Early‐Season Headspace Volatiles from Apple and Their Effect on the Apple Blossom Weevil Anthonomus pomorum

Apple volatiles emitted at early phenological stages are little investigated, although they may influence behavior of early‐season pests. The apple blossom weevil Anthonomus pomorum is a herbivore pest of orchards in Europe. It colonizes apple trees in early season and oviposits into developing flower buds, often leading to economic damage. Using in situ radial diffusive sampling and thermal desorption, followed by GC/MS analysis, headspace volatiles from apple twigs with flower buds at three early phenological tree stages were identified and quantified. The volatile blend consisted of 13 compounds for the first, and increased to 15 compounds for the third phenological stage sampled. These blends included benzenoids, terpenes, and derivatives of fatty acids. A recombined synthetic blend served as the odor source in a still‐air dual‐choice olfactometer bioassay, in which individual male and female weevils were tested. Results from this behavioral test document an attraction of both sexes to odors of their host plant, suggesting that apple volatiles emitted in early season serve as olfactory cues for host location of A. pomorum in the field.     

Induced emissions of apple fruit volatiles by the codling moth: changing patterns with different time periods after infestation and different larval instars

The changes in the emission of volatiles from mature apple fruits in response to larval feeding by the codling moth (Cydia pomonella) under laboratory conditions are reported. A time course experiment investigated the emission of volatiles throughout the period of larval development following infestation. The volatiles consisted mainly of esters, a few aldehydes, and the terpene alpha-farnesene. Infested apples emitted the same compounds as healthy apples. The quantities of volatiles released were much higher for infested as compared to healthy fruits for an initial three day period. Following this period there was a decrease in volatile emissions (days 6-9), eventually declining back to the levels emitted from healthy apples or below by 9-21 days after infestation. In a separate experiment, the volatile emissions from healthy and artificially damaged fruits were compared to those from herbivore damaged fruits for each of the five larval instars of C. pommonella. The results from the discriminant analysis indicate that the most effective induction of volatiles occurred when fruits were infested with first instar larvae. Induction by first instar larvae was generally higher than after infestation by later instars, and for most compounds it also exceeded the emission from artificially damaged fruits.     

Parasitoid exploitation of the seasonal variation in host plant volatile emission for herbivore location

Volatile compounds from the apple, Malus domestica Borkh. (Rosaceae), change considerably as the season progresses, and this is successfully exploited by the female codling moth Cydia pomonella L. (Lepidoptera, Tortricidae), as it searches for oviposition sites. In this study, we investigated the effect of seasonal emissions of apple fruit volatiles on the host location behaviour of a parasitoid of the codling moth larvae, Hyssopus pallidus (Askew) (Hymenoptera: Eulophidae). In dual choice olfactory bioassays, the behaviour of the parasitoid in response to apple cues was observed over the complete 2003 growing season. Our results show that codling moth infested apples evoked a strong response from the parasitoid at the beginning of the season, until July. Then, attraction dropped drastically, increasing again at the beginning of August. At the end of the growing season, just prior to harvest, infested apples hardly evoked any behavioural response. Interestingly enough, mid‐season emissions of healthy apples were per se attractive to the parasitoid, and even preferred over volatiles from infested apples. Simultaneous volatile collections from healthy apples on twigs in the field were analysed throughout the season, showing that the overall quantity of headspace volatiles peaks at the beginning of June and mid‐August. The seasonal volatile emission is correlated with the behaviour of the parasitoid during the fruit ripening stage. The results are discussed in relation to the use of H. pallidus as a potential biocontrol agent, in order to enhance current integrated pest management (IPM) programs.     

How rainfall, relative humidity and temperature influence volatile emissions from apple trees in situ

Headspace volatiles from apple-bearing twigs were collected in the field with a Radiello sampler during three different diurnal periods over the complete fruit growing season. Analyses by thermal desorption-GC-MS identified a total of 62 compounds in changing quantities, including the terpenoids alpha-pinene, camphene, beta-pinene, limonene, beta-caryophyllene and (E,E)-alpha-farnesene, the aldehydes (E)-2-hexenal, benzaldehyde and nonanal, and the alcohol (Z)-3-hexen-1-ol. The variations in emission of these plant odours were statistically related to temperature, humidity and rainfall in the field. Remarkably, rainfall had a significant positive influence on changes in volatile release during all three diurnal periods, and further factors of significance were temperature and relative humidity around noon, relative humidity in the late afternoon, and temperature and relative humidity during the night. Rainfall was associated consistently with an increase in the late afternoon in terpene and aldehyde volatiles with a known repellent effect on the codling moth, one of the key pests of apple fruit. During the summer of 2003, a season characterized by below-average rainfall, some postulated effects of drought on trees were tested by establishing correlations with rainfall. Emissions of the wood terpenes alpha-pinene, beta-pinene and limonene were negatively correlated with rainfall. Another monoterpene, camphene, was only detected in this summer but not in the previous years, and its emissions were negatively correlated with rainfall, further supporting the theory that drought can result in higher formation of secondary metabolites. Finally, the two green leaf volatiles (E)-2-hexenal and (Z)-3-hexen-1-ol were negatively correlated with rainfall, coinciding well with the expectation that water deficit stress increases activity of lipoxygenase. To our knowledge, this work represents the first empirical study concerning the influence of abiotic factors on volatile emissions from apple trees in situ.     

白星花金龟对寄主植物挥发物的电生理反应

[目的]白星花金龟Protaetia brevitarsis是我国经济上重要的农业害虫.本研究拟鉴定苹果、葡萄和玉米3种果实在两种状态(健康与受白星花金龟成虫为害)下的挥发物中能引起白星花金龟雌成虫触角电生理反应的挥发物成分.[方法]采用动态顶空吸附法收集苹果、葡萄和玉米3种果实的挥发物,用气相色谱-触角电位联用技术(...     

Comparative analysis of peach and pear fruit volatiles attractive to the oriental fruit moth,Cydia molesta

Cydia molesta is an important pest of peach and pear fruits late in growing season. We identified and quantified volatiles from immature and mature fruits of peach and pear using gas chromatography-mass spectrometry. Antennally, active compounds were identified by gas chromatography-electroantennogram detection and were further tested in the field. Consistent electroantennographic activity was obtained for nine compounds from headspace collections of the mature fruits of two peach and two pear varieties. Esters were the major components, but qualitative and quantitative differences were found among four odor profiles. Blends mimicking pear fruit volatiles were more attractive to both sexes than blends mimicking peach fruit volatiles in both orchards. Our finding indicated that mixtures mimicking peach and pear fruit volatiles attracted both females and males of C. molesta, and these host volatiles may also be involved in the host switch behavior from peach to pear in mid-late season.     

Apple and peach fruit volatiles and the apple constituent butyl hexanoate attract female oriental fruit moth, Cydia molesta, in the laboratory

Volatiles emitted from immature and mature peach and apple fruits were all attractive to mated female oriental fruit moth, Cydia molesta (Busck), in a dual choice arena. Females did not discriminate between odours emitted by these two major host plants. The same natural blends were behaviourally ineffective for virgin females. A major component of apple fruit volatiles, butyl hexanoate, also attracted female C. molesta. Mated females were attracted to two medium dosages, while virgin females responded positively to the lowest of the five dosages tested. The time course of the captures of the moths shows a diurnal activity cycle known from the field. The possible implications of a semiochemical which attracts females are discussed in the context of previous findings that gravid females may immigrate from peaches into apple orchards particularly in the later phase of the season.     

Response of female oriental fruit moth to volatiles from apple and peach trees at three phenological stages

The attractiveness of peach ( Prunus persica L. Batsch) and apple ( Malus domestica L. Borkh.) (both Rosaceae) tissue to gravid female oriental fruit moth, Grapholita (=  Cydia ) molesta (Busck) (Lepidoptera: Tortricidae), was assessed at three distinct stages throughout the growing season using a dual-choice bioassay. Plant material offered to the female moths consisted of a plant shoot in early spring, before fruit became available. Later, it consisted of a leaf-bearing twig and a fruit, either alone or in combination. The level of attraction of the female moths to the various plant tissues varied substantially over time and according to the plant species. Before fruit became available, female moths were significantly attracted to peach as well as to apple shoots. During the early fruit growth stage, moths were attracted to a leaf-bearing twig originating from a peach tree, but not to that from an apple tree. In peach, it was the vegetative tissue that accounted for the attraction, whereas in apple, it was the reproductive tissue (a developing fruit). During the late fruit growth stage, both peach fruit and apple fruit were highly attractive, whereas a twig with leaves from either an apple or a peach tree was neither attractive nor repellent to the female moths. This changing female olfactory response to volatiles emitted by vegetative tissue and fruits from the two host plant species with progressing season is discussed with respect to the biology and the dispersal of this moth species.     

Behavioral and electrophysiological responses of the parasitic wasp Psyttalia concolor (Szépligeti) (Hymenoptera: Braconidae) to Ceratitis capitata-induced fruit volatiles

Psyttalia concolor (Szépligeti) is a koinobiont larval-pupal endoparasitoid of many Tephritidae of great economic importance, such as the medfly, Ceratitis capitata (Wiedemann). In several species of parasitoids it has been demonstrated that the mated females are strongly attracted by specific volatiles from insect-damaged plants. Yet the role of olfactory cues deriving from C. capitata-infested fruits on the female’s decision during the P. concolor host location was poorly investigated. In the present study, the responses of P. concolor females to either healthy or C. capitata-infested fruits was studied through behavioral assays. Volatiles emitted by healthy and infested fruits were SPME-sampled and analyzed by gas chromatography–mass spectrometry (GC–MS). The attractiveness of the identified volatiles was assessed and their electrophysiological activity was analyzed through gas-chromatography coupled with electroantennography (GC-EAD). P. concolor preferred infested peaches and apples over healthy ones, either when visual and olfactory or only olfactory cues were given. Nine compounds were found as exclusive of infested peaches, with respect to healthy ones, and seven of them evoked electrophysiological responses. In apples only quantitative changes in volatile emissions were observed after the medfly infestation. The emissions of 1-butyl butylate, 1-hexyl acetate and 1-butyl esanoate increased in infested apples, whereas 1-hexyl (E)-2-methyl butenoate decreased significantly. Among apple volatiles, 1-butyl butylate, 2-methyl-1-butyl acetate, 1-hexyl acetate, 2-methyl-1-butyl 2-methylbutanoate, 1-butyl hexanoate and 1-hexyl (E)-2-methyl butenoate elicited responses in female antennae. Synthetic blends reproducing the odors emitted by infested peaches and apples elicited strong attraction towards P. concolor females. For both fruits, the blend attractiveness was mainly due to some specific electrophysiological active chemicals: ethyl octanoate, decanal and 4-decanolide for peach, and 1-butyl butylate and 1-butyl hexanoate for apple. The responses induced by the identified fruit volatiles to P. concolor females allow us to suppose that they play a role as short-range attractants during host location.     

Plant volatiles mediate attraction to host and non-host plant in apple fruit moth, Argyresthia conjugella

Plant volatiles mediate host finding in insect herbivores and lead to host fidelity and habitat‐specific mating, generating premating reproductive isolation and facilitating sympatric divergence. The apple fruit moth, Argyresthia conjugella Zeller (Lepidoptera: Argyresthiidae), is a particularly suitable species to study the cues and behavioural mechanisms leading to colonization of a new host: it recurrently oviposits on the non‐host plant, apple Malus domestica Borkh. (Rosaceae), where the larvae cannot complete their development. The larval host of the apple fruit moth (Lepidoptera, Argyresthiidae), is rowan Sorbus aucuparia L. (Rosaceae). Fruit setting in rowan, however, fluctuates strongly over large areas in Scandinavia. Every 2–4 years, when too few rowanberries are available for egg laying in forests, apple fruit moth females oviposit instead on apple in nearby orchards, but not on other fruits, such as pear or plum. This poses the question of which cues mediate attraction to rowan and apple, and how apple fruit moth discriminates rowan from apple. Chemical analysis and antennal recordings showed that 11 out of 15 rowan volatiles eliciting an antennal response in A. conjugella females co‐occur in rowan and apple headspace, in a different proportion. In the field, A. conjugella was attracted to several of these plant volatiles, especially to 2‐phenyl ethanol, methyl salicylate, and decanal. Addition of anethole to 2‐phenyl ethanol had a strong synergistic effect, the 1 : 1 blend is a powerful attractant for A. conjugella males and females. These results confirm that volatiles common to both plants may account for a host switch in A. conjugella from rowan to apple. Some of the most attractive compounds, including 2‐phenyl ethanol, anethole, and decanal, which have been found in several apple cultivars, were not present in the headspace of the apple cultivar, Aroma, which is also susceptible to attack by A. conjugella. This supports the idea that the odour signal from apple is suboptimal for attraction of A. conjugella, but is nonetheless sufficient for attraction, during times when rowan is not available for egg laying.     

Volatiles from apple (Malus domestica) eliciting antennal responses in female codling moth Cydia pomonella (L.) (Lepidoptera: Tortricidae): effect of plant injury and sampling technique

The antennal responses of codling moth females, Cydia pomonella, to volatiles from apple branches with green fruits were recorded by electroantennography coupled to gas chromatography. The antennae strongly responded to 4,8-dimethyl-1,3(E),7-nonatriene, linalool, beta-caryophyllene, (E)-beta-farnesene, germacrene D, (Z,E)-alpha-farnesene, (E,E)-alpha-farnesene and methyl salicylate. These compounds were all present in volatile collections on Porapak Q from both living and cut branches. Analysis by the solid phase microextraction technique (SPME) showed that the emission of some electrophysiologically active compounds increased after branches had been cut, especially 4,8-dimethyl-1,3(E),7-nonatriene, linalool and (E,E)-alpha-farnesene. The identification of apple volatiles eliciting antennal responses is the first step towards the identification of compounds mediating host-finding and oviposition in codling moth females.     

Electrophysiological and behavioral response of the oriental fruit moth <Emphasis Type="Italic">Grapholita</Emphasis><Emphasis Type="Italic">molesta</Emphasis> (Busck, 1916) (Lepidoptera: Tortricidae) to the volatiles of apple fruits at different stages of development

The oriental fruit moth, Grapholita molesta, occurs in Southern Brazil throughout the year, and migrates from peach to apple orchards. Because moths rely on volatile organic compounds (VOCs) during the host-location process, variations in the emission of these compounds during fruit maturation can influence the time of infestation and preference of the moths for a particular genotype. The aim of this work was to identify VOCs emitted by the apples “Eva” and “Gala” at different stages of development and to determine the behavioral and electrophysiological responses of G. molesta to these compounds. For this purpose, VOCs from immature, maturing, and mature fruits of both cultivars were collected and analyzed by gas chromatography and gas chromatography-mass spectroscopy. The response of the antennae of virgin males and females and mated females to volatiles released by the three fruit stages was registered by gas chromatography coupled to an electroantennography. A dual-choice behavioral test for the different combinations of insect groups and fruit stage was also performed. Amongst the volatiles released by mature fruits, twelve compounds elicited a response. The antennae of the oriental fruit moth did respond to isoamyl hexanoate and α-farnesene emitted by “Eva” maturing fruits. In general, virgin females did not respond to volatiles in olfactometer bioassays and mated females were attracted to volatiles released by mature fruits. Our results show that the variation in the emission during the maturation of fruits can influence the orientation of G. molesta.     

A putative PhODO1 homologous MYB transcription factor gene, MdMYBB, is not involved in the regulation of aroma volatile biosynthesis in apple

To get deeper insight on the molecular mechanism underlying production of volatile compounds in apple (Malus domestica Borkh.), we performed the isolation and expression analysis of one R2R3-type MYB gene named MdMYBB. The amino acid sequence and the structural features of MdMYBB highly resembled those of PhODO1, which is a key regulator for floral scent biosynthesis in petunia. The expression of MdMYBB was repressed concomitantly with the inhibition of ethylene production, which regulates the volatile synthesis in apple. However, MdMYBB expression was not detected in the flesh from nearly ripened apple fruits, although the detection of exogenous volatiles had actually occurred in the same portion. In addition, overexpression of MdMYBB did not cause any induction of the volatile compounds in the transgenic tobacco lines. Thus, the features of MdMYBB were not in accordance with the aroma volatile emission, unlike the case of PhODO1, suggesting that MdMYBB may not be involved in the regulation of the biosynthesis for apple aroma volatiles. On the basis of the specific expression patterns, we discussed possible physiological roles of MdMYBB in apple.     

Visualization of Fruit Odor by Photoluminescence

Photoluminescence with visible emission spectrum was observed and visualized at the surface of certain fruits. This photoluminescence is associated with vapors of natural organic volatiles (odorants) emitted from the fruit surface. The photoluminescence spectra of various fruits (apple, pear, kiwi, and strawberry) were measured in vivo using a number of fluorometric methods. Fruit aging was found to be accompanied by modification of the photoluminescence spectrum shape and a noticeable increase in the photoluminescence intensity. Laser photoluminescence microscopy in vapors of fruit extracts and artificial compounds was used to assess the contribution of various substances to natural odor emission of fruits. The results of this study show that fluorometry of odors is a promising method for studying fruits and other objects.     

Visualization of fruit scent using photoluminescence

Photoluminescence with visible emission spectrum was observed and visualized at the surface of certain fruits. This photoluminescence is associated with vapors of natural organic volatiles (odorants) emitted from the fruit surface. The photoluminescence spectra of various fruits (apple, pear, kiwi, and strawberry) were measured in vivo using a number of fluorimetric methods. Fruit aging was found to be accompanied by modification of the photoluminescence spectrum shape and a noticeable increase in the photoluminescence intensity. Laser photoluminescence microscopy in vapors of fruit extracts and artificial compounds was used to assess the contribution of various substances to natural odor emission of fruits. The results of the study show that fluorimetry of odors is a promising method for studying fruits and other objects.     

Lycopene accumulation affects the biosynthesis of some carotenoid-related volatiles independent of ethylene in tomato

For elucidating the regulatory mechanism of ethylene on carotenoid-related volatiles (open chain) compounds and the relationship between lycopene and carotenoid-related volatiles,transgenic tomato fruits in which ACC synthase was suppressed were used.The transgenic tomato fruit showed a significant reduction of lycopene and aroma volatiles with low ethylene production.6-methyl-5-hepten-2-one,6-methyl-5-hepten-2-ol and geranylacetone,which were suspected to be lycopene degradation products,were lower than those in wild type tomato fruits.In order to identify whether lycopene accumulation effects the biosynthesis of some carotenoid-related volatiles independent of ethylene in tomato or not,the capability of both wild type and transgenic tomato fruits discs to convert lycopene into carotenoid-related volatiles was evaluated.The data showed that external lycopene could convert into 6-methyl-5-hepten-2-one and 6-methyl-5-hepten-2-ol in vivo,Indicating that the strong inhibition of ethylene production had no effect on enzymes in the biosynthesis pathway of some carotenoid-related volatiles.Therefore,in ACS-suppression transgenic tomato fruits,the low levels of 6-methyl-5-hepten-2-one,6-methyl-5-hepten-2-ol was due to decreased lycopene accumulation,not ethylene production.Ethylene only affected the accumulation of lycopene,and then indirectly influenceed the level of lycopene-related volatiles.     

Induction of volatile emissions from ripening apple fruits infested with Cydia pomonella and the attraction of adult females

We studied the effect of herbivory by Cydia pomonella L. (Lepidoptera: Tortricidae) first instar larvae on the volatile emissions from immature apples collected at the end of August. Volatiles from infested, artificially damaged, and healthy fruit were collected using solid-phase microextraction and analysed using combined gas chromatography mass spectrometry. The results show that neonate C. pomonella larvae induce the release of E--ocimene from immature fruit compared to healthy or artificially damaged fruit. Seven further compounds, a terpene, an alcohol, four aldehydes, and a ketone, were consistently found in all treatments. There were no significant differences amongst these latter compounds, although the amount of the terpene E,E--farnesene was three to five times higher from the infested as compared to healthy fruits. Headspace volatiles were also collected on Tenax to test for behavioural responses to the volatiles. Volatile collections from infested fruit attracted mated adult females in a Y-tube olfactometer, whereas no preference was evinced for volatiles from healthy or artificially damaged fruits when compared to a solvent control.     

Role of Ethylene in the Biosynthetic Pathways of Aroma Volatiles in Ripening Fruit

During the past decade, fruit aroma biosynthetic pathways were established in some climacteric fruits, such as tomato, apple, and melon. Inhibition of ethylene biosynthesis or its action in these fruits can reduce the production of fruit volatiles. Furthermore, ethylene partially regulates expression of a few important enzyme genes in fruit volatile biosynthetic pathways. The aim of this review is to bring together recent advances for understanding the regulatory role of ethylene in the biosynthesis of aroma volatiles in some fruits.     

Effect of suppression of ethylene biosynthesis on flavor products in tomato fruits

To elucidate the role of ethylene in the production of flavor compounds by tomato fruits, wild-type tomato (Lycopersicon esculentum L., cv. Lichun) and its transgenic antisense LeACS2 line with suppressed ethylene biosynthesis were used. The metabolism of individual sugars was ethylene-independent. However, citric acid and malic acid were under ethylene regulation. The content of these acids was higher in transgenic tomato fruits and returned to normal level after transgenic fruits were treated with ethylene. Because most of amino acids, which are important precursors of volatiles, were shown to be correlated with ethylene, we surmise that amino acid-related aroma volatiles were also affected by ethylene. Headspace analysis of volatiles showed a significant accumulation of aldehydes in wild-type tomato fruits during fruit ripening and showed a dramatic decrease in most aroma volatiles in transgenic tomato fruits as compared with wild-type fruits. The production of hexanal, hexanol, trans-2-heptenal, cis-3-hexanol, and carotenoid-related volatiles, except β-damascenone and β-ionone, was inhibited by suppression of ethylene biosynthesis. No remarkable differences were observed in the concentrations of cis-3-hexenal and trans-2-hexenal between transgenic and wild-type tomato fruits, indicating these two volatiles to be independent of ethylene. Thus, there are various regulation patterns of flavor profiles in tomato fruits by ethylene. Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 1, pp. 92–101. The text was submitted by the authors in English. Both authors equally contributed to this work.     

Response of female Cydia molesta (Lepidoptera: Tortricidae) to plant derived volatiles

Peach shoot volatiles were attractive to mated female oriental fruit moth, Cydia molesta (Busck), in a dual choice arena. No preference was observed between leaf odours from the principle host plant, peach, and the secondary host plant, apple. Twenty-two compounds were identified in headspace volatiles of peach shoots using gas chromatography-mass spectrometry. Green leaf volatiles accounted for more than 50% of the total emitted volatiles. A bioassay-assisted fractionation using different sorbent polymers indicated an attractant effect of compounds with a chain length of 6-8 carbon atoms. The major compounds of this fraction were tested either singly or in combinations for behavioural response of females. Significant bioactivity was found for a three-component mixture of (Z)-3-hexen-1-yl acetate, (Z)-3-hexen-1-ol and benzaldehyde in a 4:1:1 ratio. This synthetic mixture elicited a similar attractant effect as the full natural blend from peach shoots as well as the bioactive fraction.