Arabidopsis thaliana expressing PbBSMT, a gene encoding a SABATH-type methyltransferase from the plant pathogenic protist Plasmodiophora brassicae, show leaf chlorosis and altered host susceptibility.

The plant pathogenic protist Plasmodiophora brassicae causes clubroot disease of Brassicaceae. This biotrophic organism can down-regulate plant defence responses. The previously characterised P. brassicae PbBSMT methyltransferase has substrate specificity for salicylic, benzoic and anthranilic acids. We therefore propose a role for the methylation of SA in attenuating plant defence response in infected roots as a novel strategy for intracellular parasitism. We overexpressed PbBSMT under the control of an inducible promoter in Arabidopsis thaliana and performed physiological, molecular and phytopathological analyses with the transgenic plants under control and induced conditions in comparison to the wild type. Upon induction, transcription of PbBSMT was associated with: (1) strong leaf phenotypes from anthocyanin accumulation and chlorosis followed by browning; (2) increased plant susceptibility after infection with P. brassicae that was manifested as more yellow leaves and reduced growth of upper plant parts; and (3) induced transgenic plants were not able to support large galls and had a brownish appearance of some clubs. Microarray data indicated that chlorophyll loss was accompanied by reduced transcription of genes involved in photosynthesis, while genes encoding glucose metabolism, mitochondrial functions and cell wall synthesis were up-regulated. Our results indicate a role for PbBSMT in attenuation of host defence responses in the roots by metabolising a plant defence signal.

The potential of L. scoparium, K. robusta and P. radiata to mitigate N-losses in silvopastural systems.

Silvopastoral systems aim to enhance economic, cultural and social principles by sustainably combining forest management with agriculture. In these typically high-nitrogen (N) environments, plant species selection can profoundly influence N fluxes. For grazed pastures, plants may be exposed to urine patches that have received the equivalent of up to 1000 kg N ha-1. We aimed to determine the growth and N fluxes in three potential trees that may be used in silvopastoral systems: L. scoparium, K. robusta and P. radiata. Plants were grown in a greenhouse lysimeter experiment, with controlled irrigation and temperature and exposed to N at rates of 200 kg ha-1 equiv. for 15 weeks, followed by the addition of 800 kg ha-1 N equiv, to simulate a urine patch. Urea produced a positive growth response of all plant species. Treatments containing L. scoparium and K. robusta leached lower amounts of nitrate (NO3-) (2 kg ha-1 NO3-) compared to P. radiata (53 kg ha-1). Measurements of N2O over 20 days after the application of 800 kg N ha-1 indicated an inhibitory effect of L. scoparium and K. robusta on denitrification, hence loss of N via N2O. Both L. scoparium and K. robusta demonstrated that they have potential to reduce N-losses in silvopastural systems, while producing valuable biomass.

Response of Leptospermum scoparium, Kunzea robusta and Pinus radiata to contrasting biowastes.

The myrtaceae family has a cosmopolitan distribution and includes the Australasian native species Leptospermum scoparium (manuka) and Kunzea robusta (kanuka), which are of economic interest for the production of high-value honey and essential oils. Potentially, these species could be established on low-fertility or degraded soils that have been amended with biowastes, including biosolids and sawdust. We aimed to determine the effect of these biowastes on nitrate leaching and the growth and chemical composition of these plant species compared to Pinus radiata (pine), a common forestry species. The addition of biosolids (1250kgNha-1 equiv.) increased the total dry biomass of manuka, kanuka, and pine by 117, 90, and 86% respectively. Mixing sawdust with biosolids stimulated growth of manuka (52%), kanuka (121%) but not pine. Biosolids increased plant uptake of N, P, and trace elements, but not to levels of concern. Nitrate leaching from all treatments was negligible (<2kgha-1).

Production of Biomass Crops Using Biowastes on Low-Fertility Soil: 1. Influence of Biowastes on Plant and Soil Quality.

Land application of biosolids to low-fertility soil can improve soil quality by increasing concentrations of macronutrients and trace elements. Mixing biosolids with sawdust could reduce the risks of contaminant accumulation posed by rebuilding soils using biosolids alone. We aimed to determine the effects of biosolids and biosolids-sawdust on the plant quality and chemical composition of sorghum, rapeseed, and ryegrass. Plants were grown in a greenhouse over a 5-mo period in a low-fertility soil amended with biosolids (1250 kg N ha), biosolids-sawdust (0.5:1), or urea (200 kg N ha). Biosolids application increased the biomass of sorghum, rapeseed, and ryegrass up to 14.0, 11.9, and 4.1 t ha eq, respectively. Mixing sawdust with biosolids resulted in a growth response similar to biosolids treatments in rapeseed but nullified the effect of biosolids in sorghum. Urea fertilization provided insufficient nutrients to promote rapeseed growth and seed production, whereas seed yields after biosolids application were 2.5 t ha. Biosolids and biosolids-sawdust application enhanced plant quality by increasing element concentrations, especially Zn, and potentially toxic elements (Cd, Cr, Ni) did not exceed food safety standards. An application of 50 t ha of biosolids, equivalent to 1250 kg N ha, did not exceed current soil limits of Cu, Zn, and Cd and hence was effective in rebuilding soil without accumulating contaminants. The effect of mixing sawdust with biosolids varies with plant species but can further enhance plant nutrient quality in biomass and seeds, especially P, Cu, Zn, Mn, Fe, S, and Na.

Production of Biomass Crops Using Biowastes on Low-Fertility Soil: 2. Effect of Biowastes on Nitrogen Transformation Processes.

Increasing production of biowastes, particularly biosolids (sewage sludge), requires sustainable management strategies for their disposal. Biosolids can contain high concentrations of nutrients; hence, land application can have positive effects on plant growth and soil fertility, especially when applied to degraded soils. However, high rates of biosolids application may result in excessive nitrogen (N) leaching, which can be mitigated by blending biosolids with other biowastes, such as sawdust. We aimed to determine the effects of biosolids and sawdust on growth and N uptake by sorghum, rapeseed, and ryegrass as well as N losses via leaching. Plants were grown in a greenhouse over a 5-mo period in a low-fertility soil amended with biosolids (1250 kg N ha), biosolids-sawdust (0.5:1), or urea (200 kg N ha). Urea application increased biomass production of sorghum and ryegrass but proved insufficient for rapeseed on low-fertility soil. Biosolids application increased plant N concentrations in ryegrass and rapeseed and increased N uptake into the seeds of sorghum, increasing seed quality. Biosolids application did result in lower N leaching compared with urea, irrespective of plant species, and N leaching was unaffected by mixing the biosolids with sawdust. There was an indication of biological nitrification inhibition in the rhizosphere of sorghum. Rapeseed had similar growth and N uptake into biomass in biosolids and biosolids-sawdust treatments and hence was the most promising species with regard to recycling fresh sawdust in combination with high rates of biosolids on low-fertility soil.

Native aphids of New Zealand--diversity and host associations.

At least 15 species of aphids are now recognised as New Zealand natives and most of these are very likely to be endemic. Most native aphids belong in the subfamily Aphidinae (Aphidini), with a possible single species in Aphidinae-Macrosiphini, at least two in Neophyllaphidinae and one in Taiwanaphidinae. With one exception, native aphids are restricted to a single host plant genus, and these hosts are from 13 genera and 12 plant families in the Pinales and Angiospermae-Eudicotyledonae, suggesting that the aphids are a remnant fauna. No known native aphids have host plants from the Pteridophyta or Angiospermae-Monocotyledonae, with the possible exception of two possibly native species extracted from native tussock grassland turfs. Most host plant genera have some degree of Gondwanan distribution, but only two indigenous species are found on large forest trees and only one host is deciduous. Native aphids have been recorded from sea level to the subalpine zone, reflecting their host plant distributions. Sexual reproduction, followed by several parthenogenetic generations on the same host plant, appears to be the norm for most species. Eggs appear to be used for surviving winter conditions in some species and summer conditions in others. Native aphid distribution and abundance varies with five species considered to be scarce, one species localised, two species sparse and three relatively common based on current knowledge.

A phylogenetic analysis of the SSU rRNA from members of the Plasmodiophorida and Phagomyxida.

The Plasmodiophorida and Phagomyxida are orders of zoosporic eukaryotes of uncertain phylogenetic affinities. We have obtained ribosomal DNA sequence information from several species in these groups. Phylogenetic analyses of these sequences have confirmed that they form a monophyletic grouping that clusters most closely with a rhizopod assemblage consisting of sarcomonads and chlorarachneans. The Phagomyxida sequences appear distant enough from those of the Plasmodiophorida to justify a separate order. The relationships between the Plasmodiophorida remain unclear, with species from the same genus showing significant degrees of divergence.

A prospective, longitudinal study on patients with trigeminal neuralgia who underwent radiofrequency thermocoagulation of the Gasserian ganglion.

Outcome after radiofrequency thermocoagulation in patients with trigeminal neuralgia was assessed in a prospective, longitudinal study. Forty-eight consecutive patients with chronic facial pain presenting for surgery to a neurosurgeon were studied. Patients were assessed preoperatively by an independent clinician both clinically, and with the use of two questionnaires: the McGill Pain Questionnaire (MPQ) and the Hospital Anxiety and Depression (HAD) scale. From these assessments, two groups of patients were identified: 31 with pure trigeminal neuralgia (TN group) and 17 with trigeminal neuralgia together with atypical facial pain and mixed trigeminal neuralgia (MTN group). All underwent radiofrequency thermocoagulation at the level of the Gasserian ganglion. Patients were reviewed by the same clinician 3 months later and then followed up by a self-administered questionnaire at 6 months, 1 year, 2 years and 3 years. The mean follow-up time was 30+/-12 months. The mean time to recurrence of pain was 40 months for the TN group and 36 months for the MTN group. Depression and anxiety dropped more significantly post-operatively in the TN group than the MTN group. TN group were more satisfied with their outcome, complained of fewer complications and were more willing to have repeat surgery if necessary than patients in MTN group. The number and severity of complications varied at different time points. Careful selection of patients for surgery using objective assessments will decrease morbidity and improve satisfaction. Physical morbidity and recurrence rates are insufficient to gauge outcomes. Psychological, sociological and patients' views must be included in evaluations.

Specific Polymerase Chain Reaction Primers for the Detection of Plasmodiophora brassicae in Soil and Water.

ABSTRACT The development of specific oligonucleotide primers for Plasmodiophora brassicae has led to a nested polymerase chain reaction (PCR) detection method for P. brassicae in soil and water. Initially, the PCR was used to amplify a section of the rDNA repeat. The PCR products were sequenced and the data used to design primers that were directed at the ribosomal RNA genes and internal transcribed spacer regions. Specificity was tested against more than 40 common soil organisms, host plants, and spore suspension contaminants, as well as P. brassicae isolates from around Australia and the world. Sensitivity was determined to be 0.1 fentograms (fg; 10(-15) g) for pure template and as low as 1,000 spores per g of potting mix. In soil, P. brassicae was detected in all soils where the inoculum was sufficient to result in clubroot symptoms. Also outlined is a simple method of DNA extraction from soil.

Culture of 91/2-day rat embryos in human serum supplemented and unsupplemented with rat serum.

We attempted to grow explanted 91/2-day rat embryos in vitro in human serum but were unable to demonstrate normal growth as has been previously described. The embryos were consistently smaller, anaemic, and frequently deformed. These abnormalities were overcome by supplementing the human serum with 10% rat serum. The latter possibly provides yet unidentified essential growth factors required by rat embryos that are not present in human serum.