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Plants of the genus Lavandula are thought to be rarely infected by viruses. To date, only alfalfa mosaic virus, cucumber mosaic virus, tobacco mosaic virus, and tomato spotted wilt virus have been reported in this host. In this study, we identified for the first time raspberry ringspot virus (RpRSV) and phlox virus M (PhlVM) in lavender using herbaceous indexing, enzyme-linked immunosorbent assay, and high-throughput sequencing. Nearly complete genome sequences for both viruses were determined. Phylogenetic and serological characterizations suggest that the obtained RpRSV isolate is a raspberry strain. A preliminary survey of 166 samples indicated RpRSV was spread only in the lavender cultivar 'Grosso', while PhlVM was detected in multiple lavender cultivars. Although RpRSV raspberry strain may have spread throughout Auckland and nearby areas in New Zealand, it is very likely restricted to the genus Lavandula or even to the cultivar 'Grosso' due to the absence or limited occurrence of the nematode vector. Interestingly, all infected lavender plants, regardless of their infection status (by RpRSV, PhlVM, or both) were asymptomatic. RpRSV is an important virus that infects horticultural crops including grapevine, cherry, berry fruits, and rose. It remains on the list of regulated pests in New Zealand. RpRSV testing is mandatory for imported Fragaria, Prunus, Ribes, Rosa, Rubus, and Vitis nursery stock and seeds for sowing, while this is not required for Lavandula importation. Our study revealed that lavender could play a role not only as a reservoir but also as an uncontrolled import pathway of viruses that pose a threat to New Zealand's primary industries.
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Lavandula , Enfermedades de las Plantas , Lavandula/virología , Lavandula/química , Enfermedades de las Plantas/virología , Nueva Zelanda , Filogenia , Genoma Viral/genética , Nepovirus/genética , Nepovirus/aislamiento & purificación , Nepovirus/fisiología , Nepovirus/clasificación , Virus de Plantas/genética , Virus de Plantas/aislamiento & purificación , Virus de Plantas/fisiologíaRESUMEN
The FMR1 premutation (PM:55-199 CGG) is associated with fragile X-associated tremor/ataxia syndrome (FXTAS) and when maternally transmitted is at risk of expansion to a hypermethylated full mutation (FM: ≥ 200 CGG) that causes fragile X syndrome (FXS). We describe a maternally transmitted PM (77 CGG) that was passed to a son (103 CGG), and to a daughter (220-1822 CGG), who were affected with FXTAS and FXS, respectively. The male with the PM showed low-level mosaicism for normal size of 30 and 37 CGG. This male had two offspring: one female mosaic for PM and FM (56, 157, >200 CGG) and another with only a 37 CGG allele detected in multiple tissues, neither with a clinical phenotype. The female with the 37 CGG allele showed normal levels of FMR1 methylation and mRNA and passed this 37 CGG allele to one of her daughters, who was also unaffected. These findings show that post-zygotic paternal retraction can lead to low-level mosaicism for normal size alleles, with these normal alleles being functional when passed over two generations.
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Proteína de la Discapacidad Intelectual del Síndrome del Cromosoma X Frágil , Síndrome del Cromosoma X Frágil , Alelos , Metilación de ADN , Femenino , Proteína de la Discapacidad Intelectual del Síndrome del Cromosoma X Frágil/genética , Síndrome del Cromosoma X Frágil/diagnóstico , Síndrome del Cromosoma X Frágil/genética , Humanos , Masculino , Mutación , Expansión de Repetición de TrinucleótidoRESUMEN
A novel virus, tentatively named "sweetbriar rose curly-top associated virus" (SRCTaV), was identified in sweetbriar rose (Rosa rubiginosa) using high-throughput sequencing. The complete genome sequence of SRCTaV was determined and characterized. Phylogenetic analysis revealed that SRCTaV is closely related to members of the genus Waikavirus.
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Rosa , Waikavirus , Secuencia de Bases , Genoma Viral , Secuenciación de Nucleótidos de Alto Rendimiento , Sistemas de Lectura Abierta , Filogenia , Enfermedades de las Plantas , Virus Satélites , Waikavirus/genéticaRESUMEN
We report the complete genome sequence of a novel virus isolated from Nandina domestica 'Firepower' in Auckland, New Zealand. It was mechanically transmitted to Nicotiana species, although all of these infections were symptomless. The complete genome of the new virus is 8892 nucleotides (nt) long, excluding the 3' poly(A) tail, contains three open reading frames (ORF), and is most closely related to citrus leaf blotch virus (CLBV) Actinidia isolate (CLBV-Act; 72% nt sequence identity), a member of the genus Citrivirus. Replicase and coat proteins, encoded by genome ORFs 1 and 3 respectively, shared 81-83% and 76-79% amino acid (aa) sequence identity, respectively, with CLBV-Act. Computer-based analysis suggests that this novel virus is the result of recombination between CLBV-Act and an unknown virus, highlighting the importance of this phenomenon for betaflexivirus evolution.
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Berberidaceae/virología , Flexiviridae/genética , Secuencia de Aminoácidos , Secuencia de Bases , Flexiviridae/clasificación , Flexiviridae/fisiología , Genoma Viral/genética , Especificidad del Huésped , Nueva Zelanda , Sistemas de Lectura Abierta , Filogenia , Enfermedades de las Plantas/virología , ARN Viral/genética , Recombinación Genética , Proteínas Virales/genéticaRESUMEN
PurposeTo describe our experience of offering simultaneous genetic carrier screening for cystic fibrosis (CF), fragile X syndrome (FXS), and spinal muscular atrophy (SMA).MethodsCarrier screening is offered through general practice, obstetrics, fertility, and genetics settings before or in early pregnancy. Carriers are offered genetic counseling with prenatal/preimplantation genetic diagnosis available to those at increased risk.ResultsScreening of 12,000 individuals revealed 610 carriers (5.08%; 1 in 20): 342 CF, 35 FXS, 241 SMA (8 carriers of 2 conditions), approximately 88% of whom had no family history. At least 94% of CF and SMA carriers' partners were tested. Fifty couples (0.42%; 1 in 240) were at increased risk of having a child with one of the conditions (14 CF, 35 FXS, and 1 SMA) with 32 pregnant at the time of testing. Of these, 26 opted for prenatal diagnosis revealing 7 pregnancies affected (4 CF, 2 FXS, 1 SMA).ConclusionThe combined affected pregnancy rate is comparable to the population risk for Down syndrome, emphasizing the need to routinely offer carrier screening. The availability of appropriate genetic counseling support and a collaborative approach between laboratory teams, genetics services, health professionals offering screening, and support organizations is essential.
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Fibrosis Quística/epidemiología , Fibrosis Quística/genética , Síndrome del Cromosoma X Frágil/epidemiología , Síndrome del Cromosoma X Frágil/genética , Tamización de Portadores Genéticos , Atrofia Muscular Espinal/epidemiología , Atrofia Muscular Espinal/genética , Adulto , Australia/epidemiología , Fibrosis Quística/diagnóstico , Femenino , Síndrome del Cromosoma X Frágil/diagnóstico , Frecuencia de los Genes , Tamización de Portadores Genéticos/métodos , Pruebas Genéticas , Humanos , Masculino , Tamizaje Masivo , Persona de Mediana Edad , Atrofia Muscular Espinal/diagnóstico , Embarazo , Diagnóstico Prenatal , Prevalencia , Adulto JovenRESUMEN
Zoe McDonald, BSc, was omitted from the list of article coauthors. Her name should have been included as the seventh author, following Clare Elizabeth Hunt. Her affiliation is Victorian Clinical Genetics Services, Parkville, Victoria, Australia. The authors regret the error.
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We report the complete genome sequence of a novel virus, tentatively named "actinidia seed-borne latent virus" (ASbLV), isolated from Actinidia chinensis in Auckland, New Zealand. The complete genome of ASbLV is 8,192 nucleotides long, excluding the 3' poly(A) tail, contains four open reading frames, and is most closely related to Caucasus prunus virus (56% nucleotide sequence identity), a member of the genus Prunevirus. Based on the demarcation criteria of the family Betaflexiviridae, ASbLV is a new member of the genus Prunevirus.
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Actinidia/virología , Flexiviridae/genética , Genoma Viral , Filogenia , Análisis de Secuencia de ADN , Nueva Zelanda , Sistemas de Lectura Abierta , Enfermedades de las Plantas/virología , ARN Viral/genéticaRESUMEN
BACKGROUND: Detection and preventing entry of exotic viruses and viroids at the border is critical for protecting plant industries trade worldwide. Existing post entry quarantine screening protocols rely on time-consuming biological indicators and/or molecular assays that require knowledge of infecting viral pathogens. Plants have developed the ability to recognise and respond to viral infections through Dicer-like enzymes that cleave viral sequences into specific small RNA products. Many studies reported the use of a broad range of small RNAs encompassing the product sizes of several Dicer enzymes involved in distinct biological pathways. Here we optimise the assembly of viral sequences by using specific small RNA subsets. RESULTS: We sequenced the small RNA fractions of 21 plants held at quarantine glasshouse facilities in Australia and New Zealand. Benchmarking of several de novo assembler tools yielded SPAdes using a kmer of 19 to produce the best assembly outcomes. We also found that de novo assembly using 21-25 nt small RNAs can result in chimeric assemblies of viral sequences and plant host sequences. Such non-specific assemblies can be resolved by using 21-22 nt or 24 nt small RNAs subsets. Among the 21 selected samples, we identified contigs with sequence similarity to 18 viruses and 3 viroids in 13 samples. Most of the viruses were assembled using only 21-22 nt long virus-derived siRNAs (viRNAs), except for one Citrus endogenous pararetrovirus that was more efficiently assembled using 24 nt long viRNAs. All three viroids found in this study were fully assembled using either 21-22 nt or 24 nt viRNAs. Optimised analysis workflows were customised within the Yabi web-based analytical environment. We present a fully automated viral surveillance and diagnosis web-based bioinformatics toolkit that provides a flexible, user-friendly, robust and scalable interface for the discovery and diagnosis of viral pathogens. CONCLUSIONS: We have implemented an automated viral surveillance and diagnosis (VSD) bioinformatics toolkit that produces improved viruses and viroid sequence assemblies. The VSD toolkit provides several optimised and reusable workflows applicable to distinct viral pathogens. We envisage that this resource will facilitate the surveillance and diagnosis viral pathogens in plants, insects and invertebrates.
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Biología Computacional , Enfermedades de las Plantas/virología , Virus de Plantas/genética , ARN de Planta/análisis , ARN Viral/análisis , Viroides/genética , Australia , Internet , Nueva Zelanda , Enfermedades de las Plantas/genética , ARN Interferente Pequeño/análisisRESUMEN
Hospitals treating childhood emergencies need instant access to records. By building data warehouses to centralize that information, they've discovered a new tool for preventing emergencies in the first place.
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Data Warehousing/métodos , Registros Electrónicos de Salud , Servicio de Urgencia en Hospital/organización & administración , Hospitales Pediátricos/organización & administración , Humanos , Almacenamiento y Recuperación de la Información , Estudios de Casos Organizacionales , Pautas de la Práctica en Medicina , Mejoramiento de la Calidad , TexasRESUMEN
A novel virus, tentatively named "opium poppy mosaic virus" (OPMV), was isolated from Papaver somniferum (opium poppy) with leaf mosaic and mottling symptoms in Auckland, New Zealand, in 2006. The virus was mechanically transmitted to herbaceous plants of several species, in which it induced local and/or systemic symptoms. No virus particles were observed by electron microscopy in the diseased P. somniferum or any of the symptomatic herbaceous plants. The complete genomic sequence of 4230 nucleotides contains four open reading frames (ORF) and is most closely related (59.3 %) to tobacco bushy top virus, a member of the genus Umbravirus. These data suggest that OPMV is a new umbravirus.
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Papaver/virología , Enfermedades de las Plantas/virología , Virus ARN/aislamiento & purificación , Datos de Secuencia Molecular , Nueva Zelanda , Sistemas de Lectura Abierta , Filogenia , Virus ARN/clasificación , Virus ARN/genéticaRESUMEN
Red clover vein mosaic virus (RCVMV) is an important virus of leguminous crops that can cause devastating losses. During a routine survey of legumes conducted on the South Island of New Zealand, RCVMV was found in mixed infections in clover plants with Alfalfa mosaic virus and White clover mosaic virus. The full-length sequence of the New Zealand isolate RCVMV-NZ from clover shared 96% nucleotide sequence identity with a chickpea isolate previously described from Washington (United States). Targeted surveys of pea, faba bean, and pasture crops showed that RCVMV-NZ is widespread on the South Island in New Zealand. This isolate is causing mild if any symptoms on experimental hosts and naturally infected plants.
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Sweet chestnut, an Asiatic tree introduced in many parts of Europe including the United Kingdom, is planted for nut production, timber, and amenity. Its major threat is the disease called blight, caused by the fungus Cryphonectria parasitica, which infects through wounds by airborne spores. Field trapping using sticky rods rotating traps was performed in an infected area in Devon (between May 2021 and April 2023). An improved dual hydrolysis Taqman probes real-time PCR was used. The number of spores was calculated by comparing the cycle threshold to the Ct of standards with known amounts of conidia or known target fragment copies cloned into a plasmid. Weekly spore counts were in the range of around 60 to approximately 8.5 × 103, with fluctuations of peaks (mainly in late summer-autumn 2021) and troughs. The effects of weather parameters were modelled, finding correlations between spore numbers and temperature, humidity, dewpoint, rainfall, wind speed, and wind duration. Additionally, an insect trapping was performed to confirm the presence/absence and quantity of C. parasitica conidia potentially phoretic on some insects by using the same molecular approach. None of the ten collected insect species harboured spores of this fungus.
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Strawberry latent ringspot virus (SLRSV) is widespread in many countries, especially in Europe. The virus was thought to be uncommon in New Zealand, having only been recorded in Prunus spp. However, this study revealed that SLRSV infects a much wider range of hosts. From 1999 to 2009, SLRSV was isolated from anemone (Anemone × hybrida), blackberry (Rubus spp.), impatiens (Impatiens walleriana), pepino (Solanum muricatum), and tibouchina (Tibouchina sp.) in the North Island of New Zealand. These SLRSV isolates were identified using electron microscopy, mechanical inoculation, enzyme-linked immunosorbent assay, and reverse-transcription polymerase chain reaction techniques. This is thought to be the first report of anemone, impatiens, pepino, and tibouchina as hosts of SLRSV. Phylogenetic analysis and host range suggest that the five newly identified New Zealand isolates belong to two distinct strains: blackberry and impatiens isolates represent one strain and the other three isolates, plus the flowering cherry isolate reported previously in New Zealand, represent another strain. Both these strains are distinct from isolates reported elsewhere in the world. The strain infecting blackberry and impatiens is especially different and produced an unusual reaction in mechanical inoculation tests on herbaceous indicators. It is postulated that SLRSV may have gone undetected on its wider host range in New Zealand due to the latent infection in some hosts. The relationship of SLRSV isolates between New Zealand and overseas and the transmission modes of this virus are also discussed.
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Biological control of Cryphonectria parasitica fungus, the causal agent of chestnut blight, by virus infection (hypovirulence) is an effective control strategy against chestnut blight in Europe and some parts of North America. The most studied mycovirus is the Cryphonectria hypovirus 1 (CHV1) type species of the Hypoviridae family. In this study, the CHV1 virus was studied within some highly infected British isolates of Cryphonectria parasitica, gained in the past through co-culture transmissions. The effects of six temperatures (5-30 °C, in 5 °C steps) on six infected isolates (three with viral strain E-5, and other three with viral strain L-18) and their respective negative non-infected controls, three isogenic virulent fungal isolates, were examined. Experiments were performed with the nine isolate types with three replicates on potato dextrose agar (PDA) with cellophane sheets per isolate and temperature. A recently developed rapid, specific, quantitative reverse transcription PCR (RT-qPCR) screening method was used. This enabled quantifying the concentration (nanograms per microliter or copy numbers) of the virus within each isolate repetition. The presence of the virus had a significant negative effect between 20 and 25 °C on the C. parasitica growth rate, which was anyway highly influenced by and positively correlated with the temperature. The temperature clearly determined the virus accumulation and its recovery from cold or heat, and the virus optimum temperature was estimated at 15-25 °C.
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Virus Fúngicos , Virus ARN , Temperatura , Enfermedades de las Plantas/microbiologíaRESUMEN
In England, Cryphonectria parasitica was detected for the first time in 2011 in a nursery and in 2016 in the wider environment. Surveys between 2017 and 2020 identified the disease at different sites in Berkshire, Buckinghamshire, Cornwall, Derbyshire, Devon, Dorset, London, West Sussex, and the island of Jersey, while the present study comprises the results of the 2020-2023 survey with findings in Derbyshire, Devon, Kent, Nottinghamshire, Herefordshire, Leicestershire, London, West Sussex, and the islands of Jersey and Guernsey. A total of 226 suspected samples were collected from 72 surveyed sites, as far north as Edinburgh and as far west as Plymouth (both of which were negative), and 112 samples tested positive by real-time PCR and isolation from 35 sites. The 112 isolates were tested for the vegetative compatibility group (VCG), mating type, and Cryphonectria hypovirus 1 (CHV1). Twelve VCGs were identified, with two of them (EU-5 and EU-22) being the first records in the UK. Both mating types were present (37% MAT-1 and 63% MAT-2), but only one mating type was present per site and VCG, and perithecia were never observed. Cryphonectria hypovirus 1 (CHV1), consistently subtype-I haplotype E-5, was detected in three isolates at a low concentration (5.9, 21.1, and 33.0 ng/µL) from locations in London, Nottinghamshire, and Devon.
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Comunicación , Aplicaciones Móviles , Humanos , Internet , Teléfono Inteligente , Envío de Mensajes de TextoAsunto(s)
Selección de Profesión , Satisfacción en el Trabajo , Personal de Enfermería , Humanos , Reino UnidoRESUMEN
Raspberry ringspot virus (RpRSV) is an important virus that infects horticultural crops including grapevine, cherry, berry fruit and rose. The genome sequences of RpRSV are highly diverse between isolates and this makes the design of a PCR-based detection method difficult. In this study, a TaqMan real-time RT-PCR assay was developed for the rapid and sensitive detection of RpRSV. Primers and probes targeting the most conserved region of the movement protein gene were designed to amplify a 229 bp fragment of RpRSV RNA-2. The assay was able to amplify all RpRSV isolates tested. The detection limit of the RpRSV target region was estimated to be 61-98 copies, depending on the RpRSV strain. The sensitivity was about 100 times greater than the conventional RT-PCR assay using the same primers as the real-time RT-PCR assay. A comparison with published conventional RT-PCR assays indicated that both published assays lacked reliability and sensitivity, as neither were able to amplify all RpRSV isolates tested, and both were at least 1000 times less sensitive than the novel TaqMan real-time RT-PCR assay. The assay can also be run as a duplex reaction with the nad5 plant internal control primers and probe to simultaneously verify the PCR competency of the samples. The amplicon obtained with the real-time RT-PCR assay is suitable for direct sequencing if it is necessary to further confirm the RpRSV identity or determine the RpRSV strain.
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Nepovirus/genética , ARN Viral/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Rubus/virología , Cartilla de ADN/genética , Límite de Detección , Nepovirus/clasificación , Enfermedades de las Plantas/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
A new disease of glasshouse-grown tomato and pepper in New Zealand has resulted in plant decline and yield loss. Affected plants are characterized by spiky, chlorotic apical growth, curling or cupping of the leaves, and overall stunting. Transmission electron microscopy revealed the presence of phloem-limited bacterium-like organisms in symptomatic plants. The strategy used to identify the bacterium involved using specific prokaryote polymerase chain reaction (PCR) primers in combination with universal 16S rRNA primers. Sequence analysis of the 16S rRNA gene, the 16S/23S rRNA spacer region, and the rplKAJL-rpoBC operon revealed that the bacterium shared high identity with 'Candidatus Liberibacter' species. Phylogenetic analysis showed that the bacterium is distinct from the three citrus liberibacter species previously described and has been named 'Candidatus Liberibacter solanacearum'. This is the first report of a liberibacter naturally infecting a host outside the Rutaceae family. A specific PCR primer pair was developed for its detection.
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BACKGROUND: Cross-sectional data suggest that changes in health insurance status are associated with expenditures. No national longitudinal analysis has examined this relationship. OBJECTIVE: To evaluate the association between changes in health insurance status and expenditures. DESIGN: Cohort analyses using the 2000 to 2003 Medical Expenditure Panel Surveys. SETTING: U.S. civilian noninstitutionalized population. PARTICIPANTS: Three 2-year cohorts that included 20,848 adults age 21 to 64 years who were stratified by insurance type (private, public, military, or none): 17,130 participants were insured in both years, 342 participants were insured in year 1 and were uninsured in year 2, 385 participants were uninsured in year 1 and were insured in year 2, and 2991 participants were uninsured in both years. Persons who were insured for longer than 2 months but less than 10 months or who switched insurance type were excluded (n = 4039). MEASUREMENTS: Annual health care expenditures (any or none; amount, contingent on any expenditure; and the difference between year 1 and year 2). RESULTS: Adjusted expenditure probabilities were similar among all participant groups while insured and were higher than those for all participant groups while uninsured: 92.1% (95% CI, 91.4% to 92.7%) in year 1 and 91.8% (CI, 90.9% to 92.5%) in year 2 for persons insured in both years, 74.2% (CI, 71.7% to 76.5%) in year 1 and 74.8% (CI, 72.1% to 77.4%) in year 2 for persons uninsured in both years, and 90.7% (CI, 87.1% to 93.4%) for persons insured in year 1 and 74.6% (CI, 69.4% to 79.2%) for persons uninsured in year 2. The pattern was also consistent for the group that was uninsured in year 1 but insured in year 2. Adjusted annual expenditures among all participant groups with insurance were similar; expenditures among participant groups without insurance were similar but were lower than those among participants with insurance. Consistent differences in expenditures between year 1 and year 2 were observed for all groups. LIMITATION: Few participants changed insurance status. CONCLUSION: Changing insurance status is associated with changes in expenditures to levels that are similar to those for persons who are continuously insured or uninsured.