Plasmopara echinaceae, a new species of downy mildew affecting cone flowers (Echinacea purpurea) in the United States.

Downy mildew is one of the most important diseases of commercial sunflower and other Asteraceae hosts, including ornamental Rudbeckia. Plasmopara halstedii has historically been identified as the causal agent of this disease, considered a complex of species affecting nearly 35 genera in various tribes. However, with the use of molecular DNA characters for phylogenetic studies, distinct lineages of P. halstedii in the Asteraceae have been identified, confirmed as distinct or segregated as new species. During August of 2022, a downy mildew was observed on potted Echinacea purpurea grown in a retail greenhouse in Jefferson County, Wisconsin, USA. Phylogenetic analyses of the cytochrome c oxidase subunit 2 (cox2) and nuclear large subunit ribosomal RNA (nc LSU rDNA) gene regions indicated these Plasmopara sp. isolates are not conspecific with P. halstedii. Based on phylogenetic evidence and new host association, the Plasmopara isolates from E. purpurea are here described as Plasmopara echinaceae. Diagnostic morphological characters for this new species were not observed when compared with other isolates of P. halstedii or other Plasmopara species found on Asteraceae hosts, and therefore a list of species-specific substitutions in the cox2 region are provided as diagnostic characters. As this study corresponds to the first observation of downy mildew in cone flowers, it is recommended to follow the required disease prevention guidelines to prevent outbreaks and the establishment of this plant pathogen in production sites. Citation: Salgado-Salazar C, Romberg MK, Hudelson B (2023). Plasmopara echinaceae, a new species of downy mildew affecting cone flowers (Echinacea purpurea) in the United States. Fungal Systematics and Evolution 12: 203-217. doi: 10.3114/fuse.2023.12.10.

First Report of Sawadaea tulasnei Powdery Mildew of Norway Maple (Acer platanoides) in Wisconsin.

Leaves affected by powdery mildew were collected from a Norway maple tree in early October 2007 in Beaver Dam, WI (Dodge County). Diseased leaves were present throughout the crown of this tree, with white mycelium in irregular and often vein-associated spots and often covering as much as 50% of the upper surfaces of leaves. Examination of the lower surfaces revealed necrosis of the areas underlying mycelium. Blades of samaras also bore white mycelium. Chasmothecia were present singly or in groups on the mycelium. Morphology of chasmothecia, including simple and bifid appendages with uncinate to circinate apices, was sufficient to identify the pathogen to the genus Sawadaea (1). Data for nuclear rDNA ITS sequence (546 bp) obtained for a specimen (GenBank Accession No. EU247884) exactly matched sequences for Sawadea tulasnei (GenBank Accession Nos. AB 193363, 478 bp; AB193385, 490 bp; AB193390 and AB193391, 546 bp). This data was 96% similar (528 of 552 nucleotides) to that of another European powdery mildew pathogen, S. bicornis (GenBank Accession No. AB193380), which is also reported to occur on maples in Idaho, Washington, and Wisconsin (2,3). A further survey revealed the same fungus on several additional nearby Norway maples along streets and in yards (including varieties with both darkly colored and variegated leaves), but on these trees very few leaves were affected and usually less than 5% of the upper leaf surfaces bore mycelium. This pathogen was not observed on leaves of either red (A. rubrum) or silver maples (A. saccharinum) examined in the same area. S. tulasnei was previously known in North America only by collections in New York, Ohio, and Montreal, Canada (4), but our observation indicates that the geographic distribution of this pathogen is probably much broader and overlaps with that of S. bicornis. Specimens from Beaver Dam, WI have been deposited in the U.S. National Fungus Collections (BPI 878273). References: (1) U. Braun. The Powdery Mildews (Erysiphales) of Europe. Gustav Fischer Verlag, Jena-Stuttgart, New York, 1995. (2) C. Nischwitz and G. Newcombe. Plant Dis. 87:451, 2003. (3) G. Stanosz et al. Plant Dis. 91:636, 2007. (4) J. Weiland and G. Stanosz. Plant Dis. 90:830, 2006.

First Report of Sudden Death Syndrome of Soybean in Wisconsin.

In August of 2006, soybean (Glycine max (L.) Merr.) plants collected from Columbia, Dane, Green Lake, Walworth, Jefferson, and Waushara counties in southern Wisconsin exhibited symptoms typical of sudden death syndrome (SDS) caused by Fusarium virguliforme O'Donnell & Aoki [synonym F. solani (Mart.) Sacc. f. sp. glycines] (1). Foliar symptoms ranged from chlorotic spots to severe interveinal chlorosis and necrosis. Taproots of symptomatic plants were necrotic and stunted and stems exhibited a light tan discoloration, but never the dark brown discoloration typical for brown stem rot, a disease with similar foliar symptoms. Isolations from root and crown tissue of symptomatic plants were made using one-quarter-strength potato dextrose agar (PDA) amended with 100 ppm of streptomycin. Slow-growing, white-to-cream fungal colonies with blue and turquoise sporodochia were observed. Spores produced in sporodochia grown on PDA ranged in size from 32.5 to 70 µm long (average 53.1 µm) and 3 to 6 µm wide (average 4.4 µm) and with 3-5 septa (mode of 3). Isolates were characteristic of F. virguliforme based on colony morphology, spore morphology and size, and the absence of microconidia (3). The identity of F. virguliforme was confirmed by PCR amplification and DNA sequencing of the ITS, BT1, Act, and EF1B regions. All isolate sequences exhibited single nucleotide polymorphisms that matched the sequences of these regions of F. virguliforme. Koch's postulates were conducted to confirm that the causal agent of the observed symptoms was F. virguliforme. Inoculum of single-spore isolates was produced on sterilized sorghum seed. After 14 days of incubation at 20 to 22°C and a 12-h photoperiod, the sorghum seed was assayed to determine colonization incidence by transferring seeds to PDA. In all trials, sorghum seed was 100% infested. Infested sorghum seeds (35) were placed in potting soil at 2 cm beneath each seed of the susceptible soybean cv. Williams 82 (4). Noninfested sorghum seed was used for a noninoculated control. Three trials were performed, each using 15 replicates of several fungal isolates and 15 replicates of the noninoculated control. Plants were grown in water baths located in a greenhouse (trial 1) and in a growth chamber (trial 2) and both maintained at an average temperature of 25°C with a 14-h photoperiod (2). The third trial was conducted in the growth chamber without a water bath with the same temperature and light regimen. In all environments, inoculated plants developed chlorotic spots 14 days after planting. After 21 days, symptoms progressed to a range of chlorotic mottling to interveinal chlorosis and necrosis. Foliar and root symptoms that resembled those on the original plant samples infected with F. virguliforme appeared on 88% of inoculated plants. Isolates that resembled the original F. virguliforme were recovered from 75% of inoculated plants and from 88% of plants showing symptoms. No symptoms were observed and no isolates were recovered from noninoculated plants. There was a statistically significant difference between inoculated and control plants (P < 0.001) based on the presence of symptoms and isolation success using the Goodman χ2 analysis. The confirmation of the presence of SDS in five counties suggests that the disease is widespread in Wisconsin and could become a serious threat to soybean production in the future. References: (1) T. Akoi et al. Mycoscience 46:162, 2005. (2) R. Y. Hashmi et al. Online publication. doi:10.1094/PHP-2005-0906-01-RS. Plant Health Progress, 2005. (3) K. W. Roy et al. Plant Dis. 81:259, 1997. (4) J. C. Rupe et al. Can. J. Bot. 79:829, 2001.

Lymphocytotoxic anti-LewisbH antibody.

We wish to report strong lymphocyte crossmatch incompatibility attributable to anti-Lea antibody. The prospective renal transplant recipient was Le(a-b-) and her serum contained potent anti-Lea and anti-LebH antibodies. Despite the fact that were able to demonstrate greater B lymphocyte than T lymphocyte Lewis(a+) antigenicity, this serum was capable of causing greater than 80% cytotoxicity of Le(a+) and Le(b+)O or A2 whole lymphocyte populations. Antibody activity was completely inhibited by Lewis substance. This inhibition was specific and did not interfere with HLA antibody activity.

Detection and description of spatial patterns of bacterial brown spot of snap beans using cyclic samples.

ABSTRACT Snap bean plants within seven-row segments that ranged from 65 to 147 m were sampled, using a cyclic sampling plan. In the cyclic sampling plan, only 6 of every 31 plants were sampled, but sampled plants were spaced such that pairs of plants that were 1, 2, 3, 4,..., 1,525 plants apart could be identified within each sample. Every leaflet on every sampled plant was assessed for bacterial brown spot, and the proportion of disease leaflets per plant was determined. Arcsine square-root-transformed disease incidence values were analyzed for spatial patterns by autocorrelation and spectral analyses. Disease patterns were detected at several different scales within a single snap bean row, at distances that ranged from 20 to 100 m. Approximately 23 to 53% of the disease variability in the samples could be described by sine and cosine curves, indicating a substantial component of regularity in the disease patterns. Possible origins for these regular patterns, including cultural practices and seed infestation, are discussed.

First Report of Brown Root Rot of Alfalfa Caused by Phoma sclerotioides in Wisconsin.

Brown root rot (BRR) has been associated with winterkill of alfalfa (Medicago sativa L.) in the temperate regions of North America where winters are severe (1). Although suspected, BRR has not been associated with winterkill of alfalfa in the upper Midwestern United States. Alfalfa plants exhibiting symptoms resembling those induced by the causal agent Phoma sclerotioides G. Preuss ex Sacc. were collected from fields in Marinette, Pierce, and Marathon counties in Wisconsin during the spring and early summer of 2003. Symptoms included stunting and decline in 1- to 3-year-old plants that were slow to break dormancy in the early spring. Roots frequently exhibited dark brown lesions or were entirely decayed. Advanced lesions often formed dark bands around the circumference of tap and secondary roots. Beaked pycnidial structures typical of P. sclerotioides were also observed on many samples with advanced lesions. Plants with symptoms of BRR were also observed in Clark, Langlade, Lincoln, Oconto, Shawno, Taylor, and Wood counties. Several lesion areas of tissue on the tap and lateral roots of each sample were excised with a sterile scalpel. Total DNA was extracted using the Fast DNA kit (Bio 101, Carlsbad, CA). In addition, soil samples were collected in the root rhizosphere of symptomatic plants from four fields in two counties. Soil DNA was extracted with the Ultra-Clean DNA soil extraction kit (Mo Bio, Solana Beach, CA). DNA extractions were diluted 1:10 or 1:50, and samples were evaluated for the presence of P. sclerotioides using polymerase chain reaction (PCR)-based sequence-characterized amplified region (SCAR) markers according to the method described previously (4). Amplicons of the expected size (499 bp) were detected from alfalfa roots sampled from Marathon (4 of 4), Marinette (4 of 5), and Pierce (4 of 4) counties but not in roots from healthy controls produced in the greenhouse at Prosser, WA. PCR amplicons were also produced from all field soil samples in Marathon and Marinette counties. Proof of pathogenicity via Koch's postulates for this host-pathogen system was not attempted because of the extensive time period required (1). However, characteristic beaked pycnidia were present, and the pathogen was identified using PCR on DNA from roots symptomatic of BRR. Detection of BRR has been limited in the United States to Wyoming (2), but has been thought to occur in other states with severe winters (3). To our knowledge, this is the first report of P. sclerotioides in Wisconsin. References: (1) J. G. N. Davidson. Brown root rot. Pages 29-31 in: Compendium of Alfalfa Diseases. 2nd ed. D. L. Stuteville and D. C. Erwin, eds. The American Phytopathological Society, St. Paul, MN, 1990. (2) F. A. Gray et al. Pages 27-28 in: Proc. 10th Western Alfalfa Improv. Conf., 1997. (3) C. R. Hollingsworth et al. Can. J Plant Pathol. 25:215, 2003. (4) R. C. Larsen et al. Plant Dis. 86:928, 2002.

Dissimilar effects of ultraviolet light on HLA-D and HLA-DR antigens.

Since HLA-DR antigens are closely related to HLA-D antigens, we studied the effect of low-dose ultraviolet irradiation on the ability of human lymphocytes to induce a proliferative response in mixed lymphocyte culture (HLA-D) and on the serologic reactivity of B lymphocyte HLA-DR antigens. We found (1) no alteration in the serologic definition of DR antigens despite the abrogation of the induction of proliferative responses in mixed lymphocyte culture, (2) no evidence that ultraviolet light (UVL) stimulated suppressor cells contributed to the loss of allostimulation, and (3) no evidence that the abrogation of the induction of proliferative responses could be modified by increasing the number of UVL exposed lymphocytes in mixed lymphocyte culture. These findings suggest that HLA-D and HLA-DR antigens are different molecularly, but are also consistent with the concept that certain active metabolic processes are required of stimulator cells for allostimulation and that these processes are UVL sensitive.