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Bulletin of Botanical Research ›› 2022, Vol. 42 ›› Issue (3): 512-520.doi: 10.7525/j.issn.1673-5102.2022.03.021

• Technique and Method • Previous Articles    

Establishment of Agrobacterium rhizogenes-mediated Genetic Transformation System of Eucalyptus urophylla × E. grandis

Ping LUO1,2, Haonan ZHANG1,3, Jianmin XU1, Bing HU1, Xiaoping WANG1, Guangyou LI1, Chunjie FAN1()   

  1. 1.State Key Laboratory of Tree Genetics and Breeding,Key Laboratory of State Forestry and Grassland Administration on Tropical Forestry,Research Institute of Tropical Forestry,Chinese Academy of Forestry,Guangzhou 510520
    2.Nanjing Forestry University,Nanjing 210037
    3.State Key Laboratory of Tree Genetics and Breeding,Northeast Forestry University,Harbin 150040
  • Received:2021-12-03 Online:2022-05-20 Published:2022-05-20
  • Contact: Chunjie FAN E-mail:fanchunjie@caf.ac.cn
  • About author:LUO Ping(1997—),female,postgraduate,forest genetics and breeding.
  • Supported by:
    Special Fund Project for Basic Scientific Research Business Expenses of Chinese Academy of Forestry(CAFYBB2020ZB004);Guangdong Foundation for Basic and Applied Basic Research(2020A1515011059)


Agrobacterium rhizogenes-mediated genetic transformation systemplays an important role in the identification of functional genes. To establish Agrobacterium rhizogenes-mediated genetic transformation system in Eucalyptus, the different strains infected the leaves and stems of Eucalyptus urophylla × E. grandis were used to select the appropriate A. rhizogenes strains and explant types, and the effects of Agrobacterium concentration and infection time on hairy root induction were explored. The results showed that MSU440 was the optimal A. rhizogenes strain and leaves was the preferred explants for hairy root induction. The highest hairy root induction rate reached 81.0% with the average root length of hairy roots was 3.23 cm. The leaves were infected by MSU440 which grown to a bacterial density of 0.3(OD600) for 30 min, then they were co-cultured for 48h and transferred the selection medium supplemented with 20 mg·L-1 Kanamycin. The exogenous genes were stably integrated in the genome of Eucalyptus hair roots which confirmed by PCR analyses and GUS staining, and the transformation efficiency reached 20.2%. In our study, an A. rhizogenes-mediated genetic transformation system of Eucalyptus was preliminarily established, which laid a foundation for gene function identification and further transgenic breeding of Eucalyptus.

Key words: Eucalyptus, Agrobacterium rhizogenes, hairy root, transgene, transformation

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