Search

Blackleg disease caused by the fungus Leptosphaeria maculans is the most important disease of canola worldwide. The impact of this disease on the development of the Australian canola industry, particularly over the last 20 years, is discussed. Deployment of a range of disease control measures has resulted in a thriving canola industry with production now approaching 4million tonnes annually. Discoveries about disease mechanisms and key plant and fungal genes are described. Analysis of the L. maculans genome sequence has enabled an understanding of how fungal populations can evolve rapidly to overcome disease resistance bred into canola cultivars.

Leptosphaeria maculanscauses blackleg of Brassica napus(canola), manifesting in symptoms including leaf lesions, stem canker, and root rot. Root rot is an extension of the stem canker phase of the disease resulting from foliar infection; however, the role of root rot in grain yield loss has not been investigated. Field experiments were conducted in south-eastern Australia to determine the efficacy of fungicides and host resistance to stem canker for the control of root rot, along with the association between root rot severity and grain yield. Fungicides applied as a seed dressing (fluquinconazole) or coated on fertiliser (flutriafol) reduced root rot severity by 16 and 41%, respectively. Root rot severity was 78% less in a B. napuscultivar with an Australian Blackleg Rating (ABR) of 9.0 (highly resistant to stem canker) than in a moderately resistant cultivar (ABR 5.5), and was also reduced in Brassicaspecies with good resistance to stem canker. Root rot caused little or no additional reduction in yield or harvest index of individual B. napusplants above that caused by stem canker. Individual upright plants with greater than 80% of the stem cross-section blackened had significantly reduced seed yield; however, this was influenced by rainfall during the grain-filling period. This study shows that current management strategies for stem canker caused by L. maculansalso control root rot in B. napus, as although the symptoms in roots are severe, these symptoms have no additional effect on yield.

Abstract Oviposition deterrence is common in many insects as an evolutionary mechanism to reduce subsequent larval competition. We investigated a suspected case of oviposition deterrence by the paropsine chrysomelid, Chrysophtharta bimaculata. In paired choice tests, gravid females were found to prefer ovipositing on host leaves without conspecific eggs, confirming the presence of an apparent oviposition deterrence mechanism. Washing egg batches in water, hexane, or ethanol did not change this preference, suggesting that a soluble marking pheromone was not involved. Furthermore, it is unlikely that a plant-derived oviposition deterring substance is produced as beetles showed no significant oviposition preference between leaves which had been oviposited upon, but then had the eggs removed, and those that had never been oviposited upon. In trials using artificial leaves and mimic egg batches, “leaves” with “egg batches” placed near the tip of the leaf (the preferred site of oviposition in this species) were significantly less likely to be laid upon than artificial leaves where mimic eggs were placed away from the tip. In combination, the results strongly infer that oviposition deterrence in C. bimaculata is due to the mechanical blocking of the oviposition site by the first laid egg-batch, rather than a specific oviposition deterring cue. The apparent oviposition deterrence in this insect may well be an outcome or evolutionary effect of oviposition-site selection, rather than a clear adaptive mechanism to decrease larval competition.

Two new cost-effective trap designs for capturing arthropod flower visitors are described. Small flower visitors such as Thysanoptera and Diptera, hardly noticeable during direct flower observations, were dominant in trap catches. These trapping techniques combined with direct observations allow a more comprehensive picture of the flower visitors. Cet article décrit le fonctionnement de deux nouveaux types de pièges bon marché pour capturer les visiteurs de fleurs. Ces nouveaux systèmes permettent de piéger des espèces miniatures visiteuses de fleurs comme les thysanoptères et diptères non identifiables lors des observations directes des fleurs. De telles techniques de piégeage en combinaison avec des observations directes permettent une évaluation plus complète de cette faune.

Due to the large increase of canola production in Australia, current blackleg cultural control recommendations (extended rotation length and isolation distance from canola stubble) are not adhered to by farmers in many canola-producing regions. Canola crops are increasingly being sown in short rotation and, in many instances, adjacent to paddocks containing canola stubble. In this study, the level of disease in commercial canola crops was determined for different rotations and distances from canola stubble. There was a strong relationship between the presence of canola stubble from the previous year (6-month-old stubble) and distance to current canola crops, but no relationship between the presence of older (18–42 month old) stubble and distance to current canola crops. Blackleg severity was highest where canola crops had been sown adjacent to 6-month-old canola stubble, with the level of blackleg severity decreasing markedly in the first 100 m. Disease severity then generally declined up to 500 m. Plants 500–1000 m from 6-month-old stubble had similar levels of blackleg infection. Blackleg severity was similar between canola crops sown into 18-month-old canola stubble (short rotation) and crops sown into paddocks that had no history of canola for at least the previous 3 years (long rotation). Based on these findings, we recommend that canola crops should be sown at distances greater than 100 m and preferably 500 m from last season's canola stubble, rather than extending rotation length between crops.

Abstract: Volatiles released from glucosinolates (predominantly 2-propenyl glucosinolate) in tissues of Brassica juncea and Wasabi japonica are toxic to the blackleg fungus, Leptosphaeria maculans, in vitro. However, there is no relationship between blackleg resistance (in stems or cotyledons) and the level of 2-propenyl glucosinolate in seed or shoot tissue of B. juncea. A representative isolate from each of two classes of Australian isolates of L. maculans was tested: those which cause stem cankers on both B. napus and B. juncea (e.g. isolate MI), and those which cause stem cankers on B. napus but not on B. juncea (e.g, isolate C13). Even in lines of B. juncea in which levels of 2-propenyl glucosinolate were as low as those in B. napus, isolate C13 was still unable to cause stem cankers. This suggests that either 2-propenyl glucosinolate does not play a major role in blackleg disease resistance, or that isolates of L. maculans able to attack B. juncea can detoxify or evade the toxic effects of hydrolysis products from this glucosinolate.

Self-recognition between pollen and stigma during pollination in Brassica oleracea is genetically controlled by the multiallelic self-incompatibility locus (S). We describe the S receptor kinase (SRK) gene, a previously uncharacterized gene that resides at the S locus. The nucleotide sequences of genomic DNA and of cDNAs corresponding to SRK predict a putative transmembrane receptor having serine/threonine-specific protein kinase activity. Its extracellular domain exhibits striking homology to the secreted product of the S-locus glycoprotein (SLG) gene and is connected via a single pass transmembrane domain to a protein kinase catalytic center. SRK alleles derived from different S-locus genotypes are highly polymorphic and have apparently evolved in unison with genetically linked alleles of SLG. SRK directs the synthesis of several alternative transcripts, which potentially encode different protein products, and these transcripts were detected exclusively in reproductive organs. The identification of SRK may provide new perspectives into the signal transduction mechanism underlying pollen recognition.

Flagellation of Salmonella typhimurium was found to require a functional pathway for ubiquinone biosynthesis as well as growth in the presence of appropriate carboxylic acids. Induction of flagellation by carboxylic acids was shown to induce incorporation of p-hydroxybenzoic acid into polyprenylphenol. Constitutive flagellation was found to correlate with constitutive incorporation of p-hydroxybenzoic acid into polyprenylphenol. A novel pathway for polyprenyl p-hydroxybenzoic acid decarboxylation to polyprenylphenol was implicated in flagellation of S. typhimurium.

Flagellation of Salmonella typhimurium was found to require a functional pathway for ubiquinone synthesis as well as growth in the presence of aliphatic or aromatic carboxylic acids. Selection for constitutive flagellation eliminated the requirement for growth in the presence of added carboxylic acids.

The carbohydrate component of Glycoprotein 2 (12% carbohydrate) from rye-grass (Lolium perenne) pollen has saccharide sequences that contribute to its antigenicity. Radioimmunoassay inhibition tests show that the antiserum to this glycoprotein cross-reacts with a number of other plant glycoproteins. In contrast, antiserum to another glycoprotein from rye-grass pollen, Glycoprotein 1 (5% carbohydrate), does not cross-react with any of the test glycoconjugates. Treatment of glycoproteins with sodium metaperiodate (0.02 M, 4 degree C, 6 h, in the dark) causes the loss of their ability to cross-react antigenically with Glycoprotein 2, and a loss of capacity to bind 125I-labelled concanavalin A. The cross-reactivity of this plant glycoprotein with other glycoconjugates imposes limitations on the interpretation of ultrastructural studies aimed at localizing a particular glycoprotein to a cellular site by using fluorescent or ferritin-labelled antisera. A radioimmunoassay inhibition technique for quantitative determination of the amounts of antigens in plants is also described.

Two glycoproteins have been purified from a buffer extract of rye-grass (Lolium perenne) pollen. Both migrated as single bands on sodium dodecyl sulphate/polyacrylamide gels. Glycoprotein 1 (0.8 mg/g of pollen) had a apparent mol.wt. of 33 000 and contained 95% protein and 5% carbohydrate. The monosaccharides glucose, galactose, mannose, arabinose and N-acetylglucosamine were present in the proportions 3:3:1:2:1. Glycoprotein 2 (0.4 mg/g of pollen) had an apparent mol. wt. of 68000 and contained 88% protein and 12% carbohydrate. The monosaccharides glucose, galactose, mannose, fucose, xylose, arabinose and N-acetylglucosamine were present in the proportions 4:7:13:5:8:6:6. This glycoprotein bound concanavalin A and Lotus tetragonolobus (asparagus pea) lectin. Radioallergosorbent (RAST) inhibition tests showed that Glycoprotein 1 is an effective allergen, whereas Glycoprotein 2 has less allergenic activity. A method for performing both lectin-binding assays and RAST inhibition tests using microtitre trays is described.

Germ tubes of both virulent and avirulent isolates ofLeptosphaeria maculanspenetrate leaves ofBrassica junceacultivar Stoke through stomatal pores. There is a spectrum of responses at different times after inoculation, ranging from necrosis of guard or mesophyll cells, to the presence of small non-sporulating colonies and large sporulating colonies. InBrassica napus, the same spectrum of events also occurred in compatible and incompatible interactions withL. maculans. However, in incompatible interactions inB. junceaonly one response occurred —rapid necrosis of guard cells associated with the arrest of hyphal growth at the stomatal pore. Injection of pycnidiospores of an avirulent isolate into the mesophyll layer ofB. juncealeaves, leads to the expression of resistance responses such as deformation of hyphae, accumulation of dark coloured material and of callose. This suggests thatB. junceacan express resistance responses in the mesophyll layer in addition to those of the stomata.

A retrospective study on 190 children with acute lymphoblastic leukaemia and marrow relapse on therapy demonstrated a universally poor prognosis with a high risk of extramedullary leukaemia. 49.1% of children achieved a second remission, the median duration of haematological remission being 97 days. The median duration of survival was 157 days, with no survivors beyond 2 years 3 months from relapse. Children with high white blood counts at diagnosis, those relapsing early and older children had a particularly poor prognosis. Children who achieved a first remission with difficulty and those receiving regular vincristine and prednisolone in their remission were less likely to achieve a second remission. Those who failed to go back into remission with the more commonly used drugs were not usually responsive to other drugs.

Two major glycoproteins have been localized in sectioned grains of ryegrass pollen by direct and indirect immunofluorescence methods using Fluorescein isothiocyanate (FITC)-labelled IgC fractions of antisera. These glycoproteins are the major allergen Group 1 allergen, and a principal antigen Antigen A. Four methods of fixation were employed: freeze-drying, methanol, 2.5% glutaraldehyde and 4% paraformaldehyde for 1 h at 4°C. The post-embedding staining technique of immunocytochemistry was used: anthers were embedded directly, or after dehydration, in JB-4 plastic resin and antibody reacted with sectioned pollen. The effects of these fixatives on the antibody combining sites of the antigens were quantified by a solid phase radioimmunoassay using [125I]protein A to measure antibody binding. Glutaraldehyde was the only fixative to significantly depress antibody binding of both Antigen A and Group 1 allergen to their homologous antisera. This radioimmunoassay was modified to reyeal that FITC conjugation to either antibody did not impair antigen binding. In mature pollen, these antigens were located in the cytoplasm and in the complex wall. In developing grains early in the maturation period, specific fluorescence was concentrated at the periphery of the cytoplasm.

The cellular sites of the glycoproteins Group 1 allergen (glycoprotein 1) and Antigen A (glycoprotein 2) in mature ryegrass pollen have been investigated by immunoelectron microscopy. Radioimmunoassays confirm previous findings of cross-reactivity between the purified glycoprotein antigens at the high immunoglobulin G (IgG) concentrations used for localization. Freeze-drying of anthers followed by anhydrous processing has been employed because of the water solubility and mobility of the glycoproteins. A double-embedding technique has been developed. This involves, first, embedding anthers in the water-soluble plastic resin JB-4, sectioning and incubating in ferritin-labelled antisera by the indirect method. The sections are then embedded in Spurr's resin for ultra-thin sectioning. Both glycoproteins are found in the following sites: (1) exine and intine wall layers; (2) pollen cytoplasm; (3) the orbicules and anther loculus; and (4) the anther cuticle. In the exine arcades and surface and in the anther loculus, the ferritin label is bound to pollenkitt. The finding that the glycoproteins are in similar sites is predictable in view of the cross-specificity of the antisera. The extent of antibody penetration of the plastic sections has been examined; labelling is confined to cut grains and absent from intact grains.

Mutants of Escherichia coli lacking ubiquinone or heme have been tested for motility and found to be essentially immotile. The loss of motility is identified with the loss of flagellum synthesis.

The specificities of the soluble and membrane aspartate-binding activities were compared with each other and with the specificity of aspartate chemotaxis and were found to be distinct. The soluble aspartate-binding protein was purified to homogeneity and had a molecular weight of 30,000. The dissociation constant was 10(-6) M for aspartate, and the protein bound glutamate, cysteic acid, and 2-amino-3-phosphonopropionate. Aspartate transport was inhibited by cysteic acid.

Cytochemical staining methods and immunofluorescence techniques have been used to follow the release of proteins, including the allergen Antigen E, from whole pollen of ragweeds (Ambrosia spp.) and Cosmos bipinnatus (Compositae). These proteins have been detected in 2 sites in the wall: in the inner cellulosic intine layer especially at the germinal apertures, and in the cavea and the cavities of the outer exine layer, the sexine. It was found that the release of exine-associated proteins, including the allergenic fraction, begins within 2 8 of contact with aqueous media, the loss being mainly through the micropores of the tectum at the bases of the sexine spines. The intine proteins were emitted more slowly, discharge beginning at the colpi after 1 min of moistening. So far as can be judged from the methods used, most of the mobile protein lost from the pollen of Cosmos on hydration comes from the exine and is therefore sporophvtic in origin, while in the ragweeds the bulk comes from the intine in the neighbour-hood of the germinal apertures, and so is gametophytic.