Development of Protoplast-Based Gene Editing System for Areca Palm
Hao Nie,
Saira Batool,
Yin Min Htwe,
Xiaomeng Fang,
Dapeng Zhang,
Peng Shi,
Zhiying Li,
Mingjun Ma,
Hanlu Su,
Qun Yu,
Xiangman He,
Yong Wang
Affiliations
Hao Nie
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Saira Batool
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Yin Min Htwe
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Xiaomeng Fang
National Nanfan Research Institute, Chinese Academy of Agricultural Sciences, Sanya 572000, China
Dapeng Zhang
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Peng Shi
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Zhiying Li
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Mingjun Ma
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Hanlu Su
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Qun Yu
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Xiangman He
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Yong Wang
National Key Laboratory for Tropical Crop Breeding/Coconut Research Institute/Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572025, China
Areca palm (Areca catechu L.) is an economically significant crop in tropical and subtropical regions. However, an efficient transformation and gene editing system for genetic improvement has still not been established. In this study, protoplasts were isolated from juvenile leaves, followed by PEG-mediated transformation and gene editing targeting the areca palm AcPDS via the CRISPR/Cas9 system. High yield (9.08 × 106 cells/g FW) and viability (91.57%) protoplasts were isolated successfully by digestion for 5 h in an enzyme solution. Transformation efficiency (11.85%) was obtained through PEG-mediated transformation (incubation for 30 min in the mixture containing 40% PEG-4000, 400 mM CaCl2, 30 µg of plasmid DNA, and 100 µL of protoplasts). Furthermore, subcellular localization was established by the cotransformation of GFP and pNLS-mCherry in the protoplasts. Moreover, the editing efficiency (2.82%) of AcPDS using the CRISPR/Cas9 system was detected by Hi-TOM sequencing. This study established an efficient transformation and gene editing system based on protoplasts in areca palm, which will be beneficial for gene function verification and genetic improvement in areca palm and other tropical palm crops.
