Is the existence of a mutualistic relationship between Lobesia botrana and Botrytis cinerea well-founded?

Some studies have highlighted benefits for Lobesia botrana by adding Botrytis cinerea mycelium to an artificial larval diet and have suggested a mutualistic relationship between the two organisms on grapevine, hypothesizing that fungal sterols were the nutritional factor involved. Because the nutritional quality of an artificial diet should be similar to grapes to allow extrapolation of the results to the field conditions, in the current study L. botrana larval performance was compared when larvae were fed on grapes (berries) or two artificial diets either with or without enrichment with B. cinerea. Based on sterol analysis, the two artificial diets had high cholesterol content, but relative to berries showed comparable and low phytosterol contents, respectively (high- and low-phytosterol, HPh, and LPh). While larval fitness on the HPh diet was similar to berries, the LPh diet led to higher mortality and worse larval performance. The addition of the fungus compensated for the shortage in the LPh diet but did not improve the HPh diet. Supplementing the LPh diet with linoleic acid, which is supplied also from B. cinerea, partially improved larval performance. In a field experiment, females did not show any egg-laying preferences towards naturally botrytized bunches. The positive effect of B. cinerea on the moth's next generation that is reported in the literature could be a consequence of fungus developed inside berry tunnels bored by larvae. Therefore, based on our data and previous reports the existence of a mutualistic relationship between L. botrana and B. cinerea is not well-founded.     

Assessing the influence of inflorescence traits on the susceptibility of grape to vine moths

In north‐eastern Italy during 1994–2006, studies were carried out on the susceptibility of grapevine cultivars to the first generations of the European vine moth [Lobesia botrana (Den. and Shiff.); Lep., Tortricidae] and the European grape berry moth [Eupoecilia ambiguella (Hb.); Lep., Cochylidae]. In five different years, the larval population density of both moth species and the larval age composition of L. botrana were recorded on 11 grapevine cultivars (Cabernet Sauvignon, Carmenère, Chardonnay, Pinot Gris, Merlot, Refosco dal Peduncolo Rosso, Rhine Riesling, Sauvignon Blanc, Terrano, Tocai Friulano, Verduzzo Friulano), grown in the same vineyard. The influence of inflorescence traits on these demographic parameters was also evaluated. On a 5‐year average, L. botrana significantly prevailed over E. ambiguella in nine of the 11 cultivars. Chardonnay and Pinot Gris were the most infested cultivars. Age composition of L. botrana larvae varied with cultivar type, being older on Chardonnay and Tocai Friulano than the other cultivars. Inflorescence earliness and hairiness explained the majority of the variability in cultivar susceptibility. In particular, the larval population level of the two vine moths was positively correlated with inflorescence earliness and negatively correlated with inflorescence hairiness. Lobesia botrana larval age composition was negatively correlated with inflorescence hairiness. Inflorescence earliness and hairiness could be used to predict in each grape‐growing area which cultivars are potentially more infested in the first generation. Inflorescences without pubescence, favouring an older age composition of first generation larvae, could induce an earlier second generation.     

A methyltransferase essential for the methoxypyrazine‐derived flavour of wine

Methoxypyrazines are a family of potent volatile compounds of diverse biological significance. They are used by insects and plants in chemical defence, are present in many vegetables and fruit and, in particular, impart herbaceous/green/vegetal sensory attributes to wines of certain varieties, including Cabernet Sauvignon. While pathways for methoxypyrazine biosynthesis have been postulated, none of the steps have been confirmed genetically. We have used the F₂ progeny of a cross between a rapid flowering grapevine dwarf mutant, which does not produce 3‐isobutyl‐2‐methoxypyrazine (IBMP), and Cabernet Sauvignon to identify the major locus responsible for accumulation of IBMP in unripe grape berries. Two candidate methyltransferase genes within the locus were identified and one was significantly associated with berry IBMP levels using association mapping. The enzyme encoded by this gene (VvOMT3) has high affinity for hydroxypyrazine precursors of methoxypyrazines. The gene is not expressed in the fruit of Pinot varieties, which lack IBMP, but is expressed in Cabernet Sauvignon at the time of accumulation of IBMP in the fruit. The results suggest that VvOMT3 is responsible for the final step in methoxypyrazine synthesis in grape berries and is the major determinant of IBMP production.     

Effects of grapevine bunch exposure to sunlight on berry surface temperature and Lobesia botrana (Lepidoptera: Tortricidae) egg laying,hatching and larval settlement

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Relative performance of European grapevine moth (<Emphasis Type="Italic">Lobesia botrana</Emphasis>) on grapes and other hosts

The European grapevine moth, Lobesia botrana is a major grapevine pest, but despite the abundance of vineyards it is a generalist and uses either grapes or alternative species. Given the abundance and predictability of grape, L. botrana could be expected to have evolved towards monophagy. In order to understand why this species remains polyphagous, we hypothesized that larvae reared on rare wild host plants should have higher fitness than those reared on the more abundant grape host. For this, we compared larval performance and several life history traits on three alternative host plants (Daphne gnidium, Olea europaea, Tanacetum vulgare) and three Vitaceae (Vitis vinifera), two cultivars and one wild species (Ampelopsis brevipedunculata), and two control groups raised on either a low or a high nutritive value medium. Alternative hosts are more suitable than Vitaceae for the reproductive performance of L. botrana: larval mortality and development time was reduced, while pupal weight, growth rate, female longevity, female fecundity, duration of laying and mating success were increased. High quality food ingested by larvae promotes higher adult body weight and enhances female reproductive output. This suggests that alternative hosts provide greater nutritional value for L. botrana than Vitaceae. The use of alternative host plants could thus be maintained in the host range because they offer L. botrana a better fitness than on the Vitaceae. This could typically represent an advantage for moths behaving in plant diversity grape landscapes.     

2-Methoxy-3-isobutylpyrazine in grape berries and its dependence on genotype

2-Methoxy-3-isobutylpyrazine (MIBP) contributes a bell pepper aroma to many grape cultivars and has a reported aroma threshold of ～2 ng L(-1) in water. The purpose of this study was twofold: (1) develop a procedure using headspace solid phase micro-extraction combined with GC-MS in the selected ion monitoring mode (HS-SPME-GC-MS-SIM) for analysis of MIBP in grape berries, and (2) determine the location of MIBP biosynthesis in grapevines by approach grafting clusters of Vitis vinifera L. cvs Cabernet Sauvignon and Muscat blanc onto each other. The soluble solids and pH of the grape juice/homogenate matrix from different grape berry developmental stages influenced the method precision; therefore, quantification via the method of standard addition was used. Using our developed method, the limit of detection (LOD) and limit of quantitation (LOQ) of MIBP were 0.1 ng L(-1) and 2 ng L(-1), respectively, measured in a model juice and non-MIBP containing Chardonnay juice. Spiked recoveries averaged between 91% and 112% in Cabernet Sauvignon and Pinot noir homogenates and the overall relative standard deviation was less than 10%. The method was used to analyze MIBP in 29 grape cultivars and in fruit from clusters grafted to Cabernet Sauvignon or Muscat vines. Quantifiable levels were found only in Cabernet franc, Cabernet Sauvignon, Merlot, Sauvignon blanc and Semillon, providing information on the genetic connection for the occurrence of MIBP in grapes. No MIBP was detected in the berries of Muscat blanc clusters grafted onto Cabernet Sauvignon vines when sampled at fruit maturity. MIBP was detected in all berries of Cabernet Sauvignon regardless the graft configuration. The data indicate that MIBP or its precursors originate in the berry and its formation depends upon grape genotype.     

Measurements of reflectance and fluorescence spectra for nondestructive characterizing ripeness of grapevine berries

In vivo reflectance and fluorescence spectra from berry skins of a white (Riesling) and red (Cabernet Sauvignon) grapevine variety were measured during a ripening season with a new CMOS radiometer instrument. Classical reference measurements were also carried out for a sugar content of the berry juice [°Brix] and pigment contents (chlorophyll a and b, carotenoids, anthocyanins) from methanol extracts of the berry skin. We showed that the colours and the spectra analysed from them could be taken as an unambiguous indicator of grapevine ripening. Reflectance spectra, which were affected by the content of pigments (chlorophylls and anthocyanins), effects of surface (wax layers), and tissue structure (cell size) of the berries well correlated (R² = 0.89) with the °Brix measurements of the berries. The fast data acquisition of both reflectance and fluorescence spectra in one sample with our radiometer instrument made it superior over the time-consuming, traditional, and mostly destructive chemical analysis used in vine-growing management.     

Diet affects male gonad maturation,female fecundity,and larval development in the granivorous ground beetle Anisodactylus punctatipennis

Abstract 1. Granivory is a specialised food habit in the Carabidae and is considered to have evolved from ancestral carnivory. Despite recent investigations, the mechanisms underlying this peculiar feeding shift have not been sufficiently elucidated. In particular, no studies have examined the effects of diet composition on male reproductive traits. 2. This study examined male gonad maturation, female fecundity, and larval development under different diets (insect larvae, mixed seeds, and insect larvae + mixed seeds) for Anisodactylus punctatipennis (Coleoptera: Carabidae). 3. Whereas sperm‐bundle length, the durations of larval stages, and adult weight did not differ among diets, the weights of seminal substance‐producing organs, fecundity, and larval survival were higher with diets containing seeds compared to the pure‐animal diet. These findings provide the first conclusive evidence for granivory in Anisodactylus. 4. This study is the first that demonstrates the effects of diet on male reproductive traits in a granivorous carabid. The results were consistent with similar observations for fecundity and larval performance in this and other studies. Thus, in addition to the female and larval traits, the dietary effects on male reproductive traits may also have played an important role in the evolution of granivory in the Carabidae.     

A seed parasitoid wasp prevents berries from changing their colour,reducing their attractiveness to frugivorous birds

1. Frugivorous and seed‐feeding insects may alter the traits of fruits, such as shape and size, which may influence fruit attractiveness to frugivorous birds. Consequently, trait‐mediated interactions may occur in systems where plants, seed‐dispersing frugivorous vertebrates, and frugivorous or seed‐feeding insects interact. We investigated colour manipulation in Ilex integra Thunb. berries caused by the seed parasitoid wasp Macrodasyceras hirsutum Kamijo and how that manipulation relates to fruit attractiveness for frugivorous birds. 2. In winter, the colour of I. integra berries varied from green to red, but most berries were greenish, indicating that the berries were immature. Berry dissection indicated that the number of live parasitoid larvae present within each berry was closely related to berry colour – the greater the number of live larvae, more intense is the green colour of the berry. However, the wasp larvae did not modify the shape or size of the berries. More than 98% of berries that were protected from the insects by gauze bags ripened and turned red. In the present study, berries with unfertilised seeds alone turned red. Field‐feeding preference tests showed that the brown‐eared bulbul Hypsipetes amaurotis Temminck preferred red berries to green berries. 3. We demonstrated that the seed parasitoid wasp manipulates the berry colour, but not its shape or size, in a density‐dependent manner. Because green berries suffered less from bird foraging, we believe that this colour manipulation helps the wasps to avoid being killed by the birds. The present study indicates that manipulation by wasps may reduce the level of mutualism between the tree and seed‐dispersing birds.     

Endophytes of Grapevine Flowers,Berries, and Seeds: Identification of Cultivable Bacteria,Comparison with Other Plant Parts,and Visualization of Niches of Colonization

Endophytic bacteria can colonize various plants and organs. However, endophytes colonizing plant reproductive organs have been rarely analyzed. In this study, endophytes colonizing flowers as well as berries and seeds of grapevine plants grown under natural conditions were investigated by cultivation as well as by fluorescence in situ hybridization. For comparison, bacteria were additionally isolated from other plant parts and the rhizosphere and characterized. Flowers, fruits, and seeds hosted various endophytic bacteria. Some taxa were specifically isolated from plant reproductive organs, whereas others were also detected in the rhizosphere, endorhiza or grape inflo/infructescence stalk at the flowering or berry harvest stage. Microscopic analysis by fluorescence in situ hybridization of resin-embedded samples confirmed the presence of the isolated taxa in plant reproductive organs and enabled us to localize them within the plant. Gammaproteobacteria (including Pseudomonas spp.) and Firmicutes (including Bacillus spp.) were visualized inside the epidermis and xylem of ovary and/or inside flower ovules. Firmicutes, mainly Bacillus spp. were additionally visualized inside berries, in the intercellular spaces of pulp cells and/or xylem of pulp, but also along some cell walls inside parts of seeds. Analysis of cultivable bacteria as well as microscopic results indicated that certain endophytic bacteria can colonize flowers, berries, or seeds. Our results also indicated that some specific taxa may not only derive from the root environment but also from other sources such as the anthosphere.     

Organ-specific transcription of putative flavonol synthase genes of grapevine and effects of plant hormones and shading on flavonol biosynthesis in grape berry skins

In order to investigate the control mechanism of flavonol biosynthesis of grapevine, we obtained five genomic sequences (FLS1 to FLS5) of putative flavonol synthase genes from Vitis vinifera cv. Cabernet Sauvignon. The mRNA of five FLSs accumulated in flower buds and flowers, while the mRNA of FLS2, FLS4, and FLS5 accumulated in small berry skins and then decreased toward veraison. At the ripening stage, the mRNA of only FLS4 and FLS5 accumulated again. This change in mRNA accumulation did not contradict the flavonol accumulation in the berry skins. Shading of the berries completely inhibited the increase in flavonol content and mRNA accumulation of FLS4, but did not affect the mRNA accumulation of FLS5. The effects of light and plant hormones on flavonol accumulation were different from those on anthocyanin accumulation. Thus flavonol biosynthesis appears to be under a different control system from that of anthocyanin biosynthesis.     

Does natural larval parasitism of Lobesia botrana (Lepidoptera: Tortricidae) vary between years, generation, density of the host and vine cultivar?

Populations of European grapevine moth Lobesia botrana Denis & Schiffermüller and its larval parasitoids were studied for two consecutive years on an experimental insecticide-free vineyard in France planted with adjacent plots of five grape cultivars (Merlot, Cabernet Franc, Sauvignon, Cabernet Sauvignon and Sémillon) using a natural L. botrana population during the first year, and a natural population supplemented with artificially inoculated individuals during the second year. Levels of natural populations of larval parasitoids were measured by their parasitism rate. The ichneumonid Campoplex capitator Aubert was the most common species collected from L. botrana larvae. Its incidence was higher during the spring compared to summer. The overall parasitism rate found on the experimental vineyard varied from 23% in 2000 to 53% in 2001, and was mainly due to C. capitator. Parasitism was not affected by the grape cultivar on which the host developed but was positively correlated with the host density, per bunch or per stock, suggesting that among the five grape cultivars tested, C. capitator females probably do not discriminate between hosts feeding on different grape cultivars, but rather the densities of L. botrana larvae.     

Maternal diet and larval diet influence survival skills of larval red drum Sciaenops ocellatus

Larval red drum Sciaenops ocellatus survival, turning rate, routine swimming speed, escape response latency and escape response distance were significantly correlated with essential fatty‐acid (EFA) concentrations in eggs. Of the five traits that varied with egg EFA content, two (escape response latency and routine swimming speed) were significantly different when larvae were fed enriched diets compared with the low fatty‐acid diet, indicating that the larval diet can compensate for some imbalances in egg composition. Turning rate during routine swimming and escape response distance, however, did not change when larvae predicted to have low performance (based on egg composition) were fed an enriched diet, indicating that these effects of egg composition may be irreversible. Escape response distances and survival rates of larvae predicted to perform well (based on egg composition) and fed highly enriched diets were lower than expected, suggesting that high levels of EFA intake can be detrimental. Altogether, these results suggest that both maternal diet, which is responsible for egg EFA composition, and larval diet may play a role in larval survivorship and recruitment.     

Why do larvae of Utetheisa ornatrix penetrate and feed in pods of Crotalaria species? Larval performance vs. chemical and physical constraints

Larvae of Utetheisa ornatrix (L.)(Lepidoptera: Arctiidae) are found mainly inside unripe pods of several alkaloid‐bearing Crotalaria (Fabaceae) species. Although eggs are laid on the leaves, the larvae are usually found feeding on unripe seeds in the pods. In this work, we investigated the selective pressures that could explain why U. ornatrix larvae feed primarily on unripe pods with seeds and not on leaves. Our results showed that larval survivorship in the laboratory was unaffected by feeding on leaves or unripe seeds, and that larval development up to the pupal stage was better in larvae that fed on unripe seeds, although perforating unripe pods to reach seeds was costly in terms of survivorship. Females were also heavier when fed on unripe seeds, but there was no significant difference in the fecundity of females fed either of the two diets. Feeding on unripe seeds in pods had other benefits for U. ornatrix, including a lower predation rate for larvae that fed inside compared to larvae that fed outside the pods. Similarly, adults derived from larvae that fed on unripe seeds were preyed upon less frequently by the orb‐weaving spider Nephila clavipes than were adults that fed on leaves. The latter benefit may be closely related to the high concentration of pyrrolizidine alkaloids in unripe seeds, which is about five times more than in leaves. These alkaloids are sequestered by the larvae and transferred to adults, which then become chemically protected. However, this chemical defence does not protect the larvae against ants such as Ectatomma quadridens and Camponotus crassus. Pods with unripe seeds that confer physical protection to larvae and pyrrolizidine alkaloids that confer chemical protection to adults limit the use of leaves by U. ornatrix larvae.     

Changes in protein abundance during powdery mildew infection of leaf tissues of Cabernet Sauvignon grapevine (Vitis vinifera L.)

A comparative analysis of differentially expressed proteins in a susceptible grapevine (Vitis vinifera ‘Cabernet Sauvignon’) during the infection of Erysiphe necator, the causal pathogen of grapevine powdery mildew (PM), was conducted using iTRAQ. The quantitative labeling analysis revealed 63 proteins that significantly changed in abundance at 24, 36, 48, and 72 h post inoculation with powdery mildew conidiospores. The functional classification of the PM‐responsive proteins showed that they are involved in photosynthesis, metabolism, disease/defense, protein destination, and protein synthesis. A number of the proteins induced in grapevine in response to E. necator are associated with the plant defense response, suggesting that PM‐susceptible Cabernet Sauvignon is able to initiate a basal defense but unable to restrict fungal growth or slow down disease progression.     

Volatiles that encode host-plant quality in the grapevine moth

Plant volatiles are signals used by herbivorous insects to locate host plants and select oviposition sites. Whether such volatiles are used as indicators of plant quality by adult insects in search of host plants has been rarely tested. We tested whether volatiles indicate plant quality by studying the oviposition of the grapevine moth Lobesia botrana on the grapevine plant Vitis vinifera. Host plants were infected with a variety of microorganisms, and larval fitness was correlated to the infected state of the substrate. Our results show an oviposition preference for volatiles that is significantly correlated with the fitness of the substrate. The chemical profiles of the bouquets from each V. vinifera–microorganism system are clearly differentiated in a PCA analysis. Both the volatile signal and the quality of the plant as larval food were affected by the introduction of microorganisms. Our study represents a broad approach to the study of plant–insect interactions by considering not only the direct effect of the plant but also the effect of plant–microorganism interactions on insect population dynamics.