Mutation of fungal endoglucanases into glycosynthases and characterization of their acceptor substrate specificity

Humicola insolens mutant Cel7B E197A is a powerful endo-glycosynthase displaying an acceptor substrate specificity restricted to β-d-glucosyl, β-d-xylosyl, β-d-mannosyl and β-d-glucosaminyl in +1 subsite. Our aim was to extend this substrate specificity to β-d-N-acetylglucosaminyl, in order to get access to a wider array of oligosaccharidic structures obtained through glycosynthase assisted synthesis. In a first approach a trisaccharide bearing a β-d-N-acetylglucosaminyl residue was docked at the +1 subsite of H. insolens Cel7B, indicating that the mutation of only one residue, His209, could lead to the expected wider acceptor specificity. Three H. insolens Cel7B glycosynthase mutants (H209A, H209G and H209A/A211T) were produced and expressed in Aspergillus oryzae. In parallel, sequence alignment investigations showed that several cellulases from family GH7 display an alanine residue instead of histidine at position 209. Amongst them, Trichoderma reesei Cel7B, an endoglucanase sharing the highest degree of sequence identity with Humicola Cel7B, was found to naturally accept a β-d-N-acetylglucosaminyl residue at +1 subsite. The T. reesei Cel7B mutant nucleophile E196A was produced and expressed in Saccharomyces cerevisiae, and its activity as glycosynthase, together with the H. insolens glycosynthase mutants, was evaluated toward various glycosidic acceptors.     

Inhibitory effects of different quantitative compositions of Hymenaea leaf resins on a generalist herbivore Spodoptera exigua

The hypotheses that quantitative variation in leaf resins in the leguminous genus Hymenaea may partially be a response to insect predation was tested in feeding experiments with the generalist herbivore, the beet armyworm, Spodoptera exigua Hübn (Noctuidae). Leaf resins of all Hymenaea species are comprised of essentially the same suite of sesquiterpene hydrocarbons, but discrete quantitative patterns have been classified as Compositional Types based on the amounts of major components. In this work pure extracts of leaf resins of Type I (high - and β-selinene), Type II (intermediate amounts of -and β-selinene and caryophyllene) and Type III (high caryophyllene) were incorporated into an artificial diet for the insects at 1 and 3.2% (dry wt). Resin Type treatments produced differential dose-dependent effects on growth rate (lower larval weights and increased time to pupation) and in mortality. More significant inhibitory effects occurred in Compositional Types with a predominance of a single compared (i.e. Types I and III). Significantly higher mortality occurring in Type III treatments suggests that caryophllene may have higher potential toxicity than - and β-selinene for S. exigua. These experiments indicate that feeding by generalist herbivores could be a factor determining quantitative compositional variation: (a) among populations; (b) during the development of leaves; (c) in the spatial distribution within the leaves; and (d) between parent tree and seedling progeny.     

Chemotaxonomic significance of flavonoids and phenolic acids in the Hieracium rohacsense group (Hieracium sect. Alpina; Lactuceae, Compositae)

Five apomictic taxa from the Hieracium rohacsense group were studied for their phenolic constituent composition. The following substances represent dominant compounds in the leaves: chlorogenic acid, 3,5-dicaffeoylquinic acid, luteolin 7-O-β- -glucopyranoside, luteolin 4′-O-β- -glucuronopyranoside and apigenin 4′-O-β- -glucuronopyranoside. Within the group only quantitative differences were found, luteolin 7-O-glucoside being the most important chemotaxonomic marker. Each taxon has its own specific quantitative pattern, invariable within the taxon. Based on these characteristic profiles, H. rohacsense can be distinguished from a closely related and still undescribed taxon from Mt. Pip Ivan. The proportion of luteolin 7-O-glucoside to apigenin 4′-O-glucuronoside also clearly separates the individuals of two morphologically close species—H. ratezaticum and H. pseudocaesium, which corresponds to a few slight but recognisable morphological and phenological characteristics. The ontogenetic stage of leaf development and seasonal variation are also important factors, which must be taken into consideration, as the quantity of the substances changes during leaf ontogeny and with season.     

Implications of variation in resin composition among organs, tissues and populations in the tropical legume Hymenaea

Variation in leaf blade, petiole and primary stem resins, composed of sesquiterpene hydrocarbons, is analysed in two contrasting species of the tropical legume Hymenaea. Five populations of the New World H. courbaril, spanning a wide range of ecosystems, are compared with a population of the disjunct African H. verrucosa. Resins in petiole and primary stem tissue are similar, but differ significantly in total composition from leaf blade tissues. The major components, caryophyllene, - and β-selinene and β-copaene vary most significantly among the tissues, but all compounds vary at highly significant levels among populations. The variation patterns in resin composition among the leaf blade and petiole/primary stem tissues are put into a larger context of comparison with the primarily diterpenoid patterns in secondary stem and pod tissues. Although the comparatively minor quantitative differences in the sesquiterpene systems could be attributed solely to developmental and physiological differences among the tissues and populations, the total weight of accumulating evidence regarding quantitative variation and demonstrated toxic and deterrent properties of sesquiterpene resins to insect herbivores leads us to hypothesize a possible role of differential predator pressures.     

Carotenoids of Heterosigma akashiwo: A chemosystematic contribution

The presence of C₃₇-norcarotenoids (peridinin and probably pyrrhoxanthin, together 87% of total carotenoids) and the carotenoid pattern in general, including dinoxanthin, diatoxanthin and β,β-carotene, but no fucoxanthin, strongly suggest that H. akashiwo is a dinoflagellate and not a chrysophyte.     

In vitro culture of the seagrass Halophila decipiens

The seagrass Halophila decipiens Ostenfeld was grown axenically in a culture medium consisting of 20% artificial seawater, f/4 nutrients (except that glutamic acid was the nitrogen source), and 1% sucrose (w:v). The culture medium was adjusted to pH 5.0. A root–rhizome layer was created by solidifying a portion of the medium with 0.9% agar (w:v) and 1% activated charcoal (w:v). The rhizome layer also contained the following vitamins: 0.5 mg l⁻¹ nicotinic acid, 0.5 mg l⁻¹ pyridoxine, 0.5 mg l⁻¹ biotin, 0.5 mg l⁻¹ cyanocobalamin and 0.1 mg l⁻¹ of thiamine HCl. The liquid overlay (without vitamins or charcoal) was poured onto the agar-solidified root–rhizome layer. Growth of H. decipiens was not improved by the addition of the auxins indoleacetic acid (IAA), indolebutyric acid (IBA) or naphthaleneacetic acid (NAA) at either of the tested concentrations (10 and 50 μM). At a concentration of 10 μM, the cytokinins 6-(γ,γ-dimethylallylamino) purine (2iP) and benzylaminopurine (BA) stimulated shoot and branch production compared to controls with no cytokinins. Among the tested nitrogen sources, growth was best on 1.7 mM glutamic acid. Cultures grown on 1.7 mM NH₄Cl showed the same growth rates as those grown on glutamic acid, but the leaves were smaller and curled, suggesting incipient ammonium toxicity. Use of nitrate or urea led to mortality of the cultures. Long term axenic culture of H. decipiens appears to require the added vitamins. Hence, H. decipiens is the first seagrass known to need exogenous vitamins. Cultures of H. decipiens died when grown suspended in liquid cultures or in a biphasic medium system without activated charcoal in the root–rhizome layer. The use of more highly charged κ-carrageenan could not replace the use of activated charcoal and agar in the root–rhizome layer.     

Settlement of the serpulid polychaete Hydroides elegans (Haswell) on the arborescent bryozoan Bugula neritina (L.): evidence of a chemically mediated relationship

The tube building polychaete Hydroides elegans Haswell was found living attached to colonies of the arborescent bryozoan Bugula neritina (L.) in Port Shelter, Hong Kong. Field data collected during the period of January through May 1996, showed that H. elegans density reached 77.6 individuals of H. elegans per g wet weight of B. neritina. Density of H. elegans on B. neritina at depths from the surface to 0.5 m was lower than that at depths below 1 m. In January–March, when there were no H. elegans settling on PVC plates or found on natural substrata, numbers on B. neritina were ca. 5 per g wet weight. H. elegans settled on B. neritina and grew rapidly as mean diameter of tubes increased from 605 μm in February to 936 μm in March. In laboratory experiments, larvae of H. elegans settled and metamorphosed on branches of B. neritina and on the bottom of dishes containing B. neritina leachate. Compounds extracted from the leachate of B. neritina induced 74% of H. elegans larvae to metamorphose at a concentration of 16 μg/ml seawater, compared to 5% in dishes containing only filtered seawater (controls). Metabolites from the leachate of B. neritina which were bound to amberlite XAD-2, indicating they are lipophilic in nature, induced over 70% metamorphosis in H. elegans larvae at 56 μg/ml seawater. A biofilm from one of four strains of bacterial isolates associated with the surface of B. neritina induced low levels of metamorphosis in H. elegans larvae, while other bacterial isolates were detrimental to the survival of juvenile H. elegans. Field experiments further demonstrated that H. elegans settled preferentially on Phytagel discs embedded with whole extracts of B. neritina over control Phytagel discs. Metabolites from B. neritina deterred feeding on alginate pellets by assemblages of local fishes in field assays. Metabolites originating from B. neritina, bacteria colonizing B. neritina, and the complex structure of B. neritina contributed to the recruitment of H. elegans to B. neritina surfaces. Hydroides elegans may gain a refuge from predation by associating with B. neritina colonies both from its structural and chemical attributes.     

Embryonic inheritance of the chromatin organisation of the imprinted H19 domain in mouse spermatozoa

Insulin-like growth factor 2 (Igf 2) and H19 genes are oppositely imprinted and as such have been most extensively studied imprinted genes both genetically and at the molecular level. Imprints of the H19 gene, being established during spermatogenesis, are epigenetically transmitted to the somatic cells of the embryo. Current hypotheses attempting to explain the allele-specific silence of the H19 gene include DNA methylation and chromatin condensation. In order to understand the molecular basis of H19 epigenesis, it is crucial to identify the markings in the chromatin organising the imprinted domain in spermatozoa. Using Micrococcal nuclease (MNase), DNase I and Methidiumpropyl-EDTA. iron II (MPE·Fe(II)) as chromatin probes, we demonstrate that in mouse epididymal spermatozoa, at least 4 kb DNA upstream of the H19 ‘cap’ site, containing the imprinted and differentially methylated domain (DMD), is heterochromatic. The cleavage sites in this domain (−2 to −4 kb) exhibit ~425 bp periodicity. This structure is maintained in the paternal allele of normal embryos and is disrupted at −2.2, −2.65 and at −3.5 kb in embryos maternally disomic for the distal end of chromosome 7 (MatDp 7). The hypersensitive sites in chromatin precisely register the MPE·Fe(II) cleavage sites in chromosomal DNA. Therefore, the DNA sequences in the imprinted domain constrain the chromatin structure in a way similar to that of 1.688 g/cm³ Drosophila satellite chromatin. In addition, we find that condensation of the paternal allele correlates with methylation-dependent alteration in the structure of DNA sequences in DMD. These results suggest that CpG-methylation induces localised changes in DNA conformation and these facilitate consequent remodelling of chromatin thereby allowing the paternal and maternal H19 alleles to be distinguished.     

A semi-synthetic diet for rearing Dipha aphidivora (Lepidoptera: Pyralidae), a promising predator of woolly aphid in sugarcane

Dipha aphidivora (Meyrick), a lepidopteran predator of sugarcane woolly aphid (SWA) Ceratovacuna lanigera Zehntner was successfully reared for three successive generations, for the first time on a freeze-dried beef liver-based larval semi-synthetic diet. We compared biological parameters viz., larval survival, adult weight, pre-and oviposition period, fertility, fecundity and female longevity of D. aphidivora reared on the semi-synthetic diet with the predators reared on SWA. Development time of larvae reared from first instar to pupation was 20.6 days on the semi-synthetic diet and 12 days on SWA, while survival of the larvae to adults was 61.8 and 91.8% on larval semi-synthetic diet and SWA, respectively. Fecundity recorded from semi-synthetic diet (41 eggs/female) was significantly less than those produced on SWA (58 eggs/female). However, fertility and longevity of the predators reared on SWA and semi-synthetic diet did not differ significantly. The study revealed the possibility of rearing D. aphidivora larvae using synthetic diet.     

蛇足石杉离体培养物形态变化与石杉碱甲积累的研究

通过蛇足石杉(Huperzia serrata)离体培养研究不同培养基对培养物细胞特征、总DNA特征、生物增长量和石杉碱甲含量的影响。结果表明,Sh、W和Shx三种培养基中,培养物分化成叶状体、根状体和苔藓状体3种不同的形态,三者的叶细胞拟由单细胞出芽成串丛生发育成组织,苔藓状体和根状体比叶状体的细胞表面有更多的根和假根生长;总DNA也存在差异,35、45和55天的苔藓状体均出现DNA梯带,表明形态变异可能与细胞凋亡相关;叶状体、根状体和苔藓状体的生物量增长分别为(1 788±31)%、(833±27)%和(1 963±52)%,石杉碱甲的含量分别为71.7±1.54、20.1±0.82和0μg·L–1(P0.01)。上述结果表明,培养基成分变化可以使相同的基因型材料生成不同的形态,且生长速度和代谢产生石杉碱甲的量等都发生明显变化,叶状体是产生石杉碱甲的最好形态。     

Suppressed apoptosis in the inflamed gastric mucosa of Helicobacter pylori-colonized iNOS-knockout mice

Deregulated cell turnover in Helicobacter pylori (H. pylori)-colonized gastric mucosa has been suggested to be linked to the gastric carcinogenesis pathway. We previously reported attenuation of apoptosis and enhancement of cellular proliferation in the H. pylori-colonized gastric mucosa of Mongolian gerbils as compared to that in mice, which might reflect a specific link between H. pylori colonization and carcinogenesis in the Mongolian gerbils; the difference between the two strains could be attributable to differences in the host genetic background. Inducible-type nitric oxide synthase (iNOS) is thought to participate in not only the inflammatory response, but also in the regulation of gastric mucosal cell turnover in H. pylori-colonized gastric mucosa. Thus, the present study was designed to examine gastric leukocyte activation and epithelial cell apoptosis in the gastric mucosa following H. pylori inoculation in iNOS-knockout mice. Methods: iNOS-knockout mice (iNOS^−/−) and their iNOS^+/+ littermates were orally inoculated with the Sydney strain of H. pylori (SS1, 10⁸ colony-forming units [CFU]). H. pylori infection was confirmed by microaerobic bacterial culture. The stomach of each mouse was evaluated 14 weeks and 30 weeks after the inoculation. Gastric mucosal accumulation of polymorphonuclear leukocytes (PMN) was assessed by determining the myeloperoxidase (MPO) activity and histological score based on the updated Sydney system. The level of apoptosis was determined by estimation of the cytoplasmic levels of mono- and oligonucleosomes and by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling method. Results: The SS1-inoculated mice showed persistent H. pylori colonization for 12 weeks. While gastric mucosal PMN infiltration increased following SS1 inoculation in both iNOS^+/+ and iNOS^−/−strains, enhanced DNA fragmentation was observed in only SS1-colonized iNOS^+/+ mice, and not in the iNOS^−/− mice. In conclusion, although the recruitment of PMN in response to H. pylori was evoked even in the gastric mucosa of iNOS^−/− mice, epithelial cell apoptosis induced by H. pylori was attenuated in this strain. These data suggest that iNOS may play an important role in promoting apoptosis in the H. pylori-infected inflamed gastric mucosa, and that persistent inflammation without apoptosis in iNOS^−/− mice with H. pylori infection may be linked to preneoplastic transformation.     

Flavonoids of five Hieracium species of British Columbia

Five Hieracium species were examined for flavonoid constituents: H. albertinum, H. albiflorum, H. cynoglossoides, H. gracile and H. umbellatum. The major compounds present in all species were mono- and diglycosides, of apigenin and luteolin. Chrysoeriol occurred per se in several species. Quercetin-3-O-glucoside was seen only in H. umbellatum. Species specific patterns of flavonoid glycosides were evident in all five taxa; H. albertinum and H. cynoglossoides showed the greatest similarity which reflects their morphological similarity. Limited examination of artificial hybrids showed additivity of flavonoid glycoside patterns.     

Oocysts of Neospora caninum, Hammondia heydorni, Toxoplasma gondii and Hammondia hammondi in faeces collected from dogs in Germany

Faecal samples of 24,089 dogs were examined coproscopically in two veterinary laboratories in Germany between March 2001 and October 2004. In 47 dogs, oocysts of 9–14 μm size were found. Their morphology was similar to those of Hammondia heydorni and Neospora caninum. Samples of 28 of these dogs were further examined by inoculation into gerbils: seven isolates induced a specific antibody response against antigens of N. caninum NC-1 tachyzoites. This response suggests that the isolates contained N. caninum. In addition to H. heydorni (12 times isolated), Toxoplasma gondii occysts (twice) and Hammondia hammondi oocysts (twice) were observed in dog faeces. The latter findings suggest that coprophagia with a subsequent intestinal passage by dogs plays a role in the dissemination of coccidian parasites for which cats are definitive hosts. Five of the seven N. caninum (NC-GER2, NC-GER3, NC-GER4, NC-GER5, NC-GER6) and the two T. gondii isolates (TG-dgGER1, TG-dgGER2) were successfully passaged into cell culture and are now available for detailed characterization. In contrast to oocysts of other parasites, N. caninum oocysts were predominantly found between January and April (Fisher exact; P=0.038). In the sera of dogs shedding N. caninum, no reactions against the immunodominant antigens with apparent molecular weights of 19, 29, 30, 33 and 37 kDa of N. caninum tachyzoites were observed 3–5 weeks after shedding. However, the animals recognized a 152-kDa N. caninum antigen. Compared with those identified as H. heydorni, T. gondii or H. hammondi, N. caninum oocyst isolates were significantly smaller in length with the 75th percentiles ≤10.7 μm when measured in concentrated sucrose solution and smaller length–width ratios with the 75th percentiles ≤1.06. It may thus be possible to develop criteria for a preliminary identification of N. caninum in dog faeces based on the oocyst morphology.     

Response to helminth infection of sheep selected for resistance to Haemonchus contortus

Lines of Merino sheep selected for increased (IRH) and decreased (DRH) resistance to Haemonchus contortus were compared with an unselected (CH) line, after approximately four generations of selection. Measurements were recorded on 69 IRH, 47 DRH and 84 CH animals. Following artificial challenge with H. contortus, the IRH line had significantly (P < 0.001) lower faecal egg counts than the CH and DRH lines (2730,12,720 and 17,400 epg, respectively). Significant differences (P<0.05) were found between all lines in the minimum packed cell volumes during artificial infection (25.7, 22.0 and 20.3%) and in faecal egg counts after natural infection (140, 3590 and 8750 epg). Differences were also recorded (P<0.05) following artificial challenge with Trichostrongylus colubriformis (490, 840 and 1340 epg). On a percentage basis, faecal egg counts in the IRH line deviated less from the CH line following artificial infection with T. colubriformis (42%) than with H. contortus (79%). The reverse was true for the DRH line (60 and 37%, respectively). Differences in egg output of this magnitude should have a marked effect on requirements for anthelmintic treatment, rate of development of drug resistance and level of pasture contamination when the lines are grazed separately.     

Discovery of an MIT-like atracotoxin family: spider venom peptides that share sequence homology but not pharmacological properties with AVIT family proteins

This project identified a novel family of six 66–68 residue peptides from the venom of two Australian funnel-web spiders, Hadronyche sp. 20 and H. infensa: Orchid Beach (Hexathelidae: Atracinae), that appear to undergo N- and/or C-terminal post-translational modifications and conform to an ancestral protein fold. These peptides all show significant amino acid sequence homology to atracotoxin-Hvf17 (ACTX–Hvf17), a non-toxic peptide isolated from the venom of H. versuta, and a variety of AVIT family proteins including mamba intestinal toxin 1 (MIT1) and its mammalian and piscine orthologs prokineticin 1 (PK1) and prokineticin 2 (PK2). These AVIT family proteins target prokineticin receptors involved in the sensitization of nociceptors and gastrointestinal smooth muscle activation. Given their sequence homology to MIT1, we have named these spider venom peptides the MIT-like atracotoxin (ACTX) family. Using isolated rat stomach fundus or guinea-pig ileum organ bath preparations we have shown that the prototypical ACTX–Hvf17, at concentrations up to 1 μM, did not stimulate smooth muscle contractility, nor did it inhibit contractions induced by human PK1 (hPK1). The peptide also lacked activity on other isolated smooth muscle preparations including rat aorta. Furthermore, a FLIPR Ca²⁺ flux assay using HEK293 cells expressing prokineticin receptors showed that ACTX–Hvf17 fails to activate or block hPK1 or hPK2 receptors. Therefore, while the MIT-like ACTX family appears to adopt the ancestral disulfide-directed β-hairpin protein fold of MIT1, a motif believed to be shared by other AVIT family peptides, variations in the amino acid sequence and surface charge result in a loss of activity on prokineticin receptors.     

Integrated Management of Two-spotted Spider Mite Tetranychus urticae on Hops using Hop ß-acids as an Antifeedant Together with the Predatory Mite Phytoseiulus persimilis

The effects on population development of two-spotted spider mite Tetranychus urticae by sprays of hop β-acids, a repellent and antifeedant of T. urticae, together with an inoculative release of the predatory mite Phytoseiulus persimilis were compared on tall and dwarf hops. The combination of the semiochemical with the predator was more effective than either factor singly on both hop types. The highest numbers of spider mites and their eggs were recorded on untreated plots. The sprays of β-acids slowed the rate of increase of the pest and contributed more to the effect of the combined treatment early in the season when the predator was establishing itself on the crop. By harvest time, the predator was the more influential of the two factors.     

Encapsulation of Haematococcus pluvialis using chitosan for astaxanthin stability enhancement

Astaxanthin is receiving commercial interest due to its use as a preferred pigment in aquaculture feeds. Its antioxidant activity is approximately 100 times higher than that of β-tocopherol, and can be used as a potential agent against cancer. Astaxanthin can easily be degraded by thermal or oxidative processes during the manufacture and storage. In this study, astaxanthin and its biological activity were protected against oxidative environmental conditions by encapsulating the homogenized cells in chitosan. Haematococcus pluvialis were formed into beads, which were then coated with 5 layers of chitosan film, resulting in chitosan-algae capsules that have a mean diameter of 0.43 cm and the total film thickness of approximately 100 μm. No significant loss in the amount of astaxanthin content in H. pluvialis was found due to the process of encapsulation. However, approximately 3% loss of antioxidant activity of the H. pluvialis was observed after encapsulation. The results of stability under different storage conditions showed that although encapsulation caused 3% loss of antioxidant activity, the longer term stability of the dried algae biomass, beads, and capsules indicated that encapsulation of H. pluvailis in chitosan film was capable of protecting the algae cells from oxidative stresses.     

北亚热带地带性森林土壤温室气体通量对土地利用方式改变和降水减少的响应

弄清土地利用和降水变化对林地土壤主要温室气体(CO₂、CH₄和N₂O)排放通量变化的影响, 是准确评估森林土壤温室气体排放能力的重要基础。该研究以常绿落叶阔叶混交林原始林、桦木(Betula luminifera)次生林和马尾松(Pinus massoniana)人工林为对象, 采用静态箱-气相色谱法研究了3种土地利用方式(常绿落叶阔叶混交林原始林、桦木次生林和马尾松人工林)和降水减少处理状况下森林土壤CO₂、CH₄和N₂O通量排放特征, 并探讨了其环境驱动机制。研究结果表明: 原始林土壤CH₄吸收通量显著高于次生林和人工林, 次生林CH₄吸收通量显著高于人工林土壤。人工林土壤CO₂排放通量显著高于原始林和次生林土壤。次生林土壤N₂O排放通量高于原始林和人工林, 但三者间差异不显著。降水减半显著抑制了3种不同土地利用方式下林地土壤CH₄吸收通量; 降水减半处理对原始林和次生林土壤CO₂排放通量均具有显著的促进作用, 而对人工林土壤CO₂排放通量具有显著的抑制作用; 降水减半处理促进了原始林和人工林林地土壤N₂O排放而抑制了次生林林地土壤N₂O排放。原始林和次生林林地土壤CH₄吸收通量随土壤温度升高显著增加, CH₄吸收通量与土壤温度均呈显著相关关系; 原始林、次生林和人工林土壤CO₂和N₂O排放通量与土壤温度均呈显著正相关关系; 土壤湿度抑制了次生林和人工林土壤CH₄吸收通量, 其CH₄吸收通量随土壤湿度增加显著减少; 原始林土壤CO₂排放通量与土壤湿度呈显著正相关关系。自然状态下, 原始林土壤N₂O排放通量与土壤湿度呈显著正相关关系, 原始林和次生林土壤N₂O排放通量与硝态氮含量呈显著相关关系。研究结果表明全球气候变化(如降水变化)和土地利用方式的转变将对北亚热带森林林地土壤温室气体排放通量产生显著的影响。     

Host rearing is a bottleneck for classical biological control of the cherry bark tortrix: A comparative analysis of artificial diets

This paper describes a comparative analysis of the suitability of three artificial diets for the development of the cherry bark tortrix (CBT), Enarmonia formosana Scopoli (Lepidoptera: Tortricidae), to simplify the rearing process for this species and its potential classical biological control agents. The three diets tested included (1) a pinto bean-based diet modified specifically for the CBT, (2) the diet for codling moth, Cydia pomonella Linnaeus (Lepidoptera: Tortricidae), and (3) the Singh general-purpose diet. Survival from first instar to the pupal stage was very low on the pinto bean, codling moth, and Singh general-purpose diets (29, 0, and 0%, respectively). Survival was consistently greater, yet still low, for larvae that were reared through the first instar on bark and subsequently transferred to the codling moth or Singh general-purpose diets (5 and 32%, respectively). In comparison, larvae started on the pinto bean diet as second instars had a survival rate of 90%, only slightly below that of sibling larvae from the cherry bark control group (100%). Larval development time was fastest on cherry bark (36±2?days), differing significantly from that on the pinto bean diet (started as first instars: 58±2?days; started as second instars: 46±2?days), but not from the development time of larvae on the Singh general-purpose diet (44±3?days). Pupal weights were greatest for specimens from the Singh general-purpose diet (14.9±0.5?mg) and lowest for those from the pinto bean diet (started as first instar: 12.3±0.6?mg; started as second instar: 12.1±0.4?mg). Pupal weights from cherry bark were intermediate (13.5±0.6?mg). Early mortality, resulting primarily from rejection of the diet, remains to be the critical impediment in CBT rearing. It is therefore suggested that a phagostimulant from cherry bark be identified and included in an artificial diet shown to be nutritionally suitable, such as the Singh general-purpose diet or the pinto bean diet.