Lopes Anabela, Phillips Alan J L, Alves Artur
Biology Department, CESAM, University of Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal.
University of Lisbon, Faculty of Sciences, Biosystems and Integrative Sciences Institute (BioISI), Campo Grande, 1749-016 Lisbon, Portugal.
Fungal Biol. 2017 Apr;121(4):394-404. doi: 10.1016/j.funbio.2016.08.011. Epub 2016 Sep 9.
The genus Neofusicoccum includes species with wide geographical and plant host distribution, some of them of economic importance. The genus currently comprises 27 species that are difficult to identify based on morphological features alone. Thus, species differentiation is based on phylogenetic species recognition using multigene genealogies. In this study, we characterised the mating type genes of Neofusicoccum species. Specific primers were designed to amplify and sequence MAT genes in several species and a PCR-based mating type diagnostic assay was developed. Homothallism was the predominant mating strategy among the species tested. Furthermore, the potential of mating type gene sequences for species delimitation was evaluated. Phylogenetic analyses were performed on both MAT genes and compared with multigene genealogies using sequences of the ribosomal internal transcribed spacer region, translation elongation factor 1-alpha and beta-tubulin. Phylogenies based on mating type genes could discriminate between the species analysed and are in concordance with the results obtained with the more conventional multilocus phylogenetic analysis approach. Thus, MAT genes represent a powerful tool to delimit cryptic species in the genus Neofusicoccum.
新壳梭孢属包含一些地理分布广泛、寄主植物多样的物种，其中一些具有经济重要性。该属目前包括27个物种，仅根据形态特征很难对它们进行鉴定。因此，物种分化是基于使用多基因谱系的系统发育物种识别。在本研究中，我们对新壳梭孢属物种的交配型基因进行了表征。设计了特异性引物，用于扩增几个物种中的MAT基因并进行测序，并开发了基于PCR的交配型诊断分析方法。同宗配合是所测试物种中的主要交配策略。此外，还评估了交配型基因序列在物种界定方面的潜力。对MAT基因进行了系统发育分析，并使用核糖体内部转录间隔区、翻译延伸因子1-α和β-微管蛋白的序列与多基因谱系进行了比较。基于交配型基因的系统发育可以区分所分析的物种，并且与使用更传统的多位点系统发育分析方法获得的结果一致。因此，MAT基因是界定新壳梭孢属中隐性物种的有力工具。
