The emission of corrosive vapours by wood. Hot - water - extracted O - acetylated hemicelluloses from sweet chestnut (Castanea sativa) and wych elm (Ulmus glabrau) and a discussion of O-acetyl-group changes occurring in these woods during incubation at 48° and 100% relative humidity

1. O-Acetylated polysaccharides were obtained from green wood of both sweet chestnut and wych elm by treatment of the residue remaining after dimethyl sulphoxide extraction with water at 98 degrees . This gives a mixture of polysaccharides containing xylose, galactose, glucose and uronic acids. Analysis of these and their fractionated products suggest that only xylans in green sweet chestnut and green wych elm are O-acetylated. 2. The isolated O-acetylated xylans are not representative of the total O-acetylated xylans occurring in sweet chestnut and wych elm. 3. Application of the method developed by Bouveng for the location of O-acetyl groups to all four O-acetylated xylans obtained in this series of investigations by dimethyl sulphoxide extraction showed that those from sweet chestnut and wych elm, under the same conditions of incubation, lost: 74.2 and 43.4% of acetyl groups respectively, at C-2; 58.0 and 28.5% of acetyl groups respectively at C-3; 41.8 and 82.2% of acetyl groups respectively at C-2 and C-3. 4. A consideration of electronic and steric factors indicates that there does not appear to be a purely chemical reason for the difference in loss of O-acetyl groups between sweet chestnut and wych elm. It is suggested that the location of O-acetylated xylans in the wood cell walls and the presence of extractive may play some part in this difference.     

Genetic diversity of the sweet chestnut (<Emphasis Type="Italic">Castanea sativa</Emphasis> Mill.) in Central Europe and the western part of the Balkan Peninsula and evidence of marron genotype introgression into wild populations

The sweet chestnut (Castanea sativa Mill.) is a widely spread and important multipurpose tree species in the Mediterranean area, which has played an important role in human history. Natural events, such as glaciations, and human influence played significant roles in the distribution and genetic makeup of the sweet chestnut. In order to better understand how natural and human-mediated past events affected the current genetic diversity and structure of the sweet chestnut, we analysed populations from Central Europe and the western part of the Balkan Peninsula, utilizing ten polymorphic nuclear microsatellite markers. The study revealed the existence of three genetically and, to a large extent, geographically distinct and well-defined groups of sweet chestnut populations. Two not entirely separated groups of populations were detected in the northern part of the studied area and one in the southern. Our results indicate that the genetic structure of sweet chestnut populations in Central Europe and the western part of the Balkan Peninsula is the result of both natural colonization events and significant and lengthy human impact. Furthermore, it has been proven that the gene flow between cultivated/grafted trees’ and wild chestnut stands can influence their genetic structure. However, our results reveal that cultivated-to-wild introgression in the sweet chestnut is dependent on the close proximity of chestnut orchards and naturally occurring populations.     

Development of the Dutch elm disease epidemic in southern England, 1971-6

The current epidemic of Dutch elm disease was studied by recording the fate of individual hedgerow elms (Ulmus procera) in five plots in the West Midlands, and by analysing data from successive Forestry Commission surveys of non-woodland elms in 234 plots in southern England. Ninty-five percent of the individual trees died between May 1972 and September 1975. The average infection rate (r) was found to be 1 -35 during the period when the proportion of disease, x, increased from 0–16 to 0–42. In the plots of the main survey the average infection rate was 0–65 and the cumulative loss increased from 6 to 62% between 1971 and 1976, with little evidence that the course of the epidemic was influenced by variations in the weather from year to year. These infection rates are as high as those recorded in Dutch elm disease epidemics elsewhere in the world. The infection rate in English elm was higher than in either the wych elm or the heterogeneous ‘smooth-leaved elm’. The study of English elm in four geographical areas of southern Britain showed that there was an initial drop in infection rate until x = 0–12, when a steady infection rate obtained in all four areas, ranging from 0–56 in the Midlands to 0–76 in the south-east. It is concluded that the epidemic is likely to continue at a high rate until most non-woodland elm have died. Most trees which survive are likely to be smooth-leaved elm in East Anglia. Few communities in southern England have been able to practice vigorous sanitation control programmes, but data from two, in East Sussex and Brighton, are analysed and the effect on disease progress discussed.     

The type-specific substance from Pneumococcus type 11A(43)

1. The type-specific substance from Pneumococcus type 11A(43) is a polymer containing d-glucose, d-galactose, glycerol, phosphate and O-acetyl in the approximate molecular proportions 2:2:1:1:2. 2. Removal of the O-acetyl groups with ammonia gave a compound no longer active towards type 11A antiserum. 3. Treatment of S.11A with sodium borohydride, followed by hydrolysis with alkali yielded a phosphorus-free polysaccharide, whose structure was studied by methylation and by degradation with periodate. 4. Examination of S.11A and its de-O-acetyl derivative by periodate oxidation led to the partial structure (XI) for the type-specific substance, which thus has several features in common with S.18.     

Pathogen induced disturbance and succession in temperate forests: Evidence from a 100-year data set in southern Sweden

Major tree species are declining in many temperate forests due to changing disturbance regimes, including invasive pests and pathogens. We examined the interaction of secondary succession and Dutch elm disease in the Swedish temperate forest reserve Dalby Söderskog, based on five tree surveys made between 1909 and 2011. The forest is characterized by the coexistence of four major European tree species: wych elm (Ulmus glabra), European ash (Fraxinus excelsior), European beech (Fagus sylvatica) and pedunculate oak (Quercus robur). After protection of the forest in 1918, lack of disturbance mainly favoured elm, while the oak population declined due to mortality of old oaks and lack of regeneration. Dutch elm disease has caused high and continuous elm mortality since 1988. As a result, increased light availability at the forest floor favoured abundant regeneration of ash, beech, and lately also oak. The recent arrival of an invasive fungal pathogen causing ash dieback may once again change the course of succession. Open space emerging from loss of elm and ash in forest reserves may be used by reserve managers to favour oak regeneration and biodiversity of semi-open woodlands once lost during succession to closed forest. We conclude that winners and losers change places as an effect of invasive pathogens, resulting in unexpected successions and both losses and gains in valuable ecological niches and habitat structures in temperate broadleaf forests.     

Antigenic bacterial polysaccharides. 36. A study of the structure of the O-specific polysaccharide chain of the lipopolysaccharide of Pseudomonas fluorescens IMV 2763 (biostrain G)

Chemical and serological characterization of the Pseudomonas fluorescens IMV 2763 (biovar G) lipopolysaccharide was carried out. The O-specific polysaccharide chain of the lipopolysaccharide is composed of D-mannose, 6-deoxy-L-talose, N-acetyl-D-galactosamine and O-acetyl groups in the ratio of approximately 2:1:1:1. The polysaccharide is branched and a half of residues of 6-deoxytalose and monosubstituted mannose carry O-acetyl groups. On the basis of methylation, partial acid hydrolysis and 13C NMR analysis it was concluded that the repeating unit of the polysaccharide has the following structure: (formula; see text)     

The potential value of the seaweed Ceylon moss (Gelidium amansii) as an alternative bioenergy resource

Sea weed (Ceylon moss) possesses comparable bioenergy production potential to that of land plants. Ceylon moss has high content of carbohydrates, typically galactose (23%) and glucose (20%). We have explored the possibility of sodium chlorite in Ceylon moss pretreatment that can ultimately increase the efficiency of enzymatic saccharification. In an acidic medium, chlorite generates ClO₂ molecules that transform lignin into soluble compounds without any significant loss of carbohydrate content and this procedure is widely used as an analytical method for holocellulose determination. Sodium chlorite-pretreated samples resulted in glucose yield up to 70% with contrast of only 5% was obtained from non-pretreated samples.The efficiency of enzymatic hydrolysis is significantly improved by sodium chlorite pretreatment, and thus sodium chlorite pretreatment is potentially a very useful tool in the utilisation of Ceylon moss biomass for ethanol production or bioenergy purposes.     

Feeding by Scolytus bark beetles to test for differently susceptible elm varieties

Dutch elm disease (DED), caused by the fungi Ophiostoma ulmi and O. novo‐ulmi, has reduced elm populations severely in Europe and North America. Breeding programmes are in action to find less susceptible elm varieties suitable for re‐establishing elm stands. Bark beetles, mainly Scolytus spp., are the only known natural vectors of DED. During twig feeding, beetles transfer Ophiostoma spores to healthy elms. Thus, less palatable elms should run a lower risk of DED infections. In feeding preference bioassays, we offered twigs from elms exhibiting different degree of susceptibility to O. novo‐ulmi, together with non‐host trees to Scolytus beetles. Scolytus multistriatus preferred wych elm, Ulmus glabra, to 100% in two‐choice tests, whereas S. laevis did not discriminate between a tolerant and a susceptible variety of field elm, U. minor. We suggest that the feeding assay is useful as a low‐tech method in breeding programmes for evaluating the suitability of promising elm genotypes to vector insects.     

Sap flow-based quantitative indication of progression of Dutch elm disease after inoculation with Ophiostoma novo-ulmi

Key message

The rate of progression of Dutch elm disease can be continuously and quantitatively estimated from sap flow measurements.

Abstract

Response of sap flow to inoculation with Ophiostoma novo-ulmi, a causal agent which causes vascular mycosis called Dutch elm disease, was studied in a field experiment comprised of 4-year-old wych elm trees (Ulmus glabra). Sap flow was measured on inoculated trees using the trunk heat balance method with external heating (EMS 62, Czech Republic) throughout the experiment. The first detectable symptoms of reduction in sap flow occurred 6 days after inoculation and all inoculated trees died within 16 days. Our experiment confirmed the ability of O. novo-ulmi to quickly kill young elm trees. The disease progressed faster than in previous experiments utilizing O. ulmi. To the best of our knowledge, this is the first experiment using sap flow measurements on trees inoculated by O. novo-ulmi. The trunk heat balance sap flow method is an effective non-invasive tool for continuous quantitative monitoring of the progression of vascular tree diseases, and show increased potential for field and greenhouse studies on changes in xylem hydraulic conductivity in a wide range of broadleaved and coniferous tree species.     

Microbial degradation of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in soils.

The microbial degradation of tensile test pieces made of poly(3-hydroxybutyrate) [P(3HB)] or a copolymer of 90% 3-hydroxybutyric acid and 10% 3-hydroxyvaleric acid was studied in soils incubated at a constant temperature of 15, 28, or 40 degrees C for up to 200 days. In addition, hydrolytic degradation in sterile buffer at temperatures ranging from 4 to 55 degrees C was monitored for 98 days. Degradation was measured through loss of weight (surface erosion), molecular weight, and mechanical strength. While no weight loss was recorded in sterile buffer, samples incubated in soils were degraded at an erosion rate of 0.03 to 0.64% weight loss per day, depending on the polymer, the soil, and the incubation temperature. The erosion rate was enhanced by incubation at higher temperatures, and in most cases the copolymer lost weight at a higher rate than the homopolymer. The molecular weights of samples incubated at 40 degrees C in soils and those incubated at 40 degrees C in sterile buffer decreased at similar rates, while the molecular weights of samples incubated at lower temperatures remained almost unaffected, indicating that molecular weight decrease is due to simple hydrolysis and not to the action of biodegrading microorganisms. The degradation resulted in loss of mechanical properties. From the samples used in the biodegradation studies, 295 dominant microbial strains capable of degrading P (3HB) and the poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer in vitro were isolated and identified.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chemical and structural analysis of the capsular polysaccharide from Escherichia coli O9:K28(A):H- (K28 antigen)

The structure of the capsular polysaccharide from Escherichia coli O9:K28(A):H- (K28 antigen) has been determined by using the techniques of methylation, periodate oxidation, and partial hydrolysis. N.m.r. spectroscopy (1H and 13C) was used to establish the nature of the anomeric linkages. O-Acetyl groups were determined spectrophotometrically and were located using methyl vinyl ether as a protective reagent. The polysaccharide is comprised of repeating units of the tetrasaccharide shown (three-plus-one type) with 70% of the fucosyl residues carrying an O-acetyl substituent. (formula; see text) This structure resembles that of E. coli K27 and has the structural pattern of Klebsiella K54 polysaccharide.     

Effect of nutrition of parasites with nectar of melliferous plants on parasitism of the elm bark beetles (Col., Scolytidae)

In the forest of Kljestevica, the elm is attacked by four species of elm bark beetles: Scolytus multistriatus (Marsh.), Scolytus scolytus (Fab.), Scolytus pygmaeus (Fab.) and Scolytus ensifer (Rich.) (Col., Scolytidae). Scolytus multistriatus and S. pygmaeus are the most numerous species. Elm bark beetle is parasitized by four species of parasites: Dendrosoter protuberans (Nees), Ecphylus silesiacus (Ratz.), Coeloides scolyticida (Wesm.) (Hym., Braconidae) and Entedon leucogramma (Ratz.) (Hym., Eulophidae). Ecphylus silesiacus and D. protuberans are the most numerous species. The nectar of mustard ( Sinapis alba L., Brassicaceae) flowers, sweet basil ( Stachys recta L., Lamiaceae) flowers and of wild carrot ( Daucus carota L., Umbelliferae), has an important effect on the number of parasites and on the parasitism of the elm bark beetles. Wild carrot is the most attractive plant to the parasites for additional nutrition, especially for D. protuberans . Mustard and sweet basil flowers are suitable for additional nutrition of E. silesiacus imagos. On meadow flowers growing spontaneously in forest clearings, the lowest number of parasite imagos was collected by an entomological catcher, and the lowest percentage of parasitized larvae and eggs of the elm bark beetle was recorded in elm control catch trunks that were not near the cultivated melliferous plants.     

The distribution of a typhlocybine leafhopper, Ribautiana ulmi (Homoptera: Cicadellidae) on a specimen wych elm tree

ABSTRACT.

• 1 Higher numbers of feeding and resting Ribautiana ulmi (L.) were found on the more highly illuminated areas of the canopy of a wych elm tree, Ulmus glabra Huds. cv. Camperdownii.
• 2 Higher numbers were also found on the basal leaves of each branch when compared with the distal leaves, and on exposed areas of individual leaves when compared with the shaded areas which they overlapped.
• 3 Leaf thickness, tannin concentrations and leaf toughness were greater, and water content lower, in leaves from exposed areas of the canopy than from shaded ones.
• 4 R.ulmi feeds selectively on the contents of the palisade mesophyll cells, and therefore was assumed to ingest the highest available levels of tannins.

     

Micropropagation of mature wych elm (<Emphasis Type="Italic">Ulmus glabra</Emphasis> Huds.)

Explants of mature vigorous donor trees of wych elm (Ulmus glabra Huds.) that had not been previously exposed to Dutch elm disease were investigated for the influence of phytohormones and media on shoot multiplication rates and organogenic capacity. The regenerates were micropropagated from cultures that originated from 15-year-old progeny of plus trees. Two plus trees aged over 70 years showed recalcitrant responses. Thidiazuron in combination with 6-benzylaminopurine (BAP) induced a significantly higher number of shoots per explant than the most optimal BAP treatment (5.88 vs. 3.05 shoots). Woody plant medium and Dubovský minimal medium had no significant effects on shoot formation and multiplication rates. All plantlets raised in vitro were phenotypically normal and successfully hardened to ex vitro conditions. Two experimental field plots with 3-year-old in vitro-propagated trees were established.Abbreviations DED: Dutch elm disease - BAP: 6-Benzylaminopurine - IBA: Indole-3-butyric acid - TDZ: Thidiazuron - WPM: Woody Plant Medium - DM: Dubovský Minimal Medium Communicated by D. Bartels     

Restoration of threatened species: a noble cause for transgenic trees

Some of the first applications of transgenic trees in North America may be for the conservation or restoration of threatened forest trees that have been devastated by fungal pathogens or insect pests. In some cases, where resistance has yet to be found in the natural population of a tree species, incorporating genes from other organisms may offer the only hope for restoration. In others, transgenics may play a role as part of an integrated approach, along with conventional breeding or biocontrol agents. American chestnut (Castanea dentata) was wiped out as a canopy species by a fungal disease accidentally introduced into the United States around 1900. Similarly, American elm (Ulmus americana) virtually disappeared as a favored street tree from Northeastern U.S. cities after the introduction of the Dutch elm disease fungus in the 1940s. In both cases, progress has been made toward restoration via conventional techniques such as selection and propagation of tolerant cultivars (American elm) or breeding with a related resistant species (American chestnut). Recently, progress has also been made with development of systems for engineering antifungal candidate genes into these “heritage trees.” An Agrobacterium-leaf disk system has been used to produce transgenic American elm trees engineered with an antimicrobial peptide gene that may enhance resistance to Dutch elm disease. Two gene transfer systems have been developed for American chestnut using Agrobacterium-mediated transformation of embryogenic cultures, setting the stage for the first tests of potential antifungal genes for their ability to confer resistance to the chestnut blight fungus. Despite the promise of transgenic approaches for restoration of these heritage trees, a number of technical, environmental, economic, and ethical questions remain to be addressed before such trees can be deployed, and the debate around these questions may be quite different from that associated with transgenic trees developed for other purposes.     

Isolation and characterization of sialate 9(4)-O-acetylesterase from influenza C virus

An esterase was isolated from influenza C virus with a specific activity from 1.7-5 U/mg protein, and its substrate specificity was tested with various naturally occurring O-acylated sialic acids, synthetic carbohydrate acetates, and other esters. The enzyme hydrolyses only acetic acid esters at significant rates. The non-natural substrates 4-methyl-umbelliferyl acetate, 4-nitrophenyl acetate, and alpha-naphthyl acetate are cleaved at highest hydrolysis rates, followed by the natural substrate N-acetyl-9-O-acetylneuraminic acid. The esterase also acts on N-glycoloyl-9-O-acetylneuraminic acid and, much slower, on N-acetyl-4-O-acetylneuraminic acid; N-acetyl-7-O-acetylneuraminic acid is not hydrolysed. 2-Deoxy-2,3-didehydro-N-acetyl-9-O-acetylneuraminic acid is also a substrate for this enzyme, however, 6-O-acetylated N-acetylmannosamine and glucose are not. Esterification of the carboxyl function of sialic acids strongly reduces or prevents esterase action on O-acetyl groups. The carboxyl ester is not hydrolysed. The relative cleavage rates also depend on the type of the non-sialic acid part of the molecule. N-Acetyl-9-O-acetylneuraminic acid as component of sialyllactose and rat serum glycoprotein shows hydrolysis rates close to the free form of this sugar, while acetyl ester groups of bovine submandibular gland mucin and rat erythrocytes are hydrolysed at slower rates. Gangliosides and 4-O-acetylated glycoproteins are no substrates for the purified enzyme. A slow hydrolysis is observed by incubation of 9-O-acetylated GD1a with intact influenza C viruses. As other natural acetyl esters (acetyl-CoA and acetylthiocholine iodide) are not hydrolysed, the enzyme can be classified as sialate 9(4)-O-acetylesterase (EC 3.1.1.53).     

Structural studies on the O-antigen of Aeromonas salmonicida

Lipopolysaccharide from a strain of Aeromonas salmonicida salmonicida was isolated from cells by the aqueous phenol method in 2.3% yield (based on dry weight of bacteria). Hydrolysis of the lipopolysaccharide in 1% acetic acid afforded O-polysaccharide (19% by weight), core-oligosaccharide (12.2%) and lipid A (44.6%). Analysis indicated that 3-deoxy-D-manno-2-octulosonic acid was absent from the lipopolysaccharide and that no low-molecular-weight compounds were released by the mild hydrolysis. The O-polysaccharide had the monosaccharide composition of rhamnose, glucose and N-acetylmannosamine in molar ratio of 1.0:1.58:0.83. 75% of the N-acetylmannosamine residues were substituted at position 4 by O-acetyl groups. Hydrolysis of the methylated polysaccharide proved to be both difficult and dependent on the method of hydrolysis chosen, in all cases a partially methylated disaccharide of rhamnose and N-acetylmannosamine was identified in the hydrolysate. Methylation analysis, periodate oxidation and proton magnetic resonance analysis were used to confirm the structure of the repeating unit as: (formula; see text).     

Antigenic polysaccharides of bacteria. 16. The structure of O-specific polysaccharide chain of Pseudomonas aeruginosa 025 (Wokatsch) lipopolysaccharide

O-Specific polysaccharide built up of trisaccharide repeating units containing 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid (ManNAcAmA), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid (Man(NAc)2A), N-acetyl-D-fucosamine (FucNAc), and O-acetyl group was obtained on mild acid hydrolysis of P. aeruginosa O25 (Wokatsch classification) lipopolysaccharide. Basing on de-O-acetylation of polysaccharide with aqueous triethylamine accompanied by hydrolysis of acetamidino group to acetamido group, as well as on the 1H and 13C NMR data, the following structure of the repeating unit of the polysaccharide was established: (Formula: see text) P. aeruginosa O25 polysaccharide has the same carbohydrate skeleton as that of P. aeruginosa O3a,b (Lányi classification) and differs from the latter only by the presence of the O-acetyl group at position 4 of N-acetylfucosamine.     

The acid and enzymic hydrolysis of O-acetylated sialic acid residues from rabbit Tamm–Horsfall glycoprotein

Rabbit Tamm-Horsfall glycoprotein and bovine submaxillary glycoprotein were both found to contain sialic acid residues which are released at a slow rate by the standard conditions of acid hydrolysis. These residues are also resistant to neuraminidases from Vibrio cholerae and Clostridium perfringens. This behaviour was attributed to the presence of O-acetylated sialic acid, since the removal of O-acetyl groups by mild alkaline treatment normalized the subsequent release of sialic acid from rabbit Tamm-Horsfall glycoprotein by acid and by enzymic hydrolysis. Determination of the O-acetyl residues in rabbit Tamm-Horsfall glycoprotein indicated that on average two hydroxyl groups of sialic acid are O-acetylated, and these were located on the polyhydroxy side-chain of sialic acid or on C-4 and C-8. These findings confirm the assumption that certain O-acetylated forms of sialic acid are not substrates for bacterial neuraminidases. Several explanations have been suggested to explain the effect of O-acetylation of the side-chain on the rate of acidcatalysed hydrolysis of sialic acid residues.     

Development of microsatellite markers in peach [ Prunus persica (L.) Batsch] and their use in genetic diversity analysis in peach and sweet cherry ( Prunus avium L.)

We report the sequence of 41 primer pairs of microsatellites from a CT-enriched genomic library of the peach cultivar 'Merrill O'Henry'. Ten microsatellite-containing clones had sequences similar to plant coding sequences in databases and could be used as markers for known functions. For microsatellites segregating at least in one of the two Prunus F(2) progenies analyzed, it was possible to demonstrate Mendelian inheritance. Microsatellite polymorphism was evaluated in 27 peach and 21 sweet cherry cultivars. All primer pairs gave PCR-amplification products on peach and 33 on cherry (80.5%). Six PCR-amplifications revealed several loci (14.6%) in peach and eight (19.5%) in sweet cherry. Among the 33 single-locus microsatellites amplified in peach and sweet cherry, 13 revealed polymorphism both in peach and cherry, 19 were polymorphic only on peach and one was polymorphic only on cherry. The number of alleles per locus ranged from 1 to 9 for peach and from 1 to 6 on sweet cherry with an average of 4.2 and 2.8 in peach and sweet cherry, respectively. Cross-species amplification was tested within the Prunus species: Prunus avium L. (sweet cherry and mazzard), Prunus cerasus L. (sour cherry), Prunus domestica L. (European plum), Prunus amygdalus Batsch. (almond), Prunus armeniaca L. (apricot), Prunus cerasifera Ehrh. (Myrobalan plum). Plants from other genera of the Rosaceae were also tested: Malus (apple) and Fragaria (strawberry), as well as species not belonging to the Rosaceae: Castanea (chestnut tree), Juglans (walnut tree) and Vitis (grapevine). Six microsatellites gave amplification on all the tested species. Among them, one had an amplified region homologous to sequences encoding a MADS-box protein in Malus x domestica. Twelve microsatellites (29.3%) were amplified in all the Rosaceae species tested and 31 (75.6%) were amplified in all the six Prunus species tested. Thirty three (80.5%), 18 (43.9%) and 13 (31.7%) gave amplification on chestnut tree, grapevine and walnut tree, respectively.