文献类型：期刊文献

英文题名：An Efficient Agrobacterium-Mediated Transformation Method for Hybrid Poplar 84K (Populus alba x P. glandulosa) Using Calli as Explants

作者：Wen, Shuang-Shuang[1] Ge, Xiao-Lan[1] Wang, Rui[1] Yang, Hai-Feng[2] Bai, Yu-E.[2] Guo, Ying-Hua[1] Zhang, Jin[3] Lu, Meng-Zhu[1,3] Zhao, Shu-Tang[1] Wang, Liu-Qiang[1,4]

第一作者：Wen, Shuang-Shuang

通信作者：Wang, LQ[1];Wang, LQ[2]

机构：[1]Chinese Acad Forestry, Res Inst Forestry, State Key Lab Tree Genet & Breeding, Key Lab Tree Breeding & Cultivat State Forestry A, Beijing 100091, Peoples R China;[2]Inner Mongolia Agr Univ, Coll Forestry, Hohhot 010019, Peoples R China;[3]Zhejiang A&F Univ, Coll Forestry & Biotechnol, State Key Lab Subtrop Silviculture, Hangzhou 311300, Peoples R China;[4]Nanjing Forestry Univ, Coinnovat Ctr Sustainable Forestry Southern China, Nanjing 210037, Peoples R China

年份：2022

卷号：23

期号：4

外文期刊名：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000770650900001)】；

基金：This work was supported by the National Key Research and Development Program of China (2021YFD2200205) and National Natural Science Foundation of China (31971620).

语种：英文

外文关键词：Agrobacterium; hybrid poplar; callus; plant transformation; regeneration

摘要：A highly efficient Agrobacterium-mediated transformation method is needed for the molecular study of model tree species such as hybrid poplar 84K (Populus alba x P. glandulosa cv. '84K'). In this study, we report a callus-based transformation method that exhibits high efficiency and reproducibility. The optimized callus induction medium (CIM1) induced the development of calli from leaves with high efficiency, and multiple shoots were induced from calli growing on the optimized shoot induction medium (SIM1). Factors affecting the transformation frequency of calli were optimized as follows: Agrobacterium concentration sets at an OD600 of 0.6, Agrobacterium infective suspension with an acetosyringone (AS) concentration of 100 mu M, infection time of 15 min, cocultivation duration of 2 days and precultivation duration of 6 days. Using this method, transgenic plants are obtained within approximately 2 months with a transformation frequency greater than 50%. Polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR) and beta-galactosidase (GUS) histochemical staining analyses confirmed the successful generation of stable transformants. Additionally, the calli from leaves were subcultured and used to obtain new explants; the high transformation efficiency was still maintained in subcultured calli after 6 cycles. This method provides a reference for developing effective transformation protocols for other poplar species.