First Report of Colletotrichum fioriniae Infecting Hop (Humulus lupulus) in Michigan.

In July of 2020, a hop (Humulus lupulus L.) grower in Berrien County, Michigan submitted 'Chinook' leaf samples to MSU Plant & Pest Diagnostics. The leaves were covered in small, tan colored lesions, with a small chlorotic halo with an approximate diameter of 5 mm. The grower reported that foliar lesions were in the lower 2 m of the fully developed hop canopy. Disease incidence and severity were estimated at approximately 20% and 5 to 10%, respectively. After incubation at 100% relative humidity, acervuli with orange spore masses and a few setae were present. A pure culture was obtained from these sporulating lesions using water agar. The isolate was hyphal tipped onto potato dextrose agar (PDA) and stored in a glycerol-salt solution at -80o C (isolate CL001) (Miles et al. 2011). On PDA, cultures displayed gray growth on the top of the colony and a red color on the underside of the Petri dish. After 14 days, acervuli with no setae appeared exuding orange conidial masses on the surface of the culture. Conidia were hyaline, aseptate, smooth-walled and rounded at the ends and measured on average 15.89 µm (13.81 to 16.91 µm) × 7.26 (6.82 to 8.41 µm) (n = 20). The color and size of the conidia matched other descriptions of C. acutatum sensu lato (Damm et al. 2012). Four loci (ITS/515 bp - OQ026167, GAPDH/238 bp - OQ230832, CHS1/228 bp -OQ230830, and TUB2/491 bp - OQ230831) were amplified from isolate CL001 (using the primers ITS1/ITS4, GDF1/GDR1, CSH-79f/CHS-354R, and T1/Bt-2b, respectively) and had 100% pairwise identity with C. fioriniae 125396 (JQ948299, JQ948629, JQ948960, JQ949950, respectively, Damm et al. 2012). The GAPDH, CSH1, and TUB2 sequences from isolate CL001 were trimmed, concatenated and aligned with 31 different members of Colletotrichum acutatum sensu lato and C. gloesporioides 356878 (Damm et al. 2012; Kennedy et al. 2022). The alignment was then used to produce a maximum likehood phylogenetic tree using Geneious Prime (Biomatters Ltd.) with the PHYML add on using the HKY + G model (G = 0.34) (Guindon et al. 2010). Isolate CL001 had the closest similarity to C. fioriniae with a bootstrap value of 100. Pathogenicity tests were performed on 2 month-old 'Chinook' hop plants. Twelve plants were inoculated with 50 ml of a conidial suspension (7.95 x 106 conidia/ml) of isolate CL001 (n = 6) or water (n = 6) using a spray bottle until runoff. Inoculated plants were sealed in clear plastic bags and grown in a greenhouse at 21o C with a photoperiod of 14 h. After 7 days, lesions appeared on the hop plants inoculated with CL001, but no symptoms appeared on the water inoculated hop plants. Lesions with a chlorotic halo were observed but they were smaller than field lesions and no setae were present (approx. 1 mm in diam.). Leaves were surface sterilized (0.3% sodium hypochlorite solution for 15 s and then rinsed three times) and the leading margin of the lesions or healthy tissue (water control) were placed on 1% ampicillin amended PDA. Fungal isolates on PDA morphologically matched C. fioriniae were recovered from all CL001-inoculated plants. No C. fioriniae isolates were recovered from the water-inoculated plants. Based on conidial morphology, the four loci, and the phylogenetic tree, isolate CL001 was identified as C. fioriniae. This is the first report of Colletotrichum fioriniae (syn = Glomerella acutata var. fioriniae Marcelino & Gouli) infecting common hop and further investigation is needed to determine if management is needed for this pathogen.

Complete Genome Sequence of the Ice-Nucleation-Active Pseudomonas syringae Strain MUP20, Isolated from Frost-Damaged Wheat (Triticum aestivum cv. Scepter) in Western Australia.

Pseudomonas syringae MUP20 was isolated from Western Australian frost-damaged wheat. The MUP20 complete genome contained a 6,045,198-bp single circular chromosome with a GC content of 59.03%. IMG/M genome annotation identified 5,245 protein-coding genes, 1 of which encoded an ice nucleation protein containing 20 occurrences of a highly repetitive PF00818 domain.

Complete Genome Sequence of the Ice-Nucleation-Active Pseudomonas syringae pv. pisi Isolate MUP32, Isolated from Frost-Damaged Pea (Pisum sativum subsp. arvense cv. Dundale) in New South Wales.

The genome of Pseudomonas syringae MUP32, which was isolated from frost-damaged pea in New South Wales, Australia, is tripartite and contains a circular chromosome (6,032,644 bp) and two plasmids (61,675 and 54,993 bp). IMG/M genome annotation identified 5,370 protein-coding genes, one of which encoded an ice-nucleation protein with 19 repetitive PF00818 domains.

Complete Genome Sequence of the Ice-Nucleation-Active Pseudomonas syringae Strain MUP17, Isolated from the Frost-Damaged Barley Cultivar Hordeum vulgare cv. La Trobe.

Pseudomonas syringae MUP17 was isolated from Western Australian frost-damaged barley. The MUP17 complete genome contained a 5,850,185-bp single circular chromosome with a GC content of 59.12%. IMG/M genome annotation identified 5,012 protein-coding genes, 1 of which encoded an ice-nucleation protein containing 19 occurrences of a highly repetitive PF00818 domain.

Quantitative Expression and Co-Localization of Wnt Signalling Related Proteins in Feline Squamous Cell Carcinoma.

Feline oral squamous cell carcinoma (FOSCC) is an aggressive neoplasm in cats. Little is known about the possible molecular mechanisms that may be involved in the initiation, maintenance and progression of FOSCC. Wnt signalling is critical in development and disease, including many mammalian cancers. In this study, we have investigated the expression of Wnt signalling related proteins using quantitative immunohistochemical techniques on tissue arrays. We constructed tissue arrays with 58 individual replicate tissue samples. We tested for the expression of four key Wnt/ß-catenin transcription targets, namely Cyclin D1 (CCND1 or CD1), FRA1, c-Myc and MMP7. All antibodies showed cross reactivity in feline tissue except MMP7. Quantitative immunohistochemical analysis of single proteins (expressed as area fraction / amount of tissue for normal vs tumor, mean ± SE) showed that the expression of CD1 (3.9 ± 0.5 vs 12.2 ± 0.9), FRA1 (5.5 ± 0.6 vs 16.8 ± 1.1) and c-Myc (5.4 ± 0.5 vs 12.5 ± 0.9) was increased in FOSCC tissue by 2.3 to 3 fold compared to normal controls (p<0.0001). By using a multilabel, quantitative fluorophore technique we further investigated if the co-localization of these proteins (all transcription factors) with each other and in the nucleus (stained with 4',6-diamidino-2-phenylindole, DAPI) was altered in FOSCC compared to normal tissue. The global intersection coefficients, a measure of the proximity of two fluorophore labeled entities, showed that there was a significant change (p < 0.01) in the co-localization for all permutations (e.g. CD1/FRA1 etc), except for the nuclear localization of CD1. Our results show that putative targets of Wnt signalling transcription are up-regulated in FOSCC with alterations in the co-localization of these proteins and could serve as a useful marker for the disease.