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An efficient root transformation system for CRISPR/Cas9-based analyses of shoot-root communication in cucurbit crops.
Horticulture Research  (IF6.793),  Pub Date : 2022-01-20, DOI: 10.1093/hr/uhab082
Shouyu Geng,Hamza Sohail,Haishun Cao,Jingyu Sun,Zhi Chen,Lijian Zhou,Wenbo Wang,Runwen Ye,Li Yang,Zhilong Bie

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.