Cultivo dirigido de microorganismos mediante metatranscriptómica

UNLABELLED: The vast majority of bacterial species remain uncultured, and this severely limits the investigation of their physiology, metabolic capabilities, and role in the environment. High-throughput sequencing of RNA transcripts (RNA-seq) allows the investigation of the diverse physiologies from uncultured microorganisms in their natural habitat. Here, we report the use of RNA-seq for characterizing the metatranscriptome of the simple gut microbiome from the medicinal leech Hirudo verbana and for utilizing this information to design a medium for cultivating members of the microbiome. Expression data suggested that a Rikenella-like bacterium, the most abundant but uncultured symbiont, forages on sulfated- and sialated-mucin glycans that are fermented, leading to the secretion of acetate. Histological stains were consistent with the presence of sulfated and sialated mucins along the crop epithelium. The second dominant symbiont, Aeromonas veronii, grows in two different microenvironments and is predicted to utilize either acetate or carbohydrates. Based on the metatranscriptome, a medium containing mucin was designed, which enabled the cultivation of the Rikenella-like bacterium. Metatranscriptomes shed light on microbial metabolism in situ and provide critical clues for directing the culturing of uncultured microorganisms. By choosing a condition under which the desired organism is rapidly proliferating and focusing on highly expressed genes encoding hydrolytic enzymes, binding proteins, and transporters, one can identify an organism's nutritional preferences and design a culture medium. IMPORTANCE: The number of prokaryotes on the planet has been estimated to exceed 10(30) cells, and the overwhelming majority of them have evaded cultivation, making it difficult to investigate their ecological, medical, and industrial relevance. The application of transcriptomics based on high-throughput sequencing of RNA transcripts (RNA-seq) to microorganisms in their natural environment can provide investigators with insight into their physiologies under optimal growth conditions. We utilized RNA-seq to learn more about the uncultured and cultured symbionts that comprise the relatively simple digestive-tract microbiome of the medicinal leech. The expression data revealed highly expressed hydrolytic enzymes and transporters that provided critical clues for the design of a culture medium enabling the isolation of the previously uncultured Rikenella-like symbiont. This directed culturing method will greatly aid efforts aimed at understanding uncultured microorganisms, including beneficial symbionts, pathogens, and ecologically relevant microorganisms, by facilitating genome sequencing, physiological characterization, and genetic manipulation of the previously uncultured microbes.

Resumen tomado de PubMed (NCBI) para el registro citado, en el idioma original. La versión de referencia es la de la publicación original.

El resumen se deja sin traducir para conservar la redacción original de los autores.

Los autores caracterizaron por RNA-seq el metatranscriptoma del microbioma intestinal de Hirudo verbana y lo emplearon para diseñar un medio de cultivo. Los datos sugirieron que la bacteria similar a Rikenella, el simbionte más abundante y no cultivado, aprovecha glicanos de mucinas sulfatadas y sialiladas y libera acetato; tinciones histológicas fueron compatibles con esas mucinas en el epitelio del buche. Un medio con mucina permitió cultivarla, lo que, según los autores, facilita el cultivo dirigido de microorganismos no cultivados.

Explore cómo esta investigación se conecta con la práctica clínica

Más estudios revisados por pares en la categoría «Genómica y proteómica».

• Leech Salivary Proteome and Transcriptome: 434 Deduced Protein Sequences — Liu Z, Tong X, Su Y, et al. (2019)
• Medicinal Leech Draft Genome — Anticoagulant Gene Survey — Kvist S, Manzano-Marín A, de Carle D, Trontelj P, Siddall ME (2020)
• La terapia con sanguijuelas (Hirudo medicinalis) atenúa las lesiones testiculares inducidas por isquemia/reperfusión testicular en un modelo animal — Davoodi F et al. (2021)
• Nueva proteína anticoagulante de la sanguijuela medicinal — Manuvera V et al. (2024)
• AIF-1 y RNASET2 desempeñan papeles complementarios en la respuesta inmunitaria innata de la sanguijuela medicinal — Baranzini N et al. (2019)
• Los péptidos antimicrobianos de la sanguijuela medicinal carentes de toxicidad representan una estrategia alternativa prometedora para combatir patógenos resistentes a antibióticos — Grafskaia E et al. (2019)
• Perspectivas estructurales sobre la actividad trombolítica de la destabilasa de la sanguijuela medicinal — Marin E et al. (2023)
• El genoma de la sanguijuela medicinal (Whitmania pigra) y un estudio genómico comparativo para la exploración de ingredientes bioactivos — Tong L et al. (2022)
• El SNC de la sanguijuela medicinal como modelo para estudios de exosomas en la comunicación cruzada entre microglía y neuronas — Raffo-Romero A et al. (2018)
• Transcriptomic profiling and the first spatial expression analysis of candidate genes in the salivary gland of the East Asian medicinal leech, Hirudo nipponia — Kim J et al. (2024)

Ver toda la investigación analizada por la ASH →