Phytophthora Root Rot: Assessing the potential threat to Australia's oldest national park

Summary   Royal National Park, Australia's oldest national park, is a significant reserve for conservation of the flora and fauna that are characteristic of the Hawkesbury Sandstone in New South Wales. Since at least 1974, Phytophthora Root Rot (caused by Phytophthora cinnamomi ) has been known to occur in the Park, but there is no knowledge of the extent of infestation or the potential impacts of the disease within the Park. This study investigated the distribution of the pathogen within Royal National Park at two scales: a systematic survey by vegetation type, and a targeted survey of populations of Waratah and Spear Grass-tree. These two species are known to be susceptible to Phytophthora Root Rot and are therefore potential indicators of the impact of the pathogen on vegetation in Royal National Park. Phytophthora cinnamomi was recovered from all vegetation types sampled but most commonly in heathland/open scrub vegetation. The pathogen was easily recovered from sites containing Spear Grass-tree, but was not isolated from any sites containing Waratah. Because of the widespread distribution of P. cinnamomi , we conclude that hygiene measures will be of little use to prevent the spread of the pathogen within Royal National Park. Monitoring of the occurrence and spread of disease symptoms in plants and applying phosphite to protect susceptible rare or threatened flora may be the most appropriate management options.     

Antifeedant and insecticidal properties of azadirachtin to the European Corn Borer, Ostrinia nubilalis

Azadirachtin, a triterpenoid substance from the neem tree, Azadirachta indica A. Juss., Meliaceae, was evaluated for its antifeedant and insecticidal properties to the European corn borer Ostrinia nubilalis Hübner. The 50% protective concentration for larval feeding on treated corn disks (PC₅₀) was 3.5 ppm for neonate larvae and 24 ppm for third instar larvae. Azadirachtin severely reduced larval growth at 1 and 10 ppm in a meridic diet and eventually caused 100% mortality at 10 ppm and 90% at 1 ppm. At 0.1 ppm azadirachtin showed no significant effect on larvae but altered the sex ratio in favor of male moths during adult emergence and arrested oviposition of female moths.Nutritional indices for naive and habituated third instar larvae suggested that reduction of growth was due to toxicity as shown by a lowered efficiency of conversion in the azadirachtin treated groups, rather than changes in digestibility. No clear evidence of tolerance was observed in the habituated group. The data suggest that at 10 ppm azadirachtin is an effective botanical insecticide for control of O. nubilalis.

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Résumé On a examiné les propriétés anti-appétantes et insecticides de l'azadirachtine, une substance triterpénoïde provenant d'Azadirachta indica envers la pyrale du maïs Ostrinia nubilalis. La concentration effective pour causer 50% d'inhibition de l'alimentation larvaire (PC₅₀) sur des disques foliaires a été de 3.5 ppm pour les larves de premier stade et de 24 g/8 pour celle de troisième stade. L'azadirachtine a sévèrement réduit la croissance larvaire à des concentrations de 1 et 10 ppm dans une diète méridique et, éventuellement, a causé 100% de mortalité à 10 ppm et 90% à 1 ppm. A 0.1 ppm l'azadirachtine n'a pas eu d'effet significatif sur les larves maïs a modifié l'indice de masculinité en faveur des mâles au moment de l'émergence et a empêché l'oviposition chez les femelles. Les indices nutritionnels pour les larves naïves de troisième âge et celles ayant subi une accoutumance suggèrent que la réduction de croissance est le résultat d'une intoxication comme le démontre une efficacité réduite de transformation chez les groupes traités à l'azadirachtine, plutôt que des changements de digestibilité. Il n'y a pas d'évidence claire de tolérance dans le groupe ayant subi l'accoutumance. Les résultats suggèrent qu'à 10 ppm l'azadirachtine est un insecticide botanique efficace envers O. nubilalis.

     

Family physician registration at locally produced short courses.

Physicians who do not take advantage of short courses that are offered within their community may miss the opportunity to learn about new services and consultants as well as to validate new information or practices with trusted colleagues. The registration patterns at short courses of all 505 family physicians in Calgary were assessed to determine whether the sociodemographic characteristics of attendees differed from those of nonattendees and whether the sociodemographic data were predictive of attendance or nonattendance. Four variables were predictive of attendance or nonattendance: certification status with the College of Family Physicians of Canada (CFPC), year of graduation, sex and hospital affiliation. Physicians who had attended four or more courses were more likely to be graduates of Canadian or US schools, to have graduated after 1969, and to be women, certificants of the CFPC and affiliated with a hospital. Universities, hospitals and professional associations planning continuing education must determine if the needs of nonattending physicians are being met through other means or if new strategies are required to ensure that these physicians have opportunities for upgrading their knowledge. Practitioners who frequently attend short courses can help planners ensure that programming is relevant.     

Structure and evolution of the 4.5-5S ribosomal RNA intergenic region from Glycine max (soya bean).

The nucleotide sequence for the 4.5-5S ribosomal DNA region from the chloroplastids of soya beans was determined as the basis of further comparative studies on the structure and evolution of this intergenic region. Comparisons with other plant sequences as well as equivalent sequences in eubacteria suggest that the longer internal transcribed spacer regions of plants have evolved, at least in part, by DNA sequence duplications and that the presence of the 4.5S rRNA in chloroplast may result from the accidental acquisition of a RNA maturation site during the evolution of longer internal transcribed spacer regions. Estimates of the secondary structures also indicate only a very limited retention of structural features and suggest that the primary role of the intergenic sequences may be to bring processed sites into close proximity.     

Plasmid mediated mutagenesis of a cellular gene in transfected eukaryotic cells.

NIH3T3 cells are widely used in transformation assays and readily take up transfected DNA. A system has been devised using NIH3T3 cells to measure the mutagenic effect of transfected DNA on recipient cell genes. NIH3T3 cells can be mutated to 6-thioguanine resistance at a frequency which suggests that at least a portion of the cells have only one functional copy of the HGPRT gene. They have a low spontaneous background mutation frequency (approximately 1 X 10(-7)). Transfection of three different plasmids into NIH3T3 cells induced 6-thioguanine resistant mutants at frequencies ranging from 3 to 11 fold above background. The mutant phenotype is stable and reversion frequencies of several mutants are less than or equal to 1 X 10(-7). Southern blot analysis of the HGPRT gene in several mutants showed that 4 of 26 mutants (15.4%) had detectable alterations in the structure of the HGPRT gene. Interestingly 3 of the 4 mutants showing rearrangements were obtained by transfection of the HSV-2 morphological transforming region.     

Extended N-terminal sequencing of proteins of archaebacterial ribosomes blotted from two-dimensional gels onto glass fiber and poly(vinylidene difluoride) membrane

Previously uncharacterized proteins from intact ribosomes and ribosomal subunits of the extreme halophile Halobacterium marismortui (Haloarcula marismortui) were isolated and separated by high-resolution two-dimensional electrophoresis (2DE). N-Terminal amino acid sequences of 14 of these acidic large-subunit proteins were obtained by direct blotting of the separated proteins from two-dimensional electrophoresis gels to sequencer-stable supports followed by excision of the protein spots and sequencing. Furthermore, long internal sequences were obtained by in situ enzymatic cleavage of halobacterial proteins in gel pieces obtained from two-dimensional gels followed by electrophoretic separation of the fragments, blotting, and sequencing. Precautions are outlined for avoidance of N-terminal blockage of proteins, and the preparation and selection of suitable supports for obtaining extended N-terminal sequences are described. The results suggest that when prior fractionation is carried out to enrich for cell organelles, subcellular components of cells, or cell membranes, it is routinely possible to obtain numerous N-terminal sequences from one or a few 2DE gels of such fractions. Our results also indicate that, with appropriate precautions, proteins are routinely obtainable from 2DE gels in a form suitable for both N-terminal and internal sequence determination and show no detectable evidence for N-terminal blockage or destruction or modification of labile amino acid residues.     

Fourier-transform infrared and Raman spectroscopic evidence for the incorporation of cinnamaldehydes into the lignin of transgenic tobacco (Nicotiana tabacum L.) plants with reduced expression of cinnamyl alcohol dehydrogenase

Xylem from stems of genetically manipulated tobacco plants which had had cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) activity down-regulated to a greater or lesser degree (clones 37 and 49, respectively) by the insertion of antisense CAD cDNA had similar, or slightly higher, lignin contents than xylem from wild-type plants. Fourier-transform infrared (FT-IR) microspectroscopy indicated that down-regulation of CAD had resulted in the incorporation of moieties with conjugated carbonyl groups into lignin and that the overall extent of cross-linking, particularly of guaiacyl (4-hydroxy-3-methoxyphenyl) rings, in the lignin had altered. The FT-Raman spectra of manipulated xylem exhibited maxima consistent with the presence of elevated levels of aldehydic groups conjugated to a carbon-carbon double bond and a guaiacyl ring. These maxima were particularly intense in the spectra of xylem from clone 37, the xylem of which exhibits a uniform red coloration, and their absolute frequencies matched those of coniferaldehyde. Furthermore, xylem from clone 37 was found to have a higher content of carbonyl groups than that of clone 49 or the wild-type (clone 37: clone 49: wild-type; 2.4:1.6:1.0) as measured by a degradative chemical method. This is the first report of the combined use of FT-IR and FT-Raman spectroscopies to study lignin structure in situ. These analyses provide strong evidence for the incorporation of cinnamaldehyde groups into the lignin of transgenic plants with down-regulated CAD expression. In addition, these non-destructive analyses also suggest that the plants transformed with antisense CAD, in particular clone 37, may contain lignin that is less condensed (cross-linked) than that of the wild-type. Received: 27 May 1996 / Accepted: 30 July 1996     

Vacuolar reticulum in oomycete hyphal tips: An additional component of the Ca2+Regulatory system?

Cultures of Achlya sp., Phytophthora cinnamomi, Saprolegnia diclina, S. ferax, and S. parasitica, treated with 6-carboxyfluorescein diacetate solution, accumulate 6-carboxyfluorescein in a reticulate system of fine tubules. The network shows longitudinal polarity within the hyphae, tubules being finest toward the hyphal tips. In more mature subapical regions the network is connected with large vacuoles that also accumulate 6-carboxyfluorescein. A morphologically similar system has also been identified in freeze-substituted hyphae of S. ferax. The network is considered to be vacuolar, but differs from the tubular vacuole system of true fungi in that tubules are less motile, more frequently branched, and do not alternate with clusters of spherical vacuoles. The appearance of the network resembles patterns of calcium-sensitive dye staining and it is suggested that the vacuolar reticulum in the tip region of oomycete hyphae may act as a Ca2+ sink. The tubular reticulum in oomycetes is very fragile and can be shown with 6-carboxyfluorescein in only those hyphal tips with a motility and organelle distribution characteristic of growing hyphae with normal morphology. Diverse abnormal hyphae show a range of other fluorochrome localizations. These include large irregular compartments filled with fluorochrome, and fluorescent cytoplasm with organelles and vacuoles standing out in negative contrast. These localizations in abnormal hyphae are correlated with other structural changes indicative of damage. Special care is required in experiments with oomycetes to avoid such artefacts of localization. Copyright 1997 Academic Press. Copyright 1997 Academic Press