Development of an Agrobacterium-mediated CRISPR/Cas9 system in pea (Pisum sativum L.)
Guan Li,
Rong Liu,
Rongfang Xu,
Rajeev K. Varshney,
Hanfeng Ding,
Mengwei Li,
Xin Yan,
Shuxian Huang,
Juan Li,
Dong Wang,
Yishan Ji,
Chenyu Wang,
Junguang He,
Yingfeng Luo,
Shenghan Gao,
Pengcheng Wei,
Xuxiao Zong,
Tao Yang
Affiliations
Guan Li
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Rong Liu
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Rongfang Xu
Key Laboratory of Rice Genetic Breeding of Anhui Province, Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei 230031, Anhui, China
Rajeev K. Varshney
State Agricultural Biotechnology Centre, Centre for Crop and Food Innovation, Food Futures Institute, Murdoch University, Murdoch, WA 6150, Australia
Hanfeng Ding
Institute of Crop Germplasm Resources, Shandong Academy of Agricultural Sciences / Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Jinan 250100, Shandong, China
Mengwei Li
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Xin Yan
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Shuxian Huang
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Juan Li
Key Laboratory of Rice Genetic Breeding of Anhui Province, Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei 230031, Anhui, China
Dong Wang
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Yishan Ji
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Chenyu Wang
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Junguang He
Zhejiang Xinnan Chemical Industrial Group Co., Ltd., Hangzhou 311300, Zhejiang, China
Yingfeng Luo
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
Shenghan Gao
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
Pengcheng Wei
Key Laboratory of Rice Genetic Breeding of Anhui Province, Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei 230031, Anhui, China; Corresponding authors.
Xuxiao Zong
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; Corresponding authors.
Tao Yang
National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; Corresponding authors.
Pea (Pisum sativum L.) is an annual cool-season legume crop. Owing to its role in sustainable agriculture as both a rotation and a cash crop, its global market is expanding and increased production is urgently needed. For both technical and regulatory reasons, neither conventional nor transgenic breeding techniques can keep pace with the demand for increased production. In answer to this challenge, CRISPR/Cas9 genome editing technology has been gaining traction in plant biology and crop breeding in recent years. However, there are currently no reports of the successful application of the CRISPR/Cas9 genome editing technology in pea. We developed a transient transformation system of hairy roots, mediated by Agrobacterium rhizogenes strain K599, to validate the efficiency of a CRISPR/Cas9 system. Further optimization resulted in an efficient vector, PsU6.3-tRNA-PsPDS3-en35S-PsCas9. We used this optimized CRISPR/Cas9 system to edit the pea phytoene desaturase (PsPDS) gene, causing albinism, by Agrobacterium-mediated genetic transformation. This is the first report of successful generation of gene-edited pea plants by this route.
