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Emission abatement cyclone performance is improved by increasing collection effectiveness or decreasing energy consumption. The object of this study was to quantify the pressure drop and fine particulate (PM2.5) collection of 1D3D cyclones (H = 4Dc, h = 1Dc) at inlet velocities from 8 to 18 m s− 1 (Stk = 0.7–1.5) using heterogeneous particulate as a test material at inlet concentrations from 3 to 75 g m− 3. Cyclone exhaust was passed through filters. Laser diffraction particle size distribution analysis was used to estimate PM2.5 emissions. Response surface models showed a strong correlation between cyclone pressure loss (Euler number) and inlet velocity and predicted a 46% reduction in pressure loss for a 25% reduction in inlet velocity (Stokes number). The model for PM2.5 emissions was less definitive and, surprisingly, predicted a 31% decrease in PM2.5 emissions when operating 25% below the design inlet velocity. Operating below the design inlet velocity (at a lower Stokes number) to reduce pressure losses (Euler number) would reduce both the financial and the environmental cost of procuring electricity. The unexpected co-benefit suggested by these trials was that emission abatement may improve at the same time, though other empirical trials have shown emissions to be independent of inlet velocity and Stokes number.

Published in Agron. J. 107:799–810 (2015) doi:10.2134/agronj14.0017 Available freely online through the author-supported open access option. Copyright © 2015 by the American Society of Agronomy, 5585 Guilford Road, Madison, WI 53711. All rights reserved. No part of this periodical may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. ABSTRACT

Protein is the second most abundant constituent in the rice grain next to starch. Association analysis for protein concentration in brown rice was performed using a “mini-core” collection, which represents the germplasm diversity found in the USDA rice world collection. Protein concentration was determined in replicated trials conducted in two southern U.S. locations, and association mapping was performed by using 157 genomewide DNA markers. Protein concentration ranged from 5.4 to 11.9% among the 202 accessions. Protein variation owing to accession and accession × location interaction were highly significant. Ample variation was seen within each subpopulation by ancestry, as well as within the 14 geographic regions where the accessions originated. Accessions from Eastern Europe had the highest level of protein. Ten markers on eight chromosomes were significantly associated with protein concentration. Five of these markers occurred near known protein precursor genes or quantitative trait loci, an...

A set of introgression lines (ILs) containing chromosomal segments from O. rufipogon (IRGC 105491), a wild relative of O. sativa, in the genetic background of an elite US variety, cv. Jefferson, was developed to confirm the performance of six yield-enhancing quantitative trait loci (QTL). Fifty BC3F3 ILs containing homozygous O. rufipogon introgressions at each of the target QTL regions, and as few background introgressions as possible, were selected for evaluation of yield and 14 yield-related traits in field studies conducted over 2 years at four locations in the southern USA. Performance of the IL families was compared with three commercial inbreds and one hybrid variety. IL families carrying introgressions from the low-yielding wild parent at the QTL yld2.1 and yld6.1 yielded 27.7 and 26.1 % more than Jefferson, respectively. IL yld2A, which possesses yld2.1, also performed well under alternate wetting and drying conditions in two field locations. After the first year of field trials, 10 of the top-performing BC3F4 families, representing five of the QTL targets, were genotyped with an Illumina 1,536 assay to define the size and location of the wild introgressions. BC3F4 families with the fewest background introgressions were backcrossed to Jefferson and selfed. The resulting BC4F2 families were screened with targeted single nucleotide polymorphism assays to identify individuals carrying homozygous introgressions across the target QTL. Twelve ILs, representing each of the six QTL targets, have been submitted to the Genetic Stocks Oryza Collection for studies on transgressive variation and as interspecific pre-breeding lines.

Bovine babesiosis, also known as cattle fever, is a tick-borne protozoal disease foreign to the United States. It was eradicated by eliminating the vector species, Rhipicephalus (Boophilus) annulatus and Rhipicephalus (Boophilus) microplus, through the efforts of the Cattle Fever Tick Eradication Program (CFTEP), with the exception of a permanent quarantine zone (PQZ) in south Texas along the border with Mexico. Keeping the U.S. free of cattle fever ticks in a sustainable manner is a critical national agricultural biosecurity issue. The efficacy of a Bm86-based anti-tick vaccine commercialized outside of the U.S. was evaluated against a strain of R. annulatus originated from an outbreak in Texas. Vaccination controlled 99.9 and 91.4% of the ticks 8 weeks and 5.5 months after the initial vaccination, respectively. Computer modeling of habitat suitability within the PQZ typically at risk of re-infestation with R. annulatus from Mexico predicted that at a level of control greater than 40%, eradication would be maintained indefinitely. Efficacy and computer modeling data indicate that the integration of vaccination using a Bm86-based anti-tick vaccine with standard eradication practices within the northwestern half of the PQZ could incentivize producers to maintain cattle on pasture thereby avoiding the need to vacate infested premises. Implementing this epidemiologically proactive strategy offers the opportunity to prevent R. annulatus outbreaks in the U.S., which would represent a significant shift in the way the CFTEP operates.

False smut (Ustilaginoidea virens) and kernel smut (Neovossia horrida) are diseases of rice (Oryza sativa) that reduce both grain yield and quality. Susceptible rice cultivars are in widespread use on production acreage in the United States, and the effects from crop management practices on smut control are poorly understood. We studied the long-term effects of crop rotation, soil tillage, and fertility level on rice smut severity. The highest levels of false smut observed in this study were on cultivars grown in rotation with soybean, on traditionally tilled soils, with high fertilizer treatments. The highest levels of kernel smut were observed in a rice-soybean rotation with winter wheat grown between summer crops. These rotations are commonly used in rice-growing regions of the southern United States. Using combinations of crop rotation, soil tillage, and fertility rate, several alternative crop-management practices were identified that provided effective control of smuts in susceptible rice cultivars. The most effective method for controlling both false smut and kernel smut was in 3-year rotations of rice, soybean, and corn. Regardless of rotation order or tillage and fertility treatments within the rotations, rotating out of rice for 2 years was the most effective approach for smut control.

Sheath blight, caused by the fungal pathogen Rhizoctonia solani, is one of the most important and widely distributed diseases capable of infesting numerous crops including rice (Oryza sativa L.). Resistant germplasm with wide variation is essential for controlling this disease via breeding efforts, and genetic background helps design breeding strategies. We used the micro-chamber method to evaluate the USDA rice core collection, including 1794 accessions from 114 countries, for sheath blight resistance. Seventy-two molecular markers were used for genotyping. Compared with 'Jasmine 85', which has been recognized as one of the most resistant germplasm accessions, 52 accessions were significantly more resistant at the 5% level of probability, and of these 17 were more resistant at the 1% level of probability. The resistant accessions originated from 26 countries in nine geographic regions and are diversified for 13 phenotypic characteristics. The resistant accessions all belonged to the cultivated species O. sativa. Genetic analysis using 72 molecular markers revealed that 45 resistant accessions (87%) were indica type, which were further classified in two groups. Three accessions were identified as aus, two as aromatic, and one each as temperate japonica and tropical japonica. Breeders could use these findings to choose sheath blight resistant accessions for cultivar improvement.

Rice (Oryza sativa L.) head-rice yield (HR) is a key export and domestic quality trait whose genetic control is poorly understood. With the goal of identifying genomic regions influencing HR, quantitative-trait-locus (QTL) mapping was carried out for quality-related traits in recombinant inbred lines (RILs) derived from crosses of common parent Cypress, a high-HR US japonica cultivar, with RT0034, a low-HR indica line (129 RILs) and LaGrue, a low-HR japonica cultivar (298 RILs), grown in two US locations in 2005-2007. Early heading increased HR in the Louisiana (LA) but not the Arkansas (AR) location. Fitting QTL-mapping models to separate QTL main and QTL × environment interaction (QEI) effects and identify epistatic interactions revealed six main-effect HR QTLs in the two crosses, at four of which Cypress contributed the increasing allele. Multi-QTL models accounted for 0.36 of genetic and 0.21 of genetic × environment interaction of HR in MY1, and corresponding proportions of 0.25 and 0.37 in MY2. The greater HR advantage of Cypress in LA than in AR corresponded to a genomewide pattern of opposition of HR-increasing QTL effects by AR-specific effects, suggesting a selection strategy for improving this cultivar for AR. Treating year-location combinations as independent environments resulted in underestimation of QEI effects, evidently owing to lower variation among years within location than between location. Identification of robust HR QTLs in elite long-grain germplasm is suggested to require more detailed attention to the interaction of plant and grain development parameters with environmental conditions than has been given to date.

Plant canopy temperature is used in many studies of plant/environment interactions and non-contact measurement is often made with radiometric surface thermometers commonly referred to as infrared thermometers. Industrial-quality infrared thermocouples are widely available and often used in agricultural research. While research on canopy temperature has provided management tools for production agriculture, the high cost of the industrial-quality infrared thermocouples has limited their adoption and use in production agriculture settings. Our objective was to evaluate a low-cost consumer-quality infrared thermocouple as a component of a wireless thermal monitoring system designed for use in a production agriculture setting. The performances of industrial-quality and low-cost consumer-quality sensors were compared under controlled constant temperature and under field conditions using both grass and cotton canopies. Results demonstrate that under controlled constant-temperature the two types of infrared thermocouples were ''significantly the same'' at 10^oC, 20^oC and 30^oC and ''significantly not the same'' at 40^oC and 50^oC. Across the temperature range tested, the consumer-quality infrared thermocouples temperature reading was closer to the thermocouple reading than the industrial-quality infrared thermocouples. A field comparison of industrial-quality and consumer-quality infrared thermocouple sensors monitoring a grass canopy and a cotton canopy indicated that the two types of sensors were similar over a 13-35^oC range. The measurement of temperature made with two types of sensors would not differ significantly. Based on these results we conclude that the lower-cost consumer-quality infrared thermometers are suitable for use in production agricultural applications.

Percent milling yield is an economically important trait of commercial rice because it largely determines the price that farmers receive for their crop. Analyzing 22 trait variables including milling yield, grain dimensions, chemistry and appearance, we identified 43 quantitative trait loci (QTLs) in a long grain japonica by long grain japonica cross. We report one QTL explaining 20% of the variation in brown rice recovery; two QTLs explaining 14% and 13% of the variation in milled rice recovery; and one QTL explaining 14% of the variation in head rice (HR) recovery. QTLs for the proportion of pre-broken brown rice kernels, seed density, amylose content, and kernel whiteness and chalkiness were found in the same region as the HR QTL. QTLs explaining up to 54% of the variation in grain shape measurements were identified and mapped to areas independent from those identified for milling yield. Analyses of grain appearance traits identified two QTLs for chalk in brown rice and one in head rice, and a QTL explaining up to 33% of the variance in green kernel area. Our results confirm previous findings on the multigenically complex nature of milling yield.

The effect dirt-floored broiler houses have on the underlying native soil, and the potential for contamination of the ground water by leaching under the foundation, is an understudied area. This study examines alterations in fifteen quantitative soil parameters (Ca, Cu, electrical conductivity, Fe, K, Mg, Mn, Na, NO3, organic matter, P, pH, S, soil moisture and Zn) in the underlayment of a newly constructed dirt-floored broiler house over the first two years of production (Native through Flock 11). The experiment was conducted near NW Robertson County, Texas, where the native soil is a fine, smectitic thermic Udertic Paleustalfs and the slopes range from zero to three percent. Multiple samples were collected from under each of three water and three feed lines the length of the house, in a longitudinal study during February 2008 through August 2010. To better define the relationship between the soil parameters and sampling times, a canonical discriminant analysis approach was used. The soil profiles assembled into five distinctive clusters corresponding to time and management practices. Results of this work revealed that the majority of parameters increased over time. The management practices of partial and total house clean-outs markedly altered soil profiles the house underlayment, thus reducing the risk of infiltration into the ground water near the farm. This is important as most broiler farms consist of several houses within a small area, so the cumulative ecological impact could be substantial if not properly managed.

Taking advantage of the high frequency of conjugation tube formation in mating mixtures and α-pheromone-treated a / a cells derived from saturation phase cultures of opaque cells of Candida albicans , 56 up-regulated and 30 down-regulated genes were identified employing microarray and Northern analyses. Combining these results with previous profiling studies of pheromone-induced cells, a more comprehensive transcript profile was developed for comparison with Saccharomyces cerevisiae . This comparison revealed the following: (i) that while a majority of mating-associated genes are regulated similarly between the two species, a significant minority are regulated dissimilarly; (ii) that filamentation genes are uniquely up-regulated and opaque-specific genes uniquely down-regulated during C. albicans mating; and (iii) that a newly identified class of genes is selectively down-regulated in opaque, but not white, cells that have entered saturation phase in a growth culture and then are up-regulated by pheromone. The observations that opaque cells are uniquely mating competent, that saturation phase facilitates mating, and that a newly identified group of genes is down-regulated only in opaque cells that have entered saturation phase led us to hypothesize that entering saturation phase may be requisite for mating. A test of this hypothesis revealed, however, that cells, whether in the exponential or saturation phase, may simply have to be in G 1 of the cell cycle to respond to pheromone and that the response includes G 1 arrest. These results add to the lists of similarities and dissimilarities between the mating processes of C. albicans and S. cerevisiae and underscore the unique regulation of filamentation and switching genes in the C. albicans mating process.

The gene encoding yeast-enhanced green fluorescent protein (GFP) was placed under control of ALS gene promoters in Candida albicans. The PALS-GFP reporter strains were validated using various techniques including a new real-time RT-PCR assay to quantify ALS gene expression. The PALS-GFP reporter strains were grown in media that promoted yeast or germ tube forms, and the resulting fluorescence was measured by flow cytometry. In addition to results that indicate differences in ALS gene expression due to growth medium, growth stage and developmental programme, new data show large differences in transcriptional level among the ALS genes. Expression of ALS1 was associated with transfer of the PALS1-GFP strain to fresh growth medium. ALS3 expression increased markedly when germ tubes were visible microscopically and ALS7 expression exhibited a transient peak between 2 and 3 h following inoculation into fresh YPD medium. Transcription from the ALS1 and ALS3 promoters was strongest among those tested and contrasted markedly with the weaker promoter strength at the ALS5, ALS6, ALS7 and ALS9 loci. These weaker transcriptional responses were also observed using real-time RT-PCR measurements on wild-type C. albicans cells. Assuming a positive correlation between transcriptional level and protein production, these results suggest that some Als proteins are abundant on the C. albicans cell surface while others are produced at a much lower level.

Candida albicansstrain SC5314 contains twoALS3alleles, which differ in sequence with respect to the number of copies of the 108 bp tandem repeat sequence within the central domain of the coding region. One allele (ALS3(12)) has 12 tandem repeat copies while the other (ALS3(9)) has 9 copies. Wild-typeC. albicans(ALS3(12)/ALS3(9)) and those containing variousALS3alleles (ALS3(12)/als3Δ(9),als3Δ(12)/ALS3(9) andals3Δ(12)/als3Δ(9)) were assayed for adhesion to monolayers of cultured vascular endothelial and pharyngeal epithelial cells. These assays showed obvious adhesive function for the larger Als3p protein, compared to a minor contribution to adhesion from the smaller protein. These functional differences in strain SC5314 prompted examination ofALS3allelic diversity across the five major genetic clades ofC. albicans. This analysis focused on the number of copies of the tandem repeat sequence within the central domain of the coding region and showed a range of alleles encoding from 6 to 19 tandem repeat copies. Clades differed with respect to prevalentALS3alleles and allele distribution, but were similar for the mean number of tandem repeat copies perALS3allele. Analysis of allelic pairing showed clade differences and the tendency forC. albicansstrains to encode one longer and one shorterALS3allele. The allelic variability observed forALS3and its functional consequences observed in strain SC5314 highlight the importance of understanding ALS allelic diversity in order to draw accurate conclusions about Als protein function.

Summary Determining the ploidy of plant germplasm is a necessary step in breeding or genetic studies in species. The purpose of this research was to determine the presence of ploidy level differentiation of hairy vetch (Vicia villosd) germplasm. Flow cytometry and root tip chromosome squashing methods were employed to assess 45 accessions labeled V. villosa available through the USDA germplasm collection. Flow cytometry determined that 43 of the accessions were 2C, one accession was 4C, and one accession was 6C. Analysis of accessions by root tip chromosome counts indicated that all accessions were diploid. The 2C accession contains 14 chromosomes and their chromosomes were approximately one-half and one-third in size as compared to the chromosomes of the 4C and 6C accessions, respectively. The 4C accession was observed to have 16 chromosomes and the 6C accession was observed to have 14 chromosomes. The large-scale differences in DNA amounts were due to chromosomal size variability as opposed to ploidy differences. This revealed the incidence of species misidentification of these two V. villosa accessions to be Vicia pannonica. All the V. villosa accessions were observed to be diploid and have similar DNA amounts. Flow cytometry proved to be useful in the efficient assessment of these accessions.

Flow cytometry was used to characterize isolates of Phialophora gregata, the causal organism of brown stem rot (BSR) in soybean, using the fluorescence intensity of YOYO-1 stained conidia. The conidia were derived from isolates of P. gregata isolated from field-grown soybean vascular tissue from two locations in Wisconsin. At each location, four soybean cultivars were grown. Two cultivars were susceptible to BSR and two cultivars were resistant to BSR. Ninety-six isolates, one per plant, were characterized for DNA content. The stained conidia were analyzed with an EPICS XL (Coulter Corporation, Hialeah, Florida, USA) flow cytometer cell sorter with CICERO software (San Rafael, CA, USA). The isolates differed in their mean DNA content, ranging from 78.0 to 116.8 arbitrary units. Statistical analysis showed a significant difference in the DNA content variability of the isolates between the two locations. A notable difference in DNA content variability was observed between the isolates collected from the resistant cultivars between the two locations. However, no significant variation occurred in the DNA content of the isolates collected from susceptible cultivars within or between locations. The observed variation among the isolates is hypothesized to be due to the occurrence of separate populations of P. gregata and selection for mean DNA content that could colonize specific plant genotypes.

With the aim of identifying rice (Oryza spp.) germplasm having enhanced grain nutritional value, the mineral nutrient and trace element concentrations (or ionome) of whole (unmilled) grains from a set of 1763 rice accessions of diverse geographic and genetic origin were evaluated. Seed for analysis of P, Mg, K, S, Ca, As, Cd, Co, Cu, Fe, Mn, Mo, Ni, Rb, Sr, and Zn concentrations by inductively coupled plasma mass spectrometry was produced over 2 yr in Beaumont, TX, under both flooded and unflooded watering regimes. The distributions of all element concentrations analyzed were skewed toward higher concentration. A significant portion of this ionomic variation has a genetic basis (broad sense heritabilities 0.14–0.75), indicating an ability to breed for improved grain concentration of all elements except possibly Ni. Variation in grain elemental concentrations was not strongly associated with plant height, heading time, or grain shape, suggesting these physiological factors are not of primary importance in controlling ionomic variation in rice grain. Accessions high in specific elements were sometimes found to have similar genetic or geographic origins, suggesting they share a heritable mechanism underlying their enhanced ionomes. For example, accessions with high Ca, Mg, or K were more common in the indica than in the japonica subgroup; low As was most common among temperate japonica accessions; and several lines high in Mo originated in Malaysia or adjacent Brunei.

Background Vaccination as a control method against the cattle tick, Rhipicephalus (Boophilus) microplus has been practiced since the introduction of two products in the mid-1990s. There is a need for a vaccine that could provide effective control of R. microplus in a more consistent fashion than existing products. During our transcriptome studies of R. microplus, several gene coding regions were discovered to encode proteins with significant amino acid similarity to aquaporins. Methods A cDNA encoding an aquaporin from the cattle tick, Rhipicephalus microplus, was isolated from transcriptomic studies conducted on gut tissues dissected from fully engorged adult female R. microplus. Results Bioinformatic analysis indicates this aquaporin, designated RmAQP1, shows greatest amino acid similarity to the human aquaporin 7 family. Members of this family of water-conducting channels can also facilitate the transport of glycerol in addition to water. The efficacy of this aquaporin as an antigen against the cattle tick was explored in cattle vaccine trials conducted in Brazil. A cDNA encoding a significant portion of RmAQP1 was expressed as a recombinant protein in Pichia pastoris, purified under native conditions using a polyhistidine C-terminus tag and nickel affinity chromatography, emulsified with Montanide adjuvant, and cattle vaccinated intramuscularly. The recombinant protein provided 75% and 68% efficacy in two cattle pen trials conducted in Campo Grande, Brazil on groups of 6 one year old Holstein calves. Conclusion The effectiveness of this vaccine in reducing the numbers of adult female ticks shows this aquaporin antigen holds promise as an active ingredient in cattle vaccines targeted against infestations of R. microplus. Electronic supplementary material The online version of this article (doi:10.1186/s13071-014-0475-9) contains supplementary material, which is available to authorized users.

Two rates (0.4 mg/kg body weight/d and 0.6 mg/kg body weight/d) of a daily feed-through formulation of novaluron (Novaluron 0.67% active ingredient Cattle Mix), a newer benzoylphenyl urea insecticide, were evaluated for efficacy in controlling the larval stage of horn flies, Haematobia irritans (L.), house flies, Musca domestica L., and stable flies, Stomoxys calcitrans (L.), developing in cow manure. Both rates of feed-through novaluron, delivered consecutively for 10 d, reduced adult emergence of all three species when compared with the untreated control. The presence of deformed puparia indicated that novaluron had an insect growth regulator effect on the developing fly larvae. Both of the feed-through rates evaluated resulted in 100% reduction of adult stable fly emergence after the second day of feed-through treatment. The level of control efficacy observed against these three fly species make this feed-through formulation a candidate for use in an integrated livestock pest management program, particularly in confined cattle production situations where a feed-through product could be easily administered.

Next generation sequencing (RNA-seq) technology was used to evaluate the effects of the Ligon lintless-2 (Li2) short fiber mutation on transcriptomes of both subgenomes of allotetraploid cotton (Gossypium hirsutum L.) as compared to its near-isogenic wild type. Sequencing was performed on 4 libraries from developing fibers of Li2 mutant and wild type near-isogenic lines at the peak of elongation followed by mapping and PolyCat categorization of RNA-seq data to the reference D5 genome (G. raimondii) for homeologous gene expression analysis. The majority of homeologous genes, 83.6% according to the reference genome, were expressed during fiber elongation. Our results revealed: 1) approximately two times more genes were induced in the AT subgenome comparing to the DT subgenome in wild type and mutant fiber; 2) the subgenome expression bias was significantly reduced in the Li2 fiber transcriptome; 3) Li2 had a significantly greater effect on the DT than on the AT subgenome. Transcriptional regulators and cell wall homeologous genes significantly affected by the Li2 mutation were reviewed in detail. This is the first report to explore the effects of a single mutation on homeologous gene expression in allotetraploid cotton. These results provide deeper insights into the evolution of allotetraploid cotton gene expression and cotton fiber development.

Harvest index is a measure of success in partitioning assimilated photosynthate. An improvement of harvest index means an increase in the economic portion of the plant. Our objective was to identify genetic markers associated with harvest index traits using 203 O. sativa accessions. The phenotyping for 14 traits was conducted in both temperate (Arkansas) and subtropical (Texas) climates and the genotyping used 154 SSRs and an indel marker. Heading, plant height and weight, and panicle length had negative correlations, while seed set and grain weight/panicle had positive correlations with harvest index across both locations. Subsequent genetic diversity and population structure analyses identified five groups in this collection, which corresponded to their geographic origins. Model comparisons revealed that different dimensions of principal components analysis (PCA) affected harvest index traits for mapping accuracy, and kinship did not help. In total, 36 markers in Arkansas and 28 markers in Texas were identified to be significantly associated with harvest index traits. Seven and two markers were consistently associated with two or more harvest index correlated traits in Arkansas and Texas, respectively. Additionally, four markers were constitutively identified at both locations, while 32 and 24 markers were identified specifically in Arkansas and Texas, respectively. Allelic analysis of four constitutive markers demonstrated that allele 253 bp of RM431 had significantly greater effect on decreasing plant height, and 390 bp of RM24011 had the greatest effect on decreasing panicle length across both locations. Many of these identified markers are located either nearby or flanking the regions where the QTLs for harvest index have been reported. Thus, the results from this association mapping study complement and enrich the information from linkage-based QTL studies and will be the basis for improving harvest index directly and indirectly in rice.

Sheath blight (ShB) caused by the soil-borne pathogen Rhizoctonia solani is one of the most devastating diseases in rice world-wide. Global attention has focused on examining individual mapping populations for quantitative trait loci (QTLs) for ShB resistance, but to date no study has taken advantage of association mapping to examine hundreds of lines for potentially novel QTLs. Our objective was to identify ShB QTLs via association mapping in rice using 217 sub-core entries from the USDA rice core collection, which were phenotyped with a micro-chamber screening method and genotyped with 155 genome-wide markers. Structure analysis divided the mapping panel into five groups, and model comparison revealed that PCA5 with genomic control was the best model for association mapping of ShB. Ten marker loci on seven chromosomes were significantly associated with response to the ShB pathogen. Among multiple alleles in each identified loci, the allele contributing the greatest effect to ShB resistance was named the putative resistant allele. Among 217 entries, entry GSOR 310389 contained the most putative resistant alleles, eight out of ten. The number of putative resistant alleles presented in an entry was highly and significantly correlated with the decrease of ShB rating (r = -0.535) or the increase of ShB resistance. Majority of the resistant entries that contained a large number of the putative resistant alleles belonged to indica, which is consistent with a general observation that most ShB resistant accessions are of indica origin. These findings demonstrate the potential to improve breeding efficiency by using marker-assisted selection to pyramid putative resistant alleles from various loci in a cultivar for enhanced ShB resistance in rice.

Postharvest yellowing (PHY) of rice kernels can be a major problem in the rice industry. This is especially true with high-valued specialty rice, because profit loss will be greater. The objective of this work was to determine whether a significant change occurs in the physicochemical properties (apparent amylose and protein concentrations, viscosity profile and gelatinisation temperature) as a result of induced PHY.In this study, four specialty rices (Basmati, Jasmine, Arborio and Sushi) were yellowed using a laboratory method. PHY increased apparent amylose concentration. It also significantly increased onset and peak gelatinisation temperatures. However, peak, breakdown and setback Rapid ViscoAnalyzer viscosities were decreased by PHY. Trough viscosity for Basmati and Jasmine decreased, whereas it increased for Arborio. Moisture and protein concentrations were unchanged by the yellowing process. Attempts to rehydrate the kernels after induced PHY caused them to fracture, thus making them unsuitable for their intended purpose.This study indicates that rice that has been subjected to PHY shows a reduction not only in appearance but also in cooking and processing quality, decreasing its value. However, the changes differed for each rice type, with Jasmine being affected the least.

Background Cotton fiber length is an important quality attribute to the textile industry and longer fibers can be more efficiently spun into yarns to produce superior fabrics. There is typically a negative correlation between yield and fiber quality traits such as length. An understanding of the regulatory mechanisms controlling fiber length can potentially provide a valuable tool for cotton breeders to improve fiber length while maintaining high yields. The cotton (Gossypium hirsutum L.) fiber mutation Ligon lintless-2 is controlled by a single dominant gene (Li 2 ) that results in significantly shorter fibers than a wild-type. In a near-isogenic state with a wild-type cotton line, Li 2 is a model system with which to study fiber elongation. Results Two near-isogenic lines of Ligon lintless-2 (Li 2 ) cotton, one mutant and one wild-type, were developed through five generations of backcrosses (BC5). An F2 population was developed from a cross between the two Li 2 near-isogenic lines and used to develop a linkage map of the Li 2 locus on chromosome 18. Five simple sequence repeat (SSR) markers were closely mapped around the Li 2 locus region with two of the markers flanking the Li 2 locus at 0.87 and 0.52 centimorgan. No apparent differences in fiber initiation and early fiber elongation were observed between the mutant ovules and the wild-type ones. Gene expression profiling using microarrays suggested roles of reactive oxygen species (ROS) homeostasis and cytokinin regulation in the Li 2 mutant phenotype. Microarray gene expression data led to successful identification of an EST-SSR marker (NAU3991) that displayed complete linkage to the Li 2 locus. Conclusions In the field of cotton genomics, we report the first successful conversion of gene expression data into an SSR marker that is associated with a genomic region harboring a gene responsible for a fiber trait. The EST-derived SSR marker NAU3991 displayed complete linkage to the Li 2 locus on chromosome 18 and resided in a gene with similarity to a putative plectin-related protein. The complete linkage suggests that this expressed sequence may be the Li 2 gene.

Yield is the most important and complex trait for genetic improvement in crops, and marker-assisted selection enhances the improvement efficiency. The USDA rice mini-core collection derived from over 18,000 accessions of global origins is an ideal panel for association mapping. We phenotyped 203 O. sativa accessions for 14 agronomic traits and identified 5 that were highly and significantly correlated with grain yield per plant: plant height, plant weight, tillers, panicle length, and kernels/branch. Genotyping with 155 genome-wide molecular markers demonstrated 5 main cluster groups. Linkage disequilibrium (LD) decayed at least 20 cM and marker pairs with significant LD ranged from 4.64 to 6.06% in four main groups. Model comparisons revealed that different dimensions of principal component analysis affected yield and its correlated traits for mapping accuracy, and kinship did not improve the mapping in this collection. Thirty marker–trait associations were highly significant, 4 for yield, 3 for plant height, 6 for plant weight, 9 for tillers, 5 for panicle length and 3 for kernels/branch. Twenty-one markers contributed to the 30 associations, because 8 markers were co-associated with 2 or more traits. Allelic analysis of OSR13, RM471 and RM7003 for their co-associations with yield traits demonstrated that allele 126 bp of RM471 and 108 bp of RM7003 should receive greater attention, because they had the greatest positive effect on yield traits. Tagging the QTLs responsible for multiple yield traits may simultaneously help dissect the complex yield traits and elevate the efficiency to improve grain yield using marker-assisted selection in rice.

Liu, G., Jia, Y., Correa-Victoria, F. J., Prado, G. A., Yeater, K. M., McClung, A., and Correll, J. C. 2009. Mapping quantitative trait loci responsible for resistance to sheath blight in rice. Phytopathology 99:1078-1084. Rice sheath blight (ShB), caused by the soilborne pathogen Rhizoctonia solani, annually causes severe losses in yield and quality in many rice production areas worldwide. Jasmine 85 is an indica cultivar that has proven to have a high level of resistance to this pathogen. The objective of this study was to determine the ability of controlled environment inoculation assays to detect ShB resistance quantitative trait loci (QTLs) in a cross derived from the susceptible cv. Lemont and the resistant cv. Jasmine 85. The disease reactions of 250 F5 recombinant inbred lines (RILs) were measured on the seedlings inoculated using microchamber and mist-chamber assays under greenhouse conditions. In total, 10 ShBQTLs were identified on chromosomes 1, 2, 3, 5, 6, and 9 using these two methods. The microchamber method identified four of five new ShBQTLs, one on each of chromosomes 1, 3, 5, and 6. Both microchamber and mist-chamber methods identified two ShB-QTLs, qShB1 and qShB92. Four of the ShB-QTLs or ShB-QTL regions identified on chromosomes 2, 3, and 9 were previously reported in the literature. The major ShB-QTL qShB9-2, which cosegregated with simple sequence repeat (SSR) marker RM245 on chromosome 9, contributed to 24.3 and 27.2% of total phenotypic variation in ShB using microchamber and mistchamber assays, respectively. qShB9-2, a plant-stage-independent QTL, was also verified in nine haplotypes of 10 resistant Lemont/Jasmine 85 RILs using haplotype analysis. These results suggest that multiple ShBQTLs are involved in ShB resistance and that microchamber and mistchamber methods are effective for detecting plant-stage-independent QTLs. Furthermore, two SSR markers, RM215 and RM245, are robust markers and can be used in marker-assisted breeding programs to improve ShB resistance.

Recent culture-independent studies have revealed that a healthy vaginal ecosystem harbors a surprisingly complex assemblage of microorganisms. However, the spatial distribution and composition of vaginal microbial populations have not been investigated using molecular methods. Here, we evaluated site-specific microbial composition within the vaginal ecosystem and examined the influence of sampling technique in detection of the vaginal microbiota. 16S rRNA gene clone libraries were prepared from samples obtained from different locations (cervix, fornix, outer vaginal canal) and by different methods (swabbing, scraping, lavaging) from the vaginal tracts of eight clinically healthy, asymptomatic women. The data reveal that the vaginal microbiota is not homogenous throughout the vaginal tract but differs significantly within an individual with regard to anatomical site and sampling method used. Thus, this study illuminates the complex structure of the vaginal ecosystem and calls for the consideration of microenvironments when sampling vaginal microbiota as a clinical predictor of vaginal health.

Gene expression profiles of developing cotton (Gossypium hirsutum L.) fibers from two near-isogenic lines (NILs) that differ in fiber-bundle strength, short-fiber content, and in fewer than two genetic loci were compared using an oligonucleotide microarray. Fiber gene expression was compared at five time points spanning fiber elongation and secondary cell wall (SCW) biosynthesis. Fiber samples were collected from field plots in a randomized, complete block design, with three spatially distinct biological replications for each NIL at each time point. Microarray hybridizations were performed in a loop experimental design that allowed comparisons of fiber gene expression profiles as a function of time between the two NILs. Overall, developmental expression patterns revealed by the microarray experiment agreed with previously reported cotton fiber gene expression patterns for specific genes. Additionally, genes expressed coordinately with the onset of SCW biosynthesis in cotton fiber correlated with gene expression patterns of other SCW-producing plant tissues. Functional classification and enrichment analysis of differentially expressed genes between the two NILs revealed that genes associated with SCW biosynthesis were significantly up-regulated in fibers of the high-fiber quality line at the transition stage of cotton fiber development. For independent corroboration of the microarray results, 15 genes were selected for quantitative reverse transcription PCR analysis of fiber gene expression. These analyses, conducted over multiple field years, confirmed the temporal difference in fiber gene expression between the two NILs. We hypothesize that the loci conferring temporal differences in fiber gene expression between the NILs are important regulatory sequences that offer the potential for more targeted manipulation of cotton fiber quality.

Microarrays were used to identify changes in gene expression associated with Candida albicans biofilm development. Two biofilm substrates (denture and catheter), and two C. albicans strains for each substrate, were tested to remove model- and strain-dependent variability from the overall dataset. Three biofilm developmental phases were examined: early (6 h), intermediate (12 h), and mature (48 h). Planktonic specimens were collected at the same time points. Data analysis focused primarily on gene expression changes over the time-course of biofilm development. Glycolytic and non-glycolytic carbohydrate assimilation, amino acid metabolism, and intracellular transport mechanisms were important during the early phase of biofilm formation. These early events increase intracellular pools of pyruvate, pentoses and amino acids, which prepare the biofilm for the large biomass increase that begins around 12 h of development. This developmental stage demands energy and utilizes specific transporters for amino acids, sugars, ions, oligopeptides and lactate/pyruvate. At mature phase (48 h), few genes were differentially expressed compared with the 12 h time point, suggesting a relative lack of initiation of new metabolic activity. Data analysis to assess biofilm model-specific gene expression showed more dynamic changes in the denture model than in the catheter model. Data analysis to identify gene expression changes that are associated with each strain/substrate combination identified the same types of genes that were identified in the analysis of the entire dataset. Collectively, these data suggest that genes belonging to different, but interconnected, functional categories regulate the morphology and phenotype of C. albicans biofilm.

The number of cotton (Gossypium sp.) ovule epidermal cells differentiating into fiber initials is an important factor affecting cotton yield and fiber quality. Despite extensive efforts in determining the molecular mechanisms regulating fiber initial differentiation, only a few genes responsible for fiber initial differentiation have been discovered. To identify putative genes directly involved in the fiber initiation process, we used a cotton ovule culture technique that controls the timing of fiber initial differentiation by exogenous phytohormone application in combination with comparative expression analyses between wild type and three fiberless mutants. The addition of exogenous auxin and gibberellins to pre-anthesis wild type ovules that did not have visible fiber initials increased the expression of genes affecting auxin, ethylene, ABA and jasmonic acid signaling pathways within 1 h after treatment. Most transcripts expressed differentially by the phytohormone treatment in vitro were also differentially expressed in the ovules of wild type and fiberless mutants that were grown in planta. In addition to MYB25-like, a gene that was previously shown to be associated with the differentiation of fiber initials, several other differentially expressed genes, including auxin/indole-3-acetic acid (AUX/IAA) involved in auxin signaling, ACC oxidase involved in ethylene biosynthesis, and abscisic acid (ABA) 8'-hydroxylase an enzyme that controls the rate of ABA catabolism, were co-regulated in the pre-anthesis ovules of both wild type and fiberless mutants. These results support the hypothesis that phytohormonal signaling networks regulate the temporal expression of genes responsible for differentiation of cotton fiber initials in vitro and in planta.

Published in Agron. J. 107:691 (2015) doi:10.2134/agronj14.0888 Available freely online through the author-supported open access option. Copyright © 2015 by the American Society of Agronomy, 5585 Guilford Road, Madison, WI 53711. All rights reserved. No part of this periodical may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Supplemental material available online. B. Glaz, retired; J. Wiersma, University of Minnesota, Northwest Research & Outreach Center, 2900 University of Minnesota, Crookston, MN 56716; J.A. Hernandez, University of Minnesota, 1991 Upper Buford Circle, Saint Paul, MN 55108. N. Martin, Syngenta Biotechnology, Stanton MN; and K.M. Yeater, USDA-ARS-PANRRC, Office of the Director, Fort Collins, CO 80526. All five authors comprise the Agronomy Journal Statistics Committee. Received 19 Dec. 2014. Accepted 22 Dec. 2014. *Corresponding author (barry.glaz@gmail.com). Mark Twain popularized the saying

Candida albicans is the most common etiological agent of vaginal candidiasis. Elevated host estrogen levels and the incidence of vaginal candidiasis are positively associated. Elevated estrogen levels may affect host and/or fungal cells. This study investigates the effect of 17-β-estradiol, 17-α-estradiol, ethynyl estradiol, and estriol on several C. albicans strains at concentrations ranging from 10 −5 to 10 −10 M. The addition of 17-β-estradiol or ethynyl estradiol to C. albicans cells caused an increase in the number of cells forming germ tubes and an increase in germ tube length in a dose- and strain-dependent manner. The addition of 17-α-estradiol or estriol did not have a significant effect on germ tube formation by the cultured cells. Exposure to exogenous estrogens did not significantly change the biomass of any C. albicans culture tested. The transcriptional profile of estrogen-treated C. albicans cells showed increased expression of CDR1 and CDR2 across several strain-estrogen concentration-time point combinations, suggesting that these genes are the most responsive to estrogen exposure. Analysis of strain DSY654, which lacks the CDR1 and CDR2 coding sequences, showed a significantly decreased number of germ tube-forming cells in the presence of 17-β-estradiol. PDR16 was the most highly up-regulated gene in strain DSY654 under these growth conditions. The cell biology and gene expression data from this study led to a model that proposes how components of the phospholipid and sterol metabolic pathways may interact to affect C. albicans germ tube formation and length.

The ALS (agglutinin-likesequence) gene family encodes eight large cell-surface glycoproteins. The work presented here focuses on Als2p and Als4p, and is part of a larger effort to deduce the function of each Als protein. BothALS4alleles were deleted from theCandida albicansgenome and the phenotype of the mutant strain (als4Δ/als4Δ; named 2034) studied. Loss of Als4p slowed germ tube formation of cells grown in RPMI 1640 medium and resulted in decreased adhesion ofC. albicansto vascular endothelial cells. Loss of Als4p did not affect adhesion to buccal epithelial cells, biofilm formation in a catheter model, or adhesion to or destruction of oral reconstituted human epithelium (RHE). Although deletion of oneALS2allele was achieved readily, a strain lacking the second allele was not identified despite screening thousands of transformants. The remainingALS2allele was placed under control of theC. albicans MAL2promoter to create anals2Δ/PMAL2-ALS2strain (named 2342). Real-time RT-PCR analysis of strain 2342 grown in glucose-containing medium (non-inducing conditions) showed that althoughALS2transcript levels were greatly reduced compared to wild-type cells, someALS2transcript remained. The decreasedALS2expression levels were sufficient to slow germ tube formation in RPMI 1640 and Lee medium, reduce adhesion to vascular endothelial cells and to RHE, decrease RHE destruction, and impair biofilm formation. Growth of strain 2342 in maltose-containing medium (inducing conditions) restored the wild-type phenotype in all assays. Real-time RT-PCR analysis demonstrated that in maltose-containing medium, strain 2342 overexpressedALS2compared to wild-type cells; however no overexpression phenotype was apparent. Microarray analysis revealed little transcriptional response toALS4deletion, but showed twofold up-regulation of orf19.4765 in the glucose-medium-grownals2Δ/PMAL2-ALS2strain. orf19.4765 encodes a protein with features of a glycosylated cell wall protein with similarity toSaccharomyces cerevisiaeCcw12p, although initial analysis suggested functional differences between the two proteins. Real-time RT-PCR measurement ofALS2andALS4transcript copy number showed a 2·8-fold increase inALS2expression in theals4Δ/als4Δ strain and a 3·2-fold increase inALS4expression in theals2Δ/PMAL2-ALS2strain, suggesting the potential for compensatory function between these related proteins.

The ALS (agglutinin-likesequence) gene family ofCandida albicansencodes eight cell-surface glycoproteins, some of which are involved in adherence to host surfaces. A mutational analysis of each ALS gene is currently being performed to deduce the functions of the encoded proteins and to better understand the role of these proteins inC. albicansbiology and pathogenesis. This paper describes construction of anals3/als3mutant and comparison of its phenotype to anals1/als1strain. Efforts to disruptALS3indicated that the gene could be deleted in two transformation steps, suggesting that the gene is encoded by a single locus and that theALS3-like locus,ALS8, does not exist. Strains lackingALS3orALS1did not exhibit a defect in germ tube formation when grown in RPMI 1640 medium, but theals1/als1mutant formed significantly fewer germ tubes in Lee medium. Analysis ofALS3andALS1promoter activity using green fluorescent protein (GFP) reporter strains and flow cytometry showed that when cells are placed into medium that promotes germ tube formation,ALS1is transcribed prior toALS3. Comparison of the mutant strains in adhesion assays showed that theals3/als3strain was defective in adhesion to both human umbilical vein endothelial cells (HUVEC) and buccal epithelial cells (BEC), but not to fibronectin-coated plastic plates. In contrast, theals1/als1strain showed decreased adherence to HUVEC, but adherence to BEC and fibronectin were the same as wild-type controls. Inoculation of the buccal reconstituted human epithelium (RHE) model of oral candidiasis with the mutant strains showed nearly a total lack of adhesion and epithelial destruction by theals3/als3mutant while theals1/als1strain showed only a slightly reduced degree of epithelial destruction compared to the wild-type control. Adhesion data presented here suggest that, in the assays performed, loss of Als3p affectsC. albicansadhesion more than loss of Als1p. Collectively, these results demonstrate functional similarities and differences between Als1p and Als3p, and suggest the potential for more complex interrelationships between the ALS genes and their encoded proteins.

Reports of genome size variation in soybean [Glycine max (L.) Merr.] have ranged from 40 to 0%. This wide range has resulted in doubts of the existence of intraspecific DNA variation in soybean. Eighteen soybean lines were analyzed by flow cytometry to determine their genome size. The lines were selected on the basis of diversity of origin. Intraspecific genome size variation was observed at approximately 4%. To ensure that the variation observed was reproducible and not due to technique error, the two highest and lowest genomes size accessions were reanalyzed. The order and variation observed between the high and low genome size accessions were maintained. To ensure further that the differences were reproducible, seeds from the two highest and lowest genome size accessions were planted in different locations in the USA, grown to maturity, harvested, and the seeds returned to Illinois. The harvested seed was analyzed and again the order and variation in genome size between the high and low genome size accessions were similar to the previous two analyses even though more than 1 yr had passed between the analysis. In addition, two experiments using Amaranthus palmeri S. Wats as an internal standard were conducted. In both of these experiments, the observed variation between the previously reported high and low genome size soybean lines was approximately 1 to 2%. The variation between the high and low genome size soybean lines is reproducible. The variation reported here indicates that the DNA amount variation is between I and 4%, lower than was originally reported.

For improvement of hairy vetch (Vicia villosa Roth) as a winter cover crop, it is necessary to gain insight into the magnitude of variability present in the species. This study was conducted to assess the sources of genetic and phenotypic variability of V. villosa accessions. Thirteen morphological and agronomic traits were measured on 42 populations of V. villosa and two populations of V. pannonica Crantz in field studies at Urbana, IL, for 2 yr. These measurements include initial seed weight, germination, stem length, stem width, leaf length, and leaf width in the fall and spring, winter survival, biomass, and a ratio of organic C to total N (C:N ratio) of the plants. The multivariate data set was analyzed by canonical discriminant analysis (CDA) in combination with a clustering procedure. In this analysis, the first two canonical variates were significant and accounted for 94% of the among-accession variability. The canonical variates indicated that fall and spring measurements of the leaf and stem, and C:N ratio are the most differentiating traits among the accessions. The canonical variates were used to cluster the accessions into four subgroups on the basis of the differentiating traits. Canonical discriminant analysis was useful in identifying the genetic variation and the traits that better describe the variation among hairy vetch populations. Cluster analysis was successful in differentiating the accessions into similar subgroups on the basis of the measured traits. Plant breeders can use the information on variation among Vicia accessions and focus on traits of particular significance.