成熟度对印度块菌香气成分的影响

研究成熟度对印度块菌Tuber indicum香气成分的影响,并测定成熟印度块菌的关键香气成分。将3种不同成熟度的印度块菌,以固相微萃取(SPME)技术为香气富集方法,利用气相色谱-质谱联用(GC-MS)分析其香气成分。结果表明未成熟印度块菌中仅检测出4种香气成分,中度成熟阶段检测出8种,成熟阶段检测出13种,而且成熟阶段检测出的香气成分大多都是前人报道过对块菌香气有贡献的成分;通过计算成熟块菌各香气组分的香气活度值(OAV),可知二甲基硫醚、2,3-丁二酮、3-甲基正丁醛、2-甲基正丁醛、己醛、1-辛烯-3-醇这6种物质是印度块菌的关键香气成分(OAV>1)。     

Bacteria associated with truffle‐fruiting bodies contribute to truffle aroma

Truffles, symbiotic fungi renown for the captivating aroma of their fruiting bodies, are colonized by a complex bacterial community of unknown function. We characterized the bacterial community of the white truffle Tuber borchii and tested the involvement of its microbiome in the production of sulphur‐containing volatiles. We found that sulphur‐containing volatiles such as thiophene derivatives, characteristic of T. borchii fruiting bodies, resulted from the biotransformation of non‐volatile precursor(s) into volatile compounds by bacteria. The bacterial community of T. borchii was dominated by α‐ and β‐Proteobacteria. Interestingly, all bacteria phyla/classes tested in this study were able to produce thiophene volatiles from T. borchii fruiting body extract, irrespective of their isolation source (truffle or other sources). This indicates that the ability to produce thiophene volatiles might be widespread among bacteria and possibly linked to primary metabolism. Treatment of fruiting bodies with antibacterial agents fully suppressed the production of thiophene volatiles while fungicides had no inhibitory effect. This suggests that during the sexual stage of truffles, thiophene volatiles are exclusively synthesized by bacteria and not by the truffle. At this stage, the origin of thiophenes precursor in T. borchii remains elusive and the involvement of yeasts or other bacteria cannot be excluded.     

Ascoma genotyping and mating type analyses of mycorrhizas and soil mycelia of Tuber borchii in a truffle orchard established by mycelial inoculated plants

Tuber borchii (the Bianchetto truffle) is a heterothallic Ascomycete living in symbiotic association with trees and shrubs. Maternal and paternal genotype dynamics have already been studied for the black truffles Tuber melanosporum and Tuber aestivum but not yet for T. borchii. In this study, we analysed maternal and paternal genotypes in the first truffle orchard realized with plants inoculated with five different T. borchii mycelia. Our aims were to test the persistence of the inoculated mycelia, if maternal and/or paternal genotypes correspond to inoculated mycelia and to assess the hermaphroditism of T. borchii. The mating type of each isolate as well as those of mycorrhizas, ascomata and extraradical soil mycelia was determined. Moreover, simple sequence repeat (SSR) profiles of maternal and paternal genotypes were assessed in 18 fruiting bodies to investigate the sexual behaviour of this truffle. The maternal genotypes of the fruiting bodies corresponded to those of the inoculated mycelia with only two exceptions. This confirmed that the inoculated mycelia persisted 9 years after plantation. As regards paternal partner, only two had the same genotype as those of the inoculated mycelia, suggesting hermaphroditism. Most of the new paternal genotypes originated from a recombination of those of inoculated mycelia.     

Behavioural responses in three ichneumonid pollen beetle parasitoids to volatiles emitted from different phenological stages of oilseed rape

Pollen beetles (Meligethes spp.; Coleoptera: Nitiduliae) are a major pest of oilseed rape, Brassica napus L. (Brassicaceae) in northern Europe. Phradis interstitialis Thomson, P. morionellus Holmgr., and Tersilochus heterocerus Thomson (Hymenoptera: Ichneumonidae) are among the most frequent pollen beetle parasitoids. These three species differ in temporal occurrence, as well as in preferred host stage. The behavioural responses of female parasitoids to odours from oilseed rape at bud and flowering stage were evaluated in two‐choice experiments. The role of visual stimuli was examined by combining green and yellow colours with odour stimuli. All three species were attracted to odours from the bud stage of oilseed rape. Tersilochus heterocerus was attracted to odours of flowering rape, but the two Phradis species avoided the flower odours. However, when the odours of flowering rape were combined with yellow, and odours of the bud stage were combined with green, P. interstitialis was equally attracted to both stimuli, and T. heterocerus showed an increased preference for flower odours, while no effect of colours could be found in P. morionellus. The observed differences in responses between the parasitoids may reflect differences in their biology and may be involved in the niche segregation of these often coexisting species. The volatile blends released from the two phenological stages were identified and compared. Clearly, odours can be reliable cues for differentiating between oilseed rape in the bud and flowering stage. Of 20 identified compounds, 18 were released at a significantly higher rate from flowering plants. The terpenes sabinene, myrcene, limonene, and (E,E)‐α‐farnesene were the dominant volatiles in the bud and flower headspace. A group of aromatic compounds including benzaldehyde, methyl benzoate, and phenyl acetaldehyde were mainly released from flowering rape.     

Attraction of a generalist predator towards herbivore-infested plants

The occurrence and strength of interactions among natural enemies and herbivores depend on their foraging decisions, and several of these decisions are based on odours. To investigate interactions among arthropods in a greenhouse cropping system, we studied the behavioural response of the predatory bug Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) towards cucumber plants infested either with thrips (Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae)) or with spider mites (Tetranychus urticae Koch (Acari: Tetranychidae)). In greenhouse release-recapture experiments, the predatory bug showed a significant preference for both thrips-infested plants and spider mite-infested plants over clean plants. Predatory bugs preferred plants infested with spider mites to plants with thrips. Experience with spider mites on cucumber leaves prior to their release in the greenhouse had no effect on the preference of the predatory bugs. However, this experience did increase the percentage of predators recaptured. Y-tube olfactometer experiments showed that O. laevigatus was more attracted to odours from plants infested with spider mites than to odours from clean plants. Thus, O. laevigatus is able to perceive odours and may use them to find plants with prey in more natural conditions. The consequences of the searching behaviour for pest control are discussed.     

Response of the elm leaf beetle to host plants induced by oviposition and feeding: the infestation rate matters

We investigated by olfactometry and feeding‐ and oviposition‐choice‐tests how the highly specialised elm leaf beetle, Xanthogaleruca luteola Müller (Coleoptera: Chrysomelidae), responds to conspecifically induced defences in the field elm Ulmus minor Miller (Ulmaceae). While egg deposition of the beetle induced elms to release volatiles attractive to the egg parasitoid Oomyzus gallerucae Fonscolombe (Hymenoptera: Eulophidae), feeding alone did not. In the present study, females of the elm leaf beetle showed preferences for the odours of twigs induced by low egg deposition and feeding over odours from uninfested twigs. In contrast, heavy infestation rendered elm odours less attractive to the beetles. Feeding and oviposition bioassays revealed an oviposition preference for leaves from uninfested twigs when compared to locally infested leaves. However, beetles preferred to feed upon systemically induced leaves compared to uninfested ones. The different preferences of the elm leaf beetle during host plant approach might be explained by a strategy that accounts for both gaining access to high quality nutrition and avoiding competition or parasitism.     

Sex‐specific responses of Asian citrus psyllid to volatiles of conspecific and host‐plant origin

Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is the primary vector of Candidatus Liberibacter spp. bacteria that cause citrus greening, a disease of worldwide importance. Olfactometry was employed to test responses of D. citri to odours from intact citrus plants (Mexican lime, Citrus aurantifolia, sour orange, Citrus aurantium, Marsh grapefruit, Citrus paradisi and Valencia orange, Citrus sinensis), citrus plants previously infested with D. citri, and odours of conspecifics including nymphs, adult insects of same and opposite sex, and their products (honeydew), both alone and in combination. In contrast to other studies, psyllids of both sexes were attracted to volatiles of undamaged Mexican lime leaves, whereas undamaged grapefruit attracted only females, and leaves of Valencia and sour orange did not attract either sex. All four plant species attracted female psyllids when previously infested, but only Mexican lime and sour orange‐attracted males. Thus, Citrus species appear to vary in the production of both constituitive and induced volatiles that attract adult psyllids. Volatiles emitted by nymphs did not attract either sex, but psyllid honeydew was attractive to males, likely due to female pheromone residues. Males oriented to the odour of females, whereas the reverse was not true, and neither males nor females oriented to same‐sex volatiles. The addition of conspecific cues (adults, nymphs or honeydew) did not increase female attraction to previously infested leaves, but male response was increased by the presence of adults and honeydew, regardless of plant species. Thus, female psyllids appear to orient more strongly to volatiles of plant origin, whereas males respond more strongly to cues emanating from females and conspecific excretions. These results suggest that female psyllids drive the initial colonization of host plants, whereas males orient to females and infested plants. Identification of the specific volatiles involved may permit their use in monitoring and management of this pest.     

A review of research on Chinese <Emphasis Type="Italic">Tuber</Emphasis> species

Truffles are abundant in some regions of China. Nevertheless, it was not until the 1980s that Tuber species were discovered by Chinese mycologists. In recent years, international truffle markets have shown an increasing interest in the import of Chinese Tuber. These truffles serve as a complement to European truffles due to their lower prices and their greater availability in a deficit market. However, Chinese truffles have been the subject of fraudulent commercial practices, and these could have a negative effect on truffle culture. These concerns have been behind numerous recent studies designed to obtain detailed information about Chinese Tuber species. Unfortunately, many of these studies are not published in English, and are dispersed throughout the specific local or national bibliography and proceedings of specialized truffle conferences. In view of the need to expand current knowledge of Chinese Tuber species, we present a comprehensive summary of the taxonomy, ecology, mycorrhizae, genetics, biochemistry, and cultivation of Chinese Tuber species. We also provide a synthetic taxonomy and morphological characterization of 16 Chinese Tuber species in order to assist in their verification and monitoring.     

Fine‐scale spatial genetic structure analysis of the black truffle Tuber aestivum and its link to aroma variability

Truffles are symbiotic fungi in high demand by food connoisseurs. Improving yield and product quality requires a better understanding of truffle genetics and aroma biosynthesis. One aim here was to investigate the diversity and fine‐scale spatial genetic structure of the Burgundy truffle Tuber aestivum. The second aim was to assess how genetic structuring along with fruiting body maturation and geographical origin influenced single constituents of truffle aroma. A total of 39 Burgundy truffles collected in two orchards were characterized in terms of aroma profile (SPME‐GC/MS) and genotype (microsatellites). A moderate genetic differentiation was observed between the populations of the two orchards. An important seasonal and spatial genetic structuring was detected. Within one orchard, individuals belonging to the same genet were generally collected during a single season and in the close vicinity from each other. Maximum genet size nevertheless ranged from 46 to 92 m. Geographical origin or maturity only had minor effects on aroma profiles but genetic structuring, specifically clonal identity, had a pronounced influence on the concentrations of C₈‐ and C₄‐VOCs. Our results highlight a high seasonal genetic turnover and indicate that the aroma of Burgundy truffle is influenced by the identity of single clones/genets.     

The Role of the Microbiome of Truffles in Aroma Formation: a Meta-Analysis Approach

Truffles (Tuber spp.) are ascomycete subterraneous fungi that form ectomycorrhizas in a symbiotic relationship with plant roots. Their fruiting bodies are appreciated for their distinctive aroma, which might be partially derived from microbes. Indeed, truffle fruiting bodies are colonized by a diverse microbial community made up of bacteria, yeasts, guest filamentous fungi, and viruses. The aim of this minireview is two-fold. First, the current knowledge on the microbial community composition of truffles has been synthesized to highlight similarities and differences among four truffle (Tuber) species (T. magnatum, T. melanosporum, T. aestivum, and T. borchii) at various stages of their life cycle. Second, the potential role of the microbiome in truffle aroma formation has been addressed for the same four species. Our results suggest that on one hand, odorants, which are common to many truffle species, might be of mixed truffle and microbial origin, while on the other hand, less common odorants might be derived from microbes only. They also highlight that bacteria, the dominant group in the microbiome of the truffle, might also be the most important contributors to truffle aroma not only in T. borchii, as already demonstrated, but also in T. magnatum, T. aestivum, and T. melanosporum.     

Small Mammal Mycophagy Response to Variations in Green-Tree Retention

Abstract: We studied the effects of 6 green-tree retention levels and patterns on the diets of northern flying squirrels (Glaucomys sabrinus), Townsend's chipmunks (Tamias townsendii), Siskiyou chipmunks (T. siskiyou), western red-backed voles (Myodes californicus), and southern red-backed voles (Myodes gapperi) using fecal pellet analysis. These rodents are truffle spore dispersers and prey for forest predators such as the northern spotted owl (Strix occidentalis caurina). Pretreatment diets showed differences in truffle and plant consumption among genera. Tree harvesting, especially in the 15% aggregated retention pattern, reduced frequency of Rhizopogon spores in the diet of voles, which may reflect a reduced ability of these animals to forage for Rhizopogon truffles, a decreased access to these truffles, or a reduction in Rhizopogon truffle abundance or frequency. Habitat island effects and edge effects provide conceptual frameworks for the reduction in consumption of Rhizopogon truffles by voles in green-tree aggregates. Overall, small mammal consumption of truffles showed little change in response to the treatments. Animals may be compensating for a locally declining food source by altering their foraging behavior. The long-term effect of this postulated behavioral compensation on small mammal energetics and population dynamics is unknown. Forest managers may reduce the impact of tree harvesting on these key forest ecosystem components by including green-tree aggregates within a dispersed retention matrix.     

How do sex and sexual maturation influence the response of Monochamus galloprovincialis to host odours?

Finding the right host plant for feeding and reproduction is crucial for herbivorous insects. Usually they need both visual and olfactory cues to locate and colonize their host plants, but the respective role of the two types of stimuli is difficult to disentangle. Model plants mimicking the shape and colour of natural hosts, combined with odours dispensers, are relevant tools for this purpose. We used dummy pines and cut branches of natural pines to investigate host attraction in Monochamus galloprovincialis, the insect vector of the pine wood nematode in Europe. We compared the behavioural response of males and females, sexually mature or immature, in a large climatic chamber and in an olfactometer. Immature females were significantly attracted by the sole odours released by pines. Immature males and mature males and females needed a combination of visual and olfactory cues to find the host tree. These findings suggest that beetles use both visual and chemical cues to locate pine trees but that relevant stimuli likely differ between beetles of different sex and sexual maturation status. Our results question the implementation of precautionary clear‐cuts around infested trees for limiting the spread of PWN.     

Identification and bioactivity of the growth inhibitors in Tuber spp. methanolic extracts

The methanolic extracts of fresh fruiting bodies of Tuber melanosporum, T. aestivum, T. magnatum and T. borchii were evaluated for their free fatty acids (FFAs) content to test their contribution to truffle allelopathic activity. Linoleic acid was the major FFA detected in all truffle species. In addition to linoleic acid, oleic acid and palmitic acid were the other abundant FFAs in the Tuber spp. extracts. Looking at the total production, T. melanosporum and T. aestivum synthesized significantly higher amounts of all identified FFA than T. magnatum and T. borchii. The allelopathic activity, based on the specific activity (EC₅₀) of the identified compounds, was evaluated in the growth of Arabidopsis thaliana, Lotus corniculatus, Melica ciliata and Silene vulgaris. Linoleic acid had the lowest specific activity and palmitic acid had the highest. The black truffle extracts had a higher inhibitory activity than white truffles, and this could be explained by the high relative amount of FFAs in them.     

Eco-archaeological excavation techniques reveal snapshots of subterranean truffle growth

Despite its status as a highly-prized and coveted fungi in gastronomy, many aspects of the subterranean life cycle of the Burgundy truffle (Tuber aestivum) are still unknown, because in situ observations of the formation and maturation of truffle fruitbodies remain difficult. Here, we adopted a suite of archaeological fine-scale excavating techniques to provide unique spatiotemporal snapshots of Burgundy truffle growth at three sites in southern Germany. We also recorded the relative position, fresh weight, maturity level and genotype composition of all excavated fruitbodies. Varying by a factor of thousand, the fresh weight of 73 truffle ranged from 0.1 to 103.2 g, with individual maturity levels likely representing different life cycle stages from completely unripe to fully ripe and even decaying. While only a slightly positive relationship between fruitbody weight and maturity level was found, our results suggest that genetically distinct specimens can exhibit different life cycle stages at the same period of time and under the same environmental conditions. We therefore argue that truffles are likely able to grow, mature and ripe simultaneously between early summer and late winter of the following year. Our case study should encourage further eco-archaeological truffle excavations under different biogeographic settings and at different seasons of the year to gain deeper insights into the fungi's subterranean ecology. The expected cross-disciplinary findings will help truffle hunters and farmers to improve their harvest practices and management strategies.     

Drivers of truffle biomass,community composition,and richness among forest types in the northeastern US

Truffle-producing fungi (hypogeous sporocarps) are important mycorrhizal symbionts and provide a key food source for many animals, including small mammals. To better understand truffle diversity and associations in the northeastern US, we surveyed for truffles and analyzed spores in eastern chipmunk (Tamias striatus) scat across hardwood (angiosperm-dominated), softwood (gymnosperm-dominated), and mixed forest at Bartlett Experimental Forest, New Hampshire. Truffle biomass ranged from 3.8 kg/ha in hardwood forest to 31.4 kg/ha in softwood forest and was up to 35 times greater than mushroom (epigeous sporocarp) production in softwood forest. Elaphomyces species were the most common truffle taxa in both field surveys and chipmunk scat. Scat analysis indicated that truffle richness increased over the summer and accurately reflected fruiting time, providing greater resolution of richness than field surveys alone. Basal area of eastern hemlock (Tsuga canadensis) was the primary driver of Elaphomyces biomass and was the best explanatory variable of truffle community composition. We discuss implications of hemlock loss, due to the introduced hemlock woolly adelgid (Adelges tsugae), on forest mycorrhizal communities and food webs.     

The attractiveness of different odour sources from the fruit–host complex on Leptopilina boulardi, a larval parasitoid of frugivorous Drosophila spp.

In parasitoid insects, successful offspring development depends on the female’s ability to find a suitable host. Specific recognition is often based on responses to olfactory cues, but their source and nature have rarely been determined. –This paper deals with the recognition of odours involved in host location by Leptopilina boulardi[Barbotin, Carton & Kelner-Pillault] (Hymenoptera: Eucoilidae), a larval parasitoid of Drosophila species that develops in mature fruits. The nature and origin of volatile stimuli recognized among odours of the host–fruit complex, and the effect of learning on this recognition, were investigated. Oriented responses to these odours were observed in a four-armed olfactometer and were analysed with the observer software (Noldus Information Technology). Fruit odours alone (banana and pear) were not spontaneously attractive to naive parasitoids, whereas naturally-infested bananas were highly attractive. The attraction was related to the odour that adult Drosophila left on the substrate but not to Drosophila oviposition activity or larval development. A synergism between some fruit odours (banana and pear) and the odour left by adult Drosophila on damp filter paper was observed. However, when testing a non-fruit substrate (mushroom), no synergism was observed. Thus, female L. boulardi may innately recognize host–food substrate odours associated with odours from the adult stage of their host. In addition, an oviposition experience on an infested banana allows L. boulardi females to memorise the fruit odour itself through associative learning. The adaptive significance of this process is discussed.     

Infestation of commercial bumblebee (Bombus impatiens) field colonies by small hive beetles (Aethina tumida)

Abstract.  1. The small hive beetle, Aethina tumida , is a parasite of honeybee ( Apis mellifera ) colonies native to sub-Saharan Africa and has become an invasive species. In North America the beetle is now sympatric with bumblebees, Bombus , not occurring in its native range. Laboratory studies have shown that small hive beetles can reproduce in bumblebee colonies but it was not known whether infestations occur in the field.
2. For the first time, infestation of bumblebee colonies by small hive beetles was investigated in the field. Commercial Bombus impatiens colonies ( n = 10) were installed in proximity to infested apiaries. Within 8 weeks, all colonies that were alive in the 5-week observation period ( n = 9) became naturally infested with adult small hive beetles and successful small hive beetle reproduction occurred in five colonies.
3. In four-square choice tests, the beetles were attracted to both adult bumblebee workers and pollen from bumblebee nests, suggesting that these odours may serve as cues for host finding.
4. The data indicate that bumblebee colonies may serve as alternative hosts for small hive beetles in the field. To foster the conservation of these essential native pollinators, investigations on the actual impact of small hive beetles on wild bumblebee populations are suggested.     

Host location and discrimination mediated through olfactory stimuli in two species of Encyrtidae

The role of olfactory stimuli in host detection and evaluation was studied in two encyrtid Hymenoptera. The first, Epidinocarsis lopezi De Santis, is a monophagous parasitoid of the cassava mealybug Phenacoccus manihoti Matile-Ferrero, itself feeding exclusively on cassava, Manihot esculenta Crantz. The second, Leptomastix dactylopii Howard, is a monophagous parasitoid of the Citrus mealybug, Planococcus citri Risso, but this latter is highly polyphagous. The behaviour of females of both parasitoids (attaction and locomotion) was compared in a tubular olfactometer for the odours of their respective hosts on cassava and poinsettia. Tests were made using: 1) healthy host-plant alone; 2) host-plant infested with unparasitized mealybugs; 3) unparasitized mealybugs only; 4) host-plant infested with parasitized mealybugs and 5) parasitized mealybugs only. Only E. lopezi was attracted by the odour of the host-plant alone, but both species were attracted by the odour of an infested host-plant and that of unparasitized mealybugs. The odour of parasitized mealybugs, alone or on host-plant, induced an undirected activity. The attraction of E. lopezi to the odour of the host-plant alone could be linked to the monophagous diet of its host, whereas the attraction of the two species of parasitoids to the odours of infested host-plants and unparasitized mealybugs could be due to the fact that both parasitoids are specialists. The behavioural response of both species to the odour of parasitized mealybugs revealed a new aspect in host discrimination: the identification of parasitized hosts could be partly mediated through olfactory stimuli, and not only through gustatory stimuli.     

Influence of pH on in vitro mycelial growth in three Japanese truffle species: Tuber japonicum,T. himalayense,and T. longispinosum

Truffle species indigenous to Japan include a white-colored truffle Tuber japonicum and two black-colored truffles, T. himalayense and T. longispinosum. As the fruiting bodies of these Tuber species are promising edible, studies on the artificial cultivation for these truffles are currently underway in Japan. In the present study, we investigated the influence of pH on in vitro mycelial growth in these Tuber species to determine the optimal pH conditions for the cultivation of these truffles. Mycelia of five strains from each species were cultured in modified Melin–Norkrans liquid medium at different pH values. Tuber japonicum grew well at pH 5.0 and 6.0, whereas T. himalayense and T. longispinosum grew well at pH 7.0. This results suggest that the optimal pH for mycelial growth varies among Tuber species. The growth data collected in this study can be used to design optimal pH conditions for artificial cultivation of these Japanese truffles.     

Divergent responses of two cereal aphids to previous infestation of their host plant

The role of pollen odour in resource location by the pollen beetle, Meligethes aeneus (Fabricius) (Coleoptera: Nitidulidae), a pollen-feeding insect regarded as a pest of oilseed rape, Brassica napus L., (Brassicaceae) crops, was investigated in a linear track olfactometer. Both male and female beetles were attracted to the odour of whole oilseed rape flowers, indicating that these insects can locate their host plants using floral odours as cues. The attractive odour of flowers was found to emanate from all floral parts tested: the petals/sepals, the anthers, and from pollen itself. Therefore, at least part of the attractive odour of oilseed rape flowers emanates from pollen. Beetles were more attracted to floral samples containing anthers than those without anthers when these odours were directly compared in a choice-test, and this indicates that there were detectable differences between them. Anthers and pollen may therefore release distinctive odours that are quantitatively and/or qualitatively different from the odour of the rest of the flower. These experiments support the hypothesis that pollen-seeking insects use pollen odour cues to locate this food source.