Chinese Bulletin of Botany ›› 2019, Vol. 54 ›› Issue (4): 515-521.DOI: 10.11983/CBB18257

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

Agrobacterium rhizogenes-mediated Transformation System of Spinacia oleracea

Yue Xu1,Yingping Cao2,Yu Wang2,Chunxiang Fu2,*(),Shaojun Dai1,3,*()   

  1. 1 Key Laboratory of Saline-alkali Vegetation Ecology Restoration, Ministry of Education, Alkali Soil Natural Environmental Science Center, Northeast Forestry University, Harbin 150040, China
    2 Shandong Technology Innovation Center of Synthetic Biology, Shandong Provincial Key Laboratory of Energy Genetics, Key Laboratory of Biofuels, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266000, China
    3 Development Center of Plant Germplasm Resources, College of Life Sciences, Shanghai Normal University, Shanghai 200234, China
  • Received:2018-11-30 Accepted:2019-04-08 Online:2019-07-10 Published:2020-01-08
  • Contact: Chunxiang Fu,Shaojun Dai

Abstract:

The plant can produce hairy roots after infection by Agrobacterium rhizogenes harboring Ri plasmids. Spinach (Spinacia oleracea) is a common edible vegetable, and its hairy root system has yet to be reported. In this study, A. rhizogenes LBA9402 was screened to be suitable for hairy root induction of spinach. The highest induction efficiency, 16%, was reached when the spinach stem was infected by LBA9402. Hairy roots grew vigorously on SH solid medium without exogenous hormones. The hairy roots of spinach were white and had enormous root hairs. The regeneration plant obtained from the callus redifferentiated from spinach hair roots; the rate of regeneration was 8%. In addition, Ri T-DNA and Ti T-DNA carrying rolB genes and GFP gene, respectively, were co-transferred into cells of spinach and produced transgenic hairy roots. PCR analysis and fluorescence microscopy assays showed rolB gene and GFP gene in the hairy roots of spinach and expressed stably; the rate of co-transformation was 50%.

Key words: co-transformation, hairy root, plant regeneration system, Spinacia oleracea