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An efficient root transformation system for CRISPR/Cas9-based analyses of shoot-root communication in cucurbit crops.
Horticulture Research ( IF 7.6 ) Pub Date : 2022-01-20 , DOI: 10.1093/hr/uhab082
Shouyu Geng 1 , Hamza Sohail 1 , Haishun Cao 1, 2 , Jingyu Sun 1 , Zhi Chen 1 , Lijian Zhou 1 , Wenbo Wang 1 , Runwen Ye 1 , Li Yang 1 , Zhilong Bie 1
Affiliation  

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.

中文翻译:


一种有效的根转化系统,用于基于 CRISPR/Cas9 的葫芦科作物茎根通讯分析。



葫芦科作物是研究园艺植物长距离信号传导的合适模型。尽管葫芦科植物中有数千种物质可以通过嫁接传播,但由于缺乏有效的遗传转化系统,功能研究受到了阻碍。在这里,我们报告了一种针对几种葫芦科作物的便捷高效的根转化方法,该方法将有助于功能基因和茎根串扰的研究。我们在 6 周内获得了具有完全转化的根和非转基因芽的健康植物。此外,我们将这种根转化方法与嫁接相结合,从而可以在砧木中进行基因操作。我们通过使用黄瓜 (Cucumis sativus)/南瓜 (Cucurbita moschata Duch.)(接穗/砧木)嫁接物探索耐盐机制来验证我们的系统,其中钠转运蛋白基因高亲和力 K+ 转运蛋白 1 (CmoHKT1;1) 在南瓜砧木,并通过在黄瓜根中过表达南瓜液泡膜 Na+/H+ 逆向转运蛋白基因钠氢交换器 4 (CmoNHX4)。
更新日期:2022-01-20
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