Genetic variation detected by use of the M13 “DNA fingerprint” probe in Malus,Prunus, and Rubus (Rosaceae)

Summary Recently, DNA fingerprints have been reported in a wide array of organisms. We used the M13 repeat probe on several genera and species in the angiosperm family Rosaceae. Four apple cultivars could be differentiated when any one of five restriction enzymes was used to analyze minisatellite DNA. Similarly, four individual trees of Prunus serotina (black cherry) exhibited different fingerprints with each of four enyzmes. A total of 14 Rubus (blackberries and raspberries) plants representing four species were investigated with two enzymes. Extensive inter-and intraspecific variation was found. However, some closely growing plants had identical fingerprints, probably due to their being derived through vegetative propagation.     

Development of two spotted spider mite (Acari: Tetranychidae) populations on strawberry and raspberry cultivars

Five strawberry (Fragaria sp.) and five raspberry (Rubus ideaus L.) cultivars were evaluated for resistance to two spotted spider mite (Tetranychus urticae Koch.). Two methods of assessing the development of two spotted mite populations using detached leaves were compared. The number of eggs laid and mites which developed were compared. The strawberry cvs Hapil and Pegasus had significantly greater development of two spotted mite populations than the cvs Rhapsody, Symphony and Elsanta. The raspberry cv. Joan Squires had higher populations of two spotted mite whilst the raspberry cv. Leo the least, when compared with cvs Glen Clova, Glen Moy and Glen Prosen. Differences were observed in oviposition sites and mite distribution when comparing raspberries with strawberries. The method of assessing the populations development of two spotted mite which involved maintaining the cut leaf stem in water may be of potential use for studying population dynamics of both two spotted mite and possible predators over extended periods of time.     

Rubus host range of rubus yellow net virus and its involvement with other aphid-borne latent viruses in inducing raspberry veinbanding mosaic disease

After graft inoculation with rubus yellow net virus (RYNV), 12 of 34 Rubus species and cultivars developed noticeable symptoms. R. macraei developed the most conspicuous symptoms and is recommended as an improved indicator plant. In attempts to determine the cause of raspberry veinbanding mosaic, a disease in which RYNV is involved, several European and North American red raspberry cvs were graft-inoculated with RYNV and three other aphid-borne viruses, black raspberry necrosis (BRNV), raspberry leaf mottle (RLMV) and raspberry leaf spot, singly and in all combinations. In periods of up to 4 yr, classical veinbanding mosaic symptoms developed in sensitive cvs only when they contained both RYNV and RLMV. These symptoms were intensified in plants co-infected with additional viruses. Veinbanding mosaic disease did not develop in any of 11 cvs infected with RYNV + BRNV, the combination of viruses previously assumed to be responsible for this disease in Britain and North America.     

Biotransformation of raspberry ketone and zingerone by cultured cells of Phytolacca americana

The biotransformation of raspberry ketone and zingerone were individually investigated using cultured cells of Phytolacca americana. In addition to (2S)-4-(4-hydroxyphenyl)-2-butanol (2%), (2S)-4-(3,4-dihydroxyphenyl)-2-butanol (5%), 4-[4-(beta-d-glucopyranosyloxy)phenyl]-2-butanone (19%), 4-[(3S)-3-hydroxybutyl]phenyl-beta-d-glucopyranoside (23%), and (2S)-4-(4-hydroxyphenyl)but-2-yl-beta-d-glucopyranoside (20%), two biotransformation products, i.e., 2-hydroxy-4-[(3S)-3-hydroxybutyl]phenyl-beta-d-glucopyranoside (12%) and 2-hydroxy-5-[(3S)-3-hydroxybutyl]phenyl-beta-d-glucopyranoside (11%), were isolated from suspension cells after incubation with raspberry ketone for three days. On the other hand, two compounds, i.e., (2S)-4-(4-hydroxy-3-methoxyphenyl)but-2-yl-beta-d-glucopyranoside (17%) and (2S)-2-(beta-d-glucopyranosyloxy)-4-[4-(beta-d-glucopyranosyloxy)-3-methoxyphenyl]butane (16%), together with (2S)-4-(4-hydroxy-3-methoxyphenyl)-2-butanol (15%), 4-[4-(beta-d-glucopyranosyloxy)-3-methoxyphenyl]-2-butanone (21%), and 4-[(3S)-3-hydroxybutyl]-2-methoxyphenyl-beta-d-glucopyranoside (24%) were obtained upon addition of zingerone. Cultured cells of P. americana can reduce, and regioselectively hydroxylate and glucosylate, these food ingredients to their beta-glycosides.     

Molecular and biochemical characterization of benzalacetone synthase and chalcone synthase genes and their proteins from raspberry (Rubus idaeus L.)

Two new members of the polyketide synthase (PKS) gene family (RiPKS4 and RiPKS5) were cloned from raspberry fruits (Rubus idaeus L., cv Royalty) and expressed in Escherichia coli. Characterization of the recombinant enzyme products indicated that RiPKS4 is a bifunctional polyketide synthase producing both 4-hydroxybenzalacetone and naringenin chalcone. The recombinant RiPKS4 protein, like the native protein from raspberry fruits [W. Borejsza-Wysocki, G. Hrazdina, Plant Physiol. 1996;110: 791-799] accepted p-coumaryl-CoA and ferulyl-CoA as starter substrates and catalyzed the formation of both naringenin chalcone, 4-hydroxy-benzalacetone and 3-methoxy-4-hydroxy-benzalacetone. Although activity of RiPKS4 was higher with ferulyl-CoA than with p-coumaryl-CoA, the corresponding product, 3-methoxy-4-hydroxy phenylbutanone could not be detected in raspberries to date. Sequence analysis of the genes and proteins suggested that this feature of RiPKS4 was created by variation in the C-terminus due to DNA recombination at the 3′ region of its coding sequence. RiPKS5 is a typical chalcone synthase (CHS) that uses p-coumaryl-CoA only as starter substrate and produces naringenin chalcone exclusively as the reaction product.     

Raspberry ketone-supplemented diet has no effect on fitness parameters or lure responsiveness in male melon flies,Zeugodacus cucurbitae (Diptera: Tephritidae)

The male annihilation technique (MAT) and sterile insect technique (SIT) are often used to control pestiferous tephritid fruit flies (Diptera: Tephritidae). MAT involves the deployment of traps containing a male attractant and insecticide with the goal of drastically reducing male abundance and ultimately eliminating the entire population. SIT, which involves the mass production, sterilization, and release of the target species, may also be implemented to achieve final extirpation. Generally, simultaneous implementation of MAT and SIT is counterproductive, because the presence of large numbers of male-specific traps in the environment (MAT) would greatly reduce the number of sterile males available for copulating with wild females (SIT). However, studies on the Queensland fruit fly, Bactrocera tryoni (Froggatt), indicate that concurrent use of MAT and SIT may be feasible. Sexually mature males of B. tryoni are attracted to the raspberry ketone and its synthetic analogue cue-lure. Males of B. tryoni fed raspberry-ketone-supplemented diet when newly emerged showed lower attraction to cue-lure baited traps than control males. In addition, newly emerged males provided this diet displayed accelerated sexual maturation, which would allow the early release of sterile males and reduce pre-release holding costs. Here, we examined whether the addition of raspberry ketone to the adult diet of male melon flies, Zeugodacus cucurbitae (Coquillett), produced effects similar to those observed for B. tryoni. Despite using similar methods, no significant effect of raspberry ketone-supplemented diet on time to sexual maturity, survival, mating competitiveness, or attraction to cue-lure baited traps in mass-reared Z. cucurbitae males.     

利用免疫组织化学及蛋白质组学对树莓提取物抑制大鼠肝癌机制的研究

目的：探讨树莓预防大鼠原发性肝癌增殖抑制和凋亡诱导作用,寻找树莓预防大鼠原发性肝癌的特异性蛋白质靶点。方法：利用二乙基亚硝胺（DEN）建立大鼠原发性肝癌动物模型;通过免疫组织化学方法研究树莓提取物对于大鼠原发性肝癌的预防效果和形态学变化,利用蛋白质组学研究树莓预防大鼠原发性肝癌的特异性蛋白质靶点。结果：树莓提取物能抑制PCNAJVEGF的表达,抑制细胞增殖,并诱导细胞凋亡。蛋白质组学差异分析表明：成瘤组大鼠血清在蛋白质峰2597．93M／Z,4513．88M／Z上与高剂量树莓干预组大鼠血清具有显著性差异（P〈0．05）。结论：树莓提取物可以抑制肝癌细胞PCNA的表达,从而抑制肝癌细胞的增殖;还可以诱导肝癌细胞凋亡;蛋白质峰2597．93M／Z及4513．88M／Z所表达蛋白为其特异性作用靶点。     

Further evidence that resistance in raspberry to the virus vector aphid, Amphorophora idaei, is related to the chemical composition of the leaf surface

In an approach to understand the mechanism(s) of resistance in raspberry to infestation by the aphid Amphorophora idaei, progeny plants segregating for the A. idaei resistance gene, A₁₀, were bioassayed and dichloromethane extracts from the leaf surface were examined by capillary column gas chromatography (GC). No single GC peak was detected that was present in only the resistant progeny plants. Nevertheless, thirteen compounds present in all samples were quantified and identified by mass spectrometry. They were of four major classes; straight chain hydrocarbons, acetic acid esters of long chain alcohols, tocopherols and triterpenoid compounds, including α and β amyrin. Several of these compounds were not recorded previously in raspberry leaves. Linear discriminant analysis, applied to the standardised chromatographic data in an attempt to relate chemical composition to resistance, successfully partitioned 24 of the 26 plants into resistant and susceptible types as determined by bioassay. These data provide further evidence that resistance in raspberry to A. idaei is related to the chemical composition of the leaf surface.     

Nitrogen fixation byRubus ellipticus J. E. Smith

Summary Nitrogenase activity as assayed by acetylene reduction was observed in detachedRubus ellipticus J. E. Smith root nodules collected in the field and tested under ambient conditions. The nitrogenase activity was 8.4 moles C₂H₄.gfr. wt nodule^–1.h^–1 or 24.0 moles C₂H₄.g dry wt nodule^–1.h^–1 being at a rate comparable with that measured in some other non-legumes assayed in Java at the same time under similar conditions. Nodule morphology bore little resemblance to the root nodules of other non-leguminous plants and nodule structure was different from the other rosaceous examples.The endophyte inhabiting the root nodules was actinomycetal.     

Cryopreservation of in vitro-grown shoot tips of raspberry (Rubus idaeus L.) by encapsulation–vitrification and encapsulation–dehydration

The first efficient cryopreservation procedure for in vitro-grown shoot tips of raspberry (Rubus idaeus L.) has been developed based on encapsulation–vitrification (EnVi) and encapsulation–dehydration (EnDe). EnVi resulted in higher survival (85%) and regrowth (75%) of cryopreserved shoot tips than EnDe (65 and 50%, respectively). In both cryogenic procedures, shoots regenerated from cryopreserved shoot tips without intermediary callus formation. Histological studies showed that a much larger number of meristematic cells survived following EnVi than EnDe. The EnVi procedure was applied to seven raspberry genotypes with an average survival and regrowth of 71 and 68%, respectively. Regenerated plants showed normal morphology. Results here indicate EnVi as a simple and efficient method for long-term preservation of R. idaeus germplasm.