Changes in biochemical composition of needles of ornamental dwarf spruce (Picea glauca ‘Conica’) induced by spruce spider mite (Oligonychus ununguis Jacobi, Acari: Tetranychidae) feeding

The impact of spruce spider mite (SSM) (Oligonychus ununguis Jacobi, Acari: Tetranychidae) feeding on needle compounds of young dwarf white spruce (Picea glauca ‘Conica’), important in defence against pests, was determined. It was shown that the direction and intensity of changes in chemical composition of spruce needles was related to the density of spruce spider mite population. Relative to uninfested controls, needles of trees infested by 2–6 mites per 5 cm oftwig during 8 weeks contained markedly higher concentration of soluble proteins, total phenolics and essential oil volatiles (linalool, β-phellandrene, β-myrcene, δ-3-carene, p-cymene, limonene, α-pinene, β-pinene, borneol, methyl salicylate, geranyl acetate). Feeding of a three times larger population of mites (18 specimens per 5 cm of twig) caused either reduction of concentration of those compounds, or no significant difference in comparison to the control. The results of our study show that O. ununguis at relatively low density (not exceeding 2–6 specimens per 5 cm of twig per 8 weeks) stimulates even susceptible host-plant to alter metabolism, however induced responses are suppressed as mite density increases. Findings are discussed in relation to the importance of changes in the concentration of needle primary and secondary metabolites to white spruce defence against O. ununguis.     

Stem anatomy of Rhipsalis (Cactaceae) and its relevance for taxonomy

We used paternally inherited chloroplast microsatellites (cpSSR) to study population genetic structure in the endemic and highly restricted Serbian spruce Picea omorika. Fragment size polymorphism at the five cpSSR regions that could be amplified out of the nine tested combined into only four different haplotypes in the seven populations studied. Mean total haplotypic diversity was H _T = 0.395, and mean within-population diversity was H _s = 0.279. Haplotypic variation was lower than in most conifers described so far and partitioned into two geographical groups. All northern P. omorika populations were fixed or nearly fixed for the common haplotype, while southern populations exhibited two to three haplotypes. We suggest that current P. omorika populations are shaped by extreme demographic bottleneck and random genetic drift linked to Quaternary glacial cycles. P. omorika thus belongs to the small group of genetically depauperate tree species.     

Survey of natural enemies of spider mites (Acari: Tetranychidae) in citrus orchards in eastern Spain

Field surveys were conducted from 2004 to 2007 to determine the species composition and relative abundance of natural enemies associated with colonies of either the citrus red mite, Panonychus citri, or the two spotted spider mite, Tetranychus urticae, in Valencian citrus orchards (eastern Spain). Fourteen species were recorded, six phytoseiid mites and eight insect predators. Two of them are reported for the first time on citrus in Spain and two more are first reports as predators associated with T. urticae. The community of predators associated with T. urticae and P. citri was almost identical, and the Morisita–Horn index of similarity between both natural enemy complexes was close to one, suggesting that predators forage on both pest species. Quantifying the presence of many known spider mites predators in Valencian citrus orchards is an important first step towards spider mite control. A challenge for future studies will be to establish conservation and/or augmentation management strategies for these predators, especially to improve T. urticae biological control.     

Population dynamics of Panonychus osmanthi (Acari: Tetranychidae) on two Osmanthus species

Panonychus osmanthi is a non-diapausing species of spider mite that superficially resembles P. citri. It infests Osmanthus species, which are evergreen roadside and garden trees. The population dynamics of P. osmanthi were studied on Osmanthus aurantiacus and O.×fortunei during a three-year period. Seasonal changes in P. osmanthi populations were fundamentally the same in each year, although their density differed greatly from year to year. TheP. osmanthi population was bimodal, with one peak in spring (May–June) and another in winter (November–January). Populations abruptly declined after the spring peak. Predators showed a delayed density-dependent response to changes in spider mites from spring to summer, whereas in autumn and winter, predators were few because they had entered diapause. To determine the effect of predators on the rapid decline of spider mites just after the spring peak, the predators were removed by treating the trees with a synthetic pyrethroid. As a result, spider mite density did not decline after the spring peak and remained at a high level during the June-August period when spider mite density is usually very low. This suggests that predators play an important role in the drastic decline of P. osmanthi density just after the spring peak. This revised version was published online in July 2006 with corrections to the Cover Date.     

The vitality of the Serbian spruce (Picea omorika) pollen during the long-term cryopreservation

Abstract

This article studies differences in the vitality (germination rate and pollen tube length) of fresh pollen and pollen stored at ?20 °C for 14 years of Serbian spruce (Picea omorika [Pan?i?] Purkyne). Fresh pollen was sampled from 24 trees from a Serbian spruce seed stand area on the site of Bela zemlja (43° 48′ 32″ N, 19° 44′ 25″ E) in the area of Mount Zlatibor (Serbia) in two successive years. Six nutrition media with different sucrose concentrations (0%, 5%, 10%, 15%, 20% and 25% aqueous solution) were used to assess the vitality of the pollen. Variance analysis shows significant differences among years of pollen sampling, years of cryopreservation, trees, and sucrose treatments for pollen germination for both germination rate and pollen tube length. The results indicate that climate conditions preceding the pollen maturation period had an impact on the pollen quality as well as on the pollen vitality during the years of cryopreservation. Inter-individual variability (differences between trees genetic variability) was very pronounced and enabled selection of pollinator trees that retained their initial vitality during the period of cryopreservation. This periodicity in Serbian spruce full flowering might have significant practical application in the breeding of this important species.     

Expression of a Bacillus thuringiensis cry1Ab gene in transgenic white spruce and its efficacy against the spruce budworm (Choristoneura fumiferana)

A synthetic version of the cry1Ab gene from Bacillus thuringiensis (Bt) was introduced into white spruce (Picea glauca) by microprojectile bombardment. A plasmid carrying the cry1Ab gene, driven by a ubiquitin (maize) promoter, was co-transferred with a plasmid containing the gus–nptII fusion gene as a screenable selection marker. Molecular analysis of the transgenic lines showed a high level (more than 90%) of co-integration of the cry1Ab gene with the screenable marker. A wide range of expression levels of the cry1Ab gene and corresponding endotoxin was obtained. Accumulation of the Cry1Ab protein was evaluated in embryogenic tissue, the needles of somatic seedlings and in the needles of 5-year-old field-grown trees of individual lines. Laboratory and field insect feeding trials suggest that several spruce transgenic lines were lethal to spruce budworm larvae.     

Microsatellites as DNA markers in Sitka spruce

Nine microsatellite loci were found by screening a genomic DNA library of Sitka spruce (Picea sitchensis) with the four oligonucleotide probes (TG), (CAC), (GATA) and (AT). Pairs of flanking primers were generated for seven microsatellites. Five primer pairs were used to screen up to 58 Sitka spruce clones. The five loci SStg3a, SStg4, SStg4a, SStg4c and SSgataS were found to have 15, 13, 4, 3 and 6 different length alleles respectively, and in using a combination of them almost all 58 Sitka spruce genotypes could be identified. The five primer pairs were successful in amplifying DNA from two other spruce species (Picea albutilia and Picea smithiana), while only one primer pair could amplify DNA from the pine species, Pinus sylvestris and Pinus latifolia. The inheritance of microsatellites in Sitka spruce was co-dominant Mendelian.     

Influence of epicuticular waxes from white spruce and balsam fir on feeding behavior of the eastern spruce budworm

Fourth-instar larvae of the eastern spruce bud-worm Choristoneura fumiferanaClem. (Lepidoptera: Tortricidae) were shown in two-choice feeding tests to respond differently to epicuticular waxes from different white spruce and balsam fir trees. Larvae also showed different preferences for various wax fractions obtained by separation on silicic acid.Deceased.     

Beat sampling accuracy in estimating spruce spider mite (Acari: Tetranychidae) populations and injury on juniper

The use of a standardized beat sampling method for estimating spruce spider mite, Oligonychus ununguis (Jacobi) (Acari: Tetranychidae), densities on a widely used evergreen ornamental plant species, Juniperus chinensis variety 'Sargentii' A. Henry (Cupressaceae), was examined. There was a significant positive relationship between total spruce spider mite densities and spider mite densities from beat sampling on juniper. The slope and intercept of the relationship may be used by pest managers to predict total spider mite densities on plants from beat sample counts. Beat sampling dramatically underestimates the total number of spider mites on a foliage sample. The relationships between spruce spider mite feeding injury and spider mite density estimates from beat sampling juniper foliage and total spider mite counts on foliage were also examined. There was a significant positive relationship between spruce spider mite density as estimated from beat sampling and injury to the plants. There was a similar positive relationship between the total number of spruce spider mites and injury to the plants, suggesting that a pest manager could use beat sampling counts to estimate plant injury and related thresholds. These findings have important implications to decision-making for spruce spider mite control, especially as it relates to threshold levels and determining rates of predator releases. Further assessment of the effectiveness of beat and other sampling methods across multiple spider mite- host plant associations needs to be examined to enable pest managers to select sampling plans that are feasible and reliable.     

Green algae in spruce forests in the north-east of European Russia

The green algal communities in podzolic soils under coniferous forest in N.E. European Russia were studied in areas that were subjected to different technogenic pollution intensities. Sixty-five green algal species belonging to 4 classes, 12 orders, 20 families and 27 genera were recorded. The algal communities in the investigated soils of the different type spruce phytocoenoses included from 15 to 38 taxa on one site. Indicator species were identified for background spruce forests (Chalmydomonas gelatinosa, Tetracystis aggregata, T. dissociata, Pseudopleurococcus botryoides, Myrmecia bisecta) and for aerotechnogenically polluted spruce forests (Actinochloris sphaerica). The results may be used for the monitoring of the ecological state of biota in soils under boreal forests of protected areas and spruce forests exposed to intense aerial technogenic pollution. Presented at the International Symposium Biology and Taxonomy of Green Algae V, Smolenice, June 26–29, 2007, Slovakia.     

The sensitivity of embryogenic tissue of Picea omorika (Panč.) Purk. to antibiotics

In an attempt to develop a transformation procedure for somatic embryos of Picea omorika (Pan.) Purk. using Agrobacterium, the sensitivity of non-transformed embryogenic tissues of P. omorika to antibiotics was measured. Two groups of antibiotics were tested: antibiotics commonly used to eliminate Agrobacterium from tissue culture (carbenicillin and cefotaxime), and antibiotics for the selection of transformed tissue (kanamycin, paromomycin, neomycin and hygromycin). Carbenicillin (500 mg l^–1) reduced proliferation of P. omorika embryogenic tissue by 50% as compared to the control. Cefotaxime (500 mg l^–1) was less toxic to embryogenic tissue growth (93% of the control) and is thus more suitable for the elimination of A. tumefaciens from embryogenic tissue of P. omorika. Embryogenic tissue showed severe susceptibility to kanamycin and hygromycin. Induction of secondary embryogenesis was blocked by 10 mg l^–1 of kanamycin or 2 mg l^–1 of hygromycin. The aminoglycoside analogs of kanamycin, paromomycin and neomycin inhibited embryogenic tissue induction at concentrations of 5 and 100 mg l^–1, respectively. The higher tolerance of embryogenic tissue to neomycin indicated that of the antibiotics tested neomycin may be most useful for selection of nptII-transformed embryogenic tissue of Picea omorika.     

Nematophagous fungus <Emphasis Type="Italic">Paecilomyces lilacinus</Emphasis> (Thom) Samson is also a biological agent for control of greenhouse insects and mite pests

Pathogenicity of nematophagous fungus Paecilomyces lilacinus (Thom) Samson in control of the most destructive greenhouse pests such as: greenhouse whitefly, Trialeurodes vaporariorum, glasshouse red spider mite, Tetranychus urticae, the cotton aphid, Aphis gossypii and western flower thrips, Frankliniella occidentalis was examined in laboratory and pot experiments. The fungus showed the greatest efficacy in controlling winged and wingless forms of the cotton aphid. The cotton aphid’s population was almost totally eliminated. In controlling the greenhouse whitefly, P. lilacinus was most successful when applied against nymphal growth stages (L₃-L₄). Control of the western flower thrips was most efficient against prepupal and pupal stages when the fungus was applied as a water spore suspension to the soil. When the fungus was applied at temperatures below 10 °C, it was able to reduce a glasshouse red spider mite population by 60%.     

Chemotaxonomic Implications of the n‐Alkane Composition and the Nonacosan‐10‐ol Content in Picea omorika,Pinus heldreichii,and Pinus peuce

The n‐alkane composition and the nonacosan‐10‐ol content in the needle cuticular waxes of Serbian spruce (Picea omorika), Bosnian pine (Pinus heldreichii), and Macedonian pine (Pinus peuce) were compared. The amount of nonacosan‐10‐ol in the needle waxes of P. omorika was higher than those in P. heldreichii and P. peuce. The range of n‐alkanes was also wider in P. omorika (C₁₈–C₃₅) than in P. heldreichii and P. peuce (C₁₈–C₃₃). The dominant n‐alkanes were C₂₉ in the needle waxes of P. omorika, C₂₃, C₂₇, and C₂₅ in those of P. heldreichii, and C₂₉, C₂₅, C₂₇, and C₂₃ in those of P. peuce. The waxes of P. omorika contained higher amounts of n‐alkanes C₂₉, C₃₁, and C₃₃, while those of P. heldreichii and P. peuce had higher contents of n‐alkanes C₂₁, C₂₂, C₂₃, C₂₄, and C₂₆. The principal component analysis of the contents of nine n‐alkanes showed a clear separation of the Serbian spruce populations from those of the two investigated pine species, which partially overlapped. The separation of the species was due to high contents of the n‐alkanes C₂₉ and C₃₁ (P. omorika), C₁₉, C₂₀, C₂₁, C₂₂, C₂₃, and C₂₄ (P. heldreichii), and C₂₈ (P. peuce). Cluster analysis also showed a clear separation between the P. omorika populations on one side and the P. heldreichii and P. peuce populations on the other side. The n‐alkane and terpene compositions are discussed in the light of their usefulness in chemotaxonomy as well as with regard to the biogeography and phylogeny of these rare and endemic conifers.     

Interactions between green spruce aphid (Elatobium abietinum (Walker)) and Norway and Sitka spruce under high and low nutrient conditions

1 Green spruce aphid (Elatobium abietinum) is a serious pest of spruce (Picea spp.) in north‐west Europe, causing defoliation of one‐year‐old and older needles. 2 Relationships between population development of E. abietinum, needle loss and tree growth were compared for five pure genotypes of Sitka spruce and mixed‐genotype material of Sitka and Norway spruce, grown under high and low nutrient conditions. 3 Despite wide differences in flushing date between spruce genotypes, E. abietinum populations peaked on the same date on each genotype and on the mixed‐genotype material, irrespective of nutrient supply. 4 Larger aphid populations developed on trees grown under high nutrient conditions than under low nutrients. However, more needles were lost per aphid in the low nutrient treatment and overall defoliation rates in the two nutrient treatments were similar. 5 Total aphid numbers differed significantly between Sitka spruce genotypes within nutrient treatments, but not in relation to bud‐burst or needle terpene content. Reductions in height growth caused by infestation were greater (15–44%), and related to mean aphid densities and defoliation, in the low nutrient treatment, but were smaller (11–27%) and not related to aphid density and defoliation in the high nutrient treatment. 6 Development of E. abietinum populations was similar on Norway and Sitka spruce, but Norway spruce lost fewer needles. However, the effects of infestation on tree growth were more closely related to aphid density and were similar for Norway and Sitka spruce. 7 Infestation caused a decrease in total root dry weight of Norway and Sitka spruce in proportion to the reductions observed in above‐ground growth.     

A review of gamasid mites (Parasitiformes,Mesostigmata) dwelling in the taiga of the Pechoro-Ilychskii Nature Reserve (northern Cis-Ural Region) with analysis of their assemblages in spruce forests

In the plain taiga of the Pechoro-Ilychskii Nature Reserve, 91 species of gamasid mites have been revealed. Cheiroseius (7 species), Veigaia (6), Arctoseius (5), Dendrolaelaps (5), and Dinychus (4) are the most diverse genera. European (29%), Palaearctic (27%), and Holarctic species (30%) are represented in the fauna by somewhat equal fractions, whereas cosmopolitan and semicosmopolitan species constitute about 14%. Comparison of gamasid assemblages populating certain habitats allowed revealing two distinct groupings, namely, forest and floodplain ones. Forest assemblages are characterized by high population density (220–760 ind./dm²) and by predominance of Parazercon radiatus, Zercon forsslundi, Veigaia nemorensis, Gamasellus montanus, and Trachytes spp. Floodplain assemblages are less abundant (80–130 ind./dm²) and comprise mainly meadow and hydrophilic species (A. cetratus, Cheiroseius spp. Gaeolaelaps spp., and Gamasellodes bicolor). Low diversity of the family Parasitidae (4 species) and structural peculiarities of assemblages dwelling in coniferous forests (namely, the absence of Pergamasinae and significant fractions of Veigaia, Gamesellus, Trachytes, and also of representatives of the family Zerconidae) demonstrate similarities between the mesostigmatan population of this northern European region and that of the Siberian taiga. Solitary records of such Arctic species as Arctoseius ornatus and Zerconopsis labradorensis, and of the hypoarcto-montane species Iphidinychus gaieri within the taiga belt, seem to reflect the instability of range margin. Taking into account the literary data, we recognize a complex of boreal and borealmountain species (Trachytes edleri, Dinychus septentrionalis. D. crassus, Caurozercon duplex, Neozercon smirnovi, Zercon forsslundi, and Z. zelawaiensis) in the Palaearctic fauna. Their habitats, however, are not restricted only to dark coniferous forest along. The structure of gamasid mite assemblages in mesophytic spruce forests of the Palaearctic varies strongly. By contrast to vascular plants, where many species accompany the dark coniferous taiga everywhere, there is not a single mesostigmatic mite species common to all the spruce forests analyzed, ranging from Romania in the west to Sakhalin Island in the east. On the whole, changes in gamasid mite assemblages across the spruce forest belt seem to follow a succession of meridional changes in gamasid faunas regardless of an edificatory spruce species.     

Influence of ectomycorrhizal fungi on the response of Sitka spruce and Japanese larch to forms of phosphorus

Ectomycorrhizal fungi (Paxillus involutus, Suillus grevillei and two unidentified basidiomycetes from excised Sitka spruce mycorrhizas) were isolated from stands of Sitka spruce either in monoculture or in a mixture with Japanese larch in an Irish conifer plantation. The growth of these fungi and their mycorrhizal formation in Sitka spruce and Japanese larch were examined after incubation in modified Melin-Norkrans medium containing either KH₂PO₄, Ca₃(PO₄)₂ or Fe phytate as the phosphorus (P) source. P. involutus and S. grevillei utilized all three P sources. The unidentified basidiomycetes had limited ability to utilize Fe phytate. Basidiomycete 1 showed poor growth on KH₂PO₄ whereas growth of basidiomycete 2 was low on Ca₃(PO₄)₂. Pure culture synthesis studies confirmed that P. involutus and the two basidiomycetes formed mycorrhizas with both tree species but S. grevillei was mycorrhizal only on Japanese larch. P. involutus formed more mycorrhizas in both conifers than the other fungi. Following inoculation with each of the four fungi, shoot and root dry mass of both Sitka spruce and Japanese larch seedlings was enhanced compared with uninoculated/nonmycorrhizal controls. On Fe phytate, Paxillus-inoculated Sitka spruce seedlings had the lowest primary root length and on KH₂PO₄, Suillus-inoculated Japanese larch had the greatest number of short roots. The only differences when Sitka spruce seedlings were grown in either monoculture or in a mixture with Japanese larch mycorrhizal with S. grevillei were primary root length and number of short roots after growth on media containing Fe phytate.     

Response of predatory mites with different rearing histories to volatiles of uninfested plants

The behavioural response of the predatory mite Phytoseiulus persimilis to volatiles from several host plants of its prey, spider mites in the genus Tetranychus, was investigated in a Y-tube olfactometer. A positive response to volatiles from tomato leaves and Lima bean leaves was recorded, whereas no response was observed to volatiles from cucumber leaves, or leaves of Solanum luteum and Solanum dulcamara.Different results were obtained for predators that differed in rearing history. Predators that were reared on spider mites (Tetranychus urticae) on Lima bean leaves did respond to volatiles from Lima bean leaves, while predators that had been reared on the same spider mite species but with cucumber as host plant did not respond to Lima bean leaf volatiles. This effect is compared with the effect of rearing history on the response of P. persimilis to volatile allelochemicals of prey-infested plant leaves.     

Altered host plant preference of Tetranychus urticae and prey preference of its predator Phytoseiulus persimilis (Acari: Tetranychidae, Phytoseiidae) on transgenic Cry3Bb-eggplants

The behavior of the two-spotted spider mite, Tetranychus urticae Koch and the predatory mite Phytoseiulus persimilis A.-H. was investigated in laboratory experiments with transgenic Bt-eggplants, Solanum melongena L., producing the Cry3Bb toxin and corresponding isogenic, non-transformed eggplants. In bitrophic experiments, dual-choice disc tests were conducted to reveal the effects of transgenic eggplants on host plant preference of T. urticae. Adult spider mite females were individually placed on leaf discs (2 cm diameter) and were observed during five days. Females occurred significantly more frequently on transgenic halves on which also significantly more T. urticae eggs were found. The effects of a Cry3Bb-eggplant fed prey on the feeding preference of P. persimilis were investigated in tritrophic experiments. Sixteen spider mite females, eight of which had been taken from transgenic and eight from isogenic eggplants, were offered to well-fed females of P. persimilis and numbers of respective spider mites consumed were registered 12 h later when the predators were offered new spider mites again. This procedure was repeated six times. The results revealed that predatory mites consumed significantly less Bt-fed spider mites than prey that had been raised on control eggplants. These results indicate that eggplants expressing the Cry3Bb toxin for resistance against the Colorado potato beetle are more preferred by spider mites but are less preferred by their predator P. persimilis. Possible consequences of these findings for biological control of spider mites on eggplants are discussed.     

Karyotype of Norway spruce by multicolor FISH

The chromosomes (2n = 2x = 24) of Norway spruce are very large since their size reflects the huge amount of genomic DNA (2C = 30 × 10⁹ bp). However, the identification of homologous pairs is hampered by their high degree of similarity at the morphological level. Data so far presented in the literature were not sufficient to solve all the ambiguities in chromosome identification. Several genomic Norway spruce DNA clones containing highly repetitive sequences have been identified and characterised in our laboratory. Three of them were selected for fluorescent in situ hybridization (FISH) experiments because of their strong signals and suitability for chromosome identification: PATR140 hybridized at the centromeric site of three chromosome pairs; PAF1 hybridized in six subtelomeric and two centromeric sites; 1PABCD6 co-localized with the subtelomeric sites identified by PAF1. The statistical analysis of microscopic measurements of chromosomes in combination with the FISH signals of these probes allowed the unambigous construction of Norway spruce karyotype. We also compared the karyotype of Norway spruce with that of other spruce species to infer the number and kind of rearrangements that have occurred during the evolution of these species.Communicated by D.B. Neale     

Oviposition behavior in spruce budwormChoristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae)

Epicuticular waxes of balsam fir, Abies balsamea,act as oviposition stimulants for fertilized females of the eastern spruce budworm, Choristoneura fumiferana(Clem.). Those same waxes do not stimulate unfertilized females, nor do the latter respond to nonchemical cues such as leaf texture, shape, etc. Ablation experiments suggest that the chemoreceptors responsible for the perception of waxes are located on the proboscis.