non-informative and incapable of discriminating species. Contrary to the wing size data, canonical variate analysis based on wing shape data discriminated all species with a relatively high degree of accuracy; individuals were correctly reassigned to their respective species on average 93.27% of the time. A single sample group of B. carambolae from locality 'TN Malaysia' was the only sample to be considerably different from its conspecific groups with regards to both wing size and wing shape. This sample was subsequently deemed to have been originally misidentified and likely represents an undescribed species. We demonstrate that geometric morphometric techniques analysing wing shape represent a promising approach for discriminating between morphologically cryptic taxa of the B. dorsalis species complex.
Sex pheromonal components of the tephritid fruit fly Bactrocera dorsalis (Hendel), 2-allyl-4,5-dimethoxyphenol and (E)-coniferyl alcohol, are biosynthesized from a highly potent male attractant, methyl eugenol, then sequestered and stored in the rectal gland prior to their release during courtship at dusk. These sex pheromonal components have been detected in the haemolymph and crop organ. Hence, attempts were made to separate and identify the haemolymph fractions which contained the sex pheromonal components. Identification of these bioactive fractions in methyl eugenol-fed male flies using gel filtration column chromatography and biodetection using live male flies showed two fractions as highly attractive to conspecific males. These fractions show a significant increase in protein absorbance in the elution profile of haemolymph from methyl eugenol-fed males compared with that from methyl eugenol-deprived males. The molecular mass of these bioactive fractions as determined by using gel filtration was in the peptide range of 3.3 to 5.5 kDa. Subsequent gas chromatography-mass spectrometry analyses further confirmed the presence of the pheromonal components in the bioactive fractions. The presence of these methyl eugenol-derived sex pheromonal components in specific haemolymph fractions suggests the involvement of a sex pheromone binding complex.
The Artocarpus fruit fly, Bactrocera umbrosa (Fabricius) (Diptera: Tephritidae), is an oligophagous fruit pest infesting Moraceae fruits, including jackfruit (Artocarpus heterophyllus Lamarck), a fruit commodity of high value in Malaysia. The scarcity of fundamental biological, physiological and ecological information on this pest, particularly in relation to behavioural response to phytochemical lures, which are instrumental to the success of many area-wide fruit fly control and management programmes, underpins the need for studies on this much-underrated pest. The positive response of B. umbrosa males to methyl eugenol (ME), a highly potent phytochemical lure, which attracts mainly males of many Bactrocera species, was shown to increase with increasing age. As early as 7 days after emergence (DAE), ca. 22% of males had responded to ME and over 50% by 10 DAE, despite no occurrence of matings (i.e. the males were still sexually immature). Male attraction to ME peaked from 10 to 27 DAE, which corresponded with the flies' attainment of sexual maturity. In wind-tunnel assays during the dusk courtship period, ME-fed males exhibited earlier calling activity and attracted a significantly higher percentage of virgin females compared with ME-deprived males. ME-fed males enjoyed a higher mating success than ME-deprived males at 1-day post ME feeding in semi-field assays. ME consumption also promotes aggregation behaviour in B. umbrosa males, as demonstrated in wind-tunnel and semi-field assays. We suggest that ME plays a prominent role in promoting sexual communication and enhancing mating performance of the Artocarpus fruit fly, a finding that is congruent with previous reports on the consequences of ME acquisition by other economically important Bactrocera species.
Males of Bactrocera dorsalis (Diptera: Tephritidae) are attracted strongly to and feed compulsively on methyl eugenol (1,2-dimethoxy- 4 -(2-propenyl)benzene), a highly potent male attractant. Pharmacophagy of methyl eugenol results in the production of phenylpropanoids 2-allyl-4,5-dimethoxyphenol and (E)-coniferyl alcohol that are sequestered and stored in the rectal gland prior to release as sex pheromonal components during mating at dusk. While these pheromonal components have also been detected in the hemolymph and crop of methyl eugenol-fed males, there is currently little information on the transport of these compounds from the crop to rectal gland in male B. dorsalis. Therefore, using physiological techniques such as parabiosis, rectal gland transplantation and hemolymph transfusion coupled with gas chromatography-mass spectrometry (GC-MS) analyses, we were able to ascertain and confirm the role of the hemolymph in the transport of these sex pheromonal components from the crop to the rectal gland. Further, the temporal profile of these methyl eugenol-derived bioactive compounds in the hemolymph also shows an increase with time post-methyl eugenol-feeding, i.e., 2-allyl-4,5-dimethoxyphenol attaining maximum amounts 15 min after ME consumption and decreasing thereafter, while for (E)-coniferyl alcohol-the increase and decrease are more gradual. These results further demonstrate the ability of insect hemolymph to transport many diverse forms of bioactive molecules including attractant-derived sex pheromonal components.
Bactrocera carambolae is a highly polyphagous fruit pest of agricultural importance. This study reports the bacterial communities associated with the developmental stages of B. carambolae. The microbiota of the developmental stages were investigated by targeted 16S rRNA gene (V3-V4 region) sequencing using the Illumina MiSeq. At 97% similarity, there were 19 bacterial phyla and unassigned bacteria, comprising 39 classes, 86 orders, 159 families and 311 genera. The bacterial composition varied among the specimens of developmental stage and across developmental stages as well as exuviae. Four phyla of bacteria (with relative abundance of ≥1% in at least one specimen)-Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria-were recovered from the larva, pupa, adult stages and exuviae. Proteobacteria was the predominant phylum in all the developmental stages as well as the exuviae. Enterobacteriaceae (Proteobacteria) was the predominant family in the adult flies while the family [Weeksellaceae] (Bacteroidetes) was predominant in the larval and pupal stages. Among the genera occurring in more than one developmental stage of B. carambolae, Erwinia was more abundant in the larval stage, Halomonas more abundant in adult female, Stenotrophomonas more abundant in adult male, and Chryseobacterium more abundant in the larval and pupal stages. The results indicate transmission of bacteria OTUs from immatures to the newly emerged adults, and from exuviae to the environment.
The male fruit fly attractants, cue-lure (CL) and raspberry ketone (RK), are important in pest management. These volatile phenylbutanoids occur in daciniphilous Bulbophyllum Thouar (Orchidaceae: Asparagales) orchids, along with zingerone (ZN) and anisyl acetone (AA). While these four compounds attract a similar range of species, their relative attractiveness to multiple species is unknown. We field tested these compounds in two fruit fly speciose locations in north Queensland, Australia (Lockhart and Cairns) for 8 wk. Of 16 species trapped in significant numbers, 14 were trapped with CL and RK, all in significantly greater numbers with CL traps than RK traps (at least in higher population locations). This included the pest species Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) (CL catches ca. 5× > RK), Bactrocera neohumeralis (Hardy) (Diptera: Tephritidae) and Bactrocera bryoniae (Tryon) (Diptera: Tephritidae) (CL catches ca. 3× > RK), and Bactrocera frauenfeldi (Schiner) (Diptera: Tephritidae) (in Cairns-CL catches ca. 1.6× > RK). Seven species were trapped with AA, and all were also caught in CL and RK traps in significantly greater numbers, with the exception of B. frauenfeldi. For this species, catches were not statistically different with CL, RK, and AA in Lockhart, and RK and AA in Cairns. Seven species were trapped with ZN, two at this lure only, and the remainder also with CL or RK but in significantly greater numbers. This is the first quantitative comparison of the relative attractiveness of CL, RK, AA, and ZN against multiple species, and supports the long-held but untested assumption that CL is broadly more attractive lure than RK.
Zeugodacus caudatus is a pest of pumpkin flowers. It has a Palearctic and Oriental distribution. We report here the complete mitochondrial genome of the Malaysian and Indonesian samples of Z. caudatus determined by next-generation sequencing of genomic DNA and determine their taxonomic status as sibling species and phylogeny with other taxa of the genus Zeugodacus. The whole mitogenome of both samples possessed 37 genes (13 protein-coding genes-PCGs, 2 rRNA and 22 tRNA genes) and a control region. The mitogenome of the Indonesian sample (15,885 bp) was longer than that of the Malaysian sample (15,866 bp). In both samples, TΨC-loop was absent in trnF and DHU-loop was absent in trnS1. Molecular phylogeny based on 13 PCGs was concordant with 15 mitochondrial genes (13 PCGs and 2 rRNA genes), with the two samples of Z. caudatus forming a sister group and the genus Zeugodacus was monophyletic. The Malaysian and Indonesian samples of Z. caudatus have a genetic distance of p = 7.8 % based on 13 PCGs and p = 7.0 % based on 15 mitochondrial genes, indicating status of sibling species. They are proposed to be accorded specific status as members of a species complex.
After pharmacophagy of methyl eugenol (ME), males of Bactrocera carambolae (Diptera: Tephritidae) produced (E)-coniferyl alcohol (CF) along with its endogenously synthesized pheromonal compounds. CF was shown to be released into the air by the ME-fed males only during the courtship period at dusk and attracted significantly more males and females than the ME-deprived males in wind tunnel assays. However, earlier onset of sexual attraction and a higher mating success were observed only in the wind tunnel and field cage assays on the third day posttreatment of ME. Field cage observations on the male-to-male interaction indicated that the ME-deprived males did not exhibit aggregation behavior, but that ME feeding promoted aggregation behavior in B. carambolae. Field cage observations revealed that the ME-deprived males were not only attracted to the ME-fed males, but also appeared to feed on their anal secretions. The secretions were subsequently confirmed to contain CF along with endogenously produced pheromonal compounds. Results obtained for B. carambolae were compared to those previously obtained from its sibling species, Bactrocera dorsalis, and are discussed in light of species advancement in fruit fly-plant relationships.
Bactrocera carambolae and B. papayae are major fruit fly pests and sympatric sibling species of the B. dorsalis complex. They possess distinct differences in male pheromonal components. In the 1990's, wild Bactrocera fruit flies with morphological traits intermediate between those of B. carambolae and B. papayae were often captured in traps baited with methyl eugenol (ME). Chemical analyses of rectal glands of ME-fed males revealed that the laboratory Fl, F2, and backcross hybrids possessed ME-derived sex pheromonal components ranging from that typical of B. papayae to that of B. carambolae without any specific trend, which included a combination of pheromonal components from both parental species within an individual hybrid. ME-fed hybrids without any ME-derived pheromonal components were also detected. Further chemical analysis of rectal glands from wild Bactrocera males, after ME feeding in the laboratory, showed a combination of pheromonal components similar to that found in the ME-fed, laboratory-bred hybrids. These findings present circumstantial evidence for the occurrence of a natural hybrid of the two Bactrocera species.
Bulbophyllum apertum flower (Orchidaceae) releases raspberry ketone (RK) in its fragrance, which attracts males of several fruit fly species belonging to the genus Bactrocera. Besides RK as a major component, the flower contains smaller amounts of 4-(4-hydroxylphenyl)-2-butanol, plus two minor volatile components, veratryl alcohol and vanillyl alcohol. Within the flower, the lip (labellum) had the highest concentration of RK with much smaller quantities present in petals; other flower parts had no detectable RK. Male fruit flies attracted to the flower belong to RK-sensitive species--such as Bactrocera albistragata, B. caudatus, B. cucurbitae (melon fly), and B. tau. Removal and attachment of the pollinarium to a fly's thoracic dorsum occurred when a male of B. albistragata was toppled into the floral column cavity, due to an imbalance caused by it shifting its body weight while feeding on the see-saw lip, and then freeing itself after being momentarily trapped between the lip and column. During this process, the stiff hamulus (the pollinia stalk protruding prominently towards the lip) acted as a crowbar when it was brushed downwards by the toppled fly and lifted the pollinia out of the anther. If the fly was big or long for the small triangular lip, it would not be toppled into the column cavity and would just walk across the column, during which time the pollinarium could be accidentally removed by the fly's leg, resulting in a failed transport of the pollinarium. This suggests an unstable situation, where the orchid relies only on a particular pollinator species in the complex ecosystem where many RK-sensitive species inhabit. Wild males of B. caudatus (most common visitors) captured on Bulbophyllum apertum flowers were found to sequester RK in their bodies as a potential pheromonal and allomonal ingredient. Thus, RK can act either as a floral synomone (pollinarium transported) or kairomone (accidental removal of pollinarium leading to total pollen wastage), depending on the body size of the male fruit flies visiting the flowers.
Sweet cherries, Prunus avium (L.) L., grown in the western United States are exported to many countries around the world. Some of these countries have enforced strict quarantine rules and trade restrictions owing to concerns about the potential establishment and subsequent spread of western cherry fruit fly, Rhagoletis indifferens Curran (Diptera: Tephritidae), a major quarantine pest of sweet cherry. We used 1) niche models (CLIMEX and MaxEnt) to map the climatic suitability, 2) North Carolina State University-Animal and Plant Health Inspection Service Plant Pest Forecasting System to examine chilling requirement, and 3) host distribution and availability to assess the potential for establishment of R. indifferens in areas of western North America where it currently does not exist and eight current or potential fresh sweet cherry markets: Colombia, India, Indonesia, Malaysia, Taiwan, Thailand, Venezuela, and Vietnam. Results from niche models conformed well to the current distribution of R. indifferens in western North America. MaxEnt and CLIMEX models had high performance and predicted climatic suitability in some of the countries (e.g., Andean range in Colombia and Venezuela, northern and northeastern India, central Taiwan, and parts of Vietnam). However, our results showed no potential for establishment of R. indifferens in Colombia, Indonesia, Malaysia, Taiwan, Thailand, Venezuela, and Vietnam when the optimal chilling requirement to break diapause (minimum temperature < or = 3 degree C for at least 15 wk) was used as the criterion for whether establishment can occur. Furthermore, these countries have no host plant species available for R. indifferens. Our results can be used to make scientifically informed international trade decisions and negotiations by policy makers.
Differentiation of Bactrocera papayae Drew & Hancock and Bactrocera carambolae Drew & Hancock (Diptera: Tephritidae) based on morphological characters has often been problematical. We describe here a single-nucleotide polymorphism (SNP)-based polymerase chain reaction (PCR) assay to differentiate between these two species. For detection of SNPs, fragments derived from each species were amplified using two primer pairs, COIF/COIR and UEA7/UEA10, sequenced, and aligned to obtain a contiguous 1,517-bp segment. Two new sets of primers were designed based on the 11 SNPs identified in the region. Results of the SNP-PCR test using any one of these species-specific primer sets indicate that these two species could be differentiated on basis of presence or absence of a band in the gel profile. We also tested the SNP-PCR primers on Bactrocera umbrosa F., Bactrocera cucurbitae Coquillett, Bactrocera latifrons Hendel, and Bactrocera tau (Walker) but did not detect any band in the gel, indicating the likelihood of a false positive for B. papayae is nil. This SNP-PCR method is efficient and useful, especially for immature life stages or when only adult body parts of the two species are available for identification, as encountered often in quarantine work.
The Malaysian fruit fly, Bactrocera latifrons (Hendel), is a pest of peppers (Capsicum spp.) in Thailand. A field trial was undertaken to determine whether five commonly used cultivars of C. annuum, with marked differences in morphology and pungency, varied in their susceptibility to infestation by B. latifrons. Experiments carried out in both the dry and rainy seasons showed temporal differences in the number of fruits per cultivar, but there was no effect of variety or season on the proportion of fruits attacked or the number of pupae obtained per infested fruit However, the number of dead larvae per infested fruit was significantly higher, and the percent of pupae giving rise to adults was lower for the larger sweet pepper than other cultivars tested.
The late-aged egg and third-instar life stages of laboratory-reared Malaysian fruit fly, Bactrocera latifrons (Hendel); Mediterranean fruit fly, Ceratitis capitata (Wiedemann); melon fly, B. cucurbitae Coquillett; and oriental fruit fly, B. dorsalis (Hendel), (Diptera: Tephritidae); and the third instars of wild Mediterranean fruit fly were exposed to thermal treatments. A heating block system was used to determine the thermal death kinetics of the four fruit fly species. Treatments consisted of heating the fruit fly life stages to 44, 46, 48, and 50 degrees C and holding for different times ranging from 0 to 120 min depending on the thermal mortality response and time required to obtain 100% mortality for each species and life stage. The 0.5-order kinetic model had the best fit to the survival ratio for all the treatment temperatures and was used to predict lethal times. The thermal death time (TDT) curves showed a tolerance order of Mediterranean fruit fly eggs < or = third instars at 44, 46, and 50 degrees C, third instars < or = eggs at 48 degrees C, and wild third instars < the laboratory-reared third instars. Comparison between Mediterranean fruit fly third instar thermotolerance from Hawaii and Israel showed that Israel Mediterranean fruit fly was more thermotolerant. A comparison of minimum treatment times at a given temperature required to obtain 100% mortality of laboratory-reared Malaysian, Mediterranean (Hawaii and Israel strains), melon, Mexican, and oriental fruit fly eggs or third instars and wild Mediterranean fruit fly (Hawaii strain) eggs or third instars showed that oriental fruit fly was the most thermotolerant among the third instars, and the difference in heat tolerance between third instars and eggs was negligible at 50 degrees C.
The aim of this study was to investigate the effect of gamma irradiation on survivability (adult emergence, sex ratio, adult longevity), fecundity (eggs hatchability, number of eggs produced), and morphological differences in the size of the ovary and testes of unirradiated and irradiated adults of Oriental fruit fly [Bactrocera dorsalis (Hendel)]. A dose of 100 Gy was determined as the minimum needed for inhibitory effects against B. dorsalis that would not deter adult emergence, sex ratio (male:female), and adult longevity, with 82.6 ±7.02, 1:1.09, and 107 ± 24.5, respectively. Doses from 50 to 400 Gy range did not completely prevent the adult emergence; however, the emergence decreased by increasing the radiation dose. Adult survivability significantly decreased among all the treatment groups of B. dorsalis, except for the 50 Gy and unirradiated flies. At a dose of 50 Gy, fertile females showed a significant reduction in fecundity by not producing eggs after mating with the sterile males. Meanwhile, the number of eggs laid decreased with increasing dose and no fertile egg was hatched starting at 100 Gy of irradiation. Testes and ovaries of 20-d old flies irradiated as pupae were smaller than those of control flies. In this study, 100 Gy was concluded as the minimum effective dose for the disinfestation and sterilization of B. dorsalis puparia. Results represent new findings used as a basis for sterile insect technique and quarantine programs for managing B. dorsalis, particularly in Malaysia.
Certain male fruit flies from the genera Bactrocera and Zeugodacus (Diptera: Tephritidae) actively forage for floral semiochemicals produced by some endemic Bulbophyllum orchids found in tropical and subtropical forests. These floral semiochemicals are largely classified as either phenylbutanoids (e.g., raspberry ketone (RK)) or phenylpropanoids (e.g., methyl eugenol (ME)). Zingerone (ZN) is a phenylbutanoid recently found that structurally resembles ME and RK, both of which are phytochemicals commonly used as male attractants in fruit fly control programmes. It was previously shown that feeding on ME and RK increased the mating success of certain tephritid fruit flies, specifically in B. dorsalis and B. tryoni males, respectively, through enhancement of sexual signaling. However, ZN, which acts as a metabolic enhancer to increase male courtship activity in B. tryoni, did not show the same effect. As fruit fly-phytochemical lure interactions are unique and species-specific phenomena, this study seeks to elucidate the ecological significance of ZN feeding to Zeugodacus tau in terms of sexual signaling. We demonstrate here that ZN feeding by Z. tau males enhanced female attraction and subsequent mating success by increasing male courtship, and the attractiveness of the sexual signals in both wind tunnel and semi-field cage bioassays. In addition, we also demonstrated temporal effects on male behaviour in relation to the amount of lure intake. However, feeding on ZN did not appear to affect the total time spent in copula for Z. tau. This is the first report showing an important role of ZN in increasing courtship activity as well as enhancement of sexual signaling in Z. tau males.
The oriental fruit fly, Bactrocera dorsalis (Handel) is one of the most destructive pests of fruits. The discovery of methyl eugenol (ME) as a potent male attractant for this species has led to its successful use in area-wide fruit fly control programs such as male annihilation. While the antenna is recognized as primarily responsible for male flies' detection of attractants such as ME, little is known of the involvement of the maxillary palp. Using behavioral assays involving males with intact and ablated antennae and maxillary palp structures, we seek to ascertain the relative involvement of the maxillary palp in the ability of the male fly to detect ME. In cage bioassays (distance of ≤40 cm from the source), >97% of unmodified males will normally show a response to ME. Here, we showed that 17.6% of males with their antennae ablated were still attracted to ME versus 75.0% of males with their palps ablated. However, none of the antennae-ablated males were able to detect ME over a distance of >100 cm. Furthermore, wind tunnel bioassays showed that maxillary palp-ablated males took a significantly longer time compared to unablated males to successfully detect and eventually feed on ME. These results suggest that although the antennae are necessary for detection of ME over longer distances, at shorter distances, both antennae and maxillary palps are also involved in detecting ME. Hence, those palps may play a larger role than previously recognized in maneuvering males toward lure sources over shorter ranges.
Sexually mature males of Bactrocera papayae are strongly attracted to and consume methyl eugenol (ME). Upon consumption, ME is biotransformed to two phenylpropanoids, 2-allyl-4,5-dimethoxyphenol (DMP) and (E)-coniferyl alcohol (CF), that are transported in the hemolymph, sequestered and stored in the rectal glands, and subsequently released as sex and aggregation pheromones during courtship. To date, very little work on the ultrastructure and anatomy of the rectal gland has been done, and the accumulation of phenylpropanoids in the rectal glands of males has not been observed visually. Our objectives are to describe the anatomy and fine structures of the rectal glands of males and females and to observe the accumulation of autofluorescent compounds in the rectal glands of males. The rectal glands of males and females have four rectal papillae with each papilla attached to a rectal pad. The rectal pads protrude from the rectal gland as the only surfaces of the gland that are not surrounded by muscles. The rectal papillae of ME-fed males had oil droplets and autofluorescent compounds that were absent from those of ME-deprived males. The autofluorescent compounds accumulated in the rectal sac, which is an evagination that is not found in rectal glands of females. The accumulation of these compounds increased with time and reached maximum at a day post-ME feeding and decreased thereafter. This trend is similar to the accumulation pattern of phenylpropanoids, CF and DMP in the rectal gland.