Improved resistance to root pests: final report to Grape and Wine Research & Development Corporation

There were two major aims of this project, which was jointly funded by GWRDC and CRCV2. The first aim was to identify and characterise grapevine genes involved in the plantâ s interaction with phylloxera and/or root-knot nematode. This information could be used to design strategies for engineering novel resistance to these pests in grapevine. The second aim of the project was to develop and use systems to rapidly assess candidate genes for preventing infestation of vine roots by phylloxera and/or root-knot nematode. Progress towards both goals was achieved despite premature termination of the project after 4, instead of 7 years.Project Leader: Dr. Robyn van Heeswijck until her retirement in July, 2002, then Prof. Steve Tyerman (in an administrative capacity); Author Details: Dr Tricia Franks (using text from the original application by Dr. Robyn van Heeswijck

Changes in Grape Maturity Induced by Spraying Ethanol

Three different ethanol solutions were sprayed onto Cabernet Sauvignon (Vitis vinifera L.) clusters during the ripening period: 2.5, 5 and 10% by volume in water. Controls were sprayed with water alone. Three different times of spraying were also tested: 8, 10 and 13 weeks post-flowering. One of the observed changes was a lower titratable acidity in grape samples at harvest, when the clusters were sprayed with ethanol at 10 weeks, in comparison with controls. The wines made with grapes treated with ethanol after mid-veraison, had higher ODs at 520 nm than did the controls. This may due to a combined effect of red pigment levels and acidity. In addition, following malolactic fermentation, the acidity levels of wines made with ethanoltreated grapes were slightly higher than those made with the control grapes. Spraying ethanol at 13 weeks post-flowering increased the berry weight by 10% at harvest without decreasing the °Brix value. The corresponding wines had similar degrees of alcohol. This observation was made for the first time in 2001

Ethanol triggers grape gene expression leading to anthocyanin accumulation during berry ripening

Recent studies have shown that low doses of ethanol stimulate the maturation of some fruits. The present work showed that spraying Cabernet Sauvignon grapes, with 5% ethanol at veraison enhances the anthocyanin accumulation. Veraison is the time when the berries turn from green to purple. HPLC analysis showed a marked increase in the total concentrations of the derivatives of delphinidin, cyanidin, petunidin, peonidin and malvidin from the fourth day after the ethanol treatment until harvest. This was not linked to a difference in berry weight in comparison to controls. Two distinct expression patterns were found for anthocyanin biosynthesis genes in the treated and untreated berries. For one group, consisting of chalcone synthase, flavanone-3-hydroxylase, dihydroxyflavonol-4-reductase and leucoanthocyanidin dioxygenase, the expression was inhibited or unchanged by the ethanol treatment, whereas for UDP glucose-flavonoid 3-O-glucosyltransferase (UFGT) there was a marked increase in expression from 1 to 20 days after ethanol treatment. These results suggest that the UFGT gene is a key factor in the observed anthocyanin accumulation following ethanol treatment

Evidence for host-associated clones of grape phylloxera Daktulosphaira vitifoliae (Hemiptera : Phylloxeridae) in Australia

Grape phylloxera, Daktulosphaira vitifoliae Fitch, is an important pest of grapevines (Vitis vinifera L.) (Vitaceae). Using microsatellite DNA markers it was demonstrated strong associations can exist between D. vitifoliae asexual lineages and vine host type within a vineyard. Also, in excised root bioassays, D. vitifoliae collected from three regions where different genotypic classes predominated showed host-specific differences in life table parameters of reproductive rate and intrinsic rate of increase. Lastly, comparisons of mitochondrial DNA (cytochrome oxidase I) sequences revealed that D. vitifoliae in Australia have paraphyletic origins and fall into two clades partially related to vine host usage. These findings indicate introduction of separate lineages of D. vitifoliae which have close host associations and as such, have important implications for management of this pest in Australia.A.M. Corrie, R. van Heeswijck and A.A. Hoffman

Metabolic engineering of Rhizopus oryzae for the production of platform chemicals

Rhizopus oryzae is a filamentous fungus belonging to the Zygomycetes. It is among others known for its ability to produce the sustainable platform chemicals l-(+)-lactic acid, fumaric acid, and ethanol. During glycolysis, all fermentable carbon sources are metabolized to pyruvate and subsequently distributed over the pathways leading to the formation of these products. These platform chemicals are produced in high yields on a wide range of carbon sources. The yields are in excess of 85 % of the theoretical yield for l-(+)-lactic acid and ethanol and over 65 % for fumaric acid. The study and optimization of the metabolic pathways involved in the production of these compounds requires well-developed metabolic engineering tools and knowledge of the genetic makeup of this organism. This review focuses on the current metabolic engineering techniques available for R. oryzae and their application on the metabolic pathways of the main fermentation products

Functional Analysis of the Phycomyces carRA Gene Encoding the Enzymes Phytoene Synthase and Lycopene Cyclase

Phycomyces carRA gene encodes a protein with two domains. Domain R is characterized by red carR mutants that accumulate lycopene. Domain A is characterized by white carA mutants that do not accumulate significant amounts of carotenoids. The carRA-encoded protein was identified as the lycopene cyclase and phytoene synthase enzyme by sequence homology with other proteins. However, no direct data showing the function of this protein have been reported so far. Different Mucor circinelloides mutants altered at the phytoene synthase, the lycopene cyclase or both activities were transformed with the Phycomyces carRA gene. Fully transcribed carRA mRNA molecules were detected by Northern assays in the transformants and the correct processing of the carRA messenger was verified by RT-PCR. These results showed that Phycomyces carRA gene was correctly expressed in Mucor. Carotenoids analysis in these transformants showed the presence of ß-carotene, absent in the untransformed strains, providing functional evidence that the Phycomyces carRA gene complements the M. circinelloides mutations. Co-transformation of the carRA cDNA in E. coli with different combinations of the carotenoid structural genes from Erwinia uredovora was also performed. Newly formed carotenoids were accumulated showing that the Phycomyces CarRA protein does contain lycopene cyclase and phytoene synthase activities. The heterologous expression of the carRA gene and the functional complementation of the mentioned activities are not very efficient in E. coli. However, the simultaneous presence of both carRA and carB gene products from Phycomyces increases the efficiency of these enzymes, presumably due to an interaction mechanism

The structure, sedimentology, sequence stratigraphy and tectonics of the northern Drummond and Galilee Basins, Central Queensland, Australia

The Late Devonian-Early Carboniferous Drummond Basin and overlying Late Carboniferous-Middle Triassic Galilee Basin are extensive intracratonic basins with predominantly fluvial sediment infill. They are inferred to have developed on Proterozoic and Early Palaeozoic basement similar to the adjacent Charters Towers Province and Anakie Inlier. Depositional and structural architecture of the basins have been investigated through a combination of seismic, geophysical well log and gravity data sets with lithologic information obtain from well cores, surface mapping and sedimentary petrography. Comprehensive interpretation of 750 km of seismic traverse resulted in the recognition of eleven seismic facies, several of which have distinctive internal signatures, separated by reflection boundaries that can be traced basin wide. Lithologic and geophysical logs from sixteen wells and boreholes can be matched to seismic stratigraphic units and allow the basinal infill to be described in terms of the lithostratigraphic frameworks already established in the literature. The construction of structure contours for unit boundary surfaces has identified extensional structures associated with rift phases of basinal development and the pattern of thrust and thrust fold features associated with basin inversion. Eleven new structural features that have continuity through the Drummond and Galilee Basins have been defined and other features noted in the literature have been reinterpreted based on evidence from seismic profiles. The construction of isopachs for each unit has allowed tracking of basinal infill thickness trends through time. Analysis of geophysical and lithologic logs from petroleum wells and boreholes, augmented by sections documented from surface exposure has resulted in the recognition of some 16 discrete sequences, each based on a repeating pattern of three types of non-marine systems tract that mainly reflect basinal tectonics but also the influence of climate and eustatic sea level change. Both basins share a common style of sequence development that is not reflected in the current literature. Provenance interpretation based on petrographic data from 121 thin sections representative of most of the Drummond Basin and all of the Galilee Basin suggests that the majority of basin infill was derived from a recycled cratonic source, such as the Thomson Fold Bel to the west, and less material derived from an eastern volcanic arc than previously thought. However SHRIMP-derived U-Pb age data for zircon populations from two samples broadly representative of quartz arenites prominent in the basinal successions conflict with this view. Such data for a sample from the Mount Hall Formation of the Drummond Basin indicates that its source was largely from Early to Mid Palaeozoic igneous terranes like those represented in the Charters Towers Province to the north and inferred for the Thomson Fold Belt to the west. Zircon ages of a sample of Warang Sandstone of the Galilee Basin indicates a Late Palaeozoic igneous source, with derivation largely from an eastern magmatic arc. The Drummond Basin commenced as a back-arc extensional basin, progressed through a thermal sag phase and ceased development during mild compression associated with a far-field expression of the Kanimblan Orogeny. Structural patterns show initial rift architecture, with compartments separated by newly defined transfer fault zones. Extensional faults between the transfer structures extend further through the basinal succession than previously thought. This basin developed a broad sag phase but the final stages show a foreland influence induced by Kanimblan thrust loading on its eastern margin. Basinal structure has been strongly modified by inversion in the Middle Triassic associated with the Hunter-Bowen Orogeny. The Galilee Basin commenced as a foreland basin expressing continuity with the late-stage development of the underlying Drummond Basin from which its division is arbitrary and based on historical misconceptions of Kanimblan tectonism. Thermal subsidence related to the rift phase of the Drummond Basin continued as an influence in addition to foreland subsidence. Tectonic quiescence marks a mid-stage of basinal development reflected in a regionally developed paraconformity and deposition of a basinwide coal measure sequence related to eustatic sea level rise over a stable substrate. The upper part of Galilee Basin infill reflects a foreland phase of development and records two episodes of thrust loading on the eastern margin associated with the Hunter-Bowen Orogeny. Ongoing crustal contraction during the Hunter-Bowen Orogeny resulted in inversion of the Drummond and Galilee Basins with the development of large-scale thrust dislocations and associated fault bend anticlinal structures. The Middle Triassic Clematis Group is the youngest unit that shows folding due to inversion. Much of the western parts of both basins remain relatively undisturbed apart from gentle regional pre-Middle Jurassic tilting that marked the final phase of Hunter-Bowen tectonism

Late Paleozoic to Early Mesozoic deformation in the\ud northeastern Galilee Basin, Australia

Newly recognised depocentres and structures in the northeastern Galilee Basin are associated with long-term passive thermal subsidence and periods of uplift. Evidence presented here shows that a contractional event in the mid-Carboniferous had an influence during late-stage development of the underlying Drummond Basin and early development of the Galilee Basin. An extensional event in the Late Carboniferous is documented by analysis of depositional patterns. Permian to mid-Triassic contractional events in the adjacent Bowen Basin had little influence in development of the Galilee Basin. Major inversion in the Middle Triassic reactivated structures and ended basin development