Novel role of the nitrite transporter NirC in Salmonella pathogenesis: SPI2-dependent suppression of inducible nitric oxide synthase in activated macrophages

Activation of macrophages by interferon gamma (IFN- ) and the subsequent production of nitric oxide (NO) are critical for the host defence against Salmonella enterica serovar Typhimurium infection. We report here the inhibition of IFN- -induced NO production in RAW264.7 macrophages infected with wild-type Salmonella. This phenomenon was shown to be dependent on the nirC gene, which encodes a potential nitrite transporter. We observed a higher NO output from IFN- -treated macrophages infected with a nirC mutant of Salmonella. The nirC mutant also showed significantly decreased intracellular proliferation in a NO-dependent manner in activated RAW264.7 macrophages and in liver, spleen and secondary lymph nodes of mice, which was restored by complementing the gene in trans. Under acidified nitrite stress, a twofold more pronounced NO-mediated repression of SPI2 was observed in the nirC knockout strain compared to the wild-type. This enhanced SPI2 repression in the nirC knockout led to a higher level of STAT-1 phosphorylation and inducible nitric oxide synthase (iNOS) expression than seen with the wild-type strain. In iNOS knockout mice, the organ load of the nirC knockout strain was similar to that of the wild-type strain, indicating that the mutant is exclusively sensitive to the host nitrosative stress. Taken together, these results reveal that intracellular Salmonella evade killing in activated macrophages by downregulating IFN- -induced NO production, and they highlight the critical role of nirC as a virulence gene.

Patterns of seed bank and size asymmetry of plant growth across varying sites in the invasive Lantana camara L. (Verbenaceae)

Lantana camara L. (Verbenaceae) is a weed of great significance in Australia and worldwide, but little is known about connections among components of its life history. We document over a 3-year period, the links between L. camara seed-bank dynamics and its above-ground growth, including size asymmetry in four land-use types (a farm, a hoop pine plantation and two open eucalypt forests) invaded by the weed near Brisbane, Queensland Australia. Seed-bank populations varied appreciably across sites and in response to rainfall and control measures, and they were higher (~1,000 seeds/m2) when annual rainfall was 15-30 % below the long-term yearly average. Fire reduced seed-bank populations but not the proportion germinating (6-8 %). Nearly a quarter of fresh seeds remain germinable after 3 years of soil burial. For small seedlings (<10 cm high), the expected trade-offs in two life-history traits-survival and growth-did not apply; rather the observed positive association between these two traits, coupled with a persistent seed-bank population could contribute to the invasiveness of the plant. Relationships between absolute growth rate and initial plant size (crown volume) were positively linear, suggesting that most populations are still at varying stages of the exponential phase of the sigmoid growth; this trend also suggests that at most sites and despite increasing stand density and limiting environmental resources of light and soil moisture, lantana growth is inversely size asymmetric. From the observed changes in measures of plant size inequality, asymmetric competition appeared limited in all the infestations surveyed. © 2013 Crown Copyright as represented by: Department of Agriculture, Fisheries and Forestry, Australia.

Comparative growth and biomass allocation of two varieties of cat's claw creeper, Dolichandra unguis-cati (Bignoniaceae) in Australia

Introduced as an ornamental vine, cat's claw creeper Dolichandra unguis-cati (syn. Macfadyena unguis-cati) has invaded coastal and subcoastal areas of subtropical eastern Australia. Two varieties have been indentified, one of which ('short-pod') is found throughout south-eastern Australia, while the other ('long-pod') appears to be restricted to several sites in south-eastern Queensland. We compared the growth and biomass allocation patterns of the two varieties in the field over a 22-month period to determine if a higher growth rate and/or more efficient allocation of biomass may contribute to this disparity in distribution. The long-pod variety produced greater aboveground and total biomass than the short-pod variety in both riparian and non-riparian zones. Belowground the two varieties produced a similar number of tubers and overall biomass, though the long-pod variety allocated a smaller portion of its carbon belowground. High growth rates and greater biomass allocation aboveground are characteristic of invasive species, allowing them to outcompete and crowd out existing vegetation. There was no significant site by variety interaction, an indication of consistency in variety performance across riparian and non-riparian sites. Results from our study suggest that differences in growth and biomass allocations are unlikely to have contributed to the disparity in distribution of the two varieties. Despite currently occupying a relatively small range, the long-pod variety may be a more adept invader than the short-pod variety, and could become more prevalent in the future. © 2012 CSIRO.

Developmental and growth controls of tillering and water-soluble carbohydrate accumulation in contrasting wheat (Triticum aestivum L.) genotypes: can we dissect them?

In wheat, tillering and water-soluble carbohydrates (WSCs) in the stem are potential traits for adaptation to different environments and are of interest as targets for selective breeding. This study investigated the observation that a high stem WSC concentration (WSCc) is often related to low tillering. The proposition tested was that stem WSC accumulation is plant density dependent and could be an emergent property of tillering, whether driven by genotype or by environment. A small subset of recombinant inbred lines (RILs) contrasting for tillering was grown at different plant densities or on different sowing dates in multiple field experiments. Both tillering and WSCc were highly influenced by the environment, with a smaller, distinct genotypic component; the genotypeenvironment range covered 350750 stems m(2) and 25210mg g(1) WSCc. Stem WSCc was inversely related to stem number m(2), but genotypic rankings for stem WSCc persisted when RILs were compared at similar stem density. Low tilleringhigh WSCc RILs had similar leaf area index, larger individual leaves, and stems with larger internode cross-section and wall area when compared with high tilleringlow WSCc RILs. The maximum number of stems per plant was positively associated with growth and relative growth rate per plant, tillering rate and duration, and also, in some treatments, with leaf appearance rate and final leaf number. A common threshold of the red:far red ratio (0.390.44; standard error of the difference0.055) coincided with the maximum stem number per plant across genotypes and plant densities, and could be effectively used in crop simulation modelling as a ocut-off' rule for tillering. The relationship between tillering, WSCc, and their component traits, as well as the possible implications for crop simulation and breeding, is discussed.

Effects of seed origin latitude on the timing of height growth cessation and field performance of silver birch

The aim of this thesis was to increase our knowledge about the effects of seed origin on the timing of height growth cessation and field performance of silver birch from different latitudes, with special attention paid to the browsing damage by moose in young birch plantations. The effect of seed origin latitude and sowing time on timing of height growth cessation of first-year seedlings was studied in a greenhouse experiment with seven seed origins (lat. 58º - 67ºN). Variation in critical night length (CNL) for 50 % bud set within two latitudinally distant stands (60º and 67ºN) was studied in three phytotron experiments. Browsing by moose on 5-11 -year-old silver birch saplings from latitudinally different seed origins (53º - 67ºN) was studied in a field experiment in southern Finland. Yield and stem quality of 22-year-old silver birch trees of Baltic, Finnish and Russian origin (54º - 63ºN) and the effect of latitudinal seed transfers were studied in two provenance trials at Tuusula, southern and Viitasaari, central Finland. The timing of height growth cessation depended systematically on latitude of seed origin and sowing date. The more northern the seed origin, the earlier the growth cessation and the shorter the growth period. Later sowing dates delayed growth cessation but also shortened the growth period. The mean CNL of the southern ecotype was longer, 6.3 ± 0.2 h (95 % confidence interval), than that of the northern ecotype, 3.1 ± 0.3 h. Within-ecotype variance of the CNL was higher in the northern ecotype (0.484 h2) than in the southern ecotype (0.150 h2). Browsing by moose decreased with increasing latitude of seed origin and sapling height. Origins transferred from more southern latitudes were more heavily browsed than the more northern native ones. Southern Finnish seed origins produced the highest volume per unit area in central Finland (lat. 63º11'N). Estonian and north Latvian stand seed origins, and the southern Finnish plus tree origins, were the most productive ones in southern Finland (lat. 60º21'N). Latitudinal seed transfer distance had a significant effect on survival, stem volume/ha and proportion of trees with a stem defect. The relationship of both survival and stem volume/ha to the latitudinal seed transfer distance was curvilinear. Volume was increased by transferring seed from ca. 2 degrees of latitude from the south. A longer transfer from the south, and transfer from the north, decreased the yield. The proportion of trees with a stem defect increased linearly in relation to the latitudinal seed transfer distance from the south.

Regulation of strawberry growth and development

Strawberries (Fragaria sp.) are adapted to diverse environmental conditions from the tropics to about 70ºN, so different responses to environmental conditions can be found. Most genotypes of garden strawberry (F. x ananassa Duch.) and woodland strawberry (F. vesca L.) are short-day (SD) plants that are induced to flowering by photoperiods under a critical limit, but also various photoperiod x temperature interactions can be found. In addition, continuously flowering everbearing (EB) genotypes are found. In addition to flowering, axillary bud differentiation in strawberry is regulated by photoperiod. In SD conditions, axillary buds differentiate to rosette-like structures called "branch crowns", whereas in long-day conditions (LD) they form runners, branches with 2 long internodes followed by a daughter plant (leaf rosette). The number of crown branches determines the yield of the plant, since inflorescences are formed from the apical meristems of the crown. Although axillary bud differentiation is an important developmental process in strawberries, its environmental and hormonal regulation has not been characterized in detail. Moreover, the genetic mechanisms underlying axillary bud differentiation and regulation of flowering time in these species are almost completely unresolved. These topics have been studied in this thesis in order to enhance strawberry research, cultivation and breeding. The results showed that 8-12 SD cycles suppressed runner initiation from the axillary buds of the garden strawberry cv. Korona with the concomitant induction of crown branching, and 3 weeks of SD was sufficient for the induction of flowering in the main crown. Furthermore, a second SD treatment given a few weeks after the first SD period can be used to induce flowering in the primary branch crowns and to induce the formation of secondary branches. Thus, artificial SD treatments effectively stimulate crown branching, providing one means for the increase of cropping (yield) potential in strawberry. It was also shown by growth regulation applications, quantitave hormone analysis and gene expression analysis that gibberellin (GA) is one of the key signals involved in the photoperiod control of shoot differentiation. The results indicate that photoperiod controls GA activity specifically in axillary buds, thereby determining bud fate. It was further shown that chemical control of GA biosynthesis by prohexadione-calcium can be utilized to prevent excessive runner formation and induce crown branching in strawberry fields. Moreover, ProCa increased berry yield up to 50%, showing that it is an easier and more applicable alternative to artificial SD treatments for controlling strawberry crown development and yield. Finally, flowering gene pathways in Fragaria were explored by searching for homologs of 118 Arabidopsis thaliana flowering-time genes. In total, 66 gene homologs were identified, and they distributed to all known flowering pathways, suggesting the presence of these pathways also in strawberry. Expression analysis of selected genes revealed that the mRNA of putative floral identity gene APETALA1 accumulated in the shoot apex of the EB genotype after the induction of flowering, whereas it was absent in vegetative SD genotype, indicating the usefulness of this gene product as the marker of floral initiation. The present data enables the further exploration of strawberry flowering pathways with genetic transformation, gene mapping and transcriptomics methods.

Species-site matching in mixed species plantations of native trees in tropical Australia

Mixed species plantations using native trees are increasingly being considered for sustainable timber production. Successful application of mixed species forestry systems requires knowledge of the potential spatial interaction between species in order to minimise the chance of dominance and suppression and to maximise wood production. Here, we examined species performances across 52 experimental plots of tree mixtures established on cleared rainforest land to analyse relationships between the growth of component species and climate and soil conditions. We derived site index (SI) equations for ten priority species to evaluate performance and site preferences. Variation in SI of focus species demonstrated that there are strong species-specific responses to climate and soil variables. The best predictor of tree growth for rainforest species Elaeocarpus grandis and Flindersia brayleyana was soil type, as trees grew significantly better on well-draining than on poorly drained soil profiles. Both E. grandis and Eucalyptus pellita showed strong growth response to variation in mean rain days per month. Our study generates understanding of the relative performance of species in mixed species plantations in the Wet Tropics of Australia and improves our ability to predict species growth compatibilities at potential planting sites within the region. Given appropriate species selections and plantation design, mixed plantations of high-value native timber species are capable of sustaining relatively high productivity at a range of sites up to age 10 years, and may offer a feasible approach for large-scale reforestation.

Association genetics in Corymbia citriodora subsp variegata identifies single nucleotide polymorphisms affecting wood growth and cellulosic pulp yield

Wood is an important biological resource which contributes to nutrient and hydrology cycles through ecosystems, and provides structural support at the plant level. Thousands of genes are involved in wood development, yet their effects on phenotype are not well understood. We have exploited the low genomic linkage disequilibrium (LD) and abundant phenotypic variation of forest trees to explore allelic diversity underlying wood traits in an association study. Candidate gene allelic diversity was modelled against quantitative variation to identify SNPs influencing wood properties, growth and disease resistance across three populations of Corymbia citriodora subsp. variegata, a forest tree of eastern Australia. Nine single nucleotide polymorphism (SNP) associations from six genes were identified in a discovery population (833 individuals). Associations were subsequently tested in two smaller populations (130160 individuals), validating our findings in three cases for actin 7 (ACT7) and COP1 interacting protein 7 (CIP7). The results imply a functional role for these genes in mediating wood chemical composition and growth, respectively. A flip in the effect of ACT7 on pulp yield between populations suggests gene by environment interactions are at play. Existing evidence of gene function lends strength to the observed associations, and in the case of CIP7 supports a role in cortical photosynthesis.

Cord blood 25-hydroxyvitamin D and fetal growth in the China-Anhui birth cohort study

We determined the association of cord blood 25-hydroxyvitamin D [25(OH)D] with birth weight and the risk of small for gestational age (SGA). As part of the China-Anhui Birth Cohort (C-ABC) study, we measured cord blood levels of 25(OH)D in 1491 neonates in Hefei, China. The data on maternal sociodemographic characteristics, health status, lifestyle, birth outcomes were prospectively collected. Multiple regression models were used to estimate the association of 25(OH)D levels with birth weight and the risk of SGA. Compared with neonates in the lowest decile of cord blood 25(OH)D levels, neonates in four deciles (the fourth, fifth, sixth and seventh deciles) had significantly increased birth weight and decreased risk of SGA. Multiple linear regression models showed that per 10 nmol/L increase in cord blood 25(OH)D, birth weight increased by 61.0 g (95% CI: 31.9, 89.9) at concentrations less than 40 nmol/L, and then decreased by 68.5 g (95% CI: −110.5, −26.6) at concentrations from 40 to 70 nmol/L. This study provides the first epidemiological evidence that there was an inverted U shaped relationship between neonatal vitamin D status and fetal growth, and the risk of SGA reduced at moderate concentration.

Pollination methods, stigma receptivity and pollen tube growth in Eucalyptus argophloia

Eucalyptus argophloia Blakely (Western white gum) has shown potential as a commercial forestry timber species in marginal environments of north-eastern Australia. We measured early pollination success in Eucalyptus argophloia to compare pollination methods, determine the timing of stigma receptivity and compare fresh and stored pollen. Early pollination success was measured by counting pollen tubes in the style of E. argophloia 12 days after pollination. We compared the early pollination success of 1) Artificially Induced Protogyny (AIP), one-stop and three-stop methods of pollination; 2) flowers pollinated at 2 day intervals between 2 days before and 6 days after anthesis and 3) fresh pollen and pollen that had been stored for 9 months. Our results show significantly more pollen tubes from unpollinated AIP and AIP treatments than either the one-stop pollination or three-stop pollination treatments. This indicates that self-pollination occurs in the unpollinated AIP treatment. There was very little pollen tube growth in the one-stop method indicating that the three-stop method is the most suitable for this species. Stigma receptivity in E. argophloia commenced six days after anthesis and no pollen tube growth was observed prior to this. Fresh pollen resulted in pollen tube growth in the style whereas the stored pollen resulted in a total absence of pollen tube growth. We recommend that breeding programs incorporating E. argophloia as a female parent use the three-stop pollination method, and controlled pollination be carried out at least six days after anthesis using fresh pollen.

Allied health growth: What we do not measure we cannot manage

Background Data describing the Australian allied health workforce is inadequate and so insufficient for workforce planning. National health policy reform requires that health-care models take into account future workforce requirements, the distribution and work contexts of existing practitioners, training needs, workforce roles and scope of practice. Good information on this workforce is essential for managing services as demands increase, accountability of practitioners, measurement of outcomes and benchmarking against other jurisdictions. A comprehensive data set is essential to underpin policy and planning to meet future health workforce needs. Discussion Some data on allied health professions is managed by the Australian Health Practitioner Regulation Agency; however, there is limited information regarding several core allied health professions. A global registration and accreditation scheme recognizing all allied health professions might provide safeguards and credibility for professionals and their clients. Summary Arguments are presented about inconsistencies and voids in the available information about allied health services. Remedying these information deficits is essential to underpin policy and planning for future health workforce needs. We make the case for a comprehensive national data set based on a broad and inclusive sampling process across the allied health population.

Focus on opportunities as a mediator of the relationship between business owners' age and venture growth

Combining upper echelons and lifespan theories, we investigated the mediating effect of focus on opportunities on the negative relationship between business owners' age and venture growth. We also expected that mental health moderates the negative relationship between business owners' age and focus on opportunities. Path analytic findings based on data from 84 business owners (mean age = 44, range 24-74) supported these hypotheses. Findings suggest that focus on opportunities is a psychological mechanism that links business owners' age with venture growth. Our findings also indicate that mental health helps maintain a high level of focus on opportunities with increasing age.

Monitoring of Fungal Growth and Degradation of Wood

Valko- ja ruskolahosienet tunnetaan luonnossa tehokkaimpina puun ja karikkeen lignoselluloosan lahottajina. Valkolahosienet pystyvät hajottamaan kaikkia puun osia: ligniiniä, selluloosaa ja hemiselluloosaa. Selektiivisesti ligniiniä hajottavat sienet lahottavat puusta suhteessa enemmän vaikeasti hajoavaa ligniiniä kuin selluloosaa tai hemiselluloosaa, jolloin jäljelle jää valkoista ja miltei puhdasta selluloosaa. Bioteknisissä sovelluksissa juuri selektiviiviset valkolahottajat ovat kiinnostavia. Niiden avulla voidaan puuhaketta esikäsitellä esimerkiksi paperinvalmistuksessa haitallisen ligniinin poistamiseksi. Ruskolahosienet ovat huomattavia puun, puutavaran ja puisten rakenteiden lahottajia, kuten tässä työssä käytetty Gloeophyllum trabeum (saunasieni ) ja Poria (Postia) placenta (istukkakääpä). Ruskolahosienet hajottavat puusta hemiselluloosan lisäksi selluloosaa, jolloin jää jäljelle ruskea ja jauhomaiseksi mureneva ligniini. Ruskolahosienet muovaavat ligniiniä jonkin verran. Kahden ruskolahosienen G. trabeumin ja P. placentan lisäksi tutkittiin valkolahosieniä, joista Ceriporiopsis subvermispora (karstakääpä) ja harvinainen Physisporinus rivulosus -sieni (talikääpä) hajottavat ligniiniä erittäin selektiivisesti. Phanerochaete chrysosporium on kaikkialla paljon tutkittu sieni, ja Phlebia radiata valkolahosientä (rusorypykkä) on tutkittu paljon mikrobiologian osastolla. Lisäksi tutkittiin Phlebia tremellosa -sienten (hytyrypykkä) ligninolyyttisten entsyymien tuottoa ja 14C-leimatun synteettisen ligniinin (DHP) hajotusta. P. radiata ja P. tremellosa -sienten on todettu aiemmin hajottavan ligniiniä selektiivisesti. Työssä selvitettiin miten sienten kasvua voi mitata, miten vertailukelpoisia eri mittaamismenetelmillä saadut tulokset ovat ja ilmenevätkö sienten aktiivisimmat kasvuvaiheet samaan aikaan eri menetelmillä mitattuna. Tärkeimmät tulokset olivat seuraavat havainnot: (i) P. radiata ja P. tremellosa -sienikannat tuottivat ligniini- ja mangaaniperoksidaasientsyymejä (LiP ja MnP) sekä lakkaasia, ja sienistä puhdistettiin 2-3 LiP- ja P. radiatasta yksi MnP-entsyymi; (ii) P. tremellosa -sienet hajottivat leimattua synteettistä ligniiniä (DHP) yhtä hyvin kuin paljon tutkitut P. chrysosporium ja P. radiata -sienet; (iii) puu, sienen luonnollinen kasvualusta, lisäsi valkolaho- ja ruskolahosienten demetoksylaatiota [O14CH3]-leimatusta ligniinin malliyhdisteestä 14CO2:ksi ilman puuta olleeseen alustaan verrattuna; (iv) demetoksylaatio (14CO2:n tuotto) oli normaalissa ilma-atmosfäärissä useimmiten parempi happeen verrattuna; (v) hapessa paras 14CO2:n tuotto saatiin puupalakasvatuksissa, joihin oli lisätty ravinnetyppeä tai typen lisäksi glukoosia sekä valkolaho- että ruskolahosienillä; (vi) ilmassa 14CO2:n tuotto oli puulla voimakkainta valkolahosienillä ilman lisäravinteita, kun taas G. trabeum -sienellä se oli yhtä hyvä eri alustoissa; (vii) biomassan muodostuminen rihmastojen ergosterolipitoisuuksista mitattuna oli ruskolahosienillä parempi kuin valkolahosienillä; (viii) ja biomassojen huippupitoisuudet olivat 6:lla sienellä eri suuruisia ja niiden maksimimäärien ajankohdat vaihtelivat viiden viikon kasvatusten kuluessa. Mikrobiologian osastolla Viikissä eristetty ja paljon tutkittu P. radiata -valkolahosieni oli mukana kaikissa tehdyissä kokeissa. Sienen LiP-aktiivisuus ja 14CO2:n tuotto 14C-rengas-leimatusta synteettisestä ligniinistä (DHP) korreloivat erittäin hyvin. Biomassan muodostuminen ergosterolilla määritettynä tuki hyvin entsyymiaktiivisuusmittauksilla ja isotooppikasvatuksilla saatuja tuloksia.