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Preparation of Barley AGPS2b Antibody and Its Application in Hormone Regulation Research

Author

Listed:
  • Boai Xi

    (State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China)

  • Qiyan Zhou

    (State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China)

  • Yang Guo

    (State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China)

  • Noman Shaoib

    (Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China)

  • Zhenbin Cheng

    (State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China)

  • Yan Gao

    (State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China)

  • Yajie Liu

    (State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China)

  • Hui Zhao

    (State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China)

  • Zongyun Feng

    (State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China)

  • Guowu Yu

    (State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130, China)

Abstract

ADP-glucose pyrophosphorylase (AGPase), which is a key enzyme in the starch biosynthesis pathway, plays a critical role in barley grain development. Despite its importance, the regulatory mechanisms governing AGPase expression, particularly the influence of plant hormones, remain poorly understood in barley. To address this, we identified and characterized the HvAGPS2b gene, which encodes the AGPase small subunit. The full-length HvAGPS2b gene was cloned from the barley database and expressed as a recombinant protein using the pET-30a system. Polyclonal antibodies were prepared against HvAGPS2b to facilitate detailed analysis. Our findings revealed that HvAGPS2b , as a small subunit of the rate-limiting enzyme AGPase, is integral to the later stages of grain development. Furthermore, RT-qPCR and Western blotting analyses showed that the phytohormones ABA, GA, ETH, and BR significantly upregulated the expression of AGPase small subunits. These results underscore the vital role of plant hormones in modulating AGPS2b expression, thereby influencing grain development. This study provides significant insights into the hormonal regulation of starch biosynthesis and establishes a foundation for further investigation into the functional dynamics of AGPase in barley.

Suggested Citation

  • Boai Xi & Qiyan Zhou & Yang Guo & Noman Shaoib & Zhenbin Cheng & Yan Gao & Yajie Liu & Hui Zhao & Zongyun Feng & Guowu Yu, 2024. "Preparation of Barley AGPS2b Antibody and Its Application in Hormone Regulation Research," Agriculture, MDPI, vol. 14(10), pages 1-16, September.
    • Handle: RePEc:gam:jagris:v:14:y:2024:i:10:p:1712-:d:1489007
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