Spermatogenesis generates mature male gametes and is critical for the proper transmission of genetic information between generations. However, the developmental landscapes of human spermatogenesis remain unknown. Here, we performed single-cell RNA sequencing (scRNA-seq) analysis for 2,854 testicular cells from donors with normal spermatogenesis and 174 testicular cells from one nonobstructive azoospermia (NOA) donor. A hierarchical model was established, which was characterized by the sequential and stepwise development of three spermatogonia subtypes, seven spermatocyte subtypes, and four spermatid subtypes. Further analysis identified several stage-specific marker genes of human germ cells, such as HMGA1, PIWIL4, TEX29, SCML1, and CCDC112. Moreover, we identified altered gene expression patterns in the testicular somatic cells of one NOA patient via scRNA-seq analysis, paving the way for further diagnosis of male infertility. Our work allows for the reconstruction of transcriptional programs inherent to sequential cell fate transition during human spermatogenesis and has implications for deciphering male-related reproductive disorders.

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It remains controversial whether the abnormal epigenetic modifications accumulated in the induced pluripotent stem cells (iPSCs) can ultimately affect iPSC pluripotency. To probe this question, iPSC lines with the same genetic background and proviral integration sites were established, and the pluripotency state of each iPSC line was characterized using tetraploid (4N) complementation assay. Subsequently, gene expression and global epigenetic modifications of "4N-ON" and the corresponding "4N-OFF" iPSC lines were compared through deep sequencing analyses of mRNA expression, small RNA profile, histone modifications (H3K27me3, H3K4me3, and H3K4me2), and DNA methylation. We found that methylation of an imprinted gene, Zrsr1, was consistently disrupted in the iPSC lines with reduced pluripotency. Furthermore, the disrupted methylation could not be rescued by improving culture conditions or subcloning of iPSCs. Moreover, the relationship between hypomethylation of Zrsr1 and pluripotency state of iPSCs was further validated in independent iPSC lines derived from other reprogramming systems.

Context. Mutualistic interactions between animals and plants shape the structure of plant-animal systems and, subsequently, affect plant-community structure and regeneration.Aims. To assess the effects of plant and rodent functional traits on the formation of mutualistic and predatory interactions regarding seed dispersal and predation in a warm-temperate forest.Methods. Seed scatter-hoarding and predation by six sympatric rodent species on seeds belonging to five sympatric tree species were tested under enclosure conditions. Key results. Functional traits (i.e. rodent body size and seed traits) are important to mutualism/predation in this seed-rodent system. The rodent-seed network is highly nested: large-sized rodents have mutualistic or predatory interactions with both large-and small-sized seed species, but small-sized rodents interacted with small-sized seed species only. Large seeds or seeds with hard coats enhanced mutualism and reduced predation.Conclusion. Body size of rodents and seed traits such as handling time and nutritional value are key factors in the formation of mutualistic and predatory interactions within seed-rodent systems.Implications. To promote seedling establishment in degenerated forests, introducing or protecting large-sized scatter hoarders and reducing the density of pure seed eaters are needed.Additional keywords: behavioural difference, plant-animal interactions, plant regeneration, seed dispersal.

As mutualists, seed dispersers may significantly affect mutualistic interactions and seedling recruitment of sympatric plants that share similar seed dispersers, but studies are rare. Here, we compared seed dispersal fitness in two co-occurring plant species (Armeniaca sibirica and Amygdalus davidiana) that inhabit warm temperate deciduous forest in northern China. We tested the hypothesis that seed trait-mediated selection by rodents may influence mutualistic interactions with rodents and then seedling establishment of co-occurring plant species. A. davidiana seeds are larger and harder (thick endocarps) than A. sibirica seeds, but they have similar levels of nutrients (crude fat, crude protein), caloric value and tannin. More A. sibirica seedlings are found in the field. Semi-natural enclosure tests indicated that the two seed species were both harvested by the same six rodent species, but that A. sibirica had mutualistic interactions (scatter hoarding) with four rodent species (Apodemus peninsulae, A. agrarius, Sciurotamias davidianus, Tamias sibiricus), and A. davidiana with only one (S. davidianus). Tagged seed dispersal experiments in the field indicated that more A. sibirica seeds were scatter-hoarded by rodents, and more A. sibirica seeds survived to the next spring and became seedlings. A. sibirica seeds derive more benefit from seed dispersal by rodents than A. davidiana seeds, particularly in years with limited seed dispersers, which well explained the higher seedling recruitment of A. sibirica compared with that of A. davidiana under natural conditions. Our results suggest that seed dispersers may play a significant role in seedling recruitment and indirect competition between co-occurring plant species.

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