Can arbuscular mycorrhizal fungi from non-invaded montane ecosystems facilitate the growth of alien trees?

It is generally assumed that recruitment and expansion of alien species along elevation gradients are constrained by climate. But, if plants are not fully constrained by climate, their expansion could be facilitated or hindered by other factors such as biotic interactions. Here, we assessed the composition of arbuscular mycorrhizal fungi (AMF) in soils along an elevation gradient (i.e. 900 m, 1600 m, 2200 m and 2700 m a.s.l.) through a fungal DNA meta-barcoding approach. In addition, we studied in the greenhouse the effects of AMF on growth and phosphorous (P) nutrition of seedlings of the alien trees Gleditsia triacanthos, Ligustrum lucidum and Pyracantha angustifolia cultivated in soils from those elevations, spanning the elevation at which they already form monospecific stands (below 1450 m a.s.l.) and higher elevations, above their current range of distribution in montane ecosystems of Central Argentina. For comparison, we also included in the experiment the dominant native tree Lithraea molleoides that historically occurs below 1300 m a.s.l. Arbuscular mycorrhizal fungal community composition showed strong community turnover with increasing elevation. The effects of these AMF communities on plant growth and nutrition differed among native and alien trees. While P nutrition in alien species' seedlings was generally enhanced by AMF along the whole gradient, the native species benefited only from AMF that occur in soils from the elevation corresponding to its current altitudinal range of distribution. These results suggest that AMF might foster upper range expansion of these invasive trees over non-invaded higher elevations.

Chemodiversity and Antiproliferative Activity of the Essential Oil of Schinus molle Growing in Jordan.

The leaves and unripe and fully-grown fruits of Schinus molle were collected from three geographical regions of Jordan: Amman (the Mediterranean), Madaba (Irano-Turanean), and Sahab (Saharo-Arabian). The hydrodistilled volatile oils of fresh and dried leaves and fruits were analyzed by gas chromatography-mass spectrometry (GC/MS). The actual composition of the emitted volatiles was determined using Solid Phase Micro-Extraction (SPME). α- and ß-Phellandrenes were the major components in all the analyzed samples. Quantitative differences were observed in the obtained essential oils (0.62-5.25 %). Additionally, cluster analysis was performed. Biologically, the antiproliferative activity of the essential oil, ethanol, and water extracts of the fruits and leaves was screened on Caco2, HCT116, MCF7, and T47D cell lines. The essential oil and ethanol extracts exhibited a dose-dependent inhibition of cell growth with IC50 ranging between 21 and 65 µg/mL. The water extract did not exhibit any antiproliferative activity against the investigated cell lines.

Arthropod Facilitation by Wood-Boring Beetles: Spatio-temporal Distribution Mediated by a Twig-girdler Ecosystem Engineer.

The twig-girdler beetle Oncideres albomarginata chamela (Chemsak and Giesbert) (Cerambycidae: Lamiinae) detaches branches of Spondias purpurea L. (Sapindales: Anacardiaceae) that fall on the forest floor or remain suspended on vegetation. Many wood-boring beetles also oviposit in these branches and larval development creates cavities that are abandoned when the adults emerge. The objective of this study was to evaluate the role of wood-boring beetles as facilitators by creating new habitats for arthropods, and test for vertical stratification and temporal variation of arthropods associated with S. purpurea branches that were previously engineered by O. albomarginata chamela in a tropical dry forest (TDF) in Jalisco, Mexico. In order to determine the effects of vertical strata and seasons on branch colonization by arthropods, we placed 60 branches on the forest floor (ground stratum) and 60 were placed in trees (vegetation stratum) from February to April (dry season), and from August to October 2016 (rainy season), for 240 branch samples in total. We collected 8,008 arthropods, which included 7,753 ants (14 species) and 255 nonsocial arthropods (80 species) from 13 different orders. We observed a greater arthropod abundance in the branches in the vegetation stratum in the dry season compared with the rainy season, whereas the richness and abundance of arthropods in the ground stratum were greater in the rainy season compared with the dry season. We concluded that wood-boring beetles are important habitat facilitators for arthropods, and that the vertical position of branches and the seasonal variations in TDFs differently affect the colonization of the abandoned cavities by arthropods.

Diameter distribution in a Brazilian tropical dry forest domain: predictions for the stand and species.

Currently, there is a lack of studies on the correct utilization of continuous distributions for dry tropical forests. Therefore, this work aims to investigate the diameter structure of a brazilian tropical dry forest and to select suitable continuous distributions by means of statistic tools for the stand and the main species. Two subsets were randomly selected from 40 plots. Diameter at base height was obtained. The following functions were tested: log-normal; gamma; Weibull 2P and Burr. The best fits were selected by Akaike's information validation criterion. Overall, the diameter distribution of the dry tropical forest was better described by negative exponential curves and positive skewness. The forest studied showed diameter distributions with decreasing probability for larger trees. This behavior was observed for both the main species and the stand. The generalization of the function fitted for the main species show that the development of individual models is needed. The Burr function showed good flexibility to describe the diameter structure of the stand and the behavior of Mimosa ophthalmocentra and Bauhinia cheilantha species. For Poincianella bracteosa, Aspidosperma pyrifolium and Myracrodum urundeuva better fitting was obtained with the log-normal function.

Restoration over time and sustainability of Schinus terebinthifolius Raddi.

The success of recovery programs on degraded areas is dependent on the genetic material to be used, which should present heterozygosity and genetic diversity in native and recovered populations. This study was carried out to evaluate the model efficiency to enable the recovery of a degraded area of the Lower São Francisco, Sergipe, Brazil. The target species for this study was Schinus terebinthifolius Raddi. Three populations were analyzed, the recovered area, seed-tree source population, and native tree population border established to the recovered area. The random amplified polymorphic DNA (RAPD) markers were used for diversity analysis. Genetic structure was estimated to evaluate the level of genetic variability existent in each population. There was no correlation between the spatial distribution and the genetic distances for all trees of the recovered area. The heterozygosity present in the recovered population was higher than the native tree population. The seed-tree source population presents genetic bottlenecks. Three clusters were suggested (ΔK = 3) with non-genetic structure. High intra-population genetic variability and inter-population differentiation are present. However, gene flow may also introduce potentially adaptive alleles in the populations of the recovered area, and the native population is necessary to ensure the sustainability and maintenance of the populations by allelic exchange.

Disentangling the effects of shrubs and herbivores on tree regeneration in a dry Chaco forest (Argentina).

Successful persistence of dry forests depends on tree regeneration, which depends on a balance of complex biotic interactions. In particular, the relative importance and interactive effects of shrubs and herbivores on tree regeneration are unclear. In a manipulative study, we investigated if thornless shrubs have a direct net effect, an indirect positive effect mediated by livestock, and/or an indirect negative effect mediated by small vertebrates on tree regeneration of two key species of Chaco forest (Argentina). In a spatial association study, we also explored the existence of net positive interactions from thorny and thornless shrubs. The number of Schinopsis lorentzii seedlings was highest under artificial shade with native herbivores and livestock excluded. Even excluding livestock, no seedlings were found with natural conditions (native herbivores present with natural shade or direct sunlight) at the end of the experiment. Surprisingly, seedling recruitment was not enhanced under thornless shrubs, because there was a complementary positive effect of shade and interference. Moreover, thornless shrubs had neither positive nor negative effects on regeneration of S. lorentzii. Regeneration of Aspidosperma quebracho-blanco was minimal in all treatments. In agreement with the experiment, spatial distributions of saplings of both tree species were independent of thornless shrubs, but positively associated with thorny shrubs. Our results suggest that in general thornless shrubs may have a negligible effect and thorny shrubs a net positive effect on tree regeneration in dry forests. These findings provide a conceptual framework for testing the impact of biotic interactions on seedling recruitment in other dry forests.

Initial development of the endocarp in Lithraea brasiliensis Marchand (Anacardiaceae): with taxonomic notes.

Investigation into the initial developmental stages of a given structure is fundamental for precise characterization as well as for comparative analysis in relation to other taxa when homologies are established. For the Anacardiaceae family, investigations of the initial development of the pericarp or its basic histological sites, the epicarp, mesocarp and endocarp, are relevant since these regions are of taxonomic and phylogenetic importance. The initial stages of endocarp development in Lithraea brasiliensis were studied using light microscopy. In L. brasiliensis, the fruits are of the drupe type. The endocarp originates exclusively in the epidermis of the locular cavity and is composed of only three strata. The crystalliferous layer, typical in fruits of the Anacardiaceae family, originates in the carpelar mesophyll and runs adjacent to the outermost layer of the endocarp. The endocarp in Lithraea brasiliensis is of the Anacardium type. The results of the present study are important for the identification of the Lithraea species that occur in the state of Rio Grande do Sul. In L. molleoides, the crystalliferous layer was shown to originate from the locular epidermis and this developmental aspect enables it to be differentiated from L. brasiliensis and demonstrates that there is variability of endocarp development in Lithraea.

Osmotic conditioning and shading on the germination and on the initial growth of Myracrodruon urundeuva Allemão seedlings.

The present work aimed to evaluate the osmotic conditioning and shading on the germination and on the initial growth of aroeira seedlings. The seeds were pre-imbibed in solutions with different concentrations of PEG (polyethylene glycol) and KNO3 and incubated at 10°C and 20°C temperatures, during 0 (control), 12 and 24 hours. After these periods, the seeds were dried until they reached the initial levels of humidity. After that, they were put for germinating in BOD (Biochemical Oxygen Demand) chambers, at alternated 20-30°C. The control treatment and the pre-conditionings that presented the best results in BOD germination were selected, PEG (-1.0 MPa) + KNO3 (-1.0 MPa) and KNO3 (-1.0 MPa), with pre-imbibition for 12 h and incubated in BOD at 20°C. The seeds were sowed on trays and then they were transplanted, keeping under a net covered with 50% and 70% of shading and at sunlight. The osmotic conditioning did not change the seeds germination in BOD, but the highest aerial part size was observed in PEG -1.0 MPa + KNO3-1.0 MPa treatment. The shading levels at 50% and 70% and the osmotic conditioning with PEG -1.0 MPa + KNO3-1.0 MPa offered a higher emergence in a greenhouse condition; however, at sunlight the seeds presented a better index of quality on the 145th day.

Developmental pathway from leaves to galls induced by a sap-feeding insect on Schinus polygamus (Cav.) Cabrera (Anacardiaceae).

Galling sap-feeding insects are presumed to cause only minor changes in host plant tissues, because they usually do not require development of nutritive tissues for their own use. This premise was examined through comparison of the histometry, cytometry and anatomical development of non-galled leaves and galls of Calophya duvauae (Scott) (Hemiptera: Calophyidae) on Schinus polygamus (Cav.) Cabrera (Anacardiaceae). Cell fates changed from non-galled leaves to galls during the course of tissue differentiation. C. duvauae caused changes in dermal, ground, and vascular systems of the leaves of S. polygamus. Its feeding activity induced the homogenization of the parenchyma, and the neoformation of vascular bundles and trichomes. The histometric and cytometric data revealed compensatory effects of hyperplasia and cell hypertrophy in the epidermis, with hyperplasia predominating in the adaxial epidermis. There was a balance between these processes in the other tissues. Thus, we found major differences between the developmental pathways of non-galled leaves and galls. These changes were associated with phenotypic alterations related to shelter and appropriate microenvironmental conditions for the gall inducer. The nondifferentiation of a typical nutritive tissue in this case was compared to other non-phylogenetically related arthropod gall systems, and is suggested to result from convergence associated with the piercing feeding apparatus of the corresponding gall-inducer.

The bird community in naturally regenerating Myracrodruon urundeuva (Anacardiaceae) forest in southeastern Brazil.

Conservation and restoration projects involving abandoned cattle pastures pose several challenges. Restoration ecologists recognize that natural restoration is possible, but few species can successfully colonize degraded pastures. The aim of the present study was to evaluate whether land colonized by the tree Myracrodruon urundeuva increase the amount of habitat available for bird communities in Southeastern Brazil. We used fixed point counts within a 50 m radius to compare the richness and composition of the bird community inhabiting naturally regenerated M. urundeuva with that an adjacent native forest remnant and a forest restored with planted Acacia spp. We recorded a total of 68 bird species. The M. urundeuva forest was able to provide valuable habitat for bird species, including more forest species, in comparison with the Acacia planting. These results indicate that M. urundeuva is a useful species for the ecological restoration of degraded pasture land.

Analysis of umbu (Spondias tuberosa Arruda (Anacardiaceae)) in different landscape management regimes: a process of incipient domestication?

Plant domestication is an evolutionary process guided by human groups who modify the landscape for their needs. The objective of this study was to evaluate the phenotypic variations between populations of Spondias tuberosa Arruda (umbuzeiro) when subjected to different local landscape management strategies. The influence of the landscape management system on these populations was evaluated in five identified regional units (mountains, base of mountains, pastures, cultivated areas and home gardens). Ten individuals were randomly selected from each region and subjected to morphological and chemical fruit analysis. The diversity index, based on Simpson's index, was determined for the different populations. We then evaluated the morphological differences between the individual fruits from the distinct landscape areas. We observed no significant differences in morphological diversity between the areas studied. Our data suggest that the umbuzeiro specimens in this region may be in the process of incipient domestication.

Habitat influence on antioxidant activity and tannin concentrations of Spondias tuberosa.

CONTEXT: Different habitat conditions can be responsible for the production of secondary metabolites and for the antioxidant properties of plant products. OBJECTIVE: Thus, the aim of this study was to evaluate whether the antioxidant activity and tannin concentrations in the stem bark of Spondias tuberosa Arruda (Anacardiaceae) varied with collection site. MATERIAL AND METHODS: The bark was collected from 25 individual trees, distributed in five different landscape units, as follows: agroforestry gardens, areas of pastures, maize cultivation areas, mountain areas and mountain bases, with the former 3 being considered as anthropogenic habitats, and the latter 2 considered as habitats with native coverage. The study was conducted in the rural area of the city of Altinho, Pernambuco State (Northeast Brazil). The DPPH (1,1-diphenyl-2-picrylhydrazyl) method was used to measure the antioxidant activity and tannin concentrations were evaluated by using the radial diffusion method. RESULTS: The results demonstrated that there were no significant differences among the tannin concentrations of the individuals from the native (6.27% ± 1.75) or anthropogenic areas (4.63% ± 2.55), (H = 2.24; p > 0.05). In contrast, there were significant differences (H = 5.1723; p < 0.05) among the CE50 means of the antioxidant activities of the individuals from the native (32.10 µg/ml ± 5.27) and anthropogenic areas (27.07 µg/ml ± 2.29). However, correlations between the tannin concentrations and antioxidant activity of the extracts were not observed in the native (r = 0.39; p > 0.05) or in the anthropogenic areas (r = 0.38; p > 0.05). DISCUSSION AND CONCLUSION: Because the variation of the antioxidant capacity of S. tuberosa bark was not accompanied by a variation in the tannin concentration, this property may be related to the presence of other metabolite(s).

Anatomy and genetic diversity of two populations of Schinus terebinthifolius (Anacardiaceae) from the Tibagi River basin in Paraná, Brazil.

Knowledge of the effects of flooding on plant survival is relevant for the efficiency of management and conservation programs. Schinus terebinthifolius is a tree of economic and ecological importance that is common in northeast Brazil. Flooding tolerance and genetic variation were investigated in two riparian populations of S. terebinthifolius distributed along two different ecological regions of the Tibagi River basin. Flooding tolerance was evaluated through the investigation of young plants, submitted to different flooding intensities to examine the morphological and anatomical responses to this stress. The growth rate of S. terebinthifolius was not affected by flooding, but total submersion proved to be lethal for 100% of the plants. Morphological alterations such as hypertrophied lenticels were observed in both populations and lenticel openings were significantly higher in plants from one population. Genetic analysis using DNA samples obtained from both populations showed a moderate degree of genetic variation between populations (13.7%); most of the variation was found within populations (86.3%). These results show that for conservation purposes and management of degraded areas, both populations should be preserved and could be used in programs that intend to recompose riparian forests.

Bark regeneration and tannin content in Myracrodruon urundeuva Allemão after simulation of extractive damages--implications to management.

Due to the current exploitation and consequent extinction of native medicinal plants around the world, new strategies have been proposed to address the sustainable use of this resource. Accordingly, the goal of this study is to assess the speed of bark regeneration of Myracrodruon urundeuva Allemão and to compare the tannin content before and after tissue regeneration. Twenty individuals from an area of Caatinga in Caruaru, Pernambuco State (NE Brazil), were selected. To evaluate the speed of bark regeneration, four treatments were established, according to the area of bark removed (10 × 2, 6 × 5, 8 × 5 and 10 × 5 cm), with five replicates (five individuals of M. urundeuva Allemão) and three treatments of tannin content (bark removed before damage, as regenerated tissue, and non-impacted bark or control). At the end of 23 months of monitoring the regeneration, we found that only seven of the 20 individuals analyzed did not fully heal. No significant correlation between the monthly percent regeneration and average monthly precipitation was found. The tannin content varied according to the quantified tissue (after damage, regenerated and control) as well as between damage classes (20 cm(2)-41.64 to 63.53 mg; 30 cm(2)-49.25 to 67.54 mg; 40 cm(2)-31.69 to 67.44 mg; 50 cm(2)-34.08 to 48.53 mg). Despite the variations found, there was no significant difference between the measurement periods (p > 0.05) or damage classes (p > 0.05). The results showed that the regeneration rate was higher in individuals belonging to the 10 × 2 and 10 × 5 cm groups and that there is no correlation between precipitation and regeneration speed. Tannin levels did not vary significantly in the tissues before damage and after regeneration.

Functional differences within a guild of tropical mammalian frugivores.

Many plants interact with groups of mutualist pollinators and seed dispersers. A key issue for both basic ecology and conservation is whether the different species within these guilds of mutualist animals are functionally equivalent. Comparing the relative effects of sympatric mutualists is important for understanding the evolution of multispecies mutualisms and for predicting mutualism stability in the face of anthropogenic change. However, empirical comparisons of the population-level impacts of mutualist animals on their host plant are rare, particularly for seed dispersal mutualisms in species-rich ecosystems. We compared the influence of three seed-dispersing tropical mammals, lar gibbons (Hylobates lar), sambar deer (Rusa unicolor), and red muntjac deer (Muntiacus muntjak), on the demography of a shared host tree in Thailand, Choerospondias axillaris (Anacardiaceae). Sambar and muntjac dispersed far more C. axillaris seeds than did gibbons. While sambar deposited many seeds under female tree canopies, muntjac were the only disperser to move seeds to open microhabitats, where C. axillaris seed germination, seedling survival, and initial growth are enhanced. Using stage-based population models, we assessed how disperser-specific seed dispersal, variation in the frequency of canopy gap formation, and their interaction influenced the potential population growth of C. axillaris. Large differences in dispersal quantity and small differences in dispersal quality among sambar and gibbons resulted in similar and negligible impacts on the tree's population dynamics. Muntjac, by taking some of the seeds to open microhabitats, are projected to have a greater positive impact on C. axillaris demography than either sambar or gibbons. Model comparisons of population-level species impacts may allow us to predict which ecological interactions are at risk from loss of critical species.

[Pollen dispersion and reproductive success of four tree species of a xerophytic forest from Argentina]. / Dispersión polínica y éxito reproductive de cuatro especies arbóreas de un bosque xerófito de Argentina.

The "talares" in eastern Buenos Aires province, Argentina, are coastal xerophitic forests structured by few arboreal species surrounded by a lower and moister soil matrix. We studied the reproductive parameters of the most representative arboreal species (Celtis tala, Scutia buxifolia, Jodina rhombifolia, and Schinus longifolia). Pollen dispersion was studied through floral visitor traps (biotic dispersion) and using gravimetric pollen collectors (abiotic dispersion). The reproductive success (fruit formation rate) of the focal species was studied by enclosing flowers with different mesh bags. The reproductive system varied among the different species. C. tala was anemophilous and selfcompatible. S. buxifolia was entomophilous and floral visitors dependant. J. rhombifolia was entomophylous, although spontaneous autogamy could favor reproduction in the absence of pollinators. Lastly, S. longifolia could be an ambophilous species (pollinated by insects and by the wind). This dual system may be the result of system flexibility mechanism or an evolutionary transition.

Megastigmus seed wasp damage on native Schinus terebinthifolia drupes in ecological restoration area in Brazil.

Megastigmus transvaalensis Hussey (Hymenoptera: Torymidae) parasitizes drupes of Rhus genus plants in Africa and Schinus (Anacardiaceae) in South America. This exotic wasp damages Schinus terebinthifolia Raddi drupes in native forests and ecological restoration areas in Brazil. The objective of the present study was to investigate the precipitation, temperature and relative humidity effects on M. transvaalensis flight activity, and to determine the parasitism rate and sex ratio of this wasp on S. terebinthifolia plants. The study was conducted with yellow sticky traps and S. terebinthifolia drupes collected in an ecological restoration area, from August 2014 to September 2015, in the Sorocaba municipality, São Paulo state, Brazil. Megastigmus transvaalensis populations were negatively correlated with maximum and minimum temperatures and precipitation, with population peaks at the end of May 2015, with 927 insects per evaluation (48.8 adults per trap). The M. transvaalensis sex ratio was higher in the laboratory (0.42) than in the field (0.08). The parasitism rate of S. terebinthifolia drupes by M. transvaalensis ranged from zero to 36.3% under natural environmental conditions. Megastigmus transvaalensis can be monitored with yellow sticky traps. Damage by M. transvaalensis in S. terebinthifolia drupes may decrease the germination of the seeds and the establishment of this plant in native and restoration ecological areas.

The history of Seasonally Dry Tropical Forests in eastern South America: inferences from the genetic structure of the tree Astronium urundeuva (Anacardiaceae).

Today, the Seasonally Dry Tropical Forests (SDTF) of eastern South America occur as large, well-defined nuclei (e.g. Caatinga in the northeast) and as smaller enclaves within other vegetations (e.g. Cerrado and Chaco). In order to infer the way the present SDTF distribution was attained, the genetic structure of Astronium urundeuva, a tree confined to SDTF, was assessed using two chloroplast spacers and nine microsatellite loci. Five haplotypes were identified, whose distribution was spatially structured. The distribution of the two most common and divergent haplotypes suggested former vicariance and progressive divergence due to isolation. More recent range expansions of these two lineages subsequently occurred, leading to a secondary contact at the southern limit of the Caatinga SDTF nucleus. The multilocus-Bayesian approach using microsatellites consistently identified three groups of populations (Northeast, Central and Southwest). Isolation by distance was found in Northeast and Southwest groups whereas admixture was detected in the Central group, located at the transition between Caatinga and Cerrado domains. All together, the results support the existence of range expansions and secondary contact in the Central group. This study provides arguments that favour the existence of a previously more continuous formation of SDTF in eastern South America.

Allocation to reproduction and relative reproductive costs in two species of dioecious Anacardiaceae with contrasting phenology.

BACKGROUND AND AIMS: The cost of reproduction in dioecious plants is often female-biased. However, several studies have reported no difference in costs of reproduction between the sexes. In this study, the relative reproductive allocation and costs at the shoot and whole-plant levels were examined in woody dioecious Rhus javanica and R. trichocarpa, in order to examine differences between types of phenophase (i.e. physiological stage of development). METHODS: Male and female Rhus javanica and R. trichocarpa were sampled and the reproductive and vegetative allocation of the shoot were estimated by harvesting reproductive current-year shoots during flowering and fruiting. Measurements were made of the number of reproductive and total current-year shoots per whole plant, and of the basal area increment (BAI). The numbers of reproductive and total current-year shoots per 1-year-old shoot were counted in order to examine the costs in the following year at the shoot level. KEY RESULTS: A female-biased annual reproductive allocation was found; however, the ratio of reproductive current-year shoots per tree and the BAI did not differ between sexes in Rhus javanica and R. trichocarpa. The percentage of 1-year-old shoots with at least one reproductive current-year shoot was significantly male-biased in R. trichocarpa, but not in R. javanica, indicating that there was a relative cost at the shoot level only in R. trichocarpa. The female-biased leaf mass per shoot, an indicator of compensation for costs, was only found in R. javanica. CONCLUSIONS: Relative reproductive costs at the shoot level were detected in Rhus trichocarpa, which has simultaneous leafing and flowering, but not in R. javanica, which has leafing followed by flowering. However, the costs for the whole-plant level were diminished in both species. The results suggest that the phenophase type may produce the different costs for R. javanica and R. trichocarpa through the development of a compensation mechanism.

Factors influencing the release of volatiles in Anastrepha obliqua males (Diptera: Tephritidae).

During sexual signaling, males of Anastrepha obliqua (Macquart) form leks and attract females by producing sounds and releasing volatile compounds. The objective of this study was to investigate the influence time of time of day, age, irradiation, mating status, and the presence of host fruit on the release of volatile components by laboratory-reared A. obliqua males. A. obliqua males released four compounds identified by gas chromatography coupled with mass spectrometry (GC-MS) as (E,E)-alpha-farnesene, (Z,E)-alpha-farnesene, (Z)-3-nonenol, and a farnesene isomer during calling behavior. Although males released volatiles during the entire day, the amount of volatiles was higher in the early morning and afternoon hours. Males began to emit volatiles when they were 6 d old, but only the amount of (Z)-3-nonenol was affected by age. Irradiation of flies with gamma rays from a 60Co source at a dose rate of 80 [corrected] Gy significantly reduced the amount of volatiles released during calling behavior. The amount of volatiles was lower in recently mated males than virgin males. However, there were no significant differences between virgin males and males 24 h after mating. The presence of host fruit did not affect the amount of volatiles compared with the control.