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- Viral diagnosis in cultivars of Ipomoea batatas (L.) LamPublication . Andrade, Carlos G.; Silva, Emanuel M. da; Ragonezi, Carla; Carvalho, Miguel Â. A. Pinheiro deIpomoea batatas (L.) Lam. commonly known as sweet potato, is an important staple food worldwide, mainly due to its high nutritional value and yield. However, vegetative reproduction of sweet potato makes it more susceptible to viral infections, which threatens its productivity, quality, and difficult long-term preservation in germplasm banks. Also, it can act as a virus reservoir infecting the rest of the plant accessions in the bank collections. Hence, this work aimed to screen Begomovirus, Potyvirus, and Carlavirus infections in 16 traditional sweet potato cultivars from the germplasm collection of the ISOPlexis Germplasm Bank, Madeira, Portugal. The infection prevalence by these viruses among cultivars was 81.25%, 25.00%, and 6.25%, respectively; being ISOP1011 the only accession coinfected by Potyvirus and Carlavirus. The accessions ISOP1006, ISOP1010, and ISOP1047 were also coinfected by Begomovirus and Potyvirus, highlighting their vulnerability to viral infections. The ISOP1005 and ISOP1027 accessions were the only ones not infected by any of these viruses. The analysis of the partial sequence obtained from the Carlavirus detected in the accession ISOP1011, revealed the existence of an ORF that encodes for 93 amino acids of the catalytic domain of an RNA-directed RNA polymerase related to the Tymovirus protein family, as could be confirmed by comparison with proteins stored in UniProtKB. Multiple sequence alignment with these proteins showed that Motifs A and B of the catalytic domain were conserved. The search for sequence similarity with sequences deposited in GenBank reported a high sequence identity with Sweet potato yellow mottle virus (SPYMV) and Sweet potato chlorotic fleck virus (SPCFV). However, the 9-11% discrepancy in nucleotide sequence identity and a phylogenetic analysis carried out using the maximum probability method suggests the virus isolated from ISOP1011 is a new divergent strain of the SPCFV species.
- Molecular identification and VOMs characterization of Saccharomyces cerevisiae strains isolated from Madeira region winery environmentsPublication . Castillo, Mariangie; Silva, Emanuel da; Câmara, José S.; Khadem, MahnazThe quality and typical characteristic of wines depends, among other factors, on the volatile organic metabolites (VOMs) that are biosynthesized by yeasts, mainly Saccharomyces cerevisiae species. The yeast strain influences the diversity and proportions of the VOMs produced during the fermentation process, as the genetic predisposition of the strains is a by-product of selective adaptation to the ecosystem. The present work reports the characterization of S. cerevisiae strains isolated from grape must, used in the Demarcated Region of Madeira (DRM) for winemaking. Yeast species were identified by amplification and by restriction fragment length polymorphism (RFLP) analysis of the region 5.8S-internal transcribed spacers (PCR-RFLP of 5.8S-ITS) of ribosomal DNA (rDNA). The strains identification was performed by analyzing the RFLP pattern of mitochondrial DNA (RFLP-mtDNA). The representative strains were selected for the characterization of the volatile profile through headspace solid-phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry (GC-MS) analysis. A total of 77 VOMs were identified. Higher alcohols, esters, and fatty acids were the major chemical families representing 63%, 16%, and 9%, respectively, in strain A and 54%, 23%, and 15% in strain B. The results indicate the influence of the strain metabolism in the production of VOMs, many of which probably participate in the aroma of the corresponding wines.