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RNA病毒介导的基因编辑用于番茄性状育种。

RNA virus-mediated gene editing for tomato trait breeding.

作者信息

Uranga Mireia, Aragonés Verónica, García Arcadio, Mirabel Sophie, Gianoglio Silvia, Presa Silvia, Granell Antonio, Pasin Fabio, Daròs José-Antonio

机构信息

Instituto de Biología Molecular y Celular de Plantas (IBMCP), Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València (CSIC-UPV), Avenida de los Naranjos s/n, 46022 Valencia, Spain.

出版信息

Hortic Res. 2023 Dec 19;11(1):uhad279. doi: 10.1093/hr/uhad279. eCollection 2024 Jan.

Abstract

Virus-induced genome editing (VIGE) leverages viral vectors to deliver CRISPR-Cas components into plants for robust and flexible trait engineering. We describe here a VIGE approach applying an RNA viral vector based on potato virus X (PVX) for genome editing of tomato, a mayor horticultural crop. Viral delivery of single-guide RNA into Cas9-expressing lines resulted in efficient somatic editing with indel frequencies up to 58%. By proof-of-concept VIGE of () and plant regeneration from edited somatic tissue, we recovered loss-of-function mutant progeny displaying an albino phenotype. VIGE of (), a gene involved in fruit color variation, generated mutant lines with recolored red-brown fruits and high chlorophyll levels. The obtained editing events were heritable, overall confirming the successful breeding of fruit color. Altogether, our VIGE approach offers great potential for accelerated functional genomics of tomato variation, as well as for precision breeding of novel tomato traits.

摘要

病毒诱导的基因组编辑(VIGE)利用病毒载体将CRISPR-Cas组件导入植物中,以实现强大且灵活的性状工程改造。我们在此描述一种VIGE方法,该方法应用基于马铃薯X病毒(PVX)的RNA病毒载体对主要园艺作物番茄进行基因组编辑。将单向导RNA通过病毒传递到表达Cas9的品系中,可实现高效的体细胞编辑,插入缺失频率高达58%。通过对()进行概念验证VIGE以及从编辑后的体细胞组织进行植株再生,我们获得了表现出白化表型的功能缺失突变体后代。对参与果实颜色变异的基因()进行VIGE,产生了具有重新着色的红棕色果实和高叶绿素水平的突变系。所获得的编辑事件是可遗传的,总体上证实了果实颜色的成功育种。总之,我们的VIGE方法为加速番茄变异的功能基因组学研究以及新型番茄性状的精准育种提供了巨大潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/344e/11184526/054d6734f4d3/uhad279f1.jpg

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