Exploring the Mycovirus Sclerotinia sclerotiorum Hypovirulence-Associated DNA Virus 1 as a Biocontrol Agent of White Mold Caused by Sclerotinia sclerotiorum.

Sclerotinia sclerotiorum is a necrotrophic fungal pathogen causing white mold on many important economic crops. Recently, some mycoviruses such as S. sclerotiorum hypovirulence-associated DNA virus 1 (SsHADV-1) converted S. sclerotiorum into a beneficial symbiont that helps plants manage pathogens and other stresses. To explore the potential use of SsHADV-1 as a biocontrol agent in the United States and to test the efficacy of SsHADV-1-infected United States isolates in managing white mold and other crop diseases, SsHADV-1 was transferred from the Chinese strain DT-8 to United States isolates of S. sclerotiorum. SsHADV-1 is readily transmitted horizontally among United States isolates of S. sclerotiorum and consistently conferred hypovirulence to its host strains. Biopriming of dry bean seeds with hypovirulent S. sclerotiorum strains enhanced resistance to white mold, gray mold, and Rhizoctonia root rot. To investigate the underlying mechanisms, endophytic growth of hypovirulent S. sclerotiorum in dry beans was confirmed using PCR, and the expression of 12 plant defense-related genes were monitored before and after infection. The results indicated that the endophytic growth of SsHADV-1-infected strains in plants stimulated the expression of plant immunity pathway genes that assisted a rapid response from the plant to fungal infection. Finally, application of the seed biopriming technology with SsHADV-1-infected hypervirulent strain has promise for the biological control of several diseases of wheat, pea, and sunflower.

Suitable Methods of Inoculation and Quantification of Fusarium Root Rot in Lentil.

Lentil (Lens culinaris L. subsp. culinaris) is an important grain legume grown worldwide. As its popularity grows among consumers and more acres are produced, new root rot complexes have become more prevalent. This work sought to develop methods for studying root rot caused by Fusarium avenaceum in lentil using controlled environments. The objectives were to (i) find an effective and seed-safe sterilization technique, (ii) optimize the inoculation technique and lentil growing environment, and (iii) develop visual and automated disease scoring systems. Results showed the use of detergent and a low concentration (0.1%) of NaClO (the active ingredient in bleach) maintained germinability and effectively eliminated bacterial and fungal contamination on seeds. Other treatments, such as ethanol, reduced seed germination or failed to kill pathogenic fungi such as Fusarium spp. Placing inoculum at a moderate rate of 1 × 106 spores both directly on the seed and on top of the media covering the seed improved severity scores and reduced escapes compared with placement on top of the media only. Visual severity scoring systems and diagrammatic scales were developed for scoring the cotyledon region and roots. A computer vision algorithm was designed to improve the efficiency of scoring the cotyledon region and roots for disease severity using a simple RGB camera and lightbox. Visual and computer scores were best correlated when images were visually scored on a monitor, and multiple images were averaged. The scores generated from the computer vision algorithm had better correlations with visual scores for cotyledon rot (r = 0.92 and ß1 = 0.96) than root rot (r = 0.62 and ß1 = 0.67).

Phenotypic and Genetic Characterization of the Lentil Single Plant-Derived Core Collection for Resistance to Root Rot Caused by Fusarium avenaceum.

Lentil (Lens culinaris) is a pulse crop grown for its amino acid profile, moderate drought tolerance, and ability to fix nitrogen. As the global demand for lentils expands and new production regions emerge so too have the complement of diseases that reduce yield, including the root rot complex. Although the predominant causal pathogen varies based on growing region, Fusarium avenaceum is often found to be an important contributor to disease. This study screened part of the lentil single plant-derived core collection for resistance to F. avenaceum in a greenhouse. Plants were phenotyped for disease severity using three scoring scales and the differences in biomass traits due to pathogen presence were measured. Lentil accessions varied in disease severity and differences in biomass traits were found to be correlated with each visual severity estimate (r = -0.37 to -0.63, P < 0.001), however, heritability estimates were low to moderate among traits (H2 = 0.12 to 0.43). Results of a genome-wide association study (GWAS) using single nucleotide polymorphism (SNP) markers derived from genotyping-by-sequencing revealed 11 quantitative trait loci (QTL) across four chromosomes. Two pairs of QTL colocated for two traits and were found near putative orthologs that have been previously associated with plant disease resistance. The identification of lentil accessions that did not exhibit a difference in biomass traits may serve as parental material in breeding or in the development of biparental mapping populations to further validate and dissect the genetic control of resistance to root rot caused by F. avenaceum.

Consequences of underexplored variation in biodiversity indices used for land-use prioritization.

For biodiversity protection to play a persuasive role in land-use planning, conservationists must be able to offer objective systems for ranking which natural areas to protect or convert. Representing biodiversity in spatially explicit indices is challenging because it entails numerous judgments regarding what variables to measure, how to measure them, and how to combine them. Surprisingly few studies have explored this variation. Here, we explore how this variation affects which areas are selected for agricultural conversion by a land-use prioritization model designed to reduce the biodiversity losses associated with agricultural expansion in Zambia. We first explore the similarity between model recommendations generated by three recently published composite indices and a commonly used rarity-weighted species richness metric. We then explore four underlying sources of ecological and methodological variation within these and other approaches, including different terrestrial vertebrate taxonomic groups, different species-richness metrics, different mathematical methods for combining layers, and different spatial resolutions of inputs. The results generated using different biodiversity approaches show very low spatial agreement regarding which areas to convert to agriculture. There is little overlap in areas identified for conversion using previously published indices (mean Jaccard similarity, Jw , between 0.3 and 3.7%), different taxonomic groups (5.0% < mean Jw  < 13.5%), or different measures of species richness (15.6% < mean Jw  < 33.7%). Even with shared conservation goals, different methods for combining layers and different input spatial resolutions still produce meaningful, though smaller, differences among areas selected for conversion (40.9% < mean Jw  < 67.5%). The choice of taxonomic group had the largest effect on conservation priorities, followed by the choice of species richness metric, the choice of combination method, and finally the choice of spatial resolution. These disagreements highlight the challenge of objectively representing biodiversity in land-use planning tools, and present a credibility challenge for conservation scientists seeking to inform policy making. Our results suggest an urgent need for a more consistent and transparent framework for designing the biodiversity indices used in land-use planning, which we propose here.

The effects of sub-bandgap transitions and the defect density of states on the photocurrent response of a single ZnO-coated silica nanospring.

The electrical and optoelectronic properties of nanometer-sized ZnO structures are highly influenced by its native point defects. Understanding and controlling these defects are essential for the development of high-performance ZnO-based devices. Here, an electrical device consisting of a polycrystalline ZnO-coated silica nanospring was fabricated and used to characterize the electrical and photoconductive properties of the ZnO layer using near-UV (405 nm) and sub-bandgap (532 and 633 nm) excitation sources. We observe a photocurrent response with all three wavelengths and notably with 532 nm green illumination, which is the energy associated with deep oxygen vacancies. The polycrystalline ZnO-coated silica nanospring exhibits a high responsivity of 1740 A W-1 with the 405 nm excitation source. Physical models are presented to describe the photocurrent rise and decay behavior of each excitation source where we suggest that the rise and decay characteristics are highly dependent on the energy of the excitation source and the trapping of electrons and holes in intermediate defect levels in the bandgap. The energy levels of the trap depths were determined from the photoconductive decay data and are matched to the reported energy levels of singly and doubly ionized oxygen vacancies. A phenomenological model to describe the dependence of the saturation photocurrent on excitation intensity is presented in order to understand the characteristics of the observed breaks in the slopes of the saturation photocurrent versus excitation intensity profile.

KASP Markers Reveal Established and Novel Sources of Resistance to Pea Seedborne Mosaic Virus in Pea Genetic Resources.

Pea seed-borne mosaic virus (PSbMV) is both seedborne and aphid-transmitted and can cause economic losses for pea (Pisum sativum L.) production by reducing yield through decreased seed weight and number. The P1 pathotype is especially virulent, affecting this important vegetable crop across the United States and internationally in regions of West Asia, North Africa, Europe, and Australia. Previously, two kompetitive allele-specific PCR (KASP) genotyping markers (eIF4E resistant 1 and 2) were developed and validated on P. sativum accessions identifying two PSbMV pathotype P1 resistance alleles in the eukaryotic translation initiation factor gene, eIF4E. The current study utilized these novel markers to rapidly evaluate 318 genetic resource accessions maintained as part of the United States Department of Agriculture National Plant Germplasm System's Pea Single Plant Collection (PSPC). The evaluations also included 58 commercial and other plant introduction (PI) lines that were assessed for the two eIF4E resistance alleles. All genotyping results were validated in greenhouse assays by confirmation of observable disease symptoms after inoculations and by enzyme-linked immunosorbent assays. The eIF4E resistant 1 and 2 alleles were found in 18 accessions from the PSPC, five commercial lines, and 14 other PI accessions. A single PSPC accession showed resistance to PSbMV pathotype P1 that is believed to be a novel source of resistance based on sequencing analysis of eIF4E. Sources of resistance were identified in the PSPC and in commercial cultivars that can be introgressed into breeding lines using traditional techniques to reduce the time and cost required to generate germplasm with superior disease-resistant traits.

Development and Validation of KASP Markers for the Identification of Pea seedborne mosaic virus Pathotype P1 Resistance in Pisum sativum.

As pesticides have become heavily relied on for management of insect pests vectoring economically important pathogens of vegetable crops, development of pathogen-resistant germplasm remains a promising alternative to reduce or eliminate costly and timely chemical inputs. Molecular markers can be used to rapidly identify resistant genotypes to aid breeders in advancing germplasm. This study developed two kompetitive allele-specific PCR (KASP) genotyping markers for rapid screening of Pisum sativum genotypes for resistance to Pea seedborne mosaic virus pathotype P1 (PSbMV-P1), the most economically devastating strain worldwide. The KASP markers differentiate two eIF4E PSbMV-P1-resistant allelic variants from susceptible eIF4E variants. A single nucleotide polymorphism (Resistant 1) and a 3-basepair deletion (Resistant 2) present in either of the two resistant alleles were used for marker design. Forty-four P. sativum lines previously characterized for resistance to PSbMV were inoculated with PSbMV-P1 in a greenhouse, observed for visual symptoms, assayed for virus susceptibility by enzyme-linked immunosorbent assay (ELISA), and genotyped by KASP marker analysis. The KASP markers were 100% accurate in characterizing PSbMV-P1-susceptible and PSbMV-P1-resistant genotypes when correlated with the ELISA results. The Resistant 1 marker also correlated with resistance to PSbMV pathotypes P2 and P4 completely, making this marker a new advanced tool for P. sativum breeding programs.

Dissecting the Genetic Architecture of Aphanomyces Root Rot Resistance in Lentil by QTL Mapping and Genome-Wide Association Study.

Lentil (Lens culinaris Medikus) is an important source of protein for people in developing countries. Aphanomyces root rot (ARR) has emerged as one of the most devastating diseases affecting lentil production. In this study, we applied two complementary quantitative trait loci (QTL) analysis approaches to unravel the genetic architecture underlying this complex trait. A recombinant inbred line (RIL) population and an association mapping population were genotyped using genotyping by sequencing (GBS) to discover novel single nucleotide polymorphisms (SNPs). QTL mapping identified 19 QTL associated with ARR resistance, while association mapping detected 38 QTL and highlighted accumulation of favorable haplotypes in most of the resistant accessions. Seven QTL clusters were discovered on six chromosomes, and 15 putative genes were identified within the QTL clusters. To validate QTL mapping and genome-wide association study (GWAS) results, expression analysis of five selected genes was conducted on partially resistant and susceptible accessions. Three of the genes were differentially expressed at early stages of infection, two of which may be associated with ARR resistance. Our findings provide valuable insight into the genetic control of ARR, and genetic and genomic resources developed here can be used to accelerate development of lentil cultivars with high levels of partial resistance to ARR.

Protein, weight, and oil prediction by single-seed near-infrared spectroscopy for selection of seed quality and yield traits in pea (Pisum sativum).

BACKGROUND: Pea (Pisum sativum) is a prevalent cool-season crop that produces seeds valued for their high protein content. Modern cultivars have incorporated several traits that improved harvested yield. However, progress toward improving seed quality has received less emphasis, in part due to the lack of tools for easily and rapidly measuring seed traits. In this study we evaluated the accuracy of single-seed near-infrared spectroscopy (NIRS) for measuring pea-seed weight, protein, and oil content. A total of 96 diverse pea accessions were analyzed using both single-seed NIRS and wet chemistry methods. To demonstrate field relevance, the single-seed NIRS protein prediction model was used to determine the impact of seed treatments and foliar fungicides on the protein content of harvested dry peas in a field trial. RESULTS: External validation of partial least squares (PLS) regression models showed high prediction accuracy for protein and weight (R2 = 0.94 for both) and less accuracy for oil (R2 = 0.74). Single-seed weight was weakly correlated with protein and oil content in contrast with previous reports. In the field study, the single-seed NIRS predicted protein values were within 10 mg g-1 of an independent analytical reference measurement and were sufficiently precise to detect small treatment effects. CONCLUSION: The high accuracy of protein and weight estimation show that single-seed NIRS could be used in the dual selection of high-protein, high-weight peas early in the breeding cycle, allowing for faster genetic advancement toward improved pea nutritional quality. © 2020 Society of Chemical Industry.

Confirmation of Fusarium root rot resistance QTL Fsp-Ps 2.1 of pea under controlled conditions.

BACKGROUND: Dry pea production has increased substantially in North America over the last few decades. With this expansion, significant yield losses have been attributed to an escalation in Fusarium root rots in pea fields. Among the most significant rot rotting pathogenic fungal species, Fusarium solani fsp. pisi (Fsp) is one of the main causal agents of root rot of pea. High levels of partial resistance to Fsp has been identified in plant genetic resources. Genetic resistance offers one of the best solutions to control this root rotting fungus. A recombinant inbred population segregating for high levels of partial resistance, previously single nucleotide polymorphism (SNP) genotyped using genotyping-by-sequencing, was phenotyped for disease reaction in replicated and repeated greenhouse trials. Composite interval mapping was deployed to identify resistance-associated quantitative trait loci (QTL). RESULTS: Three QTL were identified using three disease reaction criteria: root disease severity, ratios of diseased vs. healthy shoot heights and dry plant weights under controlled conditions using pure cultures of Fusarium solani fsp. pisi. One QTL Fsp-Ps 2.1 explains 44.4-53.4% of the variance with a narrow confidence interval of 1.2 cM. The second and third QTL Fsp-Ps3.2 and Fsp-Ps3.3 are closely linked and explain only 3.6-4.6% of the variance. All of the alleles are contributed by the resistant parent PI 180693. CONCLUSION: With the confirmation of Fsp-Ps 2.1 now in two RIL populations, SNPs associated with this region make a good target for marker-assisted selection in pea breeding programs to obtain high levels of partial resistance to Fusarium root rot caused by Fusarium solani fsp. pisi.

Electrical characterization of ZnO-coated nanospring ensemble by impedance spectroscopy: probing the effect of thermal annealing.

The effects of thermal annealing on the electrical properties of randomly oriented ZnO-coated nanospring ensembles were extensively investigated through AC impedance spectroscopy. Annealing the nanospring mats for an hour at 873 K in air showed significant change in ZnO morphology, reduced electrical conductivity due to the presence of grain boundaries, decreased apparent donor concentration, and faster decay of sub-band gap photocurrent. The role of the nanospring-nanospring junctions in the conduction of carriers in the ensemble was also examined, as well as evaluation of their responsiveness to thermal and optical stimulations. This work identifies the effects of heat treatment in the presence of air on the electrical properties of the nanospring ensembles, which are related to the mesoscopic morphology and interconnect within the ensemble and the properties of the ZnO coating.

Discerning the Impact of a Lithium Salt Additive in Thin-Film Light-Emitting Electrochemical Cells with Electrochemical Impedance Spectroscopy.

Light-emitting electrochemical cells (LEECs) from small molecules, such as iridium complexes, have great potential as low-cost emissive devices. In these devices, ions rearrange during operation to facilitate carrier injection, bringing about efficient operation from simple, single-layer devices. Prior work has shown that the luminance, efficiency, and responsiveness of iridium LEECs is greatly enhanced by the inclusion of small fractions of lithium salts, but much remains to be understood about the origin of this enhancement. Recent work with planar devices demonstrates that lithium additives in iridium LEECs enhance double-layer formation. However, the quantitative influence of lithium salts on the underlying physics of conventional thin-film, sandwich structure LEECs, which beneficially operate at low voltages and generate higher luminance, has yet to be clarified. Here, we use electrochemical impedance spectroscopy to discern the impact of the lithium salt concentration on double-layer formation within the device and draw correlations with performance metrics, such as current, luminance, and external quantum efficiency.

Pythium Species Associated with Damping-off of Pea in Certified Organic Fields in the Columbia Basin of Central Washington.

Organic vegetable production accounted for 19% of the total organic acreage in Washington State in 2013, with 1,700 ha of certified organic vegetable pea. However, production is challenged constantly with the threat of poor emergence after planting due to damping-off caused by Pythium spp. A survey of Pythium spp. in organic vegetable production areas of the semiarid Columbia Basin of central Washington was carried out in fall 2009 to identify species associated with damping-off during early spring planting. Of 305 isolates baited from soil sampled from 37 certified organic fields, 264 were identified to 16 Pythium spp. by sequencing the internal transcribed spacer region of ribosomal DNA. A soil DNA-CFU regression curve was developed using real-time quantitative polymerase chain reaction assays for each of the three predominant pathogenic species (Pythium abappressorium, the P. irregulare complex, and P. ultimum var. ultimum) found in soil sampled from the 37 fields. The P. irregulare complex, P. abappressorium, and P. ultimum var. ultimum were detected in 57, 78, and 100% of the fields sampled, respectively. A regression analysis was used to determine that P. ultimum var. ultimum ranged from 14 to 332 CFU/g of soil in the 37 fields, the P. irregulare complex ranged from 25 to 228 CFU/g of soil, and P. abappressorium DNA was below the quantifiable limit. In summary, P. ultimum var. ultimum was the most prevalent pathogenic Pythium sp. detected in certified organic fields in the semiarid Columbia Basin of central Washington but multiple Pythium spp. may be associated with damping-off in cool and wet, early spring planting conditions.

Characterization and Pathogenicity of Rhizoctonia and Rhizoctonia-Like spp. From Pea Crops in the Columbia Basin of Oregon and Washington.

Isolates of Rhizoctonia and Rhizoctonia-like spp. (n = 179) were baited selectively from soil and plant samples collected from irrigated pea crops in the semiarid Columbia Basin of Oregon and Washington from 2011 to 2013, and characterized to species, subspecies, and anastomosis groups (AG) based on sequences of the internal transcribed spacer region of ribosomal DNA. Rhizoctonia solani comprised 76% of all isolates, and included isolates of AG 4 (31% of all isolates), AG 2-1 (18%), AG 3 (10%), AG 8 (8%), AG 5 (5%), AG 10 (3%), and AG 9 (1%). The isolates of Ceratobasidium spp. (20%) comprised four AGs: AG K (11%), AG A (6%), AG I (2%), and AG I-like (1%). Waitea circinata isolates (4%) comprised two subspecies: W. circinata var. circinata (approximately 4%) and W. circinata var. zeae (<1%). Repeated pathogenicity tests of isolates of the 10 most frequently detected AGs and subspecies on 'Serge' pea at 15°C revealed that R. solani AG 2-1 caused the greatest reduction in pea emergence, followed by R. solani AG 4. R. solani AG 4 caused the most severe root rot, stunting, and reduction in pea seedling biomass, followed by isolates of AG 2-1. R. solani AG 8 did not affect emergence, plant height, and total biomass compared with noninoculated control plants; however, root rot caused by isolates of AG 8 was ranked the third most severe among isolates of the 10 Rhizoctonia subgroups, after that caused by isolates of AG 4 and AG 2-1. Isolates of other AGs and subspecies were either weakly virulent or nonpathogenic on pea. The most common AGs (AG 4 and AG 2-1) detected in pea fields in the Columbia Basin were also the most virulent. In a growers' pea crop grown for seed ('Prevail') planted 5 days after herbicide application and incorporation of a preceding winter wheat crop, severe stunting caused by Rhizoctonia spp. resulted in an average 75% yield loss within patches of stunted plants. In contrast, the yield of processing pea from a green pea crop of Serge did not differ significantly for plants sampled within versus outside patches of stunted plants; however, plants within patches were significantly more mature. In the Prevail seed crop, a greater frequency of R. solani AG 8 was detected than AG 2-1 or AG 4 from within patches of stunted plants, indicating that isolates of AG 8 may be associated with the root rot complex in some pea crops in the Columbia Basin.

Using changes in agricultural utility to quantify future climate-induced risk to conservation.

Much of the biodiversity-related climate change impacts research has focused on the direct effects to species and ecosystems. Far less attention has been paid to the potential ecological consequences of human efforts to address the effects of climate change, which may equal or exceed the direct effects of climate change on biodiversity. One of the most significant human responses is likely to be mediated through changes in the agricultural utility of land. As farmers adapt their practices to changing climates, they may increase pressure on some areas that are important to conserve (conservation lands) whereas lessening it on others. We quantified how the agricultural utility of South African conservation lands may be altered by climate change. We assumed that the probability of an area being farmed is linked to the economic benefits of doing so, using land productivity values to represent production benefit and topographic ruggedness as a proxy for costs associated with mechanical workability. We computed current and future values of maize and wheat production in key conservation lands using the DSSAT4.5 model and 36 crop-climate response scenarios. Most conservation lands had, and were predicted to continue to have, low agricultural utility because of their location in rugged terrain. However, several areas were predicted to maintain or gain high agricultural utility and may therefore be at risk of near-term or future conversion to cropland. Conversely, some areas were predicted to decrease in agricultural utility and may therefore prove easier to protect from conversion. Our study provides an approximate but readily transferable method for incorporating potential human responses to climate change into conservation planning.

Projected climate impacts to South African maize and wheat production in 2055: a comparison of empirical and mechanistic modeling approaches.

Crop model-specific biases are a key uncertainty affecting our understanding of climate change impacts to agriculture. There is increasing research focus on intermodel variation, but comparisons between mechanistic (MMs) and empirical models (EMs) are rare despite both being used widely in this field. We combined MMs and EMs to project future (2055) changes in the potential distribution (suitability) and productivity of maize and spring wheat in South Africa under 18 downscaled climate scenarios (9 models run under 2 emissions scenarios). EMs projected larger yield losses or smaller gains than MMs. The EMs' median-projected maize and wheat yield changes were -3.6% and 6.2%, respectively, compared to 6.5% and 15.2% for the MM. The EM projected a 10% reduction in the potential maize growing area, where the MM projected a 9% gain. Both models showed increases in the potential spring wheat production region (EM = 48%, MM = 20%), but these results were more equivocal because both models (particularly the EM) substantially overestimated the extent of current suitability. The substantial water-use efficiency gains simulated by the MMs under elevated CO2 accounted for much of the EM-MM difference, but EMs may have more accurately represented crop temperature sensitivities. Our results align with earlier studies showing that EMs may show larger climate change losses than MMs. Crop forecasting efforts should expand to include EM-MM comparisons to provide a fuller picture of crop-climate response uncertainties.

Anti-oncogenic role of the endoplasmic reticulum differentially activated by mutations in the MAPK pathway.

Dysfunction of the endoplasmic reticulum (ER) has been reported in a variety of human pathologies, including cancer. However, the contribution of the ER to the early stages of normal cell transformation is largely unknown. Using primary human melanocytes and biopsies of human naevi (moles), we show that the extent of ER stress induced by cellular oncogenes may define the mechanism of activation of premature senescence. Specifically, we found that oncogenic forms of HRAS (HRAS(G12V)) but not its downstream target BRAF (BRAF(V600E)), engaged a rapid cell-cycle arrest that was associated with massive vacuolization and expansion of the ER. However, neither p53, p16(INK4a) nor classical senescence markers--such as foci of heterochromatin or DNA damage--were able to account for the specific response of melanocytes to HRAS(G12V). Instead, HRAS(G12V)-driven senescence was mediated by the ER-associated unfolded protein response (UPR). The impact of HRAS on the UPR was selective, as it was poorly induced by activated NRAS (more frequently mutated in melanoma than HRAS). These results argue against premature senescence as a converging mechanism of response to activating oncogenes and support a direct role of the ER as a gatekeeper of tumour control.

Cutaneous Richter syndrome: report of 3 cases from one institution.

BACKGROUND: Richter syndrome (RS) is large-cell transformation of chronic lymphocytic leukemia (CLL). It commonly involves lymph nodes and bone marrow, but may rarely manifest in skin. Certain triggering factors, such as Epstein-Barr virus infection and p53 overexpression, have been implicated in the pathogenesis of RS. Here, we present 3 cases of cutaneous RS from our institution with a follow-up period of up to 8 years. OBJECTIVE: We present a series of cutaneous RS from a single institution with the longest follow-up period (up to 8 years) to date. METHODS: Clinical characteristics were collected and histopathological findings of skin biopsy specimens were analyzed. RESULTS: All 3 patients had prior CLL and later developed cutaneous RS lesions. The mean age at the diagnosis of cutaneous RS was 67 years old. The time intervals between CLL and cutaneous RS were 3 to 8 years. Skin biopsy specimens demonstrated dermal nodular or perivascular infiltrates of large B cells, showing similar immunophenotypes to the lesional cells in the original CLL. Overexpression of p53 and positive stain for Epstein-Barr virus--encoded small RNA was found in one patient. One patient remained alive 8 years after the diagnosis whereas the other two died of the disease at 5 years and 3 weeks, respectively, after the onset of cutaneous RS. LIMITATIONS: Three patients with RS were followed up for up to 8 years. CONCLUSIONS: Our findings suggested that, in contrast to extracutaneous RS, cutaneous RS generally has a less aggressive course with longer survival unless other worse prognostic factors are present.

Complex patterns of speciation in cosmopolitan "rock posy" lichens--discovering and delimiting cryptic fungal species in the lichen-forming Rhizoplaca melanophthalma species-complex (Lecanoraceae, Ascomycota).

A growing body of evidence indicates that in some cases morphology-based species circumscription of lichenized fungi misrepresents the number of existing species. The cosmopolitan "rock posy" lichen (Rhizoplaca melanophthalma) species-complex includes a number of morphologically distinct species that are both geographically and ecologically widespread, providing a model system to evaluate speciation in lichen-forming ascomycetes. In this study, we assembled multiple lines of evidence from nuclear DNA sequence data, morphology, and biochemistry for species delimitation in the R. melanophthalma species-complex. We identify a total of ten candidate species in this study, four of which were previously recognized as distinct taxa and six previously unrecognized lineages found within what has been thus far considered a single species. Candidate species are supported using inferences from multiple empirical operational criteria. Multiple instances of sympatry support the view that these lineages merit recognition as distinct taxa. Generally, we found little corroboration between morphological and chemical characters, and previously unidentified lineages were morphologically polymorphic. However, secondary metabolite data supported one cryptic saxicolous lineage, characterized by orsellinic-derived gyrophoric and lecanoric acids, which we consider to be taxonomically significant. Our study of the R. melanophthalma species-complex indicates that the genus Rhizoplaca, as presently circumscribed, is more diverse in western North American than originally perceived, and we present our analyses as a working example of species delimitation in morphologically cryptic and recently diverged lichenized fungi.