Draft Genome Sequence Resource for the Orange Rust Pathogen of Sugarcane Puccinia kuehnii.

Puccinia kuehnii is an obligate biotrophic fungal pathogen that causes orange rust of sugarcane, which is prevalent in many countries around the globe. In the United States, orange rust was first detected in sugarcane in Florida in 2007 and poses a persistent and economically damaging threat to the sugarcane industry in this region. Here, we generated the first genome assemblies for two isolates of P. kuehnii (1040 and 2143) collected in Florida in 2017 from two sugarcane cultivars, CL85-1040 and CP89-2143, respectively. These two rust genome resources will be of immense value for future genomic studies, particularly further exploration of the predicted secretomes that may help define key pathogenicity determinants for this economically important pathogen.

Eight Species of Poaceae Are Hosting Different Genetic and Pathogenic Strains of Sugarcane Mosaic Virus in the Everglades Agricultural Area.

Sugarcane mosaic virus (SCMV) was detected by reverse transcription polymerase chain reaction in eight different species of the Poaceae family in the Everglades Agricultural Area (EAA) of south Florida: broadleaf signalgrass (Urochloa platyphylla), Columbus grass (Sorghum almum), goosegrass (Eleusine indica), maize (Zea mays), sorghum (Sorghum bicolor), St. Augustine grass (Stenotaphrum secundatum), southern crabgrass (Digitaria ciliaris), and sugarcane (Saccharum interspecific hybrids). Based on their coat protein (CP) gene sequence, 62 isolates of SCMV from Florida and 29 worldwide isolates representing the known genetic diversity of this virus were distributed into eight major phylogenetic groups. SCMV isolates infecting Columbus grass, maize, and sorghum in Florida formed a unique group, whereas virus isolates infecting sugarcane in the United States (Florida and Louisiana) clustered with isolates from other countries. Based on the entire genome coding region, SCMV isolates infecting sugarcane in Florida were closest to virus isolates infecting sorghum species or St. Augustine grass. Virus isolates from Columbus grass, St. Augustine grass, and sugarcane showed different virulence patterns after mechanical inoculation of Columbus grass, St. Augustine grass, and sugarcane plants, thus proving that these isolates were different pathogenic strains. Sugarcane was symptomless and tested negative for SCMV by tissue blot immunoassay after inoculation with crude sap from SCMV-infected Columbus grass, indicating that Columbus grass was not a reservoir for SCMV infecting sugarcane in the EAA. Close CP sequence identity between isolates of SCMV from Columbus grass, maize, and sorghum suggested that the same virus strain was naturally spreading between these three plants in south Florida.

Lack of transmission of Sugarcane yellow leaf virus in Florida from Columbus grass and sugarcane to sugarcane with aphids or mites.

Sugarcane yellow leaf virus (SCYLV), the causal agent of yellow leaf disease, naturally infects at least three plant species in Florida: sugarcane (Saccharum spp.), the weed Columbus grass (Sorghum almum) and cultivated sorghum (S. bicolor). All three hosts are also colonized by the sugarcane aphid (Melanaphis sacchari), the main vector of SCYLV worldwide. To understand the high incidence of SCYLV observed in sugarcane commercial fields and in germplasm collections, we investigated the transmission efficiency of SCYLV from sugarcane and Columbus grass to sugarcane using the sugarcane aphid and a spider mite (Oligonychus grypus) that also tested positive for SCYLV in Florida. Healthy and SCYLV-infected leaf pieces of sugarcane and Columbus grass carrying viruliferous aphids or spider mites were transferred to virus-free plants of the yellow leaf susceptible sugarcane cultivar CP96-1252. Three- and 6-months post inoculation, the 108 aphid-inoculated plants of Columbus grass and the 90 mite-inoculated plants of sugarcane tested negative for SCYLV by tissue blot immunoassay (TBIA) or reverse transcription polymerase chain reaction (RT-PCR). Similar results were obtained for 162 aphid-inoculated plants of sugarcane, except for two plants that tested positive for SCYLV by TBIA and RT-PCR. In two field experiments planted with SCYLV-free and virus-infected sugarcane (cultivar CP96-1252), only 18-28% of healthy plants became infected during a 24- to 28-month period. SCYLV prevalence in these field experiments did not differ between aphicide treated and untreated plots. Incidence of M. sacchari haplotypes in the Everglades agricultural area also indicated that the predominant haplotype that is currently colonizing sugarcane was not a vector of SCYLV in Florida. Lack of virus transmission by the spider mite suggested that this arthropod only acquired the virus when feeding on infected plants but was unable to transmit SCYLV. The current vector of SCYLV in Florida remains to be identified.

Field Infection of Virus-Free Sugarcane by Sugarcane Yellow Leaf Virus and Effect of Yellow Leaf on Sugarcane Grown on Organic and on Mineral Soils in Florida.

Sugarcane yellow leaf virus (SCYLV), the causal agent of yellow leaf, is widespread in Florida. Two field trials were set up, one on organic soil and one on mineral soil, to investigate the rate and timing of sugarcane infection by SCYLV under field conditions and the effect of the virus on yield. Each trial consisted of plots planted with healthy or SCYLV-infected seed cane of two commercial cultivars. Virus prevalence varied from 83 to 100% in plots planted with infected seed cane regardless of cultivar, location, and crop season. On organic soil, plants of virus-free plots became progressively infected in plant cane and first ratoon crops. On mineral soil, healthy sugarcane became initially infected in the first ratoon crop. After three crop seasons, the highest SCYLV prevalence rates were 33 and 7% on organic and mineral soils, respectively. No significant negative effect of SCYLV on yield was found in plant cane crop regardless of cultivar and soil type. However, yield reductions in ratoon crops varied from nonsignificant to 27% depending on cultivar and soil type. Low virus prevalence observed after three crop seasons suggested that planting virus-free seed cane should limit the impact of SCYLV on sugarcane production in Florida.

Comparison of Progress of Brown Rust and Orange Rust and Conditions Conducive for Severe Epidemic Development During the Sugarcane Crop Season in Florida.

Brown rust (caused by Puccinia melanocephala) and orange rust (caused by P. kuehnii) are two major diseases of sugarcane in Florida. To better understand the epidemiology of these two rusts, disease severity and weather variables were monitored for two seasons in cultivars CL90-4725 (susceptible to brown rust and resistant to orange rust) and CL85-1040 (susceptible to orange rust and resistant to brown rust). Brown rust was most severe during mid-May to mid-July, whereas orange rust severity peaked during two periods: mid-May to early August and then November to December. Overall, disease severity was higher for orange rust than for brown rust. Maximum disease severity was correlated with the number of hours at night with an average temperature of 20 to 22.2°C for brown rust one season and orange rust both seasons. Slightly higher correlation was obtained when relative humidity above 90% was included in the number of hours at night with an average temperature of 20 to 22.2°C for brown rust but not orange rust, suggesting that leaf wetness is not a limiting factor for either disease in Florida. Epidemics of brown rust began at lower night temperatures (16.7 to 22.2°C) in one season, but epidemics of orange rust lasted longer under higher temperatures. The correlation of rust severity on recently emerged leaves with conducive temperatures recorded in 10-, 20-, or 30-day windows starting 7 days before disease assessment suggested that earlier inoculum production is needed to create severe epidemics that result in yield loss.

Baseline Sensitivity of Guignardia citricarpa Isolates from Florida to Azoxystrobin and Pyraclostrobin.

Citrus black spot (CBS), caused by Guignardia citricarpa, is an emerging disease in Florida. Fungicide applications are the main control measure worldwide. The in vitro activity and baseline sensitivity of G. citricarpa isolates to quinone outside inhibitor (QoI) fungicides (azoxystrobin and pyraclostrobin) were evaluated. The effective concentration needed to reduce mycelial growth or spore germination by 50% (EC50) was determined for 86 isolates obtained from Florida counties where CBS is found. The effect of salicylhydroxamic acid (SHAM) plus azoxystrobin and pyraclostrobin was also assessed for mycelial growth and conidial germination. The mean EC50 for mycelial growth for azoxystrobin was 0.027 µg/ml and that for pyraclostrobin was significantly lower at 0.007 µg/ml (P < 0.0001). Similarly, the mean EC50 for conidial germination for azoxystrobin was 0.016 µg/ml and that for pyraclostrobin was significantly lower at 0.008 µg/ml (P < 0.0001). There was no effect of SHAM on inhibition of mycelial growth or conidial germination by the QoI fungicides but SHAM slightly affected mycelium inhibition by pyraclostrobin. Cytochrome b was partially sequenced and three group 1 introns were found. One intron was immediately post G143, likely inhibiting resistance-conferring mutations at that site. It is surmised that the QoI resistance risk is low in the Florida G. citricarpa population.

Ocho casos de piedra blanca en población pediátrica / Eight white piedra pediatric cases

Las piedras son infecciones fúngicas de baja agresividad que comprometen la porción extrafolicular del pelo, originadas por un hongo levaduriforme del género Trichosporon spp. Se caracterizan clínicamente por la presencia de lesiones nodulares en pelos de axilas, pubis, barba y, con menor frecuencia, del cuero cabelludo. La piedra blanca predomina en hombres entre los 18 y los 35 años de edad, y aunque todo el pelo corporal puede verse afectado, la localización en la región púbica en estas edades es considerada la más frecuente; sin embargo, en los niños la afectación predomina en el cuero cabelludo. En su diagnóstico se destaca el examen directo con hidróxido de potasio de pelos y cultivo en agar Sabouraud. Se presentan ocho casos de piedra blanca en cuero cabelludo de pacientes pediátricos, residentes en el área metropolitana de Bucaramanga, Colombia. El examen fúngico demostró la presencia de Trichosporon spp. Además, se hace una revisión de los aspectos históricos, epidemiológicos, clínicos, diagnósticos y terapéuticos de la enfermedad.

Análisis de la estructura genética en una muestra poblacional de Bucaramanga, departamento de Santander / An assessment of genetic structure in a Bucaramanga, department of Santander, population sample

Introducción: El fenómeno de sub-estructura en las poblaciones ha tenido desde hace varios años un abordaje amplio, que se enfocó, entre otros, en la identificación y cuantificación de la mezcla étnica presente en estudios de mapeo asociativo, para comprobar la asociación de marcadores polimórficos en el desarrollo de enfermedades comunes complejas, como responsable de falsos positivos. No obstante el reconocimiento de este problema, no se tiene suficiente información genética en el contexto nacional ni local que permita determinar la posible diferenciación de subgrupos poblacionales en cada región en particular. Objetivo: Determinar la estructura genética en una muestra poblacional de la ciudad de Bucaramanga, a partir del análisis de 19 marcadores microsatélites autosómicos en distintos subgrupos poblacionales. Metodología: De la base de datos del Laboratorio de Genética Humana de la Universidad Industrial de Santander, se seleccionaron aleatoriamente 350 muestras de ADN, y se amplificaron 19 marcadores autosómicos Short Tandem Repeat mediante los "kits Powerplex® 16 y FFFL (Promega)".Resultados: En el análisis de equilibrio Hardy Weinberg, no se obtuvieron diferencias estadísticamente significativas en 18 de 19 marcadores Short Tandem Repeat autosómicos analizados en la población de Bucaramanga. El único marcador que mostró no estar en equilibrio Hardy Weinberg en la población de Bucaramanga fue el F13B (valor de significancia de p=0.00264, después de aplicar la corrección de Bonferroni). Discusión: Las poblaciones representadas en los seis estratos socioeconómicos mostraron alta diversidad genética intragrupos, que ratificó una alta variabilidad entre los individuos de la ciudad de Bucaramanga, acorde con el bajo valor de FST entre distintos grupos, determinado en el análisis molecular de varianza con base en frecuencias alélicas observadas para los 19 Short Tandem Repeat analizados.

Introduction: The phenomenon of substructure in the populations has been greatly analyzed for several years, and it has been focused especially on the identification and quantification of ethnic mixture present in studies of associative mapping to verify the association of polymorphic markers in the development of complex and common diseases responsible for false positives. Nevertheless, despite the recognition of this issue, there is insufficient genetic information within the national or local contexts that allow assessing the possible differentiation of population sub-groups in each particular region. Objective: To determine the genetic structure in the city of Bucaramanga through the analysis of 19 autosomal microsatellite markers in different subgroups of the population. Methodology: A total of 350 DNA samples were randomly selected from the database of the Human Genetic Laboratory at Universidad Industrial de Santander by using Epi Info version 6.04 2001. Also, 19 Short Tandem Repeat markers were amplified using "kits Powerplex® 16 and FFFL (Promega)". Results: In the Hardy Weinberg equilibrium analysis (100 steps in Markov chain and 1000 dememorization steps), no statistically significant differences in 18 out of the 19 analyzed STRs markers in the population of Bucaramanga were obtained. A unique marker that proved not present in HWE in the population of Bucaramanga was the F13B (for a significance value of p=0.00264, after applying the Bonferroni correction). Discussion: The populations represented in the six socioeconomic levels presented high genetic diversity intragroups, which ratified the high variability among the individuals in this city according to the low value of FST for different groups, determined via the molecular analysis of variance based on the allelic frequencies observed for the 19 analyzed Short Tandem Repeats.