Neolignan and flavonoid glycosides in Juniperus communis var. depressa

Two neolignan glycosides (junipercomnosides A and B) were isolated from aerial parts of Juniperus communis var. depressa along with two known neolignan glycosides and seven flavonoid glycosides. The structures of the isolated compounds were determined by spectral analysis, in particular by 2D-NMR analysis. The significance of distribution of flavonoids in the chemotaxonomy of genus Juniperus was also discussed.     

Subterranean, herbivore-induced plant volatile increases biological control activity of multiple beneficial nematode species in distinct habitats

While the role of herbivore-induced volatiles in plant-herbivore-natural enemy interactions is well documented aboveground, new evidence suggests that belowground volatile emissions can protect plants by attracting entomopathogenic nematodes (EPNs). However, due to methodological limitations, no study has previously detected belowground herbivore-induced volatiles in the field or quantified their impact on attraction of diverse EPN species. Here we show how a belowground herbivore-induced volatile can enhance mortality of agriculturally significant root pests. First, in real time, we identified pregeijerene (1,5-dimethylcyclodeca-1,5,7-triene) from citrus roots 9-12 hours after initiation of larval Diaprepes abbreviatus feeding. This compound was also detected in the root zone of mature citrus trees in the field. Application of collected volatiles from weevil-damaged citrus roots attracted native EPNs and increased mortality of beetle larvae (D. abbreviatus) compared to controls in a citrus orchard. In addition, field applications of isolated pregeijerene caused similar results. Quantitative real-time PCR revealed that pregeijerene increased pest mortality by attracting four species of naturally occurring EPNs in the field. Finally, we tested the generality of this root-zone signal by application of pregeijerene in blueberry fields; mortality of larvae (Galleria mellonella and Anomala orientalis) again increased by attracting naturally occurring populations of an EPN. Thus, this specific belowground signal attracts natural enemies of widespread root pests in distinct agricultural systems and may have broad potential in biological control of root pests.     

Volatile oils of mature and juvenile leaves of Juniperus horizontalis: Chemosystematic significance

Comparison of 39 terpenoids between young (juvenile) foliage and mature (adult) foliage from naturally growing plants of Juniperus horizontalis revealed no significant differences. Canonical variate analysis of the terpenoids of J. scopulorum and J. virginiana along with the mature foliage of J. horizontalis and co-plotting juvenile foliage showed a slight loosening of the J. horizontalis group but not enough to blur taxonomic distinctions. These results stand in sharp contrast with the previous work on J. scopulorum and appear to be due to the indeterminant growth pattern seen in J. horizontalis.     

Photosynthetic gas exchange and the stable isotope composition of leaf water: comparison of a xylem-tapping mistletoe and its host

Photosynthetic gas exchange and the stable isotopic composition of foliage water were measured for a xylem tapping mistletoe, Phoradendron juniperinum, and its host tree, Juniperus osteosperma, growing in southern Utah. The observed isotopic composition of water extracted from foliage was compared to predictions of the Craig-Gordon model of isotopic enrichment at evaporative sites within leaves. Assimilation rates of juniper were higher and stomatal conductance was lower than the values observed for the mistletoe. This resulted in lower intercellular/ ambient CO₂ values in the juniper tree relative to its mistletoe parasite. For mistletoe, the observed foliage water hydrogen and oxygen isotopic enrichment was less than that predicted by the model. In juniper, foliage water hydrogen isotopic enrichment was also lower than that predicted by the evaporative enrichment model. In contrast, the oxygen isotopic enrichment in juniper foliage water was slightly greater than that predicted for the evaporative sites within leaves. Hydrogen isotopic enrichment in mistletoe foliage shows systematic variation with stem segment, being highest near the tips of the youngest stems and decreasing toward the base of the mistletoe, where isotopic composition is close to that of stem water in the host tree. In a correlated pattern, mid-day stomatal conductance declined abruptly in mistletoe foliage of increasing age.     

Starvation induced stress and the susceptibility of the Colorado potato beetle,Leptinotarsa decemlineata,to infection by Beauveria bassiana

Starvation of second instar Colorado potato beetle larvae for 24h immediately after treatment with Beauveria bassiana conidia increased susceptibility to the pathogen and subsequent sporulation of cadavers but decreased time to larval death. In feeding studies, B. bassiana-treatment had no effect on subsequent larval development, and mortality occurred 5-6 days after treatment. Twenty-four hours of starvation alone retarded subsequent larval development but did not affect mortality. Mortality of B. bassiana-treated starvation stressed larvae occurred 4-5 days after treatment. Both B. bassiana treatment and 24h starvation significantly reduced total foliage consumption and daily weight gains. On the day of treatment, B. bassiana had no effect on the efficiency with which food was converted to biomass (ECI). ECI was not affected by B. bassiana or starvation alone on the day following treatment but was significantly affected by a combination of both. When larvae were exposed to a range of limited food quantities, ECI decreased with decreasing food availability but only extreme stress (starvation for 24h) increased susceptibility to B. bassiana. Topical application of Dacryodes excelsa resin (an antifeedant) to potato leaves caused a concentration dependent reduction in foliage consumption and weight gain by second instar larvae but did not affect larval mortality. When larvae were exposed to a fixed concentration of B. bassiana and a range of antifeedant concentrations there were significant linear relationships between 24h larval weight gain and mortality and 24h larval weight gain and sporulation. The interaction between starvation stress and the susceptibility to B. bassiana infection is discussed and its possible implications in pest management considered.     

Aerial dispersal and host plant selection by neonate Thyridopteryx ephemeraeformis (Lepidoptera: Psychidae)

Abstract.  1. Neonate evergreen bagworms, Thyridopteryx ephemeraeformis (Haworth) (Lepidoptera: Psychidae), disperse by dropping on a strand of silk, termed silking , and ballooning on the wind. Larvae construct silken bags with fragments of plant foliage. This species is highly polyphagous, feeding on more than 125 species of woody plants of 45 families. The larvae commonly infests juniper ( Juniperus spp.) and arborvitae ( Thuja spp.), but rarely feed on deciduous hosts such as maples. The hypothesis is proposed that polyphagy in T. ephemeraeformis is maintained by variation among larvae in dispersal behaviour, and time constraints on the opportunity to disperse, but patterns of host species preference result from a predisposition for larvae to settle on arborvitae and juniper but disperse from other hosts.
2. Consistent with that hypothesis, laboratory experiments revealed: (a) starved larvae varied in their tendency to disperse from paper leaf models; (b) starved larvae readily silked only during their first day; (c) larvae became increasingly sedentary the longer they were exposed to plant foliage; (d) when provided with several opportunities to silk, larvae became sedentary after exposure to arborvitae foliage, but repeatedly silked after exposure to maple ( Acer species) foliage or paper; and (e) larvae were less inclined to silk from foliage of arborvitae than from maple.
3. Field experiments supported the hypothesis by demonstrating that: (a) neonates tended to disperse from maple leaves while larvae older than 1 day tended to settle and remain; and (b) neonates were less likely to disperse from arborvitae and juniper trees than from maples.     

Chemically enhanced phytoextraction of lead-contaminated soils

The effects of the combined application of soil fungicide (benomyl) and ethylenediaminetetraacetic acid (EDTA) on lead (Pb) phytoextraction by ryegrass (Lolium perenne) were examined. Twenty-five pots of Pb-contaminated soil (200 mg Pb kg(-1)) were seeded with ryegrass and randomly arranged into the following treatments: (1) Control, (2) benomyl, (3) EDTA, (4) benomyl and EDTA (B+E), and (5) benomyl followed by an application of EDTA 14 days later (B .. . E). Chemicals were applied when plants had reached maximum growth. Plants were analyzed for foliage Pb concentration using inductively coupled argon plasma (ICAP) spectrometry. The synergistic effects of the combined benomyl and EDTA application (treatments 4 and 5) were made evident by the significantly (p < 0.05) highest foliage Pb concentrations. However, the foliage dry biomass was significantly lowest for plants in treatments 4 and 5. The bioaccumulation factor (BF) and phytoextraction ratio (PR) were highest for plants in treatment 5 followed by plants in treatment 4.     

The green peach aphid,Myzus persicae,acquires a LIPOXYGENASE5-derived oxylipin from Arabidopsis thaliana,which promotes colonization of the host plant

Oxylipins derived from lipoxygenase (LOX) activity play important roles in plant growth, development and stress response. In a recent study, we provided evidence that infestation of Arabidopsis thaliana foliage by the green peach aphid (GPA; Myzus persicae), a phloem sap-consuming insect, was promoted by plant LOX5-derived oxylipins. In comparison to the wild-type (WT) plant, GPA population was smaller on the Arabidopsis lox5 mutant. The insect spent less time feeding from the sieve element and xylem of the lox5 mutant compared with the WT plant. In addition, compared with insects feeding on the WT plant, when on the lox5 mutant, the GPA was unable to suppress an antibiotic activity that is present in Arabidopsis vascular sap. Roots are the critical source of a LOX5-derived oxylipin(s) that promotes colonization of the foliage by GPA. Here we show that the 9-hydoxy-10E, 12Z-octadecadienoic acid (9-HOD), a LOX5-derived oxylipin, accumulated in GPA that were reared on the WT, but not the lox5 mutant plant. However, 9-HOD accumulated in insects reared on lox5 mutant plants that were irrigated with 9-HOD, thus indicating that the insect ingests oxylipins from the host plant. We further demonstrate that the host plant requires LOX5 function to promote expression of the defense regulatory gene PHYTOALEXIN-DEFICIENT4 in the foliage. Taken together, our previous observations and results presented here indicate that while the host plant utilizes LOX5-dependent factors for promoting defense mechanisms, GPA has evolved to utilize plant 9-LOX-derived oxylipins as cues to facilitate infestation, thus suggesting a complex involvement of oxylipins in Arabidopsis interaction with GPA.     

SEASONAL VARIATION IN THE VOLATILE TERPENOIDS OF JUNIPERUS SCOPULORUM (CUPRESSACEAE)

Fifteen trees in a natural population of Juniperus scopulorum Sarg. were sampled at 4-week intervals throughout a year and the terpenoid constituents were extracted from the foliage. Several environmental variables were measured. Correlations with changes in volatile oil composition (computed as both relative percentages and as weights) were examined. Significant seasonal variations were discovered in both the relative percentage and weight data. These seasonal changes in the volatile oil composition were correlated with linear growth, temperature, yield of volatile oil, and dry weight of foliage. Seasonal variation in the terpenoids computed on a weight basis was found to be greater than the seasonal variation of the terpenoids computed as relative percents of the total oil. Evidence is presented which indicates that there is no stable period to sample on a weight basis, but sampling in the late fall and winter periods minimizes variation in the relative percentage data. The use of relative percentage data is thus encouraged for chemosystematic studies.     

NMR spectroscopic analysis of exopolysaccharides produced by Leuconostoc citreum and Weissella confusa

Dextrans are the main exopolysaccharides produced by Leuconostoc species. Other dextran-producing lactic acid bacteria include Streptococci, Lactobacilli, and Weissella species. Commercial production and structural analysis has focused mainly on dextrans from Leuconostoc species, particularly on Leuconostoc mesenteroides strains. In this study, we used NMR spectroscopy techniques to analyze the structures of dextrans produced by Leuconostoc citreum E497 and Weissella confusa E392. The dextrans were compared to that of L. mesenteroides B512F produced under the same conditions. Generally, W. confusa E392 showed better growth and produced more EPS than did L. citreum E497 and L. mesenteroides B512F. Both L. citreum E497 and W. confusa E392 produced a class 1 dextran. Dextran from L. citreum E497 contained about 11% alpha-(1-->2) and about 3.5% alpha-(1-->3)-linked branches whereas dextran from W. confusa E392 was linear with only a few (2.7%) alpha-(1-->3)-linked branches. Dextran from W. confusa E392 was found to be more linear than that of L. mesenteroides B512F, which, according to the present study, contained about 4.1% alpha-(1-->3)-linked branches. Functionality, whether physiological or technological, depends on the structure of the polysaccharide. Dextran from L. citreum E497 may be useful as a source of prebiotic gluco-oligosaccharides with alpha-(1-->2)-linked branches, whereas W. confusa E392 could be a suitable alternative to widely used L. mesenteroides B512F in the production of linear dextran.     

Volatile emissions of eastern hemlock,Tsuga canadensis,and the influence of hemlock woolly adelgid

The volatile emissions of eastern hemlock, Tsuga canadensis Carriere, were identified and quantified using standard and chiral gas chromatography and mass spectrometry. All of the identified compounds were monoterpenes, and included alpha-pinene, myrcene, tricyclene, camphene, alpha-phellandrene, beta-pinene, limonene, beta-phellandrene, terpinolene, and bornyl acetate. alpha-Pinene, myrcene, and camphene comprised greater than 75% by mass of the total release. Infestation by the exotic insect, hemlock woolly adelgid (HWA, Adelges tsugae Annand), resulted in an increased release rate of monoterpenes from branch tips. Release rate was negatively correlated to the amount of the branch tip sample that was new growth, suggesting that release rate is greater from previous-year foliage. Additionally the percent composition of the volatile profile is slightly altered by infestation, with alpha-pinene comprising 57% of volatiles from infested foliage and 66% from uninfested foliage.     

Oviposition preference of a Eucalyptus herbivore and the importance of leaf age on interspecific host choice

1. Paropsine chrysomelids are eucalypt folivores for which it has been hypothesized that the availability of suitable age class foliage (i.e. young foliage) is a more important determinant of host use than the physical and/or chemical characteristics of foliage from different hosts. This prediction was tested by assessing the oviposition preference of Chrysophtharta bimaculata (Olivier) on excised branches of Eucalyptus regnans F. Mueller and E. nitens (Deane & Maiden) Maiden.
2. In cage trials, and as observed in the wild, beetles oviposited preferentially on E. regnans when the branches of both eucalypt species displayed the full range of foliar developmental classes, from immature to fully expanded. However, when the branches were pruned so that both species displayed only immature and expanding leaves, no difference in oviposition between the species occurred. Further, when the branches were pruned so that E. regnans displayed only fully expanded leaves and E. nitens displayed only immature and expanding leaves, females oviposited entirely on the latter species.
3. These results support and extend the hypothesis that C. bimaculata 's ovipositional preference is dictated more by leaf age class than by the tree species from which the leaves derive. In the discussion it is proposed that a eucalypt that initiates and expands leaves rapidly will be less preferred for oviposition than one that develops and expands leaves slowly, because of the smaller area of its canopy with less sclerophyllous foliage.     

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.     

Mechanisms of enhanced cellulosic bioethanol fermentation by co-cultivation of Clostridium and Thermoanaerobacter spp

Engineering microbial consortia capable of efficient ethanolic fermentation of cellulose is a strategy for the development of consolidated bioprocessing for bioethanol production. Co-cultures of cellulolytic Clostridium thermocellum with non-cellulolytic Thermoanaerobacter strains (X514 and 39E) significantly improved ethanol production by 194-440%. Strain X514 enhanced ethanolic fermentation much more effectively than strain 39E in co-cultivation, with ethanol production in X514 co-cultures at least 62% higher than that of 39E co-cultures. Comparative genome sequence analysis revealed that the higher ethanolic fermentation efficiency in strain X514 was associated with the presence of a complete vitamin B(12) biosynthesis pathway, which is incomplete in strain 39E. The significance of the vitamin B(12)de novo biosynthesis capacity was further supported by the observation of improved ethanol production in strain 39E by 203% following the addition of exogenous vitamin B(12). The vitamin B(12) biosynthesis pathway provides a valuable biomarker for selecting metabolically robust strains for bioethanol production.     

A rationally designed monomeric variant of anthranilate phosphoribosyltransferase from Sulfolobus solfataricus is as active as the dimeric wild-type enzyme but less thermostable

The anthranilate phosphoribosyltransferase from Sulfolobus solfataricus (ssAnPRT) forms a homodimer with a hydrophobic subunit interface. To elucidate the role of oligomerisation for catalytic activity and thermal stability of the enzyme, we loosened the dimer by replacing two apolar interface residues with negatively charged residues (mutations I36E and M47D). The purified double mutant I36E+M47D formed a monomer with wild-type catalytic activity but reduced thermal stability. The single mutants I36E and M47D were present in a monomer-dimer equilibrium with dissociation constants of about 1 μM and 20 μM, respectively, which were calculated from the concentration-dependence of their heat inactivation kinetics. The monomeric form of M47D, which is populated at low subunit concentrations, was as thermolabile as monomeric I36E+M47D. Likewise, the dimeric form of I36E, which was populated at high subunit concentrations, was as thermostable as dimeric wild-type ssAnPRT. These findings show that the increased stability of wild-type ssAnPRT compared to the I36E+M47D double mutant is not caused by the amino acid exchanges per se but by the higher intrinsic stability of the dimer compared to the monomer. In accordance with the negligible effect of the mutations on catalytic activity and stability, the X-ray structure of M47D contains only minor local perturbations at the dimer interface. We conclude that the monomeric double mutant resembles the individual wild-type subunits, and that ssAnPRT is a dimer for stability but not for activity reasons.     

内生真菌角担子菌B6对连作西瓜土壤尖孢镰刀菌的影响

建立了快速定量检测土壤中角担子菌(Ceratobasidum stevensii)B6的实时定量PCR方法,同时跟踪了土壤中尖孢镰刀菌数量的动态变化,以及不同的发酵组分对开花期西瓜土壤微生物区系的影响。PCR扩增分析表明引物Cf1/Cr1有很好的特异性,能对角担子菌B6特异性扩增得到371bp的条带,对其它10株真菌不能有效扩增。使用荧光定量PCR对施加角担子菌B6的土壤总DNA扩增,结果表明将活菌B6固体发酵物施加到土壤1周后,B6的数量有一定的增殖,达到7.4 log(pg DNA/g干土),随着时间推移,数量逐渐减少,在第5周的时候低于检测限;而液体发酵液处理从一开始施加到土壤后,B6的数量就开始逐渐减少,在第4周的时候就低于检测限。施加活的B6菌4周内能够有效地控制尖孢镰刀菌的数量(尖孢镰刀菌数量维持在5×103 CFU/g干土左右),之后随着B6数量的减少尖孢镰刀菌数量大量增加。活菌B6在土壤中能够存活1个月左右,不会过度影响土著微生物区系,是一株环境友好型菌株,对土壤微环境的干扰较小。     

Further compatibility tests of the entomopathogenic fungus Lecanicillium muscarium with conventional insecticide products for control of sweetpotato whitefly, Bemisia tabaci on poinsettia plants

The effect on spore germination of the entomopathogenic fungus Lecanicillium muscarium following direct exposure for 24 h to the insecticides Majestik, Spray Oil, Agri-50E, Savona and Oberon for the control of both egg and second instar stages of the sweetpotato whitefly, Bemisia tabaci , was determined. Exposure to both Agri-50E and Oberon was followed by acceptable spore germination. Infectivity rates of L. muscarium on poinsettia foliage in the presence of dry residues of the insecticides were also investigated. No significant detrimental effects on the levels of control of B. tabaci were recorded compared with fungus applied to residue-free foliage. Sequential application of the chemicals Savona, Spray Oil and Majestik with the fungus all produced mortalities of second instar B. tabaci above 90%. Incorporation of these chemicals with L. muscarium into integrated control programs for B. tabaci is discussed.     

Life-history strategies affect aphid preference for yellowing leaves

According to the nutrient-translocation hypothesis, yellowing tree leaves are colonized by aphids at the end of the growing season owing to improved availability of nutrients in the phloem sap after chlorophyll degradation. We measured aphid densities on potted Betula pendula seedlings in a field site where a small proportion of foliage rapidly turned yellow before normal autumn coloration as a consequence of root anoxia. The number of adults and nymphs of the birch-feeding specialist aphids Euceraphis betulae, Betulaphis brevipilosa and Callipterinella tuberculata were counted from leaves on each of the 222 plants. Aphids were detected on 19 per cent of green leaves and on 41 per cent of yellow leaves. There was no indication of aphid avoidance of yellow leaves, and the number of winged (alate) viviparous E. betulae adults and their nymphs were significantly higher on yellow leaves than on green leaves, while the numbers of apterous B. brevipilosa and C. tuberculata did not differ between the leaf colour types. Our result suggests that only aphid species with alate generation during colour change can take advantage of yellowing leaves. This may explain the exceptional abundance of E. betulae compared with other aphid species on birches.     

Investigations on the destruxin production of the entomopathogenic fungus Metarhizium anisopliae

The dynamics of cyclic peptide destruxins (dtxs) produced by Metarhizium anisopliae strains V245 and V275 were monitored both on solid and in liquid media. The results showed that both strains did not produce dtxs in large-scale fermenter cultures or solid Czapek Dox (CD) agar. Production of the major dtxs A and B could be determined in both strains when grown on rice for up to 10-30 days. The main dtxs A, B, E, and E diol were detected in CD liquid culture filtrate from both strains after three days post-inoculation on. Parallel decrease of dtx E and increase of E diol in the culture medium were found, indicating that the latter is the hydrolytic product from the former. Production of dtxs A and B was significantly positively correlated. A negative correlation was observed between the production of the metabolites and pH value of the medium. The influence of different nutrient sources on dtx production was evaluated by using media with different carbon and nitrogen ratios as well as with different insect homogenates. The findings showed that the amount of dtxs A, B, and E increased with the increasing content of peptone in the medium. When insect homogenate was used as single nutrient source or added to CD medium, no toxins were detected in the culture filtrate. The potential risk posed by the toxic metabolites during mass production is discussed.     

Host-mediated effects of feeding by winter moth on the survival of Euceraphis betulae

Abstract 1. A potential host-mediated interaction between the birch-feeding aphid Euceraphis betulae Koch (Homoptera: Aphididae) and Operophtera brumata L. (Geometridae: Lepidoptera) was investigated, by measuring survival and feeding preference responses of E. betulae to leaves damaged recently by O. brumata larvae.
2. Euceraphis betulae survival was considerably lower on damaged foliage than on undamaged foliage, both in field experiments using cages and on potted saplings in laboratory tests.
3. Euceraphis betulae did not avoid damaged foliage in field experiments although they did avoid damaged foliage in laboratory preference tests using individual leaves. A link between aphid preference and performance on damaged foliage could therefore be demonstrated in the greenhouse/laboratory but not in the field. Multiple factors may influence feeding preference in the more complex field environment.
4. These results suggest that host-mediated competition may occur between O. brumata and E. betulae , especially in outbreak years when winter moth populations are very high.