Chinese Bulletin of Botany ›› 2017, Vol. 52 ›› Issue (4): 511-519.DOI: 10.11983/CBB16155

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Preparation of Protoplast and Establishment of Transient Expression System in Gerbera hybrida

Aihua Song1, Wenbin Zhang1, Shulan Sun1, Lingfei Li1,2*, Xiaojing Wang1   

  1. 1Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou 510631, China
    2Key Laboratory of Southern Subtropical Plant Diversity, Fairy Lake Botanical Garden, Shenzhen & Chinese Academy of Sciences, Shenzhen 518004, China;
  • Received:2016-07-22 Accepted:2016-10-16 Online:2017-07-01 Published:2017-05-05
  • Contact: Li Lingfei
  • About author:

    # Co-first authors

Abstract: Gerbera hybrida has been used as model plant to study the evolution and development of composite inflorescences. To establish an efficient protoplast isolation and transient transformation system in G. hybrida, leaves of cultivar ‘Shenzhen No. 5’ in vitro seedlings were used as materials and the effects of different concentrations of cellulase R10 and macerozyme were tested to screen a suitable preparation method. The best enzyme solution concentration consisted of cellulase R10 2.0% (w/v), macerozyme 0.3% (w/v), and 0.4 mol·L-1 mannitol and more than 2.2×107∙g-1 FW protoplasts with high activity were obtained after 4 h treatment under 25°C condition. Subcellular localization and protein-protein interaction assays were performed using these protoplasts and revealed 80% transformation rate. Collectively, this study established an efficient protoplast isolation and transient assay system that can provide an important platform to facilitate gene function studies in G. hybrida.