A High-Throughput Gene Silencing Approach for Studying the Interaction Between Perennial Ryegrass (\u3cem\u3eLolium Perenne\u3c/em\u3e) and the Fungal Endophyte \u3cem\u3eNeotyphodium Lolii\u3cem\u3e

Perennial ryegrass (Lolium perenne L.) and its fungal endophyte (Neotyphodium lolii) are known to establish a mutualistic association that impacts on the agronomic productivity of endophyte-infected ryegrass pastures. To study this interaction at the molecular level, a genomic resource consisting of 13,964 endophyte ESTs has been generated. However, the functions of a large proportion of these genes remain to be elucidated. Recent work has demonstrated the potential for RNA-mediated gene silencing to suppress gene expression in a sequence specific manner thus allowing for the subsequent analysis of gene function

Isolation and Characterization of Novel BTB Domain Protein Encoding Genes from Fungal Grass Endophytes

Pasture grasses belonging to the Pooideae sub-family of the Poaceae family frequently host symbiotic fungal endophytes. These include the sexual Epichloë species and the anamorphic asexual Neotyphodium species, which are thought to have evolved from Epichloë species either by the direct loss of sexual reproduction or by interspecific hybridisation. The two key temperate pasture grasses, tall fescue (Festuca arundinacea Schreb) and perennial ryegrass (Lolium perenne L.) interact with the fungal endophytes N. coenophialum and N. lolii, respectively. Large insert genomic DNA libraries are valuable resources for the discovery and isolation of genes and their regulatory sequences, for physical mapping, map-based cloning of target genes as well as for whole genome sequencing. BTB (Bric-a-brac, tram-track, broad complex) domains are highly conserved motifs of 120 amino acids in length. The domains are rich in hydrophobic amino acids, and mediate protein-protein interaction that lead to homomeric dimerisation and in some cases heteromeric dimerisation of a large number of functionally diverse proteins. The presence of BTB domains defines a large family of genes involved in various biological processes, such as the regulation of transcription, DNA binding activity and structural organisation of macromolecular structures. Genes encoding BTB domain proteins (BDP) have previously been described in viruses, yeasts, plants, nematodes, insects, fish and mammals. However, BDP genes have not as yet been described for filamentous fungi

Microarray-Based Transcriptome Analysis of the Interaction Between Perrenial Ryegrass (\u3cem\u3eLolium Perenne\u3c/em\u3e) and the Fungal Endophyte \u3cem\u3eNeotyphodium Lolii\u3c/em\u3e

Neotyphodium lolii, Neotyphodium coenophialum and Epichloë festucae are common symbiotic fungal endophytes of the temperate pasture grasses perennial ryegrass (Lolium perenne), tall fescue (Festuca arundinacea) and red fescue (Festuca rubra), respectively. A genomic resource of 13,964 expressed sequence tags (ESTs), representing 7,585 unique endophyte genes, has been established for Neotyphodium and Epichloë fungal endophytes

A Proposal for an International Transcriptome Initiative for Forage and Turf: Microarray Tools for Expression Profiling in Ryegrass, Clover and Grass Endophytes

Knowledge of the expression pattern of genes provides a valuable insight into gene function and role in determining the observed heritable phenotype. High–density cDNA and oligonucleotide microarrays represent powerful tools for transcriptome analysis to gain an understanding of gene expression patterns for thousands of genes. Internationally coordinated efforts in transcriptome analyses and sharing of microarray resources will benefit the advancement of our understanding of gene function in forage and turf species

Discovery and Functional Categorisation of Expressed Sequence Tags from Flowers of \u3cem\u3eEragrostis Curvula\u3c/em\u3e Genotypes Showing Different Ploidy Levels and Reproductive Modes

Two novel genotypes of weeping lovegrass (Eragrostis curvula) - a dihaploid strain obtained in vitro from an apomictic cultivar and a tetraploid plant derived from the dihaploid after chromosome duplication – have recently been developed. These materials represent an excellent system for the identification, through transcriptional profiling, of genes involved in diplospory and/or ploidy level gene regulation. The aim of this work was the discovery and functional classification of expressed sequence tags (ESTs) from immature inflorescences of the apomictic E. curvula cultivar Tanganyika (2n=4x=40), a dihaploid sexual strain derived from it (2n=2x=20) and a tetraploid sexual strain (2n=4x=40) obtained by colchicine duplication of the dihaploid

Metabolome Analysis of the Interaction Between Perennial Ryegrass (\u3cem\u3eLolium Perenne\u3c/em\u3e) and the Fungal Endophyte \u3cem\u3eNeotyphodium Lolii\u3c/em\u3e

Perennial ryegrass (Lolium perenne L.) and tall fescue (Festuca arundinacea Schreb.) frequently contain endophytic fungi (Neotyphodium lolii in perennial ryegrass and N. coenophialum in tall fescue). The presence of the endophyte has been shown to improve seedling vigour, persistence and drought tolerance in marginal environments as well as provide protection against some insect pests. Endophyte-infected grasses also produce a wide range of metabolites, including ergopeptine alkaloids, indole-isoprenoid lolitrems, pyrrolizidine alkaloids, and pyrrolopyrazine alkaloids. In contrast to information on alkaloids and animal toxicosis, the beneficial physiological aspects of the endophyte/grass interactions have not been well characterised. The physiological mechanisms which lead to increased plant vigour and enhanced tolerance to abiotic stresses unrelated to the reduction in pest damage to endophyte-infected grasses are unknown. Recent technological advances in metabolomics enable dynamic changes in the metabolome of an organism under varying experimental conditions to be studied. This provides opportunities for the investigation and validation of each and every detected metabolite, investigation of known metabolic pathways through searching of databases of known metabolites, molecular formula determination of unknown metabolites and creation of pathways from novel metabolites

Endophyte ASTRA: a Web-Based Resource for Neotyphodium and Epichloë EST Analysis

Large-scale gene discovery has led to the production of 13,964 expressed sequence tags (ESTs) collectively from the grass endophytes Neotyphodium coenophialum, N. lolii and Epichloë festucae

Future Directions in the Molecular Breeding of Forage and Turf

Key points Molecular breeding of forage and turf plants and their endosymbionts has entered the post-genomic era with a large amount of structural genomics information and genomic resources available for key forage and turf species and relevant model systems. A primary future challenge is the conversion of this information into useful functional knowledge for the development of molecular breeding technologies and products that address a range of high impact outcome scenarios in forage and turf. High-throughput approaches for spatial and temporal analysis, from genome to phenome, and the respective data integration in a systems biology context will be critical for the establishment of stringent gene-function correlations. Translational genomics will permit results obtained using model systems to have major impact on the understanding of the molecular basis of plant processes and direct application to the molecular breeding of forage and turf plants. These developments will be enhanced through applications of transgenesis and functionally-associated genetic markers in forage and turf molecular breeding building on genomic and post-genomic discoveries in these target species

History of adversity, health and psychopathology among prisoners: comparison between men and women

Adversity in childhood, risk behaviors and psychopathology are highly prevalent phenomena in inmate populations and have a strong impact on health. Knowing the differences in these variables between the sexes is most important in order to develop appropriate intervention strategies in a prison context. By administering the Socio-demographic and Life History Questionnaire and the Brief Symptoms Inventory, we sought to characterize adverse childhood experiences and relate them to risk behaviors and to psychopathological symptoms, and study the differences between the 65 male and 42 female detainees in Portuguese prison establishments. Men and women report a complex web of adversity in childhood. In a range of ten possible categories, a medium value of 5.05 (DP = 2.63) in total adversity for women and 2.63 (DP = 2.18) for men was encountered, with the prevalence being significantly higher within the female population (Z = -4.33; p = .000). A high prevalence of risk behaviors and psychopathological symptoms was found in both groups, the latter being higher among females. We concluded that the differences between men and women calls for in depth studies in order to provide guidelines for intervention projects in specific populations.Adversidade na infância, comportamentos de risco e psicopatologia são fenómenos muito prevalentes na população reclusa e com forte impacto na saúde. Conhecer as diferenças entre sexos, no que diz respeito a tais variáveis, é de elevada importância no sentido de adequar estraté- gias de intervenção em contexto prisional. Utilizando o Questionário Sociodemográfico e Histó- ria de Vida, o Questionário de Adversidade na Infância e o Brief Symptons Inventory, procuramos caracterizar a adversidade na infância, os comportamentos de risco e as dimensões psicopatológicas, e averiguar as diferenças entre 65 homens e 42 mulheres reclusos em estabelecimentos prisionais Portugueses. Homens e mulheres relatam um quadro complexo de adversidade na infância. Num total possível de dez categorias, verificamos uma média de adversidade total de 5.05 (DP = 2.63) para as mulheres e de 2.63 (DP = 2.18) para os homens, sendo a prevalência significativamente mais elevada junto da população feminina (Z = -4.33; p = .000). Foi ainda encontrada uma elevada prevalência de comportamentos de risco e de sintomatologia psicopatológica em ambos os grupos, sendo esta última superior nas mulheres. Concluímos que as diferenças entre sexos devem ser estudadas para guiarem a adequação dos projetos

Trauma history and depression predict incomplete adherence to antiretroviral therapies in a low income country.

As antiretroviral therapy (ART) for HIV becomes increasingly available in low and middle income countries (LMICs), understanding reasons for lack of adherence is critical to stemming the tide of infections and improving health. Understanding the effect of psychosocial experiences and mental health symptomatology on ART adherence can help maximize the benefit of expanded ART programs by indicating types of services, which could be offered in combination with HIV care. The Coping with HIV/AIDS in Tanzania (CHAT) study is a longitudinal cohort study in the Kilimanjaro Region that included randomly selected HIV-infected (HIV+) participants from two local hospital-based HIV clinics and four free-standing voluntary HIV counselling and testing sites. Baseline data were collected in 2008 and 2009; this paper used data from 36 month follow-up interviews (N = 468). Regression analyses were used to predict factors associated with incomplete self-reported adherence to ART. INCOMPLETE ART ADHERENCE WAS SIGNIFICANTLY MORE LIKELY TO BE REPORTED AMONGST PARTICIPANTS WHO EXPERIENCED A GREATER NUMBER OF CHILDHOOD TRAUMATIC EVENTS: sexual abuse prior to puberty and the death in childhood of an immediate family member not from suicide or homicide were significantly more likely in the non-adherent group and other negative childhood events trended toward being more likely. Those with incomplete adherence had higher depressive symptom severity and post-traumatic stress disorder (PTSD). In multivariable analyses, childhood trauma, depression, and financial sacrifice remained associated with incomplete adherence.\ud This is the first study to examine the effect of childhood trauma, depression and PTSD on HIV medication adherence in a low income country facing a significant burden of HIV. Allocating spending on HIV/AIDS toward integrating mental health services with HIV care is essential to the creation of systems that enhance medication adherence and maximize the potential of expanded antiretroviral access to improve health and reduce new infections