Gomes Ronald Muryellison Oliveira da Silva, da Silva Kássia Jéssica Galdino, Ferreira Leonardo Capistrano, Arantes Thales Domingos, Theodoro Raquel Cordeiro
Institute of Tropical Medicine, Universidade Federal do Rio Grande do Norte, Natal 59064-741, RN, Brazil.
Department of Biochemistry, Center of Bioscience, Universidade Federal do Rio Grande do Norte, Natal 59064-741, RN, Brazil.
J Fungi (Basel). 2023 May 30;9(6):629. doi: 10.3390/jof9060629.
The species complexes and are the causative agents of cryptococcosis. Virulence and susceptibility to antifungals may vary within each species according to the fungal genotype. Therefore, specific and easily accessible molecular markers are required to distinguish cryptic species and/or genotypes. Group I introns are potential markers for this purpose because they are polymorphic concerning their presence and sequence. Therefore, in this study, we evaluated the presence of group I introns in the mitochondrial genes and in different isolates. Additionally, the origin, distribution, and evolution of these introns were investigated by phylogenetic analyses, including previously sequenced introns for the gene. Approximately 80.5% of the 36 sequenced introns presented homing endonucleases, and phylogenetic analyses revealed that introns occupying the same insertion site form monophyletic clades. This suggests that they likely share a common ancestor that invaded the site prior to species divergence. There was only one case of heterologous invasion, probably through horizontal transfer to (VGIV genotype) from another fungal species. Our results showed that the complex has fewer introns compared to Additionally, there is significant polymorphism in the presence and size of these elements, both among and within genotypes. As a result, it is impossible to differentiate the cryptic species using a single intron. However, it was possible to differentiate among genotypes within each species complex, by combining PCRs of and introns, for species, and and introns for species.
新型隐球菌复合体和格特隐球菌复合体是隐球菌病的病原体。根据真菌基因型,每个物种内的毒力和对抗真菌药物的敏感性可能会有所不同。因此,需要特异性且易于获取的分子标记来区分隐匿种和/或基因型。I类内含子是实现这一目的的潜在标记,因为它们在存在情况和序列方面具有多态性。因此,在本研究中,我们评估了I类内含子在不同新型隐球菌分离株线粒体基因和中的存在情况。此外,通过系统发育分析研究了这些内含子的起源、分布和进化,其中包括之前已测序的基因内含子。在36个已测序的内含子中,约80.5%含有归巢内切酶,系统发育分析表明,占据相同插入位点的内含子形成单系分支。这表明它们可能有一个共同的祖先,在物种分化之前侵入了该位点。只有一例异源入侵情况,可能是通过水平转移从另一种真菌转移到新型隐球菌(VGIV基因型)。我们的结果表明,与格特隐球菌复合体相比,新型隐球菌复合体的内含子较少。此外,这些元件在基因型之间和基因型内部的存在情况和大小都存在显著的多态性。因此,不可能使用单个内含子来区分隐匿种。然而,通过结合新型隐球菌物种的内含子和内含子的PCR,以及格特隐球菌物种的内含子和内含子的PCR,可以区分每个物种复合体内的基因型。