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Article|20 Jan 2022|OPEN
An efficient root transformation system for CRISPR/Cas9-based analyses of shoot–root communication in cucurbit crops
Shouyu Geng1 ,† , Hamza Sohail1 ,† , Haishun Cao1,2 ,† , Jingyu Sun1 , Zhi Chen1 , Lijian Zhou1 , Wenbo Wang1 , Runwen Ye1 and Li Yang1 , , Zhilong Bie,1 ,
1Key Laboratory of Horticultural Plant Biology, Ministry of Education, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, China
2Institute of Facility Agriculture, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China
*Corresponding author. E-mail:,
Shouyu Geng,Hamza Sohail and Haishun Cao contributed equally to the study.

Horticulture Research 9,
Article number: uhab082 (2022)
Views: 388

Received: 25 Sep 2021
Revised: 16 Jun 2022
Accepted: 25 Nov 2021
Published online: 20 Jan 2022


Cucurbit crops are suitable models for studying long-distance signaling in horticultural plants. Although thousands of substances are graft transmissible in cucurbits, functional studies have been hampered by the lack of efficient genetic transformation systems. Here, we report a convenient and efficient root transformation method for several cucurbit crops that will facilitate studies of functional genes and shoot–root crosstalk. We obtained healthy plants with completely transformed roots and non-transgenic shoots within 6 weeks. Furthermore, we combined this root transformation method with grafting, which allowed for gene manipulation in the rootstock. We validated our system by exploring salt tolerance mechanisms using a cucumber (Cucumis sativus)/pumpkin (Cucurbita moschata Duch.) (scion/rootstock) graft in which the sodium transporter gene High-affinity K+ transporter1 (CmoHKT1;1) was edited in the pumpkin rootstock and by overexpressing the pumpkin tonoplast Na+/H+ antiporter gene Sodium hydrogen exchanger4 (CmoNHX4) in cucumber roots.