Single-cell transcriptome atlas reveals somatic cell embryogenic differentiation features during regeneration.

Understanding somatic cell totipotency remains a challenge facing scientific inquiry today. Plants display remarkable cell totipotency expression, illustrated by single-cell differentiation during somatic embryogenesis (SE) for plant regeneration. Determining cell identity and exploring gene regulation in such complex heterogeneous somatic cell differentiation have been major challenges. Here, we performed high-throughput single-cell sequencing assays to define the precise cellular landscape and revealed the modulation mode of marker genes during embryogenic differentiation in cotton (Gossypium hirsutum L.) as the crop for biotechnology application. We demonstrated that nonembryogenic calli (NEC) and primary embryogenic calli (PEC) tissues were composed of heterogeneous cells that could be partitioned into four broad populations with six distinct cell clusters. Enriched cell clusters and cell states were identified in NEC and PEC samples, respectively. Moreover, a broad repertoire of new cluster-specific genes and associated expression modules were identified. The energy metabolism, signal transduction, environmental adaptation, membrane transport pathways, and a series of transcription factors were preferentially enriched in cell embryogenic totipotency expression. Notably, the SE-ASSOCIATED LIPID TRANSFER PROTEIN (SELTP) gene dose-dependently marked cell types with distinct embryogenic states and exhibited a parabolic curve pattern along the somatic cell embryogenic differentiation trajectory, suggesting that SELTP could serve as a favorable quantitative cellular marker for detecting embryogenic expression at the single-cell level. In addition, RNA velocity and Scissor analysis confirmed the pseudo-temporal model and validated the accuracy of the scRNA-seq data, respectively. This work provides valuable marker-genes resources and defines precise cellular taxonomy and trajectory atlases for somatic cell embryogenic differentiation in plant regeneration.

Chironomidae larvae: A neglected enricher of antibiotic resistance genes in the food chain of freshwater environments.

Infection caused by pathogenic bacteria carrying antibiotic resistance genes (ARGs) is a serious challenge to human health. Water environment, including water and surface sediments, is an important repository of ARGs, and the activity of aquatic animal can affect the development of ARG pollution in the water environment. Macrobenthic invertebrates are an important component of aquatic ecosystems, and their effects on ARG development in aquatic environments remain unreported. The distribution of ARGs, including tetA gene, sul2 gene, and kan gene, in Chironomidae larvae is demonstrated in this study for the first time. The ARG distribution was related to sampling points, metal elements, and seasons. Animal models demonstrated that Chironomidae larvae enriched ARGs from water and passed them on to downstream predators in the food chain. Conjugative transfer mediated by resistant plasmids was crucial in the spread of ARG in Chironomidae larvae, and upregulated expression of trfAp gene and trbBp gene was the molecular mechanism. Escherichia in Proteobacteria and Flavobacterium in Bacteroidetes, which are gram-negative bacteria in Chironomidae larvae, are the primary host bacteria of ARGs confirmed via resistance screening and DNA sequencing of V4 region of 16S rRNA gene. Feeding experiments further confirmed that ARGs from Chironomidae larvae can be enriched in the fish gut. Research gaps in food chain between sediments and fish are addressed in this study, and Chironomidae larvae is an important enricher of ARGs in the freshwater environment.