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Isolation of active mitochondria from tomato fruit.

H S Ku1, H K Pratt, A R Spurr

  • 1Department of Vegetable Crops, University of California, Davis, California 95616.

Plant Physiology
|June 1, 1968
PubMed
Summary
This summary is machine-generated.

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This study details an improved method for isolating intact mitochondria from tomato fruit, yielding higher respiratory control ratios. The technique is also effective for cantaloupe and honeydew melon isolation.

Area of Science:

  • Plant Biology
  • Cell Biology
  • Biochemistry

Background:

  • Mitochondria are crucial for cellular respiration and energy production in plants.
  • Efficient isolation of intact mitochondria is essential for studying their function and integrity.
  • Previous methods for isolating plant mitochondria, particularly from fruit tissues, have limitations in yield and quality.

Purpose of the Study:

  • To develop and describe an improved protocol for isolating intact mitochondria from tomato fruit (Lycopersicon esculentum Mill.).
  • To ensure the isolated mitochondria exhibit high integrity and functionality, indicated by respiratory control ratio.
  • To validate the applicability of the developed method to other fruit species.

Main Methods:

  • Tomato fruit tissue was chilled and finely sliced in a specialized buffer (0.4 M sucrose, 2 mM MgCl2, 8 mM EDTA, 4 mM cysteine, 10 mM KCl, 0.5 mg/ml BSA, 50 mM Tris-HCl, pH 7.6).

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  • Buffer pH was maintained between 7.0-7.2 during tissue processing.
  • Tissue was filtered through cheesecloth, followed by differential centrifugation (2000 x g for 15 min, then 11,000 x g for 20 min).
  • Isolated mitochondria were washed with a specific sucrose-based medium.

    • Main Results:

    • The optimized method yielded homogeneous and relatively intact mitochondria from tomato fruit.
    • Electron microscopy confirmed the structural integrity of the isolated mitochondria.
    • The isolated mitochondria demonstrated a higher respiratory control ratio compared to those obtained by other reported methods.
    • The protocol was successfully applied to cantaloupe and honeydew melon fruits.

    Conclusions:

    • This improved isolation method provides high-quality mitochondria from tomato fruit, suitable for biochemical and functional studies.
    • The enhanced integrity and respiratory control ratio of isolated mitochondria advance plant organelle research.
    • The method's versatility across different melon species suggests its broad utility in fruit mitochondrial research.