文献类型：期刊文献

英文题名：An Efficient Genetic Transformation and CRISPR/Cas9-Based Genome Editing System for Moso Bamboo (Phyllostachys edulis)

作者：Huang, Biyun[1,2] Zhuo, Renying[1,2] Fan, Huijin[1,2] Wang, Yujun[1,2] Xu, Jing[1,2] Jin, Kangming[1,2] Qiao, Guirong[1,2]

第一作者：Huang, Biyun

通信作者：Qiao, GR[1];Qiao, GR[2]

机构：[1]Chinese Acad Forestry, State Key Lab Tree Genet & Breeding, Beijing, Peoples R China;[2]Chinese Acad Forestry, Res Inst Subtrop Forestry, Key Lab Tree Breeding Zhejiang Prov, Hangzhou, Peoples R China

年份：2022

卷号：13

外文期刊名：FRONTIERS IN PLANT SCIENCE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000761122800001)】；

基金：This research was funded by the National Key Research and Development Program of China (Grant No. 2021YFD2200504) and the National Non-profit Institute Research Grant of Chinese Academy of Forestry (Grant No. CAFYBB2020ZB004).

语种：英文

外文关键词：Phyllostachys edulis; plant regeneration; genetic transformation; genome editing; immature embryo culture

摘要：Moso bamboo (Phyllostachys edulis) is the most important monopodial bamboo species worldwide. Without a genetic transformation system, it is difficult to verify the functions of genes controlling important traits and conduct molecular breeding in moso bamboo. Here, we established a plant regeneration system from immature embryos. Calli were induced on MS medium added 4-6 mg.L-1 2,4-dichlorophenoxyacetic acid (2,4-D) with high efficiency (>60%). A plant growth regulator combination of 0.5 mg.L-1 1-naphthylacetic acid (NAA), 2.0 mg.L-1 6-benzylaminopurine (BAP), and 3.0 mg.L-1 zeatin (ZT) was suitable for shoot differentiation, and the shoot induction frequency was increased to 43% after 0.5 mg.L-1 abscisic acid (ABA) pretreatment. An effective antibiotic screening concentration was determined by hygromycin sensitivity test. We further optimized the Agrobacterium concentration and added vacuum infiltration for infection, which improves the transient expression efficiency. A genetic transformation system was established for the first time in moso bamboo, with the transformation efficiency of approximately 5%. To optimize genome editing, two endogenous U3 small nuclear RNA (snRNA) promoters were isolated and used to drive small guide RNA (sgRNA) expression. The results showed that the PeU3.1 promoter exhibited higher efficiency, and it was used for subsequent genome editing. Finally, homozygous pds1pds2 mutants were obtained by an efficient CRISPR/Cas9 genome-editing system. These technical systems will be conducive to gene functional validation and accelerate the molecular breeding process of moso bamboo.