Reclassification of Olsenella gallinarum as Thermophilibacter gallinarum comb. nov. and description of Thermophilibacter immobilis sp. nov., isolated from the mud in a fermentation cellar used for the production of Chinese Luzhou-flavour Baijiu.

A novel Gram-stain-positive, strictly anaerobic, elliptical, non-motile and non-flagellated bacterium, designed LZLJ-2T, was isolated from the mud in a fermentation cellar used for the production of Chinese Luzhou-flavour Baijiu. Growth occurred at 28-45 °C (optimum, 37 °C), at pH 6.0-7.0 (optimum, pH 6.0) and with concentrations of NaCl up to 2 % (w/v; optimum, 0 %). On the basis of 16S rRNA gene sequence similarity, strain LZLJ-2T belonged to the genus Thermophilibacter and was most closely related to Thermophilibacter mediterraneus Marseille-P3256T (similarity 96.9 %), Olsenella gallinarum ClaCZ62T (similarity 96.6 %) and Thermophilibacter provencensis Marseille-P2912T (similarity 96.4 %). In addition, strain LZLJ-2T had high similarity to the genus Olsenella, including Olsenella profusa DSM 13989T (similarity 94.9 %), Olsenella umbonata DSM 22620T (similarity 94.9 %), Olsenella uli ATCC 49627T (similarity 94.22 %), Tractidigestivibacter scatoligenes DSM 28304T (similarity 93.9 %) and Paratractidigestivibacter faecalis KCTC 15699T (similarity 93.25 %). Comparative genome analysis showed that orthoANI values between strain LZLJ-2T and Thermophilibacter mediterraneus Marseille-P3256T, Olsenella gallinarum ClaCZ62T, Thermophilibacter provencensis Marseille-P2912T, Olsenella profusa DSM 13989T, Olsenella umbonata DSM 22620T, Olsenella uli ATCC 49627T, Tractidigestivibacter scatoligenes DSM 28304T and Paratractidigestivibacter faecalis KCTC 15699T were 78.68, 78.99, 78.29, 73.40, 74.00, 74.30, 75.08 and 77.23 %, and the genome-to-genome distance values were respectively 22.3, 22.5, 22.4, 19.6, 20.5, 19.7, 20.5 and 21.5 %. The genomic DNA G+C content of strain LZLJ-2T was 65.21 mol%. The predominant cellular fatty acids (>10 %) of strain LZLJ-2T were C18 : 1 cis 9 (33.7 %), C14 : 0 (22.0 %) and C18 : 1 cis 9 DMA (13.5 %). d-Glucose, sucrose, mannose, maltose, lactose (weak), salicin, glycerol (weak), cellobiose and trehalose (weak) could be used by strain LZLJ-2T as sole carbon sources. Enzyme activity results showed positive reactions with valine arylamidase, leucine arylamidase, crystine arylamidase, acid phosphatase, alkaline phosphatase, esterase (C4) (weakly positive), naphthol-AS-BI-phosphohydrolase, α-glucosidase and ß-glucosidase. The major end products of glucose fermentation were lactic acid and acetic acid. It produced skatole from indole acetic acid, and produced p-cresol from modified peptone-yeast extract medium with glucose. Based on the 16S rRNA gene trees as well as the genome core gene tree, it is suggested that Olsenella gallinarum are transferred to genus Thermophilibacter as Thermophilibacter gallinarum comb. nov. Based on phenotypic, genotypic and phylogenetic data, strain LZLJ-2T is considered to represent a novel species of the genus Thermophilibacter, for which the name Thermophilibacter immobilis sp. nov. is proposed. The type strain is LZLJ-2T (=KCTC 25162T=JCM 34224T).

Anthocyanin accumulation and biosynthesis are modulated by regulated deficit irrigation in Cabernet Sauvignon (Vitis Vinifera L.) grapes and wines.

Anthocyanins contents and compositions play an important role in grape berries and wines. Grapevines are widely cultivated in arid and semi-arid areas, and water shortage restricts the development of wine industry. The aim of this work was to gain insight on the effect of regulated deficit irrigation (RDI) on the accumulation and biosynthesis of anthocyanins in Cabernet Sauvignon (Vitis Vinifera L.) grapes and wines. High-performance liquid chromatography (HPLC) was used for anthocyanins profiles analyses and real-time quantitative PCR (qRT-PCR) was used for the genes expressions measurement. The grapevines were treated with 60% (RDI-1), 70% (RDI-2), 80% (RDI-3), 100% (CK, traditional drip irrigation) of their estimated evapotranspiration (ETc) respectively. RDI treatments significantly reduced titration acid and increased pH with higher total soluble solids. RDI-1 treatment increased total anthocyanins contents in berries and wines in both two vintages. RDI-1 and RDI-2 treatments significantly increased the contents of acylated anthocyanins in berries and wines, especially Malvidin-3-acetly-glucoside. RDI treatments significantly increased non-acylated anthocyanins contents in wines, such as Delphinidin-3-gliucoside and Malvidin-3-glucoside. RDI treatments upregulated the expressions of VvPAL, VvC4H, VvCHS, VvF3'H, VvF3'5'H, VvLDOX, and VvOMT in both two vintages. Correlation analysis showed the accumulation of anthocyanins was closely related to the key genes expressions, including VvPAL, VvF3'H, VvF3'5'H etc. The present results provided direct evidence and detailed data to explain that RDI treatments regulated the accumulation of anthocyanins by regulating genes expressions in the anthocyanin synthesis pathway.

Effects of Regulated Deficit Irrigation on Amino Acid Profiles and Their Derived Volatile Compounds in Cabernet Sauvignon (Vitis vinifera L.) Grapes and Wines.

Amino acid contents and their derived volatile compositions in Cabernet Sauvignon grapes and wines after regulated deficit irrigation (RDI) were investigated during the 2015 and 2016 growing seasons in Yinchuan (NingXia, China). High-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) were used for amino acid and volatile compound analyses. Three RDI strategies were tested: 60% (RDI-1), 70% (RDI-2), and 80% (RDI-3) of grapevine estimated evapotranspiration (ETc), and 100% ETc was used as the control group (CK). RDI-treated vines had lower yields and berry weights with higher total soluble solids than the control treatment. RDI-1 increased proline levels in berries and wines. RDI-2 enhanced tyrosine and asparagine levels in wines. RDI-3 enhanced arginine, alanine, valine, leucine, and isoleucine levels in berries and wines. RDI-2 and RDI-3 increased the concentrations of 2-methyl-1-butyl acetate, benzaldehyde, 3-methyl-1-pentanol, and 3-methyl-1-butanol in wines. The accumulation of volatile compounds was closely related to the amino acid concentrations-especially isoleucine, valine, and leucine-in grapes. Our results showed that RDI treatments altered amino acid concentrations and their derived volatile compositions in wines.

Effect of regulated deficit irrigation on fatty acids and their derived volatiles in 'Cabernet Sauvignon' grapes and wines of Ningxia, China.

The effect of regulated deficit irrigation (RDI) on fatty acids and their derived volatiles in 'Cabernet Sauvignon' grapes and wines was investigated during two growing seasons in the east foot of Mt. Helan, the semi-arid area. The vines received water with 60% (RDI-1), 70% (RDI-2), 80% (RDI-3), 100% (CK, traditional drip irrigation) of their estimated evapotranspiration (ETc) respectively. RDI treatments resulted in lower yield, berry weight and titratable acidity with higher total soluble solids. RDI-1 increased the content of unsaturated fatty acids in berries and decreased the level of alcohols and esters volatiles in wines. RDI-2 and RDI-3 enhanced 1-hexanol and esters in wines in comparison with CK. The concentrations of C6 aroma compounds were closely correlated with unsaturated fatty acids (p < .05), especially linolenic acid and linoleic acid. The present results provided direct evidence and detailed data to explain the effect of RDI on grapes and wines composition regarding fatty acids and their derived volatiles.