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基于多重基因组编辑的水稻复杂性状快速定向改良。

Multiplex-genome-editing based rapid directional improvement of complex traits in rice.

机构信息

Rice Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, China.

State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops/Key Laboratory of Germplasm Innovation and Molecular Breeding of Hybrid Rice for South China, Ministry of Agriculture and Affairs, P.R. China/Incubator of National Key Laboratory of Germplasm Innovation and Molecular Breeding between Fujian and Ministry of Sciences and Technology/Fuzhou Branch, National Rice Improvement Center of China/Fujian Engineering Laboratory of Crop Molecular Breeding/Fujian Key Laboratory of Rice Molecular Breeding, Fuzhou, China.

出版信息

Plant Biotechnol J. 2024 Sep;22(9):2624-2628. doi: 10.1111/pbi.14375. Epub 2024 May 27.

Abstract

Although thousands of genes have been identified or cloned in rice (Oryza sativa) in the last two decades, the majority of them have only been separately characterized in specific varieties or single-gene modified backgrounds, thus limiting their practical application. We developed an optimized multiplex genome editing (MGE) toolbox that can efficiently assemble and stably express up to twelve sgRNA targets in a single plant expression vector. In this study, we established the MGE-based Rapid Directional Improvement (MRDI) strategy for directional improvement of complex agronomic traits in one small-scale rice transformation. This approach provides a rapid and practical procedure, encompassing sgRNA assembly, transgene-free screening and the creation of promising germplasm, by combining the precision of gene editing with phenotype-based field breeding. The MRDI strategy was used to generate the full diversity of twelve main agronomic genes in rice cultivar FXZ for the directional improvement of its growth duration and plant architecture. After applying the MRDI to FXZ, ideal plants with the desired traits of early heading date reduced plant height, and more effective panicles were generated without compromising yield, blast resistance and grain quality. Furthermore, the results of whole-genome sequencing (WGS), including the analysis of structural variations (SVs) and single nucleotide variations (SNVs) in the MGE plants, confirmed the high specificity and low frequency of unwanted mutations associated with this strategy. The MRDI breeding strategy would be a robust approach for exploring and applying crucial agronomic genes, as well as for generating novel elite germplasm in the future.

摘要

尽管在过去的二十年中,已经在水稻(Oryza sativa)中鉴定或克隆了数千个基因,但其中大多数基因仅在特定品种或单个基因修饰背景下进行了单独表征,从而限制了它们的实际应用。我们开发了一种优化的多重基因组编辑(MGE)工具盒,可以在单个植物表达载体中有效地组装和稳定表达多达十二个 sgRNA 靶标。在这项研究中,我们建立了基于 MGE 的快速定向改良(MRDI)策略,用于在小规模水稻转化中定向改良复杂的农艺性状。该方法通过将基因编辑的精确性与基于表型的田间育种相结合,提供了一种快速实用的程序,包括 sgRNA 组装、无转基因筛选和有前途的种质创造。MRDI 策略用于生成水稻品种 FXZ 的十二个主要农艺基因的全部多样性,以定向改良其生长周期和植物结构。在将 MRDI 应用于 FXZ 后,生成了具有所需性状的理想植株,即早期抽穗日期、降低株高、更有效的穗,而不影响产量、抗稻瘟病和籽粒品质。此外,全基因组测序(WGS)的结果,包括 MGE 植株中结构变异(SV)和单核苷酸变异(SNV)的分析,证实了该策略具有高度特异性和低频率的非期望突变。MRDI 育种策略将是探索和应用关键农艺基因以及未来生成新型优良种质的强大方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/63bd/11373770/1c250b5afd5d/PBI-22-2624-g001.jpg

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