The fungus Simplicillium lanosoniveum was isolated from soybean leaves infected with Phakopsora pachyrhizi, the soybean rust pathogen, in Louisiana and Florida. The fungus did not grow or become established on leaf surfaces until uredinia erupted, but when soybean rust signs and symptoms were evident, S. lanosoniveum colonized leaves within 3 days and sporulated within 4 days. Development of new uredinia was suppressed by about fourfold when S. lanosoniveum colonized uredinia. In the presence of S. lanosoniveum, uredinia became increasingly red-brown, and urediniospores turned brown and germinated at very low rates. Assays using quantitative real time polymerase chain reaction revealed that the fungus colonized leaf surfaces when plants were infected with P. pachyrhizi, either in a latent stage of infection or when symptoms were present. However, when plants were inoculated before infection, there was no increase of DNA of S. lanosoniveum, suggesting that the pathogen must be present in order for the antagonist to become established on soybean leaf surfaces. We documented significantly lower amounts of DNA of P. pachyrhizi and lower disease severity when soybean leaves were colonized with S. lanosoniveum. These studies documented the mycophilic and disease-suppressive nature of S. lanosoniveum.

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Cercospora leaf blight (CLB) of soybean, caused by Cercospora kikuchii, is a serious disease in the southern United States. A sensitive TaqMan probe-based real-time quantitative polymerase chain reaction (qPCR) assay was developed to specifically detect and quantify C. kikuchii in naturally infected soybean plants. The sensitivity was 1 pg of genomic DNA, which was equivalent to about 34 copies of genome of C. kikuchii. Using this qPCR assay, we documented a very long latent infection period for C. kikuchii in soybean leaves beginning at the V3 growth stage (as early as 22 days after planting). The levels of biomass of C. kikuchii remained low until R1, and a rapid increase was detected from the R2/R3 to R4/R5 growth stages shortly before the appearance of symptoms at R6. The efficacy of various fungicide regimens under field conditions also was evaluated over a 3-year period using this qPCR method. Our results showed that multiple fungicide applications beginning at R1 until late reproductive stages suppressed the development of C. kikuchii in leaves and delayed symptom expression. Different fungicide chemistries also had differential effects on the amount of latent infection and symptom expression during late reproductive growth stages.

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Impatiens downy mildew (Plasmopara obducens (J. Schröt.) J. Schröt. (syn Peronospora obducens) was first reported in the United States in 2004, but widespread outbreaks were observed throughout North America in 2011 (5). In June 2012, symptoms, including severe defoliation while plants retained upright stems, were observed on approximately 100 landscape impatiens (Impatiens walleriana Hook.f.) in Franklin County in central Kentucky. All plants in the landscape were affected. Plants were primarily defoliated and remaining leaves were stunted, mottled, and chlorotic with edges curled downward; no flowers were present. Under examination with a dissecting microscope, white downy fungal growth was observed. Closer examination confirmed that the growth consisted of colorless sporangiophores that were mainly unbranched, straight, and rigid (1,3). Sporangiophores consisted of apical branches attached at right angles to main axes, ranging from 67.2 to 89.9 μm long (1). Sporangia were ovoid and hyaline, measuring 11.2 to 13.3 μm × 8.2 to 10.7 μm (3). No oospores were observed. Pathogenicity tests were performed by inoculating 20 to 40 leaves on three plants each of the cvs. Dazzler and Super Elfin with suspensions of 1 × 105 sporangiophores per ml in sterile distilled water. Sporangia were obtained by washing infected leaves with sterile distilled water, and inoculations were completed by spraying leaves until runoff. Plants sprayed with sterile water served as controls. Plants were covered with black plastic bags for 48 h and then maintained under fluorescent lights for 10 days at room temperature (22 to 25°C). Sporangiophores were recovered from inoculated plants after 10 days, and morphology matched original inoculum; symptoms included chlorotic, downward curling leaves with sporulation on the undersides. Non-inoculated plants did not develop symptoms after 21 days. Molecular identification of the pathogen was conducted using three leaves from one plant from each cultivar. PCR was conducted by amplifying the large ribosomal subunit DNA using primers NL-1 and NL-4 (2). Amplicons of 762 to 691 bp were produced from diseased plant tissue that contained visible sporangiophores, and the bands were extracted from the gel and purified. Sequence results confirmed 100% similarity to accessions from Florida (GenBank Accession No. JX217746.1) and Ohio (JX142134.1) and 99% similarity to amplicons reported from Serbia (HQ246451.1) and UK (AY587558.1). This is believed to be the first report of downy mildew infecting impatiens in Kentucky. References: (1) O. Constantinescu. Mycologia 83:473, 1991. (2) W. Maier et al. Can. J. Bot 81:12, 2003. (3) P. A. Saccardo. Syllogue Fungorum 7:242, 1888. (4) S. N. Wegulo et al. Plant Dis. 88:909, 2004.