Development of an Agrobacterium-mediated CRISPR/Cas9 system in pea(Pisum sativum L.)  

作　　者：Guan Li Rong Liu Rongfang Xu Rajeev KVarshney 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 

机构地区：[1]National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081,China [2]Key Laboratory of Rice Genetic Breeding of Anhui Province,Rice Research Institute,Anhui Academy of Agricultural Sciences,Hefei 230031,Anhui,China [3]State Agricultural Biotechnology Centre,Centre for Crop and Food Innovation,Food Futures Institute,Murdoch University,Murdoch,WA 6150,Australia [4]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 [5]Zhejiang Xinnan Chemical Industrial Group Co.,Ltd.,Hangzhou 311300,Zhejiang,China [6]State Key Laboratory of Microbial Resources,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China

出　　处：《The Crop Journal》2023年第1期132-139,共8页作物学报（英文版）

基　　金：the financial support of the China Agriculture Research System of MOF and MARA-Food Legumes(CARS-08);the Agricultural Science and Technology Innovation Program(ASTIP)of the Chinese Academy of Agricultural Sciences。

摘　　要：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.

关 键 词：Agrobacterium-mediated transformation CRISPR/Cas9 PEA Genome editing 

分 类 号：S643.3[农业科学—蔬菜学]