文献类型：期刊文献

英文题名：Agrobacterium-mediated genetic transformation of the most widely cultivated superior clone Eucalyptus urophylla x E. grandis DH32-29 in Southern China

作者：Wang, Xiaoping[1,2] Chen, Shanshan[1,2,3] Zhang, Haonan[1,2,4] Luo, Ping[1,2] Zhou, Fangping[1,2] Zeng, Bingshan[2] Xu, Jianmin[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, Peoples R China;[2]Chinese Acad Forestry, Res Inst Trop Forestry, Key Lab State Forestry & Grassland Adm Trop Forest, Guangzhou, Peoples R China;[3]Northeast Forestry Univ, State Key Lab Tree Genet & Breeding, Harbin, Peoples R China;[4]Northeast Forestry Univ, Coll Life Sci, Harbin, Peoples R China

年份：2023

卷号：13

外文期刊名：FRONTIERS IN PLANT SCIENCE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000919467700001)】；

语种：英文

外文关键词：Eucalyptus; regeneration; adventitious bud; Agrobacterium tumefaciens; genetic transformation

摘要：Eucalyptus, as an economically important species for wood and paper industries, remains a challenge to genetic improvement by transgenic technology owing to the deficiency of a highly efficient and stable genetic transformation system, especially in cultivated superior clones. Eucalyptus urophylla x E. grandis clone DH32-29 is most widely planted in southern China, but it is relatively recalcitrant to adventitious bud regeneration, which blocks the establishment of a genetic transformation system. Here, an efficient adventitious bud regeneration and transformation system of Eucalyptus was established using E. urophylla x E. grandis DH32-29 as material. The in vitro leaves from microshoots that were subcultured for 20-25 days were immersed into liquid Woody Plant Medium supplemented with 0.02 mg center dot L-1 alpha-naphthaleneacetic acid (NAA) and 0.24 mg center dot L-1 forchlorfenuron [callus-inducing medium (CIM)]. After 15 days, explants were transferred to a medium containing 0.10 mg center dot L-1 NAA and 0.50 mg center dot L-1 6-benzyladenine (shoot-inducing medium, SIM) for adventitious bud induction. The highest regeneration efficiency of adventitious buds was 76.5%. Moreover, an Agrobacterium tumefaciens-mediated genetic transformation system was optimized. The leaves were precultured for 7 days and infected for 30 min with A. tumefaciens strain EHA105 grown to a bacterial density of 0.3 (OD600). After 72 h of cocultivation in the dark, leaves were transferred to CIM supplemented with 100 mg center dot L-1 cefotaxime (Cef), 100 mg center dot L-1 timentin, and 15 mg center dot L-1 kanamycin (Kan) for 15 days to induce calluses. Then, the explants were transferred to SIM supplemented with the same concentration of antibiotics, and the fresh medium was replaced every 15 days until resistant adventitious buds appeared. After inducing roots in root-inducing medium supplemented with 200 mg center dot L-1 Cef and 75 mg center dot L-1 Kan, completely transgenic plants were obtained. Using the aforementioned method, the transformation frequency can reach 1.9%. This provides a powerful approach for genetic improvement of E. urophylla x E. grandis DH32-29 and gene function analysis in Eucalyptus.