濒危植物南方红豆杉种群生命表及谱分析

Based on the investigation in Longxi Mountain National Nature Reserve and the theory of survival analysis, a static life table of Taxus chinensis var. mairei population was worked out, the curves of its survival rate, mortality rate and killing power were drawn, and the population dynamics was analyzed by spectral analysis. The results showed that the survival curve of the population appeared to be a type of Deevey-Ⅲ, and the high mortality of seeding was one of the important reasons which caused Taxus chinensis var. mairei to be endangered. The spectral analysis of the population showed that there was a marked periodic regularity in the process of natural regeneration of Taxus chinensis var. mairei.     

Functional diversifications of GhERF1 duplicate genes after the formation of allotetraploid cotton

Whole genome duplication, a prevalent force of evolution in plants, results in massive genome restructuring in different organisms. Roles of the resultant duplicated genes are poorly understood both functionally and evolutionarily. In the present study, differentially expressed ethylene responsive factors(GhERF1 s), anchored on Chr-A07 and Chr-D07 were isolated from a high-yielding cotton hybrid(XZM2)and its parents. The GhERF1 was located in the B3 subgroup of the ethylene responsive factors subfamily involved in conferring tolerance to abiotic stress Nucleotide sequence analysis of 524 diverse accessions together with quantitative real-time polymerase chain reaction analysis, elucidated that de-functionalization of GhERF1-7 A occurred due to one base insertion following formation of the allotetraploid cotton. Our quantitative trait loci and association mapping analyses highlighted a role for GhERF1-7 A in conferring high boll number per plant in modern cotton cultivars. Overexpression of GhERF1-7 A in transgenic Arabidopsis resulted in a substantial increase in the number of siliques and total seed yield. Neo-functionalization of GhERF1-7 A was also observed in modern cultivars rather than in races and/or landraces, further supporting its role in the development of high-yielding cotton cultivars. Both de-and neofunctionalization occurred in one of the duplicate genes,thus providing new genomic insight into the evolution of allotetraploid cotton species.     

Molecular and in vitro Characterization of Field Isolates of Bovine Herpesvirus-1

Bovine Herpesvirus-1 (BoHV-1) is distributed worldwide and is a major pathogen in cattle, being the causal agent of a variety of clinical syndromes. The aim of this study was to isolate and to characterize (molecular and biological characterization) BoHV-1 from 29 immunosuppressed animals. It was possible to obtain 18 isolates, each from a different animal, such as from the respiratory and reproductive tracts. In some cases the cytopathic effect was visible 12 hours post-inoculation, and became characteristic after 36-48 hours. Biological characteristics were evaluated and compared with Iowa and Colorado-1 reference strains, and differences were found in plaque size, virus titer measured by TCID50 and PFU/mL, and one step virus curves. These results showed that some isolates had a highly virulent-like behavior in vitro, compared to the reference strains, with shorter eclipse periods, faster release of virus into the supernatants, and higher burst size and viral titer. There were no differences in glycoprotein expression of BoHV-1 isolates, measured by Western blot on monolayers. Moreover, using restriction endonucleases analysis, most of the viruses were confirmed as BoHV-1.1 and just one of them was confirmed as BoHV-1.2a subtype. These findings suggest that some wild-type BoHV-1 isolates could be useful as seeds to develop new monovalent vaccines.     

Pedigree investigation of severe hemolytic disease of the newborn caused by rare anti-Jk3

The study was designed to evaluate the molecular and serological features of a newborn with severe hemolytic disease of the newborn (HDN) caused by anti-Jk3, and to further rich our understanding of Kidd subgroup genetics using pedigree analysis which is the first analysis of a Jk null phenotype in China and the first report of severe HDN caused by anti-Jk3. A female baby presented with hyperbilirubinemia (36 μmol/L) on the day of birth. Antibody screening tests using blood samples from the patient and her family indicated that the mother’s plasma contained alloantibodies against high frequency antigens and the results of direct Coombs test were all negative. Kidd phenotypes were Jk(a-b-), Jk(a-b+), and Jk(a-b+) in the mother, father, and baby, respectively. Kidd genotype was determined by PCR amplification of a single nucleotide polymorphism (838) and all family members were Jk(a-b+). Kidd gene exons 4 to 11 were sequenced to identify potential mutations.Sequencing analysis revealed that c.838 G>A and intron c.3 -78 G>A homozygosity occurred in all family members along with homozygosity and heterozygosity for c.IVS5-1G>A in the mother and newborn, respectively. In conclusion, serological and genetic analyses confirmed that the Jk(a-b-) phenotype was caused by homozygous IVS5-1G>A mutation of the Kidd gene. This result is consistent with that of a previous report and presents a useful diagnostic tool to identify HDN caused by anti-Jk3. A further study is required to identify the effect of intron 3 -78 G>A mutation on phenotype.     

1-(3-Aminopropyl)-3-butylimidazolium bromide for carboxyl group derivatization: potential applications in high sensitivity peptide identification by mass spectrometry

The cationic reagent 1-(3-aminopropyl)-3-butylimidazolium bromide(BAPI) was exploited for the derivatization of carboxyl groups on peptides.Nearly 100% derivatization efficiency was achieved with the synthetic peptide RVYVHPI(RI-7).Furthermore,the peptide derivative was stable in a 0.1% TFA/water solution or a 0.1%(v/v) TFA/acetonitrile/water solution for at least one week.The effect of BAPI derivatization on the ionization of the peptide RI-7 was further investigated,and the detection sensitivity was improved >42-fold via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS),thus outperforming the commercial piperazine derivatization approach.Moreover,the charge states of the peptide were largely increased via BAPI derivatization by electrospray ionization(ESI) MS.The results indicate the potential merits of BAPI derivatization for high sensitivity peptide analysis by MS.     

甾体1，4-脱氢和11α-羟基化反应的两种不同微生物转化

Two kinds of micro-organism, Arthrobacter sp. AX86(1,4-dehdrogenator)and Absidia sp. A28(11α-hydroxylator) were used in this experiment. Two different fermentation techniques were performed to accomplish the multiple conversional reactions for producing 16β-methyl-11α, 17α, 21-trihydroxy-1,4-pregnadiene-3,20-dione (Ⅲ) from 16β-methyl-3β, 17α,21-trihydroxy-5α-pregnane-20-one-21-acetate(1) 1)To produce product(Ⅲ)by means of a two-step fermentation method which were independently performed first by Arthrobacter and next by Abslaia, and 2)the product was obtained by a sequential fermentation system of aforesaid two micro-organisms in a single fermentor without isolation of the intermediates from the mixture. Our results showed that in both fermentation systems high yield of product was obtained. However, according to the technical simplicity, shorter duration of fermentation cycle and efficient yield of product, the second method is better than the first one.     

Characterization of a pectic polysaccharide from the leaves of Diospyros kaki and its modulating activity on lymphocyte proliferation

Pectin is a group of carbohydrate polymers constructing the primary cell walls and the middle lamella of terrestrial plants. Herein, we demonstrated the structure and immunomodulatory activity of the major pectic polysaccharide DL‐3B₂ isolated from the leaves of Diospyros kaki. Based on composition analysis, methylation analysis, two‐step acid hydrolysis, lithium‐mediated selective degradation, ¹³C NMR spectroscopy, and electrospray ionization mass spectrometry, DL‐3B₂ was found to contain an α‐1, 4‐linked galacturonic acid (GalA) backbone with some insertions of α‐1, 2‐linked rhamnose residues. The arabinan‐ and arabinogalactan‐side chains were attached to O‐4 of the rhamnose residues, whereas the linear arabinoxylan was probably linked to O‐3 of the GalA residues. Immunological tests in vitro showed that DL‐3B₂ could help stimulate lipopolysaccharide‐induced B lymphocyte proliferation, but not ConA‐induced T lymphocyte proliferation, and that the arabinose residues play a role in maintaining this immunological activity. © 2010 Wiley Periodicals, Inc. Biopolymers 93: 649–656, 2010. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com     

Partial structure of a polysaccharide from leaves of Welwitschia mirabilis

Gum-tears from the leaves of Welwitschia mirabilis contain a polysaccharide composed of arabinose, galactose and glucuronic acid as main constituents with xylose, fucose and rhamnose in smaller quantities. Periodate oxidation and permethylation studies indicated that the gum could consist of a framework of glucuronic acid residues linked 1 → 4 and galactose residues linked 1 → 6 and of short chains of arabinose, xylose, fucose and rhamnose linked 1 → 3 to both residues. All rhamnose and fucose and part of arabinose were found as non-reducing terminal units.     

Chemical characterisation of the released polysaccharide from the cyanobacterium Aphanothece halophytica GR02

The released polysaccharide from the halophilic cyanobacterium Aphanothece halophytica GR02 was separated into two main fractions byanion-exchange chromatography. The major fraction consisted of glucose,fucose, mannose, arabinose and glucuronic acid. Judging from thechromatography on Sepharose 2B, the major fraction was not furtherfractionated, and its apparent molecular weight was above 2.0 × 10⁶ Da.The minor fraction consisted of rhamnose, mannose, fucose,glucose, galactose and glucuronic acid, with traces of arabinose.Methylation and GC-MS spectrometry analyses of the major fractionrevealed the presence of 1-linked glucose, 1,3-linked glucose, 1,3-linkedfucose, 1,4-linked fucose, 1,3-linked arabinose, 1,2,4-linked mannose,1,3,6-linked mannose, 1-linked glucuronic acid and 1,3-linked glucuronicacid residues. The major fraction was thought to originate from capsularpolysaccharide. The released polysaccharides, obtained from cultures atdifferent age of culture, showed no striking variations in themonosaccharide composition and the relative proportions of themonosaccharides. However, the proportions of galactose and rhamnose inthe released polysaccharides, obtained from cultures under different salinity,were significantly different. The released polysaccharide also exhibitedgelling properties and strong affinity for metal ions.     

A one‐pot process for synthesis of mitomycin analogs catalyzed by laccase/lipase optimized by response surface methodology

To reach the excellent yield as well as environmental friendliness, an efficient one‐pot process for the synthesis of 2‐methyl‐3‐n‐butylaminoyl‐1,4‐benzoquinone, a mitomycin‐like compound by the domino reaction of 2‐methyl‐1,4‐hydroquinone and butylamine using laccase/lipase as co‐catalysts, has been developed. In this present study, the process proposed here was further improved by optimizing the relevant factors using the response surface methodology based on Box–Benkhen Design. The optimum condition that afforded the highest yield (98%) of 2‐methyl‐3‐n‐butylaminoyl‐1,4‐benzoquinone was obtained as follows: molar ratio of amines to hydroquinones 1.16:1, activity ratio of laccase to lipase 1.14:2, and reaction temperature 38.9°C. The results obtained indicate that this process may be useful as a green alternative method for higher yield production of mitomycin analogs.     

Metabolic fingerprinting to discriminate diseases of stored carrots

Volatile metabolites from headspace gas of carrot cv. Vita‐treat inoculated with water or four different pathogens Botrytis cinerea, Erwinia carotovora subsp. carotovora, Aspergillus niger and Fusarium avenaceum were profiled using gas chromatography and mass spectrometry to develop a technology to discriminate diseases. The inoculation of carrot roots with water or different pathogens released a total of 137 different volatile metabolites. Among them, 39 compounds were relatively consistent and 11 were specific to one or more diseases/inoculations. E. carotovora subsp. carotovora produced seven disease‐specific metabolites: 1‐butanol, 3‐methyl; 1‐pentanol; 1‐propanol, 2‐methyl; 2,3‐butanedione; boronic acid, ethyl; butane, 1‐methoxy‐3‐methyl; and ethane, ethoxy. Some metabolites were disease/inoculation discriminatory and were not detected in all treatments: 1,2‐dimethoxy‐ethene was common in carrots inoculated with E. carotovora subsp. carotovora and B. cinerea, while 2‐butanone, 3‐chloro‐4‐hydroxy‐1,4‐diphenyl was common in carrots inoculated with E. carotovora subsp. carotovora, F. avenaceum and water‐inoculated control. The significant mass ions, based on univariate analysis, from a total of 150 (46–195 m/z) and compounds from a total of 32 were further subjected to stepwise discriminant analysis and discriminant analysis. The models for 3 days after inoculation (DAI) were better than those for 6 DAI and 3 + 6 DAI, where up to 90% of the observations were correctly classified into respective inoculations. The disease‐discriminatory compounds from different diseases/inoculations and discriminant analysis models developed here have the potential for the early detection and discrimination of postharvest diseases of carrot cv. Vita‐treat, after validation under commercial conditions.     

Water-soluble polysaccharides from Ginkgo biloba leaves.

The water-soluble polysaccharides from dried Ginkgo biloba leaves were isolated after exhaustive extraction with organic solvents. The polysaccharide mixture could be separated into a neutral (GF1) and two acidic (GF2 and GF3) polysaccharide fractions by ion exchange chromatography. According to the Mr distribution GF1 and GF3 seemed to be homogenous, whereas GF2 could be further fractionated into two subfractions (GF2a and GF2b) by gel permeation chromatography. GF1 (Mr 23,000) showed the structural features of a branched arabinan. The main chain was composed of 1,5-linked arabinose residues and three in 12 arabinose molecules were branched via C-2 or C-3. GF2a (Mr 500,000) consisted mainly of 1,2,4-branched mannose (29%), 1,4-linked glucuronic (32%) and galacturonic (8%) acid as well as terminal rhamnose (25%). After removal of ca 70% of the terminal rhamnose the remaining polysaccharide showed a decrease in 1,2,4-branched mannose and an increase in 1,2-linked mannose indicating that at least half of the rhamnose residues were linked to mannose via C-4. GF3 (Mr 40,000) consisted of 1,4-linked galacturonic (30%) and glucuronic (16) acid, 1,3,6-branched galactose (15%), 1,2-linked (5%) and 1,2,4-branched (3.5%) rhamnose as well as 1,5-linked arabinose (11%). Rhamnose (5%) and arabinose (10%) were present as terminal groups. Mild acid hydrolysis selectively cleaved arabinose and the remaining polysaccharide showed an increased amount of 1,6-linked and terminal galactose and a decreased quantity of 1,3,6-branched galactose. These results indicated that the terminal as well as the 1,5-linked arabinose were mainly connected to galactose via C-3. The GF3 polysaccharide appeared to be a rhamnogalacturonan with arabinogalactan side chains.     

Resolution of 1‐n‐Butyl‐3‐Methyl‐3‐Phospholene 1‐Oxide With TADDOL Derivatives and Calcium Salts of O,O'‐Dibenzoyl‐(2R,3R)‐ or O,O'‐di‐p‐Toluoyl‐(2R,3R)‐tartaric Acid

The resolution methods applying (?)‐(4R,5R)‐4,5‐bis(diphenylhydroxymethyl)‐2,2‐dimethyldioxolane (“TADDOL”), (?)‐(2R,3R)‐α,α,α',α'‐tetraphenyl‐1,4‐dioxaspiro[4.5]decan‐2,3‐dimethanol (“spiro‐TADDOL”), as well as the acidic and neutral Ca²⁺ salts of (?)‐O,O'‐dibenzoyl‐ and (?)‐O,O'‐di‐p‐toluoyl‐(2R,3R)‐tartaric acid were extended for the preparation of 1‐n‐butyl‐3‐methyl‐3‐phospholene 1‐oxide in optically active form. In one case, the intermediate diastereomeric complex could be identified by single‐crystal X‐ray analysis. The absolute P‐configuration of the enantiomers of the phospholene oxide was also determined by comparing the experimentally obtained and calculated CD spectra. Chirality 26:174–182, 2014. © 2014 Wiley Periodicals, Inc.     

Antennal sugar sensitivity in the click beetle Agriotes obscurus

The occurrence of salt‐, sugar‐sensitive neurones and a mechanoreceptor neurone in the antennal hair‐like gustatory sensilla of the click beetle Agriotes obscurus L. (Coleoptera, Elateridae) is demonstrated using the electrophysiological sensillum tip‐recording technique. The stimulating effect of 13 water soluble sugars at 100 mm is tested on the neurones of these sensilla. Sucrose and fructose are the two most stimulating sugars for the sugar‐sensitive neurone, evoking almost 30 spikes s^?1 at 100 mm . The stimulating effect of arabinose, glucose, mannose, maltose and raffinose is three‐ to five‐fold lower, in the range 5.9–9.6 spikes s^?1. The remaining six sugars, xylose, galactose, rhamnose, cellobiose, trehalose and lactose, have very low (<1 spikes s^?1) or no ability to stimulate the sugar‐sensitive neurone. Concentration/response curves of the sugar‐sensitive neurone to sucrose, fructose and glucose at 0.01–100 mm overlap to a large extent in hibernating, cold reactivated and reproductively‐active beetles. A remarkable 9–50% decrease in the number of spikes evoked by 100 mm fructose and 10–100 mm sucrose occurs, however, in reproductively‐active beetles in June compared with beetles at the beginning of hibernation in October. These findings show that A. obscurus is capable of sensing a wide range sugars via their antennal gustatory sensilla.     

Mathematical modeling of soil carbon turnover in natural Podocarpus forest and Eucalyptus plantation in Ethiopia using compound specific δ13C analysis

Forest soils exhibit huge potential in storing carbon, but may also release large amounts of it if they undergo major changes in land use and environmental conditions. Biogeochemical processes controlling accumulation and release of soil organic carbon (SOC) are not yet sufficiently understood. We investigate the dynamics of SOC depending on its chemical composition below a natural forest (Podocarpus falcatus dominated) and a plantation (Eucalyptus saligna) growing on Nitisols in southern Ethiopia. Soils at the study‐site show a huge shift to less negative δ¹³C values at a depth of 20–30 cm, indicating a change from C4 savanna to C3 forest during the late Holocene. Total organic carbon (TOC), black carbon (BC), and sugars from microbial (rhamnose, fucose) and plant origin (xylose, arabinose) are subjected to compound‐specific stable isotope analysis. Turnover characteristics are calculated using a numerical advection–diffusion–decomposition model. Our measurements show significant differences in carbon storage (P<0.05) for both sites (Podocarpus 23.5 ± 3.2 kg SOC m^?3; Eucalyptus 18.6 ± 2.7 kg SOC m^?3). These differences can be explained with an initial loss of 15–26% of TOC about 50 years ago, induced by clearing the natural forest. After canopy closure, the carbon input below Eucalyptus is 15–34% less than below natural forest. At present, mean residence times (MRTs) of the investigated compounds do not differ between both stands. Sugars show the shortest MRTs in the topsoil with 2–7 years (xylose) and 5–13 years (arabinose) and have been affected the most by clear‐cutting. TOC and BC show MRTs of 13–25 years and 20–34 years, respectively. Old C4 carbon below 20 cm has merely been affected by the land use change. Contrary to expectation, our study does not indicate a pronounced recalcitrance of BC.     

VARIATIONS IN THE SUBSTITUTED 3‐LINKED MANNANS CLOSELY ASSOCIATED WITH THE SILICIFIED WALLS OF DIATOMS1

The polysaccharides from cleaned frustules of the diatoms Pinnularia viridis (Nitzsch) Ehrenberg, Craspedostauros australis Cox, Thalassiosira pseudonana Hasle et Heimdal, and Nitzschia navis‐varingica Lundholm et Moestrup were extracted with hot alkali that dissolved the silica and were characterized by constituent sugar and linkage analyses. The polysaccharides from P. viridis were investigated further by permethylation, partitioning according to solubility, desulfation, and CD₃I‐methylation. Yields of carbohydrate in the hot alkali extracts ranged from 0.9% to 1.8% w/w based on the dry weight of the silica. Mannose was the dominant sugar in the polysaccharides from all four species (54–69 mol% of constituent sugars), although 14 other monosaccharides, including neutral sugars (glucose, galactose, xylose, arabinose, rhamnose, fucose), acidic sugars (glucuronic acid, galacturonic acid, 2‐O‐methylglucuronic acid), and O‐methylated neutral sugars (2‐O‐methylrhamnose, 3‐O‐methylrhamnose, 2,3‐di‐O‐methylrhamnose, 3‐O‐methylxylose, 4‐O‐methylxylose) were also detected in varying proportions among the four samples. The polysaccharides were predominantly composed of a 3‐linked mannopyranose backbone with a prevalence of linkage and/or substitution at O‐2 of the 3‐linked mannopyranosyl residues, and they were polyanionic, bearing uronic acid residues and/or sulfate esters. There were, however, species‐specific differences in the degree and position of substitution on the mannan backbone, the type and substitution patterns of the anionic substituents, and the type and linkage patterns of sugars other than mannose. Although definitive functions for these polysaccharides in diatom biology remain uncertain, a possible role in biosilicification is discussed.