Robust CRISPR/Cas9 mediated genome editing and its application in manipulating plant height in the first generation of hexaploid Ma bamboo (Dendrocalamus latiflorus Munro)

• Published in: Plant biotechnology journal 2020-07, Vol.18 (7), p.1501-1503
• Main Authors: Ye, Shanwen, Chen, Gang, Kohnen, Markus V., Wang, Wenjia, Cai, Changyang, Ding, WenSha, Wu, Chu, Gu, Lianfeng, Zheng, Yushan, Ma, Xiangqing, Lin, Chentao, Zhu, Qiang
• Format: Article
• Language: English
• Subjects: Albinism; Bamboo; Brief Communication; Brief Communications; Budding; Cas9; Cloning; CRISPR; Gene manipulation; Genes; Genetic engineering; Genetic transformation; Genome editing; Genomes; Genomics; International trade; Molecular modelling; Mutation; Offspring; Plasmids
• ISSN: 1467-7644; 1467-7652
• DOI: 10.1111/pbi.13320

Here, we reported the generation of bamboo mutants with CRISPR/Cas9 technology by targeting one specific copy or all homoeologous genes. Since our recently established genetic transformation protocol is time‐consuming (~1.5 years; Ye et al., ), we optimized the CRISPR/Cas9 system in bamboo protoplast. The ratios of large fragment deletions (≥14 bp), insertions and combined indels were 15.9%, 2.21% and 7.82%, respectively (Figure G). Since bamboo propagates through asexual budding, those homozygote/biallelic mutations will remain in the genome of their offspring clones during breeding. sgRNA2 that perfectly targets DlmPSY1‐A1, but not DlmPSY1‐B1 or DlmPSY1‐C1 was designed to study the recognition specificity (Figure D). To our knowledge, this is the first example on controlling bamboo height through gene manipulation, which will contribute to subsequent studies on the molecular mechanisms behind the fast growth of bamboo. The homozygote mutations were obtained in the first generation of transgenic lines, which are extremely important for bamboo species due to its long vegetative growth periods.