The fungal genus Massospora (Zoopagomycota: Entomophthorales) includes more than a dozen obligate, sexually transmissible pathogenic species that infect cicadas (Hemiptera) worldwide. At least two species are known to produce psychoactive compounds during infection, which has garnered considerable interest for this enigmatic genus. As with many Entomophthorales, the evolutionary relationships and host associations of Massospora spp. are not well understood. The acquisition of M. diceroproctae from Arizona, M. tettigatis from Chile, and M. platypediae from California and Colorado provided an opportunity to conduct molecular phylogenetic analyses and morphological studies to investigate whether these fungi represent a monophyletic group and delimit species boundaries. In a three-locus phylogenetic analysis including the D1–D2 domains of the nuclear 28S rRNA gene (28S), elongation factor 1 alpha-like (EFL), and beta-tubulin (BTUB), Massospora was resolved in a strongly supported monophyletic group containing four well-supported genealogically exclusive lineages, based on two of three methods of phylogenetic inference. There was incongruence among the single-gene trees: two methods of phylogenetic inference recovered trees with either the same topology as the three-gene concatenated tree (EFL) or a basal polytomy (28S, BTUB). Massospora levispora and M. platypediae isolates formed a single lineage in all analyses and are synonymized here as M. levispora. Massospora diceroproctae was sister to M. cicadina in all three single-gene trees and on an extremely long branch relative to the other Massospora, and even the outgroup taxa, which may reflect an accelerated rate of molecular evolution and/or incomplete taxon sampling. The results of the morphological study presented here indicate that spore measurements may not be phylogenetically or diagnostically informative. Despite recent advances in understanding the ecology of Massospora, much about its host range and diversity remains unexplored. The emerging phylogenetic framework can provide a foundation for exploring coevolutionary relationships with cicada hosts and the evolution of behavior-altering compounds.

真菌属Massospora（Zoopagomycota：虫霉目），包括十多个专，性传播致病菌种可感染蝉（半翅目）全球。已知至少有两个物种在感染过程中产生精神活性化合物，这对该神秘属引起了极大的兴趣。与许多昆虫纲一样，马氏孢属的进化关系和寄主联系。尚未完全了解。收购M. diceroproctae来自亚利桑那州，M. tettigatis从智利和M. platypediae来自加利福尼亚州和科罗拉多州的真菌提供了进行分子系统发育分析和形态学研究的机会，以调查这些真菌是否代表一个单系群体并划定物种边界。在三轨迹系统发育分析包括核28S rRNA基因（28S），延伸因子1α样（的D1-D2结构域EFL），和β-微管蛋白（BTUB），Massospora在强烈支持的单系组解析基于系统发育推断的三种方法中的两种，包含四个得到充分支持的族谱排他谱系。单基因树之间的不一致：系统发育推理的两种恢复树的拓扑结构与三基因级联树（EFL）相同）或基底膜切除术（28S，BTUB）。Massospora levispora和M. platypediae菌株形成在所有分析的单个谱系和这里被作为异名M. levispora。Massospora diceroproctae是姐妹M. cicadina在所有三个单基因树木和相对极长分支到另一个上Massospora，甚至外群，这可能反映了分子进化和/或不完整的分类单元采样的加速速率。此处呈现的形态学研究结果表明，孢子测量可能在系统发育或诊断上均无意义。尽管最近在了解马氏菌的生态方面取得了进展，关于其寄主范围和多样性的许多方面仍待探索。新兴的系统发育框架可以为探索与蝉宿主的进化关系和改变行为的化合物提供基础。