Fungal dynamic changes in naturally fermented 'Kyoho' grape juice.

The 'Kyoho' grape (Vitaceae, Plantae) has large ears, plenty of flesh, and rich nutrition and is planted across a large area in China. There are few reports on this variety in winemaking, especially on the dynamic changes of fungi in the wine fermentation broth. In this study, we used the 'Kyoho' grapes as raw materials and adopted a high throughput to analyze dynamic changes in fungal species composition of the natural fermentation broth at four time points: day 1 (D1P), day 3 (D3P), day 5 (D5P), and day 15 (D15P). Changes in fungal metabolic pathways and dominant yeasts were also analyzed. A total of 78 families, 110 genera, and 137 species were detected, in the natural fermentation broth samples. Forty-nine families, 60 genera, and 72 species were found in the control check (CK). A total of 66 differential metabolic pathways were enriched; of those, 41 were up-regulated compared to CK, such as CDP-diacylglycerol biosynthesis I (PWY 5667), chitin degradation to ethanol (PWY 7118), and the super pathway of phosphatidate biosynthesis (PWY 7411). Changes in fungal metabolic pathways were in line with the dynamic changes of dominant yeast species in the whole process of fermentation. Pichia kluyveri, P. membranifaciens, and Citeromyces matritensis are the dominant species in the later stages of natural fermentation. These yeast species may play vital roles in the 'Kyoho' wine industry in the future.

Genomic Comparative Analysis of Cordyceps pseudotenuipes with Other Species from Cordyceps.

The whole genome of Cordyceps pseudotenuipes was sequenced, annotated, and compared with three related species to characterize the genome. The antibiotics and Secondary Metabolites Analysis Shell (antiSMASH) and local BLAST analysis were used to explore the secondary metabolites (SMs) and biosynthesis gene clusters (BGCs) of the genus Cordyceps. The genome-wide basic characteristics of C. pseudotenuipes, C. tenuipes, C. cicadae, and C. militaris revealed unequal genome size, with C. cicadae as the largest (34.11 Mb), followed by C. militaris (32.27 Mb). However, the total gene lengths of C. pseudotenuipes and C. tenuipes were similar (30.1 Mb and 30.06 Mb). The GC contents of C. pseudotenuipes, C. tenuipes, C. cicadae, and C. militaris genomes differed slightly (51.40% to 54.11%). AntiSMASH and local BLAST analysis showed that C. pseudotenuipes, C. tenuipes, C. cicadae, and C. militaris had 31, 28, 31, and 29 putative SM BGCs, respectively. The SM BGCs contained different quantities of polyketide synthetase (PKS), nonribosomal peptide synthetase (NRPS), terpene, hybrid PKS + NRPS, and hybrid NRPS + Other. Moreover, C. pseudotenuipes, C. tenuipes, C. cicadae, and C. militaris had BGCs for the synthesis of dimethylcoprogen. C. pseudotenuipes, C. tenuipes, and C. cicadae had BGCs for the synthesis of leucinostatin A/B, neosartorin, dimethylcoprogen, wortmanamide A/B, and beauvericin. In addition, the SM BGCs unique to C. pseudotenuipes were clavaric acid, communesin, and deoxynivalenol. Synteny analysis indicated that the scaffolds where the SM BGC was located were divided into more than 70 collinear blocks, and there might be rearrangements. Altogether, these findings improved our understanding of the molecular biology of the genus Cordyceps and will facilitate the discovery of new biologically active SMs from the genus Cordyceps using heterologous expression and gene knockdown methods.