A rapid and universal method for isolating starch granules in plant tissues

Area of Science:

• Plant Biology
• Biochemistry
• Molecular Biology

Background:

• Starch granules (SGs) are crucial for plant carbohydrate storage and function.
• Isolating high-quality SGs from diverse plant tissues is essential for understanding their roles in plant development and stress responses.
• Current methods face challenges in isolating small or low-quantity SGs from leaves and pollen.

Purpose of the Study:

• To develop a novel, efficient, and versatile method for isolating starch granules (SGs) from various plant tissues.
• To demonstrate the applicability of the new method for purifying SGs from maize seeds, pollen, and leaves.

Main Methods:

• Development of an aqueous two-phase system (ATS) using ethanol/NaH2PO4 for SG isolation.
• Separation of SGs based on differences in density, solubility, and polarity.
• Biochemical, microscopic, and proteomic analyses to assess SG purity and composition.

Main Results:

• The ATS method successfully isolated and purified SGs from maize seeds, pollen, and leaves.
• Analyses confirmed the high purity of the isolated SGs.
• Proteomic analysis revealed distinct protein profiles between seed and pollen SGs.

Conclusions:

• The developed ATS-based method is a simple, rapid, low-cost, and highly reproducible technique for SG isolation.
• This method is broadly applicable to various starch-containing plant tissues and species.
• The technique facilitates further research into SG functions and composition across different plant organs.