We here assessed the capability of the MinION sequencing approach to detect and characterize viruses infecting a water yam plant. This sequencing platform consistently revealed the presence of several plant virus species, including Dioscorea bacilliform virus, Yam mild mosaic virus and Yam chlorotic necrosis virus. A potentially novel ampelovirus was also detected by a complimentary Illumina sequencing approach. The full-length genome sequence of yam chlorotic necrosis virus was determined using Sanger sequencing, which enabled determination of the coverage and sequencing accuracy of the MinION technology. Whereas the total mean sequencing error rate of yam chlorotic necrosis virus-related MinION reads was 11.25%, we show that the consensus sequence obtained either by de novo assembly or after mapping the MinION reads on the virus genomic sequence was >99.8% identical with the Sanger-derived reference sequence. From the perspective of potential plant disease diagnostic applications of MinION sequencing, these degrees of sequencing accuracy demonstrate that the MinION approach can be used to both reliably detect and accurately sequence nearly full-length positive-sense single-strand polyadenylated RNA plant virus genomes.
The ethanol yields from lignocellulo-starch biomass (peels of sweet potato, elephant foot yam, tannia, greater yam and beet root) by fed-batch separate hydrolysis and fermentation (F-SHF) and simultaneous saccharification and fermentation (F-SSF) using Saccharomyces cerevisiae were compared. Fed-batch saccharification of steam or dilute sulphuric acid pretreated biomass enhanced the reducing sugar yield which resulted in high RS consumption, volumetric ethanol productivity and ethanol yield during the first 24 h fermentation under F-SHF mode, while continuous production and utilization of reducing sugars occurred up to 72 h in F-SSF. Dilute sulphuric acid pretreated residues under F-SHF gave higher ethanol yield (34–43 g/L) and productivity (274–346 ml/kg dry biomass) than steam pretreatment (27–36 g/L and 223–295 ml/kg respectively), while F-SSF was superior for steam pretreated peels of sweet potato, elephant foot yam and tannia giving ethanol yields from 281 to 302 ml/kg. Glucose and xylose were present in all the hydrolysates with a preponderance of glucose and fermentation resulted in significant reduction in glucose levels in both F-SHF and F-SSF. Higher levels of total soluble phenolics and hydroxymethyl furfural were observed in the hydrolysates from dilute sulphuric acid pretreatment and yeast assimilated/detoxified part of the inhibitors, while only trivial amounts of furfural were present due to the low xylose content in the hydrolysates. Continuous formation led to higher accumulation of inhibitors in F-SSF despite supplementation with the detoxification mix comprising Tween 20, polyethylene glycol and sodium borohydride. F-SHF of dilute sulphuric acid pretreated biomass could be considered as a comparatively advantageous process where only one time feeding of enzyme cocktail and yeast was adopted compared to multiple feeds of enzymes and yeast along with other additives such as detoxification mix or nutrient solution in F-SSF.
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