Krobanan Kulsumpun, Shen Wei-Chiang
Department of Plant Pathology and Microbiology, National Taiwan University, No. 1 Roosevelt Road, Sec. 4, Taipei, 10617, Taiwan.
Bot Stud. 2018 Jul 3;59(1):17. doi: 10.1186/s40529-018-0233-y.
The coprophilous ascomycete Sordaria fimicola usually reproduces sexually. Sexual differentiation in S. fimicola is accompanied by cellular and morphological changes, followed by multicellular tissue development to complete the sexual cycle. Although the morphological features of the sexual reproductive structure in S. fimicola have been well characterized, little is known about the nuclear dynamics and organization during these processes. Therefore, in this study, we successfully developed an Agrobacterium-mediated protoplast transformation protocol and generated histone H2B-mCherry-labeled S. fimicola strains. The life cycle of S. fimicola begins with germination of the ascospore and ends with the formation and discharge of new ascospores from the mature black sexual fruiting bodies, the so-called perithecia. The nuclear dynamics of the fluorescently labeled strains were examined during ascospore germination, hyphal elongation, and hyphal fusion using fluorescent microscopy.
Live imaging revealed that the nuclei in the germlings and fusion hyphae during the pre-contact interaction are located adjacent to the tip.
This is the first report of the application of a fluorescence labeling technique in S. fimicola. This application will help researchers gain a better understanding of nuclear distribution and investigate the protein-protein interaction networks during fruiting body formation for advanced molecular genetic studies in S. fimicola.
粪生子囊菌粪生粪壳菌通常进行有性繁殖。粪生粪壳菌的性别分化伴随着细胞和形态变化,随后是多细胞组织发育以完成有性周期。尽管粪生粪壳菌有性生殖结构的形态特征已得到充分表征,但对于这些过程中的核动力学和组织情况知之甚少。因此,在本研究中,我们成功开发了一种农杆菌介导的原生质体转化方案,并生成了组蛋白H2B-樱桃红标记的粪生粪壳菌菌株。粪生粪壳菌的生命周期始于子囊孢子的萌发,止于成熟黑色有性子实体(即所谓的子囊壳)中新子囊孢子的形成和释放。使用荧光显微镜检查了荧光标记菌株在子囊孢子萌发、菌丝伸长和菌丝融合过程中的核动力学。
实时成像显示,在接触前相互作用期间,芽管和融合菌丝中的细胞核位于顶端附近。
这是荧光标记技术在粪生粪壳菌中应用的首次报道。该应用将有助于研究人员更好地了解核分布,并研究子实体形成过程中的蛋白质-蛋白质相互作用网络,以进行粪生粪壳菌的高级分子遗传学研究。