文献类型：期刊文献

英文题名：Adventitious bud regeneration and Agrobacterium tumefaciens-mediated genetic transformation of Eucalyptus urophylla x E. tereticornis interspecific hybrid

作者：Wang, Xiaoping[1,2,3] Luo, Ping[1,2,3] Qiu, Zhenfei[2] Li, Xiaodan[2] Zeng, Bingshan[2] Fan, Chunjie[1,2]

第一作者：Wang, Xiaoping

通信作者：Fan, CJ[1];Fan, CJ[2]

机构：[1]Chinese Acad Forestry, State Key Lab Tree Genet & Breeding, Beijing 100091, Peoples R China;[2]Chinese Acad Forestry, Res Inst Trop Forestry, Key Lab State Forestry Adm Trop Forestry, Guangzhou 510520, Peoples R China;[3]Nanjing Forestry Univ, Nanjing 210037, Peoples R China

年份：2022

卷号：58

期号：3

起止页码：416-426

外文期刊名：IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-PLANT

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000716382400001)】；

基金：This work was supported by the Fundamental Research Funds for the Central Non-profit Research Institution of CAF (Grant No. CAFYBB2020ZB004).

语种：英文

外文关键词：Eucalyptus; Agrobacterium; Adventitious buds; Transformation

摘要：A high-efficiency regeneration and genetic transformation system is indispensable for generating desirable traits in important trees such as Eucalyptus. However, lower regeneration efficiency is common for most varieties because of the recalcitrance of this genus. Here, a stable and highly efficient in vitro organogenesis protocol and Agrobacterium-mediated genetic transformation system of Eucalyptus were developed, and transgenic plants were obtained. In this protocol, the preferred explants were the top and middle stem internodes from in vitro micro-shoots of the E. urophylla x E. tereticornis hybrid. Modified Woody Plant Medium (mWPM) containing 0.025 mg center dot L-1 thidiazuron (TDZ) and 0.10 mg center dot L-1 indole-3-butyric acid (IBA) was used to induce multiple adventitious buds that allowed 85.6% shoot formation. The binary vector pBI121 carrying the neomycin phosphotransferase II (nptII) and beta-glucuronidase (uidA) genes was applied for transformation. The preferred internodes were precultured for 0 to 3 d and infected with A. tumefaciens strain GV3101 grown to a bacterial density of 0.5 (OD600). Then, they were transferred to a co-culture medium supplemented with 50 mu M acetosyringone (AS) and co-cultured for 2 d in the dark. The transgenic adventitious buds formed in regeneration medium, which was replaced by the same medium with a 2-wk subculture interval through kanamycin selection. Using the aforementioned method, transgenic plantlets can be obtained within 3 mo with a transformation frequency of 3.8%, which was verified by polymerase chain reaction amplification (PCR) and histochemical analysis of GUS activity. The constructed genetic transformation system will lay a foundation for mining gene functions and further molecular breeding of Eucalyptus.