Long-Read Sequencing Genome Assembly of Ceratocystis fimbriata Enables Development of Molecular Diagnostics for Sweetpotato Black Rot.

Ceratocystis fimbriata is a destructive fungal pathogen of sweetpotato (Ipomoea batatas) that leads to losses at all stages of sweetpotato production. Accurate detection of C. fimbriata would allow for more efficient deployment of management tactics in sweetpotato production. To develop a diagnostic assay, a hybrid genome assembly of C. fimbriata isolate AS236 was generated. The resulting 31.7-MB assembly was near-chromosome level, with 18 contigs, 6,481 predicted genes, and a BUSCO completion score of 98.4% when compared with the fungus-specific lineage database. Additional Illumina DNA reads from C. manginecans, C. platani, and a second C. fimbriata isolate (C1421) were then mapped to the assembled genome using BOWTIE2 and counted using HTSeq, which identified 148 genes present only within C. fimbriata as molecular diagnostic candidates; 6 single-copy and 35 highly multi-copy (>40 BLAST hits), as determined through a self-BLAST-P alignment. Primers for PCR were designed in the 200-bp flanking region of the first exon for each candidate, and the candidates were validated against a diverse DNA panel containing Ceratocystis species, sweetpotato pathogens, and plants. After validation, two diagnostic candidates amplified only C. fimbriata DNA and were considered to be highly specific to the species. These genetic markers will serve as valuable diagnostic tools with multiple applications including the detection of C. fimbriata in seed, soil, and wash water in sweetpotato production.

Assessing the Role of Temperature, Inoculum Density, and Wounding on Disease Progression of the Fungal Pathogen Ceratocystis fimbriata Causing Black Rot in Sweetpotato.

In 2014, Ceratocystis fimbriata, causal agent of black rot in sweetpotato, reemerged and inflicted large financial losses on growers in the United States. Black rot continues to damage sweetpotatoes and has become a priority to the industry since then. In contrast, little is known about the biology of C. fimbriata and the epidemiology of sweetpotato black rot. In this study, effects of environmental factors such as inoculum density, RH, and temperature on sweetpotato black rot were determined. Cured sweetpotatoes were wounded with a toothpick to simulate puncture wounds, inoculated with different spore suspensions (inoculum density) (104, 105, or 106 spores/ml), and incubated under different RH (85.53, 94.09, or 97.01%) and temperature (13, 18, 23, 29, or 35°C) for 21 days. In a separate experiment, five root wounding types (cuts, punctures, abrasions, end breaks, and macerating bruises) were compared. All wounded roots were subsequently soaked in a 103 spores/ml suspension and incubated at 100% RH and 23°C for 21 days. This study found 29 and 23°C to be the optimal temperature for black rot disease development and sporulation, respectively. No pathogen growth was observed at 13 and 35°C. Increased inoculum density significantly (P < 0.0001) increased disease incidence, but increasing RH had an effect only on sporulation area. All wound types resulted in increased disease incidence and sporulation as early as 7 days postinoculation. Our results highlight the importance of characterizing factors that affect disease development for achieving successful disease management strategies. Findings from this study will be used to improve disease management for sweetpotato black rot by suggesting tighter regulation of curing and storage conditions and better postharvest handling of sweetpotato roots to avoid unnecessary wounding.

[Somatomedin C level and stimulation of growth hormone and adrenal cortex function by administration of releasing hormones and physical exertion in patients with obesity]. / Somatomedin-C-Spiegel und Stimulation von Wachstumshormon und Nebennierenrindenfunktion durch Gabe von Releasing-Hormonen und körperliche Belastung bei Patienten mit Adipositas.

In a clinical study on potential determinants underlying the impairment of growth hormone stimulation in obese human subjects, we examined in 20 otherwise healthy adult obese subjects (14 females, six males, age 18 to 40 years, body mass index greater than 29 kg/m2) the responses of growth hormone (hGH), adrenocorticotropic hormone (ACTH) and cortisol to releasing hormone stimulation (growth hormone-releasing hormone and corticotropin-releasing hormone) and the responses of hGH, ACTH, cortisol and free fatty acids (FFA) to physical exercise. Subjects with somatomedin-C levels less than or equal to 0.7 U/ml (group 1) were more obese than subjects with somatomedin-C levels greater than 0.7 U/ml (group 2) (p less than 0.01). In group 1, hGH increased by 4.3 +/- 1.2 ng/ml in response to releasing hormone administration and by 0.9 +/- 0.3 ng/ml in response to physical exercise (normal responses, increase by greater than 7 ng/ml), in group 2, hGH increased by 6.7 +/- 1.4, and 2.4 +/- 0.8 ng/ml, respectively (p less than 0.05 vs. group 1). Moreover, FFA stimulation by physical exercise was blunted in group 1 (p less than 0.05 vs. group 2). In contrast, ACTH stimulation was found increased in group 1 in comparison to group 2, particularly in response to physical exercise (p less than 0.01), and resulted in enhanced cortisol stimulation (p less than 0.05). Thus, impaired hGH stimulation in obese human subjects is not explained by an altered relationship between hGH and somatomedin-C levels.(ABSTRACT TRUNCATED AT 250 WORDS)