Odorant receptors from the light brown apple moth (Epiphyas postvittana) recognize important volatile compounds produced by plants.

Moths recognize a wide range of volatile compounds, which they use to locate mates, food sources, and oviposition sites. These compounds are recognized by odorant receptors (OR) located within the dendritic membrane of sensory neurons that extend into the lymph of sensilla, covering the surface of insect antennae. We have identified 3 genes encoding ORs from the tortricid moth, Epiphyas postvittana, a pest of horticulture.

Rapid saliva processing techniques for near real-time analysis of salivary steroids and protein.

Introduction: Point-of-care (POC) measurements using saliva samples have immense potential to assess systemic health and wellbeing, but sample viscosity and contaminants can affect analyses. We sought a portable clean-up method for whole saliva appropriate for use with POC measurement techniques such as biosensors.

Methods: Whole saliva from each of 13 male subjects was split into 5 fractions.

Analysis of expressed sequence tags from Actinidia: applications of a cross species EST database for gene discovery in the areas of flavor, health, color and ripening.

Background: Kiwifruit (Actinidia spp.) are a relatively new, but economically important crop grown in many different parts of the world. Commercial success is driven by the development of new cultivars with novel consumer traits including flavor, appearance, healthful components and convenience.

Expressed sequence tags and proteomics of antennae from the tortricid moth, Epiphyas postvittana.

Genomic and proteomic analyses of the antennae of the light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae) were undertaken to identify genes and proteins potentially involved in odorant and pheromone binding and turnover. An EST approach yielded 5739 sequences, comprising 808 contigs and 1545 singletons. InterPro and Blast analyses revealed members of families implicated in odorant and pheromone binding (PBPs, GOBPs, ABPXs and CSPs) and turnover (CXEs, GSTs, CYPs).

Identification of a cis-regulatory element by transient analysis of co-ordinately regulated genes.

Background: Transcription factors (TFs) co-ordinately regulate target genes that are dispersed throughout the genome. This co-ordinate regulation is achieved, in part, through the interaction of transcription factors with conserved cis-regulatory motifs that are in close proximity to the target genes. While much is known about the families of transcription factors that regulate gene expression in plants, there are few well characterised cis-regulatory motifs.

Extraction of proteins from plant tissues.

A basic protocol is described for extracting protein from plants. Suggestions are included for overcoming some of the common obstacles encountered (e.g.

The missing step of the L-galactose pathway of ascorbate biosynthesis in plants, an L-galactose guanyltransferase, increases leaf ascorbate content.

The gene for one postulated enzyme that converts GDP-L-galactose to L-galactose-1-phosphate is unknown in the L-galactose pathway of ascorbic acid biosynthesis and a possible candidate identified through map-based cloning is the uncharacterized gene At4g26850. We identified a putative function for At4g26850 using PSI-Blast and motif searching to show it was a member of the histidine triad superfamily, which includes D-galactose uridyltransferase. We cloned and expressed this Arabidopsis gene and the homologous gene from Actinidia chinensis in Escherichia coli and assayed the expressed protein for activities related to converting GDP-L-galactose to L-galactose-1-P.

Chirality and biosynthesis of lilac compounds in Actinidia arguta flowers.

Biosynthesis of lilac compounds in 'Hortgem Tahi' kiwifruit (Actinidia arguta) flowers was investigated by treating inflorescences with d(5)-linalool. The incorporation of the deuterium label into 8-hydroxylinalool, 8-oxolinalool, the lilac aldehydes, alcohols, and alcohol epoxides was followed by GC-MS and enantioselective GC-MS. Both (R)- and (S)-linalool were produced naturally by the flowers, but 8-hydroxylinalool, 8-oxolinalool, and the lilac aldehydes and alcohols occurred predominantly as the (S) and 5'(S)-diastereoisomers, respectively.

Unusual features of a recombinant apple alpha-farnesene synthase.

A recombinant alpha-farnesene synthase from apple (Malus x domestica), expressed in Escherichia coli, showed features not previously reported. Activity was enhanced 5-fold by K(+) and all four isomers of alpha-farnesene, as well as beta-farnesene, were produced from an isomeric mixture of farnesyl diphosphate (FDP). Monoterpenes, linalool, (Z)- and (E)-beta-ocimene and beta-myrcene, were synthesised from geranyl diphosphate (GDP), but at 18% of the optimised rate for alpha-farnesene synthesis from FDP.

Analyses of expressed sequence tags from apple.

The domestic apple (Malus domestica; also known as Malus pumila Mill.) has become a model fruit crop in which to study commercial traits such as disease and pest resistance, grafting, and flavor and health compound biosynthesis. To speed the discovery of genes involved in these traits, develop markers to map genes, and breed new cultivars, we have produced a substantial expressed sequence tag collection from various tissues of apple, focusing on fruit tissues of the cultivar Royal Gala.

Lilac alcohol epoxide: a linalool derivative in Actinidia arguta flowers.

Lilac alcohol epoxide (2-(5-methyl-5-(oxiran-2-yl)-tetrahydrofuran-2-yl)propan-1-ol), a previously unreported monoterpene, was identified in the solvent extract of the flowers of seven Actinidia arguta genotypes. The diastereomeric lilac alcohol epoxides co-occurred with the lilac aldehydes and alcohols. Another compound, the lilac diol (2-(5-(1-hydroxyethyl)-5-methyl-tetrahydrofuran-2-yl)propan-1-ol) was synthesised as part of our efforts to identify the lilac alcohol epoxide.

Maternal behaviour in sheep (Ovis aries) following administration of opioid agonists.

Aims: As kappa(kappa)-opioids have marked effects on pain threshold in sheep during oestrus, late pregnancy and after birth, a study was undertaken to determine if kappa -opioids also had other roles during these states.

Methods: Opioid agonists (GR89696, DAMAGO) were administered into either the lateral hypothalamus (LH) or amygdala of non-pregnant sheep (n = 5) or pregnant sheep (n = 15) in late pregnancy (n = 5) within 12 h of birth (n = 5) or in the third week after birth (n = 5). Cortisol, behavioural and electrocardiographic responses to an auditory stimulus were assessed with and without drug administration, as was feed consumption and suckling events.

Steroidal hormones determine sex-related differences in opioid-induced elevation of nociceptive threshold in sheep (Ovis aries).

Aims: As sex related differences in opioid-induced elevations of nociceptive thresholds have been observed in sheep, a study was undertaken to determine if puberty, oestrus, pregnancy or administration of the hormones oestrogen and testosterone had any effect on these differences.

Methods: Withdrawal latency to a nociceptive thermal source was measured in Romney cross sheep consisting of lambs (n = 6) prior to and after their first oestrus, ram lambs (n = 6) prior to and after puberty, non-oestrous ewes (n = 6), oestrous ewes (n = 6), pregnant and subsequently suckling ewes (n = 12), wethers (n = 6) and rams (n = 6). The effects of a kappa opioid agonist (GR 89696), oestradiol benzoate, testosterone and saline on this withdrawal latency were tested in all groups.

An alcohol acyl transferase from apple (cv. Royal Gala), MpAAT1, produces esters involved in apple fruit flavor.

Apple flavor is characterized by combinations of ester compounds, which increase markedly during fruit ripening. The final step in ester biosynthesis is catalyzed by alcohol acyl transferases (AATs) that use coenzyme A (CoA) donors together with alcohol acceptors as substrates. The gene MpAAT1, which produces a predicted protein containing features of other plant acyl transferases, was isolated from Malus pumila (cv.

A highly specific L-galactose-1-phosphate phosphatase on the path to ascorbate biosynthesis.

Ascorbate is a critical compound in plants and animals. Humans are unable to synthesize ascorbate, and their main source of this essential vitamin are plants. However, the pathway of synthesis in plants is yet to be established, and several unknown enzymes are only postulated to exist.

Purification and characterization of phytocystatins from kiwifruit cortex and seeds.

Kiwifruit cysteine proteinase inhibitors (KCPIs) were purified from the cortex and seeds of kiwifruit after inactivation of the abundant cortex cysteine proteinase actinidain. One major (KCPI1) and four minor cystatins were identified from Actinidia deliciosa ripe mature kiwifruit cortex as well as a seed KCPI from A. chinensis.

Actinidia arguta: volatile compounds in fruit and flowers.

More than 240 compounds were detected when the volatile components of the flowers and the fruit from several Actinidia arguta genotypes were investigated. Around 60-70 different compounds were extracted from individual tissues of each genotype. Two different methods of volatile sampling (headspace and solvent) favoured different classes of compounds, dependent upon their volatilities and solubilities in the flower or fruit matrices.

Mechanism of inactivation of ornithine transcarbamoylase by Ndelta -(N'-Sulfodiaminophosphinyl)-L-ornithine, a true transition state analogue? Crystal structure and implications for catalytic mechanism.

The crystal structure is reported at 1.8 A resolution of Escherichia coli ornithine transcarbamoylase in complex with the active derivative of phaseolotoxin from Pseudomonas syringae pv. phaseolicola, N(delta)-(N'-sulfodiaminophosphinyl)-l-ornithine.

A unified nomenclature for sap flow measurements.

A unified nomenclature for use in heat pulse measurement of sap flow is proposed. This unified nomenclature overcomes fundamental misunderstandings of the physics of heat and sap movement in wood. The nomenclature is also appropriate to other methodologies for sap flow measurement, such as heat balance methods.

What determines carbon partitioning between competing sinks?

Carbohydrate sinks have been described by their ability to attract photosynthate, denoted by sink strength, and by their priority rank ordering for supply in the presence of a reduced availability of photosynthate. Sink strength has been defined as the rate of carbohydrate flow into a sink, but this flow rate is also dependent upon supply, other sinks, and resistance to flow of the transport pathway, so it is not a property of the sink alone. It is a property of the entire system.

Disruption of virus movement confers broad-spectrum resistance against systemic infection by plant viruses with a triple gene block.

White clover mosaic virus strain O (WClMV-O), species of the Potexvirus genus, contains a set of three partially overlapping genes (the triple gene block) that encodes nonvirion proteins of 26 kDa, 13 kDa, and 7 kDa. These proteins are necessary for cell-to-cell movement in plants but not for replication. The WClMV-O 13-kDa gene was mutated (to 13*) in a region of the gene that is conserved in all viruses known to possess triple-gene-block proteins.