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通过编辑非编码区提高两个常规栽培水稻品种的细菌性条斑病抗性。

Improvement of Bacterial Blight Resistance in Two Conventionally Cultivated Rice Varieties by Editing the Noncoding Region.

机构信息

Hubei Key Laboratory of Food Crop Germplasm and Genetic Improvement, Food Crops Institute, Hubei Academy of Agricultural Sciences, Wuhan 430064, China.

College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.

出版信息

Cells. 2022 Aug 16;11(16):2535. doi: 10.3390/cells11162535.

Abstract

is a recessive pleiotropic gene that positively regulates rice disease resistance and negatively regulates rice fertility; thus, seriously restricting its rice breeding application. In this study, CRISPR/Cas9 gene-editing technology was used to delete the gene promoter partial sequence, including the pathogenic bacteria-inducible expression element. Rice with the edited promoter region lost the ability for pathogen-induced gene expression without affecting background gene expression in leaves and anthers, resulting in disease resistance and normal yield. The study also screened a family of disease-resistant and normal fertile plants in which the target sequence was deleted and the exogenous transgene fragment isolated in the T generation (transgene-free line). Important agronomic traits of the T generation rice were examined. T generation rice with/without exogenous DNA showed no statistical differences compared to the wild type in heading stage, plant height, panicles per plant, panicle length, or seed setting rate in the field. Two important conventional rice varieties, namely Kongyu131 (KY131, /) and Huanghuazhan (HHZ, /), were successfully transformed, and disease-resistant and fertile materials were obtained. Currently, these are the two important conventional rice varieties in China that can be used directly for production after improvement. Expression of the gene in the leaves of transgenic rice (KY-PD and HHZ-PD) was not induced after pathogen infection, indicating that this method can be used universally and effectively to promote the practical application of , a recessive disease-resistant pleiotropic gene, for rice bacterial blight resistance. Our study on the regulation of gene expression by editing noncoding regions of the genes provides a new idea for the development of molecular design breeding in the future.

摘要

是一个隐性多效基因,正向调控水稻抗病性,负向调控水稻育性;因此,严重限制了其在水稻育种中的应用。本研究利用 CRISPR/Cas9 基因编辑技术删除了基因启动子部分序列,包括病原菌诱导表达元件。编辑启动子区域的水稻失去了病原菌诱导基因表达的能力,而不影响叶片和花药中背景基因的表达,从而表现出抗病性和正常的产量。该研究还筛选出了一个具有抗病性和正常育性的家系,其靶序列缺失且外源转基因片段在 T 代被分离(无转基因系)。对 T 代水稻的重要农艺性状进行了检测。与野生型相比,T 代水稻在抽穗期、株高、每株穗数、穗长和结实率等方面均无显著差异。成功转化了两个重要的常规水稻品种,即空育 131(KY131,/)和黄华占(HHZ,/),并获得了抗病和育性的材料。目前,这两个品种是中国两个重要的常规水稻品种,经过改良后可以直接用于生产。转基因水稻(KY-PD 和 HHZ-PD)叶片中基因的表达在病原菌感染后没有被诱导,表明该方法可普遍有效地促进隐性抗病多效基因的实际应用,为水稻细菌性条斑病抗性的分子设计育种提供了新的思路。我们对基因非编码区进行基因编辑调控基因表达的研究,为未来分子设计育种的发展提供了新的思路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a30/9406647/2b011ec603d1/cells-11-02535-g001.jpg

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