The influence of pre- and post-zygotic barriers on interspecific Corymbia hybridization

Corymbia species from different sections hybridize readily, with some of increasing economic importance to plantation forestry. This study explores the locations of reproductive barriers between interspecific Corymbia hybrids and investigates the reproductive success of a wide taxonomic range of C. torelliana hybrid crosses. Pollen, pistil and embryo development were investigated for four C. torelliana crosses (C. torelliana, C. citriodora subsp. citriodora, C. tessellaris and C. intermedia) using fluorescent and standard microscopy to identify the locations of interspecific reproductive isolating barriers. Corymbia torelliana was also crossed with 16 taxa, representing six of the seven Corymbia sections, both Corymbia subgenera and one species each from the related genera, Angophora and Eucalyptus. All crosses were assessed for capsule and seed yields. Interspecific C. torelliana hybridization was controlled by pre-zygotic reproductive isolating barriers inhibiting pollen adhesion to the stigma, pollen germination, pollen tube growth in the style and pollen tube penetration of the micropyle. Corymbia torelliana (subgenus Blakella, sect. Torellianae) was successfully hybridized with Corymbia species from subgenus Blakella, particularly C. citriodora subsp. citriodora, C. citriodora subsp. variegata, C. henryi (sect. Maculatae) and C. tessellaris (sect. Abbreviatae), and subgenus Corymbia, particularly C. clarksoniana and C. erythrophloia (sect. Septentrionales). Attempted intergeneric hybrids between C. torelliana and either Angophora floribunda or Eucalyptus pellita were unsuccessful. Corymbia hybrids were formed between species from different sections and subgenera, but not with species from the related genera Angophora or Eucalyptus. Reproductive isolation between the interspecific Corymbia hybrid crosses was controlled by early- and late-acting pre-zygotic isolating barriers, with reproductive success generally decreasing with increasing taxonomic distance between parent species. These findings support the monophyly of Corymbia and the close relationships of infrageneric clades. The hybridizing propensity of Corymbia species provides opportunities for breeding but suggests risks of environmental gene flow. © The Author 2012. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Characterization of lignin-modifying enzymes of the selective white-rot fungus Physisporinus rivulosus

White-rot fungi are wood degrading organisms that are able to decompose all wood polymers; lignin, cellulose and hemicellulose. Especially the selective white-rot fungi that decompose preferentially wood lignin are promising for biopulping applications. In biopulping the pretreatment of wood chips with white-rot fungi enhances the subsequent pulping step and substantially reduces the refining energy consumption in mechanical pulping. Because it is not possible to carry out biopulping in industrial scale as a closed process it has been necessary to search for new selective strains of white-rot fungi which naturally occur in Finland and cause selective white-rot of Finnish wood raw-material. In a screening of 300 fungal strains a rare polypore, Physisporinus rivulosus strain T241i isolated from a forest burn research site, was found to be a selective lignin degrader and promising for the use in biopulping. Since selective lignin degradation is apparently essential for biopulping, knowledge on lignin-modifying enzymes and the regulation of their production by a biopulping fungus is needed. White-rot fungal enzymes that participate in lignin degradation are laccase, lignin peroxidase (LiP), manganese peroxidase (MnP), versatile peroxidase (VP) and hydrogen peroxide forming enzymes. In this study, P. rivulosus was observed to produce MnP, laccase and oxalic acid during growth on wood chips. In liquid cultures manganese and veratryl alcohol increased the production of acidic MnP isoforms detected also in wood chip cultures. Laccase production by P. rivulosus was low unless the cultures were supplemented with sawdust and charred wood, the components of natural growth environment of the fungus. In white-rot fungi the lignin-modifying enzymes are typically present as multiple isoforms. In this study, two MnP encoding genes, mnpA and mnpB, were cloned and characterized from P. rivulosus T241i. Analysis of the N-terminal amino acid sequences of two purified MnPs and putative amino acid sequence of the two cloned mnp genes suggested that P. rivulosus possesses at least four mnp genes. The genes mnpA and mnpB markedly differ from each other by the gene length, sequence and intron-exon structure. In addition, their expression is differentially affected by the addition of manganese and veratryl alcohol. P. rivulosus produced laccase as at least two isoforms. The results of this study revealed that the production of MnP and laccase was differentially regulated in P. rivulosus, which ensures the efficient lignin degradation under a variety of environmental conditions.

oxidation of catechol in plants iv. purification and properties of the 3,4,3',4'-tetrahydroxydiphenyl-forming enzyme system from tecoma leaves

Acetone powders prepared from leaf extracts of Tecoma stans L. were found to catalyze the oxidation of catechol to 3,4,3',4'-tetrahydroxydiphenyl. Fractionation of the acetone powders obtained from Tecoma leaves with acetone, negative adsorption of the acetone fraction with tricalcium phosphate gel, and chromatography of the gel supernatant on DEAE-Sephadex yielded a 68-fold purified enzyme with 66% recovery. The enzyme had an optimum pH around 7.2. It showed a temperature optimum of 30° and the Km for catechol was determined as 2 x 10-4 m. The purified enzyme moved as a single band on polyacrylamide gel electrophoresis. Its activity was found to be partially stimulated by Mg2+. The reaction was not inhibited by o-phenanthroline and agr,agr'-dipyridyl. The purified enzyme was highly insensitive to a range of copper-chelating agents. It was not affected appreciably by thiol inhibitors. The reaction was found to be suppressed to a considerable extent by reducing agents like GSH, cysteine, cysteamine, and ascorbic acid. The purified enzyme was remarkably specific for catechol. Catalase affected neither the enzyme activity nor the time course of the reaction. Hydrogen peroxide was not formed as a product of the reaction.

Regio- and stereoselectivity of oxidative coupling reactions of phenols : Spirodienones as construction units in lignin

Dimeric phenolic compounds lignans and dilignols form in the so-called oxidative coupling reaction of phenols. Enzymes such as peroxidases and lac-cases catalyze the reaction using hydrogen peroxide or oxygen respectively as oxidant generating phenoxy radicals which couple together according to certain rules. In this thesis, the effects of the structures of starting materials mono-lignols and the effects of reaction conditions such as pH and solvent system on this coupling mechanism and on its regio- and stereoselectivity have been studied. After the primary coupling of two phenoxy radicals a very reactive quinone me-thide intermediate is formed. This intermediate reacts quickly with a suitable nucleophile which can be, for example, an intramolecular hydroxyl group or another nucleophile such as water, methanol, or a phenolic compound in the reaction system. This reaction is catalyzed by acids. After the nucleophilic addi-tion to the quinone methide, other hydrolytic reactions, rearrangements, and elimination reactions occur leading finally to stable dimeric structures called lignans or dilignols. Similar reactions occur also in the so-called lignification process when monolignol (or dilignol) reacts with the growing lignin polymer. New kinds of structures have been observed in this thesis. The dimeric com-pounds with so-called spirodienone structure have been observed to form both in the dehydrodimerization of methyl sinapate and in the beta-1-type cross-coupling reaction of two different monolignols. This beta-1-type dilignol with a spirodienone structure was the first synthetized and published dilignol model compound, and at present, it has been observed to exist as a fundamental construction unit in lignins. The enantioselectivity of the oxidative coupling reaction was also studied for obtaining enantiopure lignans and dilignols. A rather good enantioselectivity was obtained in the oxidative coupling reaction of two monolignols with chiral auxiliary substituents using peroxidase/H2O2 as an oxidation system. This observation was published as one of the first enantioselective oxidative coupling reaction of phenols. Pure enantiomers of lignans were also obtained by using chiral cryogenic chromatography as a chiral resolution technique. This technique was shown to be an alternative route to prepare enantiopure lignans or lignin model compounds in a preparative scale.

Preparation of Industrially Important Hydroxy Acids and Diacids from 2,2-Disubstituted Propane-1,3-Diols and Linear Primary Diols by Green Chemistry Methods

Environmentally benign and economical methods for the preparation of industrially important hydroxy acids and diacids were developed. The carboxylic acids, used in polyesters, alkyd resins, and polyamides, were obtained by the oxidation of the corresponding alcohols with hydrogen peroxide or air catalyzed by sodium tungstate or supported noble metals. These oxidations were carried out using water as a solvent. The alcohols are also a useful alternative to the conventional reactants, hydroxyaldehydes and cycloalkanes. The oxidation of 2,2-disubstituted propane-1,3-diols with hydrogen peroxide catalyzed by sodium tungstate afforded 2,2-disubstituted 3-hydroxypropanoic acids and 1,1-disubstituted ethane-1,2-diols as products. A computational study of the Baeyer-Villiger rearrangement of the intermediate 2,2-disubstituted 3-hydroxypropanals gave in-depth data of the mechanism of the reaction. Linear primary diols having chain length of at least six carbons were easily oxidized with hydrogen peroxide to linear dicarboxylic acids catalyzed by sodium tungstate. The Pt/C catalyzed air oxidation of 2,2-disubstituted propane-1,3-diols and linear primary diols afforded the highest yield of the corresponding hydroxy acids, while the Pt, Bi/C catalyzed oxidation of the diols afforded the highest yield of the corresponding diacids. The mechanism of the promoted oxidation was best described by the ensemble effect, and by the formation of a complex of the hydroxy and the carboxy groups of the hydroxy acids with bismuth atoms. The Pt, Bi/C catalyzed air oxidation of 2-substituted 2-hydroxymethylpropane-1,3-diols gave 2-substituted malonic acids by the decarboxylation of the corresponding triacids. Activated carbon was the best support and bismuth the most efficient promoter in the air oxidation of 2,2-dialkylpropane-1,3-diols to diacids. In oxidations carried out in organic solvents barium sulfate could be a valuable alternative to activated carbon as a non-flammable support. In the Pt/C catalyzed air oxidation of 2,2-disubstituted propane-1,3-diols to 2,2-disubstituted 3-hydroxypropanoic acids the small size of the 2-substituents enhanced the rate of the oxidation. When the potential of platinum of the catalyst was not controlled, the highest yield of the diacids in the Pt, Bi/C catalyzed air oxidation of 2,2-dialkylpropane-1,3-diols was obtained in the regime of mass transfer. The most favorable pH of the reaction mixture of the promoted oxidation was 10. The reaction temperature of 40°C prevented the decarboxylation of the diacids.

Infrared spectroscopy as a rapid tool to detect methylglyoxal and antibacterial activity in Australian honeys

Methylglyoxal (2-oxopropanal) is a compound known to contribute to the non-peroxide antimicrobial activity of honeys. The feasibility of using infrared spectroscopy as a predictive tool for honey antibacterial activity and methylglyoxal content was assessed. A linear relationship was found between methylglyoxal content (279–1755 mg/kg) in Leptospermum polygalifolium honeys and bacterial inhibition for Escherichiacoli (R2 = 0.80) and Staphylococcusaureus (R2 = 0.64). A good prediction of methylglyoxal (R2 0.75) content in honey was achieved using spectroscopic data from the mid infrared (MIR) range in combination with partial least squares regression. These results indicate that robust predictive equations could be developed using MIR for commercial application where the prediction of bacterial inhibition is needed to ‘value’ honeys with methylglyoxal contents in excess of 200 mg/kg.

Re-examination of one-point methods of liquid limit determination

This Paper deals with the analysis of liquid limit of soils, an inferential parameter of universal acceptance. It has been undertaken primarily to re-examine one-point methods of determination of liquid limit water contents. It has been shown by basic characteristics of soils and associated physico-chemical factors that critical shear strengths at liquid limit water contents arise out of force field equilibrium and are independent of soil type. This leads to the formation of a scientific base for liquid limit determination by one-point methods, which hitherto was formulated purely on statistical analysis of data. Available methods (Norman, 1959; Karlsson, 1961; Clayton & Jukes, 1978) of one-point liquid limit determination have been critically re-examined. A simple one-point cone penetrometer method of computing liquid limit has been suggested and compared with other methods. Experimental data of Sherwood & Ryley (1970) have been employed for comparison of different cone penetration methods. Results indicate that, apart from mere statistical considerations, one-point methods have a strong scientific base on the uniqueness of modified flow line irrespective of soil type. Normalized flow line is obtained by normalization of water contents by liquid limit values thereby nullifying the effects of surface areas and associated physico-chemical factors that are otherwise reflected in different responses at macrolevel.Cet article traite de l'analyse de la limite de liquidité des sols, paramètre déductif universellement accepté. Cette analyse a été entreprise en premier lieu pour ré-examiner les méthodes à un point destinées à la détermination de la teneur en eau à la limite de liquidité. Il a été démontré par les caractéristiques fondamentales de sols et par des facteurs physico-chimiques associés que les résistances critiques à la rupture au cisaillement pour des teneurs en eau à la limite de liquidité résultent de l'équilibre des champs de forces et sont indépendantes du type de sol concerné. On peut donc constituer une base scientifique pour la détermination de la limite de liquidité par des méthodes à un point lesquelles, jusqu'alors, n'avaient été formulées que sur la base d'une analyse statistique des données. Les méthodes dont on dispose (Norman, 1959; Karlsson, 1961; Clayton & Jukes, 1978) pour la détermination de la limite de liquidité à un point font l'objet d'un ré-examen critique. Une simple méthode d'analyse à un point à l'aide d'un pénétromètre à cône pour le calcul de la limite de liquidité a été suggérée et comparée à d'autres méthodes. Les données expérimentales de Sherwood & Ryley (1970) ont été utilisées en vue de comparer différentes méthodes de pénétration par cône. En plus de considérations d'ordre purement statistque, les résultats montrent que les méthodes de détermination à un point constituent une base scientifique solide en raison du caractère unique de la ligne de courant modifiée, quel que soit le type de sol La ligne de courant normalisée est obtenue par la normalisation de la teneur en eau en faisant appel à des valeurs de limite de liquidité pour, de cette manière, annuler les effets des surfaces et des facteurs physico-chimiques associés qui sans cela se manifesteraient dans les différentes réponses au niveau macro.

Observability of conduction electron spin resonance (CESR) in superconductors

The condition for the observability of CESR in superconducting thin films is analysed taking into account the finiteness of the flux penetration depth. We have explicitly evaluated the path-dependent phase mixing factor occuring in the expression for power absorption. The calculated line width turns out to be of the order of, or larger than, the nominal resonance frequency for the experimentally realisable choice of parameters.

Application of shock waves in pencil manufacturing industry

A new shock wave generator has been designed, fabricated and tested for preservative impregnation studies into wood slats used for manufacturing pencils in the Shock Waves Laboratory, IISc, Bangalore. Series of experiments have been carried out in the laboratory to achieve satisfactory preservative impregnation into VATTA wood slats. The experiments have shown that it is indeed possible to impregnate preservatives into VATTA wood slats using shock waves and the depth of penetration and the retention of preservatives by wood slats is as good as the conventional methods. This method is expected to result in substantial reduction in the treatment process time compared to conventional methods that are currently being used by the pencil manufacturing industry.

Cadmium stress alters the redox reaction and hormone balance in oilseed rape (Brassica napus L.) leaves

In order to understand the physiological response of oilseed rape (Brassica napus L.) leaves to cadmium (Cd) stress and exploit the physiological mechanisms involved in Cd tolerance, macro-mineral and chlorophyll concentrations, reactive oxygen species (ROS) accumulation, activities of enzymatic antioxidants, nonenzymatic compounds metabolism, endogenous hormonal changes, and balance in leaves of oilseed rape exposed to 0, 100, or 200 μM CdSO4 were investigated. The results showed that under Cd exposure, Cd concentrations in the leaves continually increased while macro-minerals and chlorophyll concentrations decreased significantly. Meanwhile, with increased Cd stress, superoxide anion (O 2 • − ) production rate and hydrogen peroxide (H2O2) concentrations in the leaves increased significantly, which caused malondialdehyde (MDA) accumulation and oxidative stress. For scavenging excess accumulated ROS and alleviating oxidative injury in the leaves, the activity of enzymatic antioxidants, such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), was increased significantly at certain stress levels. However, with increased Cd stress, the antioxidant enzyme activities all showed a trend towards reduction. The nonenzymatic antioxidative compounds, such as proline and total soluble sugars, accumulated continuously with increased Cd stress to play a long-term role in scavenging ROS. In addition, ABA levels also increased continuously with Cd stress while ZR decreased and the ABA/ZR ratio increased, which might also be providing a protective role against Cd toxicity.

Oxidations of thio ketones by singlet and triplet oxygen

Oxidation of di-tert-butyl thioketone (1) and 2,2,4,4-tetramethylcyclobutylth ioketone (2) by singlet oxygen yields the corresponding sulfine and ketone; in the case of 1 the sulfine is the major product, whereas in 2 it is the ketone. 1,2,3-Dioxathietane has been suggested as the precursor for the ketones, and the zwitterionic/diradid peroxide is believed to be a common primary intermediate for both sulfine and ketone. Steric influence is felt both during primary interaction between singlet oxygen and thioketone and during the partitioning of the peroxide intermediate. Steric interaction is suggested as the reason for variations in the product distribution between 1 and 2. Singlet oxygen is also generated through energy transfer from the triplet state of thioketones. These excited states also directly react with oxygen to yield ketone.

DNP3 network scanning and reconnaissance for critical infrastructure

The Distributed Network Protocol v3.0 (DNP3) is one of the most widely used protocols to control national infrastructure. The move from point-to-point serial connections to Ethernet-based network architectures, allowing for large and complex critical infrastructure networks. However, networks and con- figurations change, thus auditing tools are needed to aid in critical infrastructure network discovery. In this paper we present a series of intrusive techniques used for reconnaissance on DNP3 critical infrastructure. Our algorithms will discover DNP3 outstation slaves along with their DNP3 addresses, their corresponding master, and class object configurations. To validate our presented DNP3 reconnaissance algorithms and demonstrate it’s practicality, we present an implementation of a software tool using a DNP3 plug-in for Scapy. Our implementation validates the utility of our DNP3 reconnaissance technique. Our presented techniques will be useful for penetration testing, vulnerability assessments and DNP3 network discovery.

The Use of Australian Honey in Moist Wound Management

This project has investigated the correlation between antimicrobial activity and the non-peroxide activity of particular Leptospermum-derived honeys. Initial investigations tested the efficacy of these honeys against one particular bacteria (Staphylococcus aureus), while later studies assessed the effect of this honey on a range of food pathogens, animal pathogens (e.g. in mastitis) and human pathogens (e.g. golden Staph). Using honeys sourced from around Australia and screened against several reference microorganisms, the anti-microbial potency of one particular Australian honey (from Leptospermum polygalifolium) was shown to be comparable to NZ Manuka honey (from L.scoparium). The results of this screening of active honeys against pathogenic bacteria has supported the registration of honey by Capilano Honey Limited as a “Drug” with the Therapeutic Goods Administration based on its antimicrobial activity.

Accounting for risk: The advent of capped conveyancing title insurance

Title insurance companies originating from America, have, in the past 15 years become part of the Australian conveyancing landscape. However for most residential freehold owners, their activities would be a mystery. A purchaser does not routinely obtain title insurance, with the companies presently focussing on servicing the mortgagee sector. While the lack of penetration in the residential purchaser market may be attributed to the consumer’s lack of knowledge, evidence from Ontario and New Zealand illustrates that title insurance is likely to become an additional cost in the conveyancing process in Australia. In this article we highlight the reasons why, and demonstrate how title insurers have, by working with the legal profession been able to subtly move the risk of responsibility for compensation for loss, (at least in the first instance) from the state to the insurer, but with the added benefit for the state and the conveyancing agents that the cost of the insurance is ultimately borne by the consumer. In New Zealand this development is being accelerated by the introduction of capped conveyancing title insurance. Whether title insurance will become part of the conveyancing process is no longer the relevant question for Australia, (it undoubtedly will), but the unknown issue is just how title insurance companies will work with conveyancing agents to infiltrate the market, and what response this infiltration will have in terms of the state’s view as to their continued role in the provision of assurance. We suggest that developments from New Zealand in relation to capped conveyancing insurance are likely to be replicated in Australia in the near future, and that the state’s role in providing an assurance fund will continue, though the state may seek to expand the areas in which the right to compensation is restricted.

Storage stability of solid rocket fuels. 2. Effect of oxidizer loadings

Ageing behaviour, leading to ballistic changes, has been studied as a function of oxidizer loading in polystyrene/ammonium perchlorate solid-propellants. The ageing studies were carried out at 100 °C in air. Change in burning rate decreased as the oxidizer loading increased from 75 to 80%. The change in thermal decomposition rates both at 230 and 260 °C also decreased as the oxidizer loading in the propellants increased. The shapes of the plots of the changes in burning rate and thermal decomposition rate (230 and 260 °C) at different storage times for different oxidizer-loaded propellants seem to be exactly similar. These results lead to the conclusion that the thermal decomposition of the propellant may be responsible for bringing about the ballistic changes during the ageing process. Infrared studies of the binder portion of the aged propellant indicate that peroxide formation takes place during the course of ageing and that peroxide formation for a particular storage time and temperature increases as the loading decreases.