Behavioral evidence for contextual olfactory‐mediated avoidance of the ubiquitous phytopathogen Botrytis cinerea by Drosophila suzukii

Herbivorous insects may benefit from avoiding the smell produced by phytopathogens infecting plant host tissue if the infected tissue reduces insect fitness. However, in many cases the same species of phytopathogen can also infect host plant tissues that do not directly affect herbivore fitness. Thus, insects may benefit from differentiating between pathogen odors emanating from food and nonfood tissues. This is based on the hypothesis that unnecessarily staying attentive to pathogen odor from nonfood tissue may incur opportunity costs associated with not responding to other important survival functions. In this study adults of Drosophila suzukii Matsumura, an invasive larval frugivore, showed reduced attraction to the odor of raspberry fruit, a food tissue, when infected with Botrytis cinerea Pers., a ubiquitous phytopathogen, in favor of odors of uninfected raspberry fruit. Moreover, D. suzukii oviposited fewer eggs on infected raspberry fruit relative to uninfected raspberry fruit. Larval survival and adult size after eclosion were significantly reduced when reared on B. cinerea‐infected raspberry relative to uninfected fruit. Interestingly, when the behavioral choice experiment was repeated using Botrytis‐infected vs. ‐uninfected strawberry leaves, a nonfood tissue, in combination with fresh raspberry fruit, odor from B. cinerea‐infected leaves did not reduce D. suzukii attraction to raspberries relative to raspberries with uninfected leaves. These behavioral results illustrate the important role context can play in odor‐mediated interactions between insects, plants and microbes. We discuss implications of our findings for developing a repellent that can be useful for the management of D. suzukii.     

The genome of black raspberry (Rubus occidentalis)

Black raspberry (Rubus occidentalis) is an important specialty fruit crop in the US Pacific Northwest that can hybridize with the globally commercialized red raspberry (R. idaeus). Here we report a 243 Mb draft genome of black raspberry that will serve as a useful reference for the Rosaceae and Rubus fruit crops (raspberry, blackberry, and their hybrids). The black raspberry genome is largely collinear to the diploid woodland strawberry (Fragaria vesca) with a conserved karyotype and few notable structural rearrangements. Centromeric satellite repeats are widely dispersed across the black raspberry genome, in contrast to the tight association with the centromere observed in most plants. Among the 28 005 predicted protein‐coding genes, we identified 290 very recent small‐scale gene duplicates enriched for sugar metabolism, fruit development, and anthocyanin related genes which may be related to key agronomic traits during black raspberry domestication. This contrasts patterns of recent duplications in the wild woodland strawberry F. vesca, which show no patterns of enrichment, suggesting gene duplications contributed to domestication traits. Expression profiles from a fruit ripening series and roots exposed to Verticillium dahliae shed insight into fruit development and disease response, respectively. The resources presented here will expedite the development of improved black and red raspberry, blackberry and other Rubus cultivars.     

The isolation of RNA from raspberry (Rubus idaeus) fruit

Previous attempts to extract high-quality, total RNA from raspberry (Rubus idaeus) fruits using published protocols have proven to be unsuccessful. Even the use of protocols developed for the extraction of RNA from other fruit tissue has resulted in low yields (1) or the isolation of degraded RNA (2). Here, we report on the development of a quick and simple method of extracting total RNA from raspberry fruit. Using this method, high yields of good quality, undegraded RNA were obtained from fruit at all stages of ripening. The RNA is of sufficient quality for northern analysis and cDNA library construction.     

Spotted Wing Drosophila Prefer Low Hanging Fruit: Insights into Foraging Behavior and Management Strategies

Spotted wing drosophila, Drosophila suzukii, is an invasive insect that attacks ripe, small fruit such as raspberries, blackberries and blueberries. Little is known about its foraging ecology and current trapping methods and monitoring systems are ineffective at commercial scales. In semi-field studies, we evaluated adult alightment and ovipositional preference within and among raspberry plants using sentinel Tangletrap-coated and clean raspberry fruit, respectively, positioned within the exterior and interior plant canopy at four different heights (60, 85, 110 and 135 cm from the base) and conducted in field cages using sexually mature adults. Alightment of adults on Tangle-trap-coated fruit indicated a preference for fruit positioned at lower heights and/or interior locations based on significantly greater numbers being captured on sentinel sticky-coated berries at the two lowest heights. Oviposition in clean raspberry fruit also yielded a similar pattern. In mark-release-recapture studies conducted in the field, spotted wing drosophila prefer sentinel sticky fruit positioned on exterior rows as they alighted on these berries in significantly greater numbers than fruit at in the central portion of the plot. Likewise, in field trials with wild fly populations, infestations were significantly greater in edge rows compared with interior rows. Collectively, our results suggest that monitoring and behaviorally based management strategies may be more effective if they target adults foraging in the lower canopy of small fruit plants located on the crop perimeter.     

Cellulase, Fruit Softening and Abscission in Red RaspberryRubus idaeusL. cv Glen Clova

The ripening of raspberry fruit (Rubus ideausL. cv Glen Clova)is associated with a climacteric rise in ethylene production.As the fruit pigments change from green to red there is a progressivesoftening, loss of skin strength and a breakdown of cell wallsin the mesocarp. An increase in cellulase (endo-1,4-ß-D-glucanase)in both drupelets and receptacles accompanies these changes.The localization of cellulase in the regions of the fruit associatedwith abscission zones suggest the enzyme may be involved infruit separation as well as softening. Rubus idaeusL; raspberry; fruit ripening; ethylene; abscission; cell wall breakdown; cellulase; endo-1,4-ß-D-glucanase     

Comparison of antioxidant capacity and phenolic compounds of berries, chokecherry and seabuckthorn

Antioxidant capacity and phenolic compounds (phenolic acids and anthocyanins) of four berry fruits (strawberry, Saskatoon berry, raspberry and wild blueberry), chokecherry and seabuckthorn were compared in the present study. Total phenolic content and total anthocyanin content ranged from 22.83 to 131.88 g/kg and 3.51 to 13.13 g/kg, respectively. 2,2-Diphenyl-1-picryhydrazyl free radical scavenging activity ranged from 29.97 to 78.86%. Chokecherry had the highest antioxidant capacity when compared with berry fruits and seabuckthorn. The highest caffeic acid, gallic acid and trans-cinnamic acid levels were found in chokecherry (6455 mg/kg), raspberry (1129 mg/kg) and strawberry (566 mg/kg), respectively. Caffeic acid was also the major phenolic acid in Saskatoon berry (2088 mg/kg) and wild blueberry (1473 mg/kg). The findings that chokecherry has very high antioxidant capacity and caffeic acid levels, are useful for developing novel value-added antioxidant products and also provide evidence essential for breeding novel cultivars of fruit plants with strong natural antioxidants.     

cDNA cloning and expression pattern of cinnamate-4-hydroxylase in the Korean black raspberry

Cinnamate-4-hydroxylase (C4H) is a key enzyme in the phenylpropanoid pathway, which is responsible for synthesizing a variety of secondary metabolites that participate in development and adaptation. In this study, we isolated a full-length cDNA of the C4H gene from the Korean black raspberry (Rubus sp.) and found that this gene existed as a single gene. By comparing the deduced amino acid sequence of Rubus sp. C4H with other sequences reported previously we determined that this sequence was highly conserved among widely divergent plant species. In addition, quantitative real time PCR studies indicated that the C4H gene had a differential expression pattern during fruit development, where gene expression was first detected in green fruit and was then remarkably reduced in yellow fruit, followed by an increase in red and black fruit. To investigate the two peaks in expression observed during fruit development and ripening, we measured the flavonoid content. The content of the major flavanol of Korean black raspberry fruits was determined to be highest at the beginning of fruit development, followed by a gradually decrease according to the developmental stages. In contrast, the content of anthocyanins during the progress of ripening was dramatically increased. Our results suggest that the C4H gene in Korean black raspberry plays a role during color development at the late stages of fruit ripening, whereas the expression of C4H gene during the early stages may be related to the accumulation of flavanols.     

Comparative analysis of quality and nutritional traits from Lonicera caerulea (Honeyberry) cultivars and other berries grown in Scotland

Lonicera caeruleabrix is a perennial shrub native to North America, Europe and Asia. It produces dark blue berries known as honeyberries or haskap berries which are produced commercially in several territories including Canada, Japan, Russia and Poland. Plants are suited to UK environments, but it is yet to be widely commercially developed in the UK. In the present work, quality and nutritional traits of six honeyberry cultivars grown in Scotland were compared with other commonly grown berry fruits (strawberry, raspberry, blueberry, blackberry, blackcurrant) to aid the identification of environmentally stable, high-quality honeyberry cultivars suitable for UK cultivation. Differences were observed in fruit quality variables (soluble solids, titratable acidity and Brix/acid ratios) between honeyberry cultivars. Three of six cultivars examined exhibited notable variation in soluble solids dependent on harvest year with ‘Aurora’ and ‘Strawberry Sensation’ having consistently high ^oBrix values. Titratable acidity exhibited cultivar differences and there was limited variation over harvest years. ‘Aurora’ exhibited consistently high ^oBrix/titratable acidity ratio reflected by high glucose and fructose content. Honeyberry fruit had good nutritional profile relative to other soft fruits with higher polyphenol and anthocyanin content than strawberry, blueberry, blackberry and raspberry, manifested in greater antioxidant capacity. The major anthocyanins in aqueous honeyberry fruit extracts were cyanidin, pelargonidin and peonidin glycosides. These findings indicate that L. caerulea represents a crop suitable for UK cultivation capable of producing high quality fruit with a valuable nutritional profile relative to other soft fruits. Cultivars exhibit significant differences in fruit quality and nutritional profile as well as harvest consistency and growers should consider this when establishing new plantations.     

The Role of Ethylene in the Shedding of Red Raspberry Fruit

The production of ethylene by red raspberry (Rubus idaeus L.cv. Glen Clova) fruit increased climacterically during development.The concentration of ethylene within green fruit was low butincreased substantially as fruit abscission and ripening commenced.The receptacle contained higher concentrations than the drupeletsat all stages measured. In the mature ripening fruit the ethyleneconcentrations were found to be physiologically significant,and would accelerate the abscission of large green non-abscisingfruit if supplied as a fumigant. The addition of ethylene toripe fruit did not accelerate abscission, probably because saturatinglevels occurred naturally within these fruit. Reduction of ethylenesynthesis rates using the inhibitor of ethylene production aminoethoxyvinylglycine(AVG) reduced the rate of abscission zone weakening which occursin detached large green fruit. The rate of ethylene productionwas found to be dependent on the supply of the precursor l-aminocyclopropane-l-carboxylicacid (ACC). This only accumulated to any extent in those ripefruit with high rates of ethylene production. Rubus idaeus, raspberry, abscission, fruit ripening, ethylene, aminocyclopropane-l-carboxylic acid     

Investigating the inoculum dynamics of Cladosporium on the surface of raspberry fruits and in the air

Raspberry production is under threat from the emerging fungal pathogenic genus Cladosporium. We used amplicon-sequencing, coupled with qPCR, to investigate how fruit age, fruit location within a polytunnel, polytunnel location and sampling date affected the fruit epiphytic microbiome. Fruit age was the most important factor impacting the fungal microbiome, followed by sampling date and polytunnel location. In contrast, polytunnel location and fruit age were important factors impacting the bacterial microbiome composition, followed by the sampling date. The within-tunnel location had a small significant effect on the fungal microbiome and no effect on the bacterial microbiome. As fruit ripened, fungal diversity increased and the bacterial diversity decreased. Cladosporium was the most abundant fungus of the fruit epiphytic microbiome, accounting for nearly 44% of all fungal sequences. Rotorod air samplers were used to study how the concentration of airborne Cladosporium inoculum (quantified by qPCR) varied between location (inside and outside the polytunnel) and time (daytime vs. nighttime). Quantified Cladosporium DNA was significantly higher during the day than the night and inside the polytunnel than the outside. This study demonstrated the dynamic nature of epiphytic raspberry fruit microbiomes and airborne Cladosporium inoculum within polytunnels, which will impact disease risks on raspberry fruit.     

Biology of the European large raspberry aphid (Amphorophora idaei): its role in virus transmission and resistance breakdown in red raspberry

1 The European large raspberry aphid Amphorophora idaei Börner is the most important vector of viral diseases afflicting commercially grown red raspberry ( Rubus idaeus L.) in Northern Europe, with European raspberry production amounting to 416 000 tonnes per annum. This review synthesizes existing knowledge on its biology and interactions with other organisms, including its host plant and the viral pathogens it vectors.
2 Information about trophic interactions with other insect herbivores and natural enemies is reviewed. Vine weevils Otiorhynchus sulcatus compromise aphid resistance in some raspberry cultivars, increasing A.   idaei abundance by 80%. Parasitoids show mixed success in parasitizing A.   idaei , although Aphidius ervi attack rates more than doubled when A.   idaei fed on a partially susceptible raspberry cultivar, compared with a resistant variety. These findings are discussed in the context of potential biological control as part of an integrated pest and disease management framework.
3  Amphorophora idaei transmits four known viruses: Black raspberry necrosis virus, Raspberry leaf mottle virus, Raspberry leaf spot virus and Rubus yellow net virus , with A.   idaei taking as little as 2 min to transmit some viruses.
4 Existing control strategies, including resistant cultivars, insecticides and eradication of disease from parent plants, are described. In particular, strong selection pressures have resulted in A .  idaei overcoming genetic resistance in many raspberry cultivars and most insecticides are now ineffective.
5 Future directions for the sustained control of A.   idaei are suggested, taking into consideration the possible effects of climate change and also changes in agronomic practices in U.K. agriculture.     

Enhancing beneficial antioxidants in fruits: a plant physiological perspective

The aim of this review is to present an outline of the physiological perspectives of beneficial antioxidant production in fruit. The drive to enhance the consumption of fruit and vegetables in the human diet is linked with positive effects of beneficial antioxidants impacting on health promotion. We briefly outline our physiological understanding of environmental processes which induce the production of reactive oxygen species and how antioxidants prevent plant cellular damage. More specifically, we describe the impact that environmental stresses, such as drought and radiation, have on the production of endogenous antioxidants and how these stresses can be incorporated into novel experimental crop growing systems to achieve high antioxidant concentrations in fruits. This includes in particular the use of irrigation application techniques and enhanced light reflectance to increase the concentrations of bioactive compounds such as ellagic acid and ascorbic acid.     

Field captures of wild melon fly (Diptera: Tephritidae) with an improved male attractant, raspberry ketone formate

Field-trapping evaluations of the new male attractant, formic acid 4-(3-oxobutyl) phenyl ester (raspberry ketone formate [RKF]) were conducted in Hawaii with wild populations of melon flies, Bactrocera cucurbitae Coquillett (Diptera: Tephritidae), to determine its activity in the field and to evaluate new plastic matrix formulations. All tests were compared with the standard melon fly attractant 4-(4-acetoxyphenyl) -2-butanone (cuelure [CL]), which is the attractant of choice for detection programs aimed at melon fly and other cuelure-responding Bactrocera fruit flies. Results of these tests over a range of doses on cotton wicks showed that at a 1-g dose raspberry ketone formate was 1.5-2 times more attractive compared with cuelure for up to 11 wk in the field. Lower doses applied on cotton wicks were less active, presumably due to hydrolysis of RKF to raspberry ketone. Raspberry ketone formate embedded in a plastic plug formulation also was field tested, and it was shown to be more attractive to male melon fly compared with cuelure. The use of this new attractant in control and detection programs is discussed.     

Evaluation of strawberry sap beetle (Coleoptera: Nitidulidae) use of habitats surrounding strawberry plantings as food resources and overwintering sites

The matrix of strawberry and alternate host crops, wooded areas, and uncultivated sections that comprises a farm landscape provides not only food resources but also habitat in both a spatial and temporal context. Reports of the strawberry sap beetle as a pest in strawberry in the northeastern United States have increased along with a trend to produce a wider diversity of fruit crops on individual farms. The three objectives of this study focused on determining which, if any, habitats outside strawberry plantings are important to consider when developing control strategies for strawberry sap beetles. First, sampling of wooded areas and multiple crops showed that strawberry sap beetles overwinter not only in wooded areas but also in blueberry and raspberry. No overwintering beetles were found in strawberry. Second, up to a 70-fold increase in mean number of strawberry sap beetles in a no-choice food source experiment indicated that considerable reproduction can occur on blueberry, cherry, raspberry, and strawberry. Third, sampling summer-bearing raspberry, peach, blueberry, and cherry in 2004 and 2005 confirmed that beetles were present, often in high densities (0.1-108.5 strawberry sap beetles/m(2)), in commercial fields with fruit or vegetable material on the ground. In summary, the beetles are able to feed, complete development, and overwinter in habitats other than strawberry. An effective integrated pest management program to control strawberry sap beetles will need to consider the type of habitat surrounding strawberry fields.     

Environmental and seasonal influences on red raspberry flavour volatiles and identification of quantitative trait loci (QTL) and candidate genes

Raspberry volatiles are important for perceptions of sensory quality, mould resistance and some have nutraceutical activities. Twelve raspberry character volatiles were quantified, 11 of them in fruit from two seasons, from plants from the Glen Moy × Latham mapping population growing in both open field and under cover (polytunnels). Effects of season and environment were examined for their impact on the content of α-ionone, α-ionol, β-ionone, β-damascenone, linalool, geraniol, benzyl alcohol, (Z)-3-hexenol, acetoin, acetic and hexanoic acids, whilst raspberry ketone was measured in one season. A significant variation was observed in fruit volatiles in all progeny between seasons and method of cultivation. Quantitative trait loci were determined and mapped to six of the seven linkage groups, as were candidate genes in the volatiles pathways.     

Post‐flowering photoperiod has marked effects on fruit chemical composition in red raspberry (Rubus idaeus)

The effect of photoperiod on nutritional quality and chemical composition of ‘Glen Ample’ red raspberry has been studied under controlled environment conditions. Long day (LD) conditions during fruit growth and maturation significantly increased fruit concentrations of ascorbic acid, total phenolics, organic acids such as malic, quinic and ellagic acids, as well as the ferric reducing activity power, while reducing the concentrations of soluble solids, sucrose and the sugar/acid ratio. The levels of total anthocyanin and oxalic acid were not significantly affected by photoperiod whereas semi‐quantitative analysis indicated a significant increase in the levels of some ellagitannins. The same responses were found whether LD was established by day‐length extension with low‐intensity light or by night interruption, thus demonstrating that the observed changes in nutritional quality of raspberry fruit was mediated through specific photoperiodic mechanism(s) independently of the daily light energy supply.     

Genomic characterisation and the detection of raspberry chromatin in polyploid Rubus

 This paper reports genomic in situ hybridization (GISH) and fluorescent in situ hybridization (FISH) data for chromosomes of raspberry (Rubus idaeus 2n=2x=14), blackberry (Rubus aggregate, subgenus Eubatus. 2n=2–12x=14–84) and their allopolyploid derivatives used in fruit breeding programmes. GISH was used to discriminate labelled chromosomes of raspberry origin from those of blackberry origin in allopolyploid hybrid plants. The raspberry chromosomes were labelled by GISH at their centromeres, and 1 chromosome was also labelled over the short arm. In one allopentaploid plant a chromosome carried a terminal signal. Karyotype analysis indicated that this is a blackberry chromosome carrying a raspberry translocation. GISH analysis of an aneuoctaploid blackberry cv ‘Aurora’ (2n=8x=58) showed that both whole and translocated raspberry chromosomes were present. The basic Rubus genome has one ribosomal DNA (rDNA) locus, and in all but one case all levels of ploidy had the expected multiples of rDNA loci. Interestingly, in the blackberry cv ‘Aurora’, there were only six sites, two less than might be predicted from its aneuoctaploid chromosome number. Our results highlight the potential of GISH and FISH for genomic designation, physical mapping and introgression studies in Rosaceous fruit crops. Received: 20 February 1998 / Accepted: 12 May 1998