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Highly efficient endosperm and pericarp protoplast preparation system for transient transformation of endosperm-related genes in wheat

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Abstract

Plant protoplasts constitute a versatile system for transient gene expression and have frequently been used in high-throughput to screen and identify functional characterization of plant genes. Wheat (Triticum aestivum L.) is one of the most important crops for our daily life. Endosperm-trait related genes are associated with grain yield or quality in wheat. However, very few studies have explored on the use of protoplasts isolated from endosperm and pericarp tissue of developing grain. In this study, endosperm tissues of developing wheat grains at 8 DPA (days post-anthesis) were collected. It was shown that, after being digested with the enzymolysis solution containing 0.714 M mannitol for 2 h, total 1.1 × 105 of intact protoplasts containing 80% vital individuals were isolated from 0.6 g samples. Pericarp protoplasts were successfully purified from wheat grains at 4 DPA using the optimized method. Transcription factor TaABI5 and amyloplast protein TaSSIIIa were transfected to the prepared protoplasts, and they were successfully localized in the nucleus and the surface of starch granule, respectively. It is an effective and reproductive method for endosperm and pericarp protoplast isolation and of great importance to further investigate gene’s functions and regulations related to endosperm development and differentiation in plants.

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Endosperm and pericarp protoplast preparation system for endosperm-related gene transformation in wheat.

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Acknowledgements

This research was supported by grants from the Natural Science Foundation of Beijing (6212001), and National Natural Science Foundation of China (31571652).

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Authors and Affiliations

  1. College of Life Sciences, Capital Normal University, No. 105, North Xi San Huan Road, Beijing, 100048, China

    Shichen Han, Ge Qu, Xiaohui Li & Feixiong Zhang

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  1. Shichen Han
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  2. Ge Qu
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  3. Xiaohui Li
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  4. Feixiong Zhang
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Contributions

FZ conceived and designed experiments. SH performed protoplast isolation and subcellular localization. GQ contributed to starch granule analysis. SH, XL and FZ wrote the paper. All authors read and approved the final manuscript.

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Correspondence to Feixiong Zhang.

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Communicated by Ewa Grzebelus.

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Han, S., Qu, G., Li, X. et al. Highly efficient endosperm and pericarp protoplast preparation system for transient transformation of endosperm-related genes in wheat. Plant Cell Tiss Organ Cult 155, 165–174 (2023). https://doi.org/10.1007/s11240-023-02561-z

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  • DOI: https://doi.org/10.1007/s11240-023-02561-z

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