ABSTRACT
BACKGROUND: Geotrichum is a genus of fungi found in different habitats throughout the world. Although Geotrichum and its related species have been extensively reclassified and taxonomically revised, it is still the target for many researches. METHODS AND RESULTS: In this study, phenotypic and molecular genetics comparisons were performed between Geotrichum candidum and Geotrichum silvicola. Mitis Salivarius Agar was used as the growing medium for the phenotypic comparison study, which was carried out at two temperatures (20-25 and 37 °C). For genotypic comparison, we compared the 18 S, ITS, and 28 S sequences of universal DNA barcode regions of both species. Important findings on the new culture media for fungal isolation were revealed by the results. The phenotypic variation between the two species' colonies, including their shapes, sizes, textures and growth rates, were strikingly different. DNA sequences of both species showed that pairwise identities of the species were 99.9% for 18 S, 100% for ITS and 99.6% for 28 S regions. CONCLUSIONS: Contrary to what is commonly seen, the results showed that 18 S, ITS and 28 S failed to discriminate the species. The first investigation into the performance of Mitis Salivarius Agar as a fungus culture medium is reported in this work, and proved its efficiency. Additionally, this is the first study to compare G. candidum with G. silvicola by means of both phenotypic and genotypic analysis.
Subject(s)
Geotrichum , Pimenta , Geotrichum/genetics , Agar , Molecular BiologyABSTRACT
Accurate identification of animal species sources in milk have become quite important due to adulteration of high-priced milk types in the dairy industry. To date, milk identification methods have mainly depended on biochemical properties or physical properties detected by spectroscopic methods. The current study aimed to develop an easy to use and sensitive DNA-based High resolution melting (HRM) assay to identify animal species and detect cross-adulteration of water buffalo, bovine, goat, sheep, camel and donkey milks. HRM compatible designed primer set, targeted mitochondrial region, successfully amplified the specific targeted region for six animal species DNA and showed a high degree of specificity based on nucleotide variations. Capillary electrophoresis analysis validated the specific amplicons and determined the amplicon lengths as 114 bp for bovine, goat, sheep, and camel, 115 bp for water buffalo, and 121 bp for donkey. HRM analysis showed a clear discrimination for water buffalo-bovine, camel-bovine and donkey-bovine adulteration down to 0.5%, and goat-sheep adulteration down to 1% in the milk admixtures. The efficacy of the method was also confirmed by its standard curve with a very high correlation coefficient In conclusion, the designed HRM assay allows for the rapid, sensitive and cost-effective authentication of milk and dairy products. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05705-3.
ABSTRACT
In this study, we determined the structure of DNA methyltransferase (DNMT) genes in apomict and sexual Boechera species and investigated the expression levels during seed development. Protein and DNA sequences of diploid sexual Boechera stricta DNMT genes obtained from Phytozome 10.3 were used to identify the homologues in apomicts, Boechera holboellii and Boechera divaricarpa. Geneious R8 software was used to map the short-paired reads library of B. holboellii whole genome or B. divaricarpa transcriptome reads to the reference gene sequences. We determined three DNMT genes; for Boechera spp. METHYLTRANSFERASE1 (MET1), CHROMOMETHYLASE 3 (CMT3) and DOMAINS REARRANGED METHYLTRANSFERASE 1/2 (DRM2). We examined the structure of these genes with bioinformatic tools and compared with other DNMT genes in plants. We also examined the levels of expression in silique tissues after fertilization by semi-quantitative PCR. The structure of DNMT proteins in apomict and sexual Boechera species share common features. However, the expression levels of DNMT genes were different in apomict and sexual Boechera species. We found that DRM2 was upregulated in apomictic Boechera species after fertilization. Phylogenetic trees showed that three genes are conserved among green algae, monocotyledons and dicotyledons. Our results indicated a deregulation of DNA methylation machinery during seed development in apomicts.
