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Related Experiment Video

Updated: Sep 4, 2025

Polysome Purification from Soybean Symbiotic Nodules
07:02

Polysome Purification from Soybean Symbiotic Nodules

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Polysome Purification from Soybean Symbiotic Nodules.

María Martha Sainz1, Carla Valeria Filippi2, Guillermo Eastman3

  • 1Laboratorio de Bioquímica, Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la República; msainz@fagro.edu.uy.

Journal of Visualized Experiments : Jove
|July 18, 2022
PubMed
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This study presents a method to analyze the soybean (Glycine max) translatome in symbiotic nodules. It isolates polyribosomes and mRNA for RNA sequencing, revealing translational regulation in this complex plant tissue.

Area of Science:

  • Plant Molecular Biology
  • Genomics
  • Proteomics

Background:

  • Understanding gene expression regulation is crucial in symbiotic interactions.
  • The soybean nodule translatome offers insights into nitrogen fixation processes.
  • Current methods may not fully capture translational control in complex plant tissues.

Purpose of the Study:

  • To establish a protocol for isolating eukaryotic polyribosomes and associated mRNAs from soybean nodules.
  • To enable the study of translational regulation in Glycine max symbiotic nodules using RNA sequencing.
  • To provide a foundation for analyzing the soybean translatome at the mRNA level.

Main Methods:

  • Homogenization of frozen soybean nodules in polysome- and RNA-preserving conditions.
  • Isolation of total RNA and polysome-associated mRNA via ultracentrifugation.

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Last Updated: Sep 4, 2025

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  • Quality control of RNA samples using capillary electrophoresis for RNA-sequencing library preparation.

    • Main Results:

    • A robust protocol for isolating high-quality RNA and polysomes from soybean nodules was developed.
    • The method allows for the simultaneous analysis of both total transcriptome and translatome.
    • Demonstrated feasibility for downstream gene expression analysis at transcript and translatome levels.

    Conclusions:

    • This protocol facilitates the study of translational regulation in soybean symbiotic nodules.
    • It provides a valuable tool for investigating gene expression dynamics in complex plant tissues.
    • Combined with RNA-seq, this method enhances our understanding of eukaryotic mRNA regulation.