Relative roles of glyceollin, lignin and the hypersensitive response and the influence of ABA in compatible and incompatible interactions of soybeans with Phytophthora sojae

The relative roles of glyceollin, lignin and the hypersensitive response (HR) in pathogen containment and restriction were investigated in soybean (Glycine max L. [Merr.]) cultivars that were inoculated with Phytophthora sojae Kaufmann and Gerdemann. Concentrations of endogenous abscisic acid (ABA) levels in etiolated soybean hypocotyls were reduced by application of the ABA biosynthesis inhibitor norflurazon or raised by exogenous ABA application. Incompatible interactions in leaves and hypocotyls were characterized by HR, phenolic and lignin deposition and glyceollin accumulation. Compatible interactions resulted in light coloured, water-soaked spreading lesions with minimal lignin deposition or glyceollin accumulation and the absence of an HR. Norflurazon treatment restricted the spread of the pathogen and increased glyceollin accumulation in compatible tissues. Exogenous ABA addition caused spreading lesions in normally incompatible interactions and reduced glyceollin accumulation. Phenolic deposition and HR were unchanged by either treatment in incompatible or compatible interactions. The uncoupling of glyceollin synthesis from the HR and phenolic and lignin deposition by ABA and norflurazon treatment showed that glyceollin is a major factor in restriction of the pathogen during these interactions.

Suppression by ABA of salicylic acid and lignin accumulation and the expression of multiple genes, in Arabidopsis infected with Pseudomonas syringae pv. tomato

Abscisic acid (ABA) has been implicated in determining the outcome of interactions between many plants and their pathogens. We had previously shown that increased concentrations of ABA within leaves of Arabidopsis induced susceptibility towards an avirulent strain of Pseudomonas syringae pathovar (pv.) tomato. We now show that ABA induces susceptibility via suppression of the accumulation of components crucial for a resistance response. Lignin and salicylic acid concentrations in leaves were increased during a resistant interaction but reduced when plants were treated with ABA. The reduction in lignin and salicylic acid production was independent of the development of the hypersensitive response (HR), indicating that, in this host-pathogen system, HR is not required for resistance. Genome-wide gene expression analysis using microarrays showed that treatment with ABA suppressed the expression of many defence-related genes, including those important for phenylpropanoid biosynthesis and those encoding resistance-related proteins. Together, these results show that resistance induction in Arabidopsis to an avirulent strain of P. syringae pv. tomato is regulated by ABA.

Lignin production and monolignol pathway gene expression is suppressed by abscisic acid during defence reactions of Arabidopsis

The phytohormone, abscisic acid (ABA) has been shown to influence the outcome of the interactions between various hosts with biotrophic and hemibiotrophic pathogens. Susceptibility to avirulent isolates can be induced by addition of low physiological concentrations of ABA to plants. In contrast, addition of ABA biosynthesis inhibitors induced resistance following challenge of plants by virulent isolates. ABA deficient mutants of Arabidopsis, such as aba1-1, were resistant to virulent isolates of Peronospora parasitica. In interactions of Arabidopsis with avirulent isolates of Pseudomonas syringae pv. tomato, susceptibility was induced following addition of ABA or imposition of drought stress. These results indicate a pivotal, albiet undefined, role for ABA in determining either susceptibility or resistance to pathogen attack. We have found that the production of the cell wall strengthening compound, lignin, is increased during resistant interactions of aba1-1 but suppressed in ABA induced susceptible interactions. Using RT-PCR and microarray analysis we have found down-regulation by ABA of key genes of the phenylpropanoid pathway especially of those genes involved directly in lignin biosynthesis. ABA also down-regulates a number of genes in other functional classes including those involved in defence and cell signalling.

Suppression by abscisic acid of lignin production and monolignol pathway gene expression in interactions of Arabidopsis with oomycete and bacterial pathogens

The phytohormone, abscisic acid (ABA) has been shown to influence the outcome of the interactions between various hosts with biotrophic and hemibiotrophic pathogens. Susceptibility to avirulent isolates can be induced in plants by addition of low physiological concentrations of ABA. In contrast, addition of ABA biosynthesis inhibitors induced resistance following challenge of plants by virulent isolates. ABA deficient mutants of Arabidopsis, such as aba1-1, were resistant to virulent isolates of Peronospora parasitica. In interactions of Arabidopsis with avirulent isolates of Pseudomonas syringae pv. tomato, susceptibility was induced following addition of ABA or imposition of drought stress. These results indicate a pivotal, albiet undefined, role for ABA in determining either susceptibility or resistance to pathogen attack. We have found that the production of the cell wall strengthening compound, lignin, is increased during resistant interactions of aba1-1 but suppressed in ABA-induced susceptible interactions. Using RT-PCR and microarray analysis we have found down-regulation by ABA of key genes of the phenylpropanoid pathway especially of those genes involved directly in lignin biosynthesis. ABA also down-regulates a number of genes in other functional classes including those involved in defence and cell signalling.

Thermal and rheological characteristics of biobased carbon fiber precursor derived from low molecular weight organosolv lignin

In the present work, electrospinnability as well as thermal, rheological, and morphological characteristics of low molecular weight hardwood organosolv lignin, as a potential precursor for carbon fiber, was investigated. Submicromter biobased fibers were electrospun from a wide range of polymer solutions with different ratios of organosolv lignin to polyacrylonitrile (PAN). Rheological studies were conducted by measuring viscosity, surface tension, and electrical conductivity of hybrid polymer solutions, and used to correlate electrospinning behavior of solutions with the morphology of the resultant electrospun composite fibers. Using scanning electron microscopy (SEM) images, the solutions that led to the formation of bead-free uniform fibers were found. Differential scanning calorimetry (DSC) analysis revealed that lignin-based fibers enjoy higher decomposition temperatures than that of pure PAN. Thermal stability of the lignin-based fibers was investigated by thermogravimetric analysis (TGA) indicating a high carbon yield of above 50% at 600 °C, which is highly crucial in the production of low-cost carbon fiber. It was also observed that organosolv lignin synergistically affects thermal decomposition of composite fibers. A significant lower activation energy was found for the pyrolysis of lignin-derived electrospun fibers compared to that of pure PAN.

Correlating the fineness and residual gum content of degummed hemp fibres

It is well known residual gum exists in degummed or retted hemp fibres. Gum removal results in improvement in fibre fineness and the properties of the resultant hemp yarns. However, it is not known what correlation if any exists between the residual gum content in retted hemp fibres and the fibre fineness, described in terms of fibre width in this paper. This study examined the mean width and coefficient of variation (CV) of fibre width of seventeen chemically retted hemp samples with reference to residual gum content. The mean and CV of fibre width were obtained from an Optical fibre diameter analyser (OFDA 100). The linear regression analysis results show that the mean fibre width is directly proportional to the residual gum content. A slightly weaker linear correlation also exists between the coefficient of variation of fibre width and the residual gum content. The strong linear co-relation between the mean of fibre width and the residual gum content is a significant outcome, since testing for fibre width using the OFDA is a much simpler and quicker process than testing the residual gum content. Scanning Electron Microscopy (SEM) reinforces the OFDA findings. SEM micrographs show a flat ribbon like fibre cross-section hence the term “fibre width” is used instead of fibre diameter. Spectral differences in the untreated dry decorticated skin samples and chemically treated and subsequently carded samples indicate delignification. The peaks at 1370 cm⁻¹, 1325 cm⁻¹, 1733 cm⁻¹, and 1600 cm⁻¹ attributed to lignin in the untreated samples are missing from the spectra of the treated samples. The spectra of the treated samples are more amine-dominated with some of the OH character lost.

Anaerobic degradation of organic materials - significance of the substrate surface area

In anaerobic degradation of substrates containing mainly particulate organic matter, solids hydrolysis is rate-limiting. In these investigations, the particle size of various substrates was reduced by comminution to support hydrolysis. Two positive effects of comminution were observed. For substrates with high fibre content, which are particularly resistant to biodegradation, a significant improvement of the degradation degree was observed as a result of comminution. Secondly, for all substrates tested, and particularly for those rich in fibres, the degradation rate of comminuted samples was significantly higher. The first reason for both effects is an increase of the sample surface area. Several methods for measuring the specific surface area of organic materials, including particle size analysis, Nitrogen-adsorption and enzyme adsorption, were used and compared for the purpose of this study, where the surface area accessible to microbial enzymes is critical. The significance of the surface area in anaerobic degradation of particulate substrates was investigated through a kinetic model where the hydrolysis rate was based on the sample surface area. Good agreements were obtained between model and experiments carried out with samples of various specific surface areas. These results reinforced the significance of the sample surface area in anaerobic degradation processes. However, other effects of comminution responsible for the increased degradation degree and degradation rate were identified and discussed. These include: the increase of dissolved compounds due to cell rupture, exposition of surface areas previously inaccessible for microbial degradation, and alteration of the sample structure such as the lignin-cellulose arrangements.

Determining residual gum content of bast fibres

The measurement of fibre quality in bast fibres is related to the amount of gum (lignin, hemicellulose, wax and pectin) left in the fibre after the retting process. Large amounts of gums present represent poor separation of fibre. Efficiency of retting can be monitored by measuring the residual gum content of the retted fibre.

This paper investigated the use of ultrasonic vibration combined with chemical retting as a pre-treatment to improve accuracy of traditional residual gum content test. Work was conducted on chemically retted hemp fibre. Pre-treatment conditions were analysed by determining the best chemical combination, chemical concentration and treatment time. Fibres were examined for successful separation using optical microscopy and optical fibre diameter analysis (OFDA). The work proposed a new method for determining the residual gum content of hemp fibre.

Interaction of Arabidopsis Thaliana with Plasmodiophora Brassicae

Plasmodiophora brassicae is a protistan pathogen that attacks roots of brassicaceous plant species causing devastating disease. Resistance is characterised by restriction of the pathogen and susceptibility by the development of severely malformed roots (‘clubroots’) and stunting of the plant that is associated with alterations in the synthesis of cytokinin and auxin hormones. We are examining the susceptible response in Arabidopsis and whether suppression of key resistance factors by the pathogen contributes to susceptibility. The interaction is being studied using a number of approaches including microscopy of the infection process and development of the pathogen within roots and host gene expression analysis. Quantitative PCR was used to confirm the timing of infection of roots and showed that infection occurred at day four and colonisation increased thereafter to high levels by 23 days after inoculation by which time roots were showing systemic abnormalities. To investigate the basis of this compatible interaction we have conducted a time course experiment following infection of a susceptible ecotype of Arabidopsis (Col-0) to examine whole genome geneexpression changes in the host. Differential gene expression analysis of inoculated versus control roots showed that a higher number of genes had altered expression levels at day four compared to that at day seven and at day ten. At day four the expression levels of several genes known to be important for recognition and signal transduction in resistant interactions and genes involved in the biosynthesis of lignin, phenylpropanoids and ethylene were suppressed. Suppression by P. brassicae of specific plant defence responses appears to be a key component of susceptibility in this system.

Latex-based nanocomposites

Nanoparticles have been widely used as filler in polymer because of their unique reinforcing effect. There are many compounding methods for nanocomposites. The recent development on latex nanocomposites, a group of special nanocomposites, is reviewed in this chapter. They include carbon black/latex nanocomposite, silica/latex nanocomposite, layered silicate/latex nanocomposite, ZnO/latex nanocomposite, carbon nanotubellatex nanocomposite, lignin/latex nanocomposite, starch/latex nanocomposite, nano-fiber/latex nanocomposite, and Chitin whiskers/latex nanocomposite. Advanced compounding techniques and the latest advance on these latex nanocomposites are described. The nanoreinforcing theories of latex nanocomposites are also studied.

Abscisic acid regulation of plant defence responses during pathogen attack

The plant hormone, abscisic acid (ABA), has previously been shown to have an impact on the resistance or susceptibility of plants to pathogens. In this thesis, it was shown that ABA had a regulatory effect on an extensive array of plant defence responses in three different plant and pathogen interaction combinations as well as following the application of an abiotic elicitor. In unique studies using ABA deficient mutants of Arabidopsis, exogenous ABA addition or ABA biosynthesis inhibitor application and simulated drought stress, ABA was shown to have a profound effect on the outcome of interactions between plants and pathogens of differing lifestyles and from different kingdoms. The systems used included a model plant and an important agricultural species: Arabidopsis thaliana (Arabidopsis) and Peronospora parasitica (a biotrophic Oomycete pathogen), Arabidopsis and Pseudomonas syringae pathovar tomato (a biotrophic bacterial pathogen) and an unrelated plant species, soybean (Glycine max) and Phytophthora sojae (a hemibiotrophic Oomycete pathogen), Generally, a higher than basal endogenous ABA concentration within plant tissues at the time of avirulent pathogen inoculation, caused an interaction shift towards what phenotypically resembled susceptibility. Conversely, a lower than basal endogenous ABA concentration in plants inoculated with a virulent pathogen caused a shift towards resistance. An extensive suppressive effect of ABA on defence responses was revealed by a range of techniques that included histochemical, biochemical and molecular approaches. A universal effect of ABA on suppression or induction of the phenylpropanoid pathway via regulation of the key entry point gene, phenylalanine ammonia-lyase (PAL), when stimulated by biotic or abiotic elicitors was shown. ABA also influenced a wide variety of other defence-related components such as: the development of a hypersensitive response (HR), the accumulation of the reactive oxyden species, hydrogen peroxide and the cell wall strengthening compounds lignin and callose, accumulation of SA and the phytoalexin, glyceollin and the transcription of the SA-dependent pathogenesis- related gene (PR-1). The near genome-wide microarray gene expression analysis of an ABA induced susceptible interaction also revealed an yet unprecedented insight into the great diversity of defence responses that were influenced by ABA that included: disease resistance like proteins, antimicrobial proteins as well as phenylpropanoid and tryptophan pathway enzymes. Subtle differences were found in the number and type of defence responses that were regulated by ABA in each type of plant and pathogen interaction that was studied. This thesis has clearly identified in plant/pathogen interactions previously unknown and important roles for ABA in the regulation of many defence responses.

Aspects of the interaction between Xanthorrhoea australis and Phytophthora cinnamomi in south-western Victoria, Australia

Diseases in natural ecosystems are often assumed to be less severe than those observed in domestic cropping systems due to the extensive biodiversity exhibited in wild vegetation communities. In Australia, it is this natural biodiversity that is now under threat from Phytophthora cinnamomi. The soilborne Oomycete causes severe decline of native vegetation communities in south-western Victoria, Australia, disrupting the ecological balance of native forest and heathland communities. While the effect of disease caused by P. cinnamomi on native vegetation communities in Victoria has been extensively investigated, little work has focused on the Anglesea healthlands in south-western Victoria. Nothing is known about the population structure of P. cinnamomi at Anglesea. This project was divided into two main components to investigate fundamental issues affecting the management of P. cinnamomi in the Anglesea heathlands. The first component examined the phenotypic characteristics of P. cinnamomi isolates sampled from the population at Anglesea, and compared these with isolates from other regions in Victoria, and also from Western Australia. The second component of the project investigated the effect of the fungicide phosphonate on the host response following infection by P. cinnamomi. Following soil sampling in the Anglesea heathlands, a collection of P, cinnamomi isolates was established. Morphological and physiological traits of each isolate were examined. All isolates were found to be of the A2 mating type. Variation was demonstrated among isolates in the following characteristics: radial growth rate on various nutrient media, sporangial production, and sporangial dimensions. Oogonial dimensions did not differ significantly between isolates. Morphological and physiological variation was rarely dependant on isolate origin. To examine the genetic diversity among isolates and to determine whether phenotypic variation observed was genetically based, Random Amplified Polymorphic DNA (RAPD) analyses were conducted. No significant variation was observed among isolates based on an analysis of molecular variance (AMQVA). The results are discussed in relation to population biology, and the effect of genetic variation on population structure and population dynamics. X australis, an arborescent monocotyledon indigenous to Australia, is highly susceptible to infection by P. cinnamomi. It forms an important component of the heathland vegetation community, providing habitat for native flora and fauna, A cell suspension culture system was developed to investigate the effect of the fungicide phosphonate on the host-pathogen interaction between X. australis and P. cinnamomi. This allowed the interaction between the host and the pathogen to be examined at a cellular level. Subsequently, histological studies using X. australis seedlings were undertaken to support the cellular study. Observations in the cell culture system correlated well with those in the plant. The anatomical structure of X australis roots was examined to assist in the interpretation of results of histopathological studies. The infection of single cells and roots of X. australis, and the effect of phosphonate on the interaction are described. Phosphonate application prior to inoculation with P. cinnamomi reduced the infection of cells in culture and of cells in planta. In particular, phosphonate was found to stimulate the production of phenolic material in roots of X australis seedlings and in cells in suspension cultures. In phosphonate-treated roots of X australis seedlings, the deposition of electron dense material, possibly lignin or cellulose, was observed following infection with P. cinnamomi. It is proposed that this is a significant consequence of the stimulation of plant defence pathways by the fungicide. Results of the study are discussed in terms of the implications of the findings on management of the Anglesea heathlands in Victoria, taking into account variation in pathogen morphology, pathogenicity and genotype. The mode of action of phosphonate in the plant is discussed in relation to plant physiology and biochemistry.

Tailoring enzymes for biofuel production with reference to Australian biomass

Manufacture of biofuels from existing biomass may provide a sustainable alternative to the extensive utilization of fossil fuels. Biomass offers environmental advantage over fossil fuels as it is a renewable energy source with low sulphur and nitrogen content and is carbon neutral over its production and utilization. Ranges of biomass are reported worldwide to be suitable raw material for bioethanol production. These can be generally classified into three groups; sucrose based (sugar cane), starch based (corn, wheat and barley) and lignocellulosic (which is mostly comprised of lignin, cellulose and hemicelluloses in grasses, wood and straw) materials. However, the limited supply of two biomass groups (sucrose and starch) will not satisfy society’s growing energy demands; thus biofuel technology based on lignocelluloses is under intense investigation. The main bottleneck in lignocellulosic biomass conversion for biofuel production is the enzymatic depolymerisation of cell wall polysaccharides into fermentable sugars. Protein engineering has recently been used to improve the performance of lignocelluloses degrading enzymes, as well as proteins involved in biofuel synthesis pathways. We have produced a recombinant enzyme that has the ability to produce monomeric sugars from a complex substrate. This presentation will summarize current efforts to develop an enzymatic treatment which would facilitate the economical processing of biomass available in Australia for bioenergy generation.

UV absorption property of bamboo

This article describes ultra violet (UV) light shielding behaviour of Australian grown bamboo (Phyllostachys pubescens). Optical reflectance showed that untreated bamboo plant has UV absorption properties. To reveal the origin of the UV absorption property, its chemical components were extracted using several polar and non-polar solvents. The extracts in most of the polar and non-polar solvents showed UV absorption property. Protic polar solvents showed better ability to extract UV absorbing chemicals than aprotic and non-polar solvents, except hexane. The chemical components of bamboo were analysed by FT-IR spectroscopy and the findings were correlated with the UV absorbance characteristics. The results confirmed that the UV absorption ability of bamboo originates from nothing but lignin. It is thus indicated that the conventional methods to manufacture bamboo fibres, such as complete degumming or viscose methods, that involve the removal of lignin, cannot retain the unique UV absorption property of bamboo plant in bamboo fibres.