RNA-Seq de Novo Assembly and Differential Transcriptome Analysis of Chaga (Inonotus obliquus) Cultured with Different Betulin Sources and the Regulation of Genes Involved in Terpenoid Biosynthesis

Chaga (Inonotus obliquus) is a medicinal fungus used in traditional medicine of Native American and North Eurasian cultures. Several studies have demonstrated the medicinal properties of chaga's bioactive molecules. For example, several terpenoids (e.g., betulin, betulinic acid and inotodiol) isolated from I. obliquus cells have proven effectiveness in treating different types of tumor cells. However, the molecular mechanisms and regulation underlying the biosynthesis of chaga terpenoids remain unknown. In this study, we report on the optimization of growing conditions for cultured I. obliquus in presence of different betulin sources (e.g., betulin or white birch bark). It was found that better results were obtained for a liquid culture pH 6.2 at 28 °C. In addition, a de novo assembly and characterization of I. obliquus transcriptome in these growth conditions using Illumina technology was performed. A total of 219,288,500 clean reads were generated, allowing for the identification of 20,072 transcripts of I. obliquus including transcripts involved in terpenoid biosynthesis. The differential expression of these genes was confirmed by quantitative-PCR. This study provides new insights on the molecular mechanisms and regulation of I. obliquus terpenoid production. It also contributes useful molecular resources for gene prediction or the development of biotechnologies for the alternative production of terpenoids

A single mega dose of vitamin D3 improves selected physical variables in vitamin D-deficient young amateur soccer players: a randomized controlled trial

This randomized controlled trial aimed to test whether vitamin D (VD) supplementation affects measures of physical performance in VD-deficient, mildly trained children. Thirty-six recreationally soccer player boys were randomly assigned to single dose (200\u2009000 IU) of VD3 (n = 19) or placebo (n = 17). Plasma 25-hydroxyvitamin D (25-OHD) was assessed and measures of physical performance (i.e., vertical and standing broad jumps, triple hop, 10-m and 30-m sprints, shuttle run) were performed before and 12 weeks after the loading dose. Mixed ANCOVA models were performed and effect size was estimated by partial eta squared (\u3b7p2). Baseline 25-OHD and physical variables were equivalent in the 2 groups. Twelve weeks after VD loading, plasma 25-OHD increased and physical variables improved only in the VD group. There was a significant interaction effects for group by time for vertical jump (F = 14.9, p = 0.001, \u3b7p2 = 0.394), triple hop jump (F = 24.2, p < 0.001, \u3b7p2 = 0.513), 10-m (F = 4.46, p = 0.046, \u3b7p2 = 0.162) and 30-m (F = 6.56, p = 0.017, \u3b7p2 = 0.222) sprints, and shuttle run (F = 13.4, p = 0.001, \u3b7p2 = 0.369). In conclusion, a single bolus of VD3 resulted in significant improvements in jumping ability, agility, and running speed in mildly trained children that are deficient in VD. The findings suggest that correcting VD deficit might be beneficial for physical performance. Novelty A mega dose of VD3 improves jumping ability, agility, and running speed in VD-deficient, mildly trained children. Effect of VD on measures of physical performance is noticeable 3 months after the loading dose