OsERF93参与调控水稻纹枯病抗性的研究

doi:10.16819/j.1001-7216.2026.250202

中国水稻科学 ›› 2026, Vol. 40 ›› Issue (1): 37-50.DOI: 10.16819/j.1001-7216.2026.250202

OsERF93参与调控水稻纹枯病抗性的研究

岳轩宇¹^,², 谢文亚¹^,²^,³^,^*(), 冯志明¹^,²^,³, 陈宗祥¹^,²^,³, 胡珂鸣¹^,², 左示敏¹^,²^,³^,^*()

¹江苏省作物基因组学和分子育种重点实验室/生物育种钟山实验室/植物功能基因组学教育部重点实验室，扬州大学农学院，江苏扬州 225009
²江苏省粮食作物现代产业技术协同创新中心/江苏省作物遗传生理重点实验室，扬州大学，江苏扬州 225009
³扬州现代种子创新研究所，江苏扬州 225600

收稿日期:2025-02-11 修回日期:2025-05-23 出版日期:2026-01-10 发布日期:2026-01-21
通讯作者: *email：smzuo@yzu.edu.cn;
email：wyxie@yzu.edu.cn
基金资助:
国家自然科学基金资助项目(32272110);国家自然科学基金资助项目(32200430);生物育种钟山实验室项目(ZSBBL-KY2023-06-3);扬州市现代农业项目(YZ2024045)

Function of OsERF93 in Regulating Resistance to Sheath Blight in Rice

YUE Xuanyu¹^,², XIE Wenya¹^,²^,³^,^*(), FENG Zhiming¹^,²^,³, CHEN Zongxiang¹^,²^,³, HU Keming¹^,², ZUO Shimin¹^,²^,³^,^*()

¹Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Agricultural College of Yangzhou University, Yangzhou 225009, China
²Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops/Jiangsu Key Laboratory of Crop Genetics and Physiology, Yangzhou University, Yangzhou 225009, China
³Yangzhou Modern Seed Innovation Institute, Yangzhou 225600, China

Received:2025-02-11 Revised:2025-05-23 Online:2026-01-10 Published:2026-01-21

摘要/Abstract

摘要：

【目的】本研究旨在解析ERF类转录因子OsERF93在水稻抗纹枯病中的功能与机制，为抗病育种提供基因资源。【方法】利用RT-qPCR、水稻原生质体转录激活等方法，分析了OsERF93的表达模式、转录调控功能和亚细胞定位位置；通过构建转基因过表达和敲除株系，研究了OsERF93调控纹枯病抗性的功能，进而比较分析了水杨酸、茉莉酸和乙烯信号相关基因在转基因系接种前后的表达差异，以及超表达转基因系与对照间的农艺性状差异。【结果】OsERF93在水稻叶片和叶鞘组织中高表达，受纹枯病菌侵染诱导表达；其编码蛋白定位于细胞核和细胞膜中，具有转录激活活性；OsERF93过表达显著增强了水稻纹枯病抗性，而敲除则显著减弱抗病性；茉莉酸合成基因OsAOS2以及部分PR基因在OsERF93过表达系和敲除系中分别明显上调和下调表达；OsERF93过表达系的株高和千粒重略低于野生型对照，但每穗粒数显著高于对照，其他性状如分蘖数、一次枝梗数和穗长与对照间无显著差异。【结论】OsERF93通过激活茉莉酸信号途径正调控水稻的纹枯病抗性，为水稻抗病育种提供了一个新的基因资源。

关键词: 水稻, 纹枯病, OsERF93, 茉莉酸, 农艺性状

Abstract:

【Objective】This study aims to elucidate the function and mechanism of the ERF transcription factor OsERF93 in rice sheath blight resistance, providing genetic resources for disease-resistant breeding.【Method】The expression pattern, transcriptional regulatory function, and subcellular localization of OsERF93 were analyzed using RT-qPCR and transcriptional activation assays in rice protoplasts. Transgenic overexpression and knockout lines were constructed to investigate the role of OsERF93 in sheath blight resistance. Expression differences of salicylic acid (SA), jasmonic acid (JA), and ethylene (ET)-related signaling genes were compared before and after inoculation in transgenic lines. Additionally, agronomic traits of overexpression lines and controls were evaluated.【Result】OsERF93 was highly expressed in rice leaves and leaf sheaths and induced by sheath blight fungus infection. The encoded protein localized to the nucleus and cell membrane, exhibiting transcriptional activation activity. Overexpression of OsERF93 significantly enhanced sheath blight resistance, while knockout lines showed reduced resistance. JA biosynthesis gene OsAOS2 and certain PR genes were upregulated in overexpression lines and downregulated in knockout lines. Overexpression lines exhibited slightly reduced plant height and thousand-grain weight compared to wild-type controls but had significantly higher grain number per panicle. No significant differences were observed in tiller number, primary branch number, and panicle length between overexpression lines and controls.【Conclusion】The results indicate that OsERF93 positively regulates rice sheath blight resistance by activating the jasmonic acid signaling pathway, offering a novel genetic resource for rice disease-resistant breeding.

Key words: rice, sheath blight, OsERF93, jasmonic acid, agronomic trait

岳轩宇, 谢文亚, 冯志明, 陈宗祥, 胡珂鸣, 左示敏. OsERF93参与调控水稻纹枯病抗性的研究[J]. 中国水稻科学, 2026, 40(1): 37-50.

YUE Xuanyu, XIE Wenya, FENG Zhiming, CHEN Zongxiang, HU Keming, ZUO Shimin. Function of OsERF93 in Regulating Resistance to Sheath Blight in Rice[J]. Chinese Journal OF Rice Science, 2026, 40(1): 37-50.

图/表 7

图1 水稻中的OsERF基因家族聚类分析与OsERF93所在亚家族蛋白序列比对 A：ERF基因家族系统发育树分析，红框标注的基因在纹枯病菌接种后的抗感材料中表现出差异性表达；B：参与响应纹枯病诱导的ERF家族基因的相关蛋白序列比对。

Fig. 1. Phylogenetic analysis of rice OsERFs gene family and protein sequence alignment of subfamily containing OsERF93 A, Phylogenetic tree analysis of the ERFs gene family, with the genes highlighted in red boxes showing differential expression in resistant and susceptible materials after inoculation with R. Solani; B, Alignment of protein sequence of ERF family genes involved in sheath blight response.

图2 OsERF93诱导表达模式分析 A：OsERF93组织特异性表达；B：OsERF93对ETH和AOA的不同响应；C：感病品种Lemont接种纹枯病菌后相对表达量变化；D：抗病品种YSBR1接种纹枯病后相对表达量变化。图中R. solani为实验组，Mock为对照组。

Fig. 2. Analysis of OsERF93 expression pattern A, Tissue-specific expression of OsERF93; B, Differential response of OsERF93 to ETH (Ethylene) and AOA (Aminocyclopropane-1-carboxylic acid); C, Relative expression levels of the susceptible variety Lemont after inoculation with sheath blight pathogen; D, Relative expression levels of the resistant variety YSBR1 after inoculation with sheath blight pathogen. In the figures, R. solani represents the treatment group, and Mock represents the control group.

图3 OsERF93转录激活活性与亚细胞定位分析 A：OsERF93在水稻细胞中的亚细胞定位，标尺=10 μm，NLS为细胞核定位信号；B：酵母双杂交实验验证转录激活活性；C：水稻原生质体瞬时转化，VP16 是转录激活对照。**表示由 P< 0.01 的t 检验确定的极显著差异。

Fig. 3. Analysis of transcriptional activation activity and subcellular localization of OsERF93 A, Subcellular localization of OsERF93 in rice cells(bar=10 μm), and NLS denoting the nuclear localization signal; B, Transcriptional activation activity verified by yeast two-hybrid assay; C, Transient transformation of rice protoplasts, with VP16 serving as the transcriptional activation control. ** indicates a highly significant difference as determined by a t-test with P< 0.01.

图4 OsERF93基因过表达系和敲除突变体的分子鉴定 A：两个OsERF93过表达株系中OsERF93的转录水平；B：OsERF93基因结构和CRISPR/Cas9靶点示意图；C：鉴定出的两个纯合oserf93突变体与野生型对照的DNA序列比对；D：CAPS标记验证oserf93突变体与野生型，引物序列见附表1，oserf93-1和oserf93-1片段长度为417 bp，野生型被酶切后片段长度分别为248 bp和169 bp；E：野生型及两个敲除系中OsERF93基因编码的氨基酸序列分析。

Fig. 4. Relative expression levels of the OsERF93 overexpression lines A, Transcript levels of OsERF93 in two OsERF93 overexpression lines; B, Schematic diagram of OsERF93 gene structure and CRISPR/Cas9 target sites; C, DNA sequence alignment of the identified homozygous oserf93 mutants with the wild-type control; D, CAPS marker validation of oserf93 mutants and wild-type, with primer sequences listed in Table S1, showing fragment lengths of 417 bp for oserf93-1 and oserf93-2, and the wild-type fragments being 248 bp and 169 bp after enzyme digestion; E, Amino acid sequence analysis of the OsERF93 gene encoded in the wild-type and two knockout lines.

图5 OsERF93超表达和敲除系的纹枯病抗性鉴定 A和B分别为离体茎秆接种鉴定中对照及OsERF93相关转基因系接种后5 d的表型照片和病斑长度统计比较；C和D分别为温室成株期植株接种鉴定中OsERF93相关转基因系接种后14 d的结果和病斑长度。**和*分别表示P< 0.01和P< 0.05 统计显著性差异水平。

Fig. 5. Resistance evaluation of OsERF93 overexpression and knock-out lines to sheath blight disease A and B respectively present the phenotypic photographs and statistical comparisons of lesion lengths at 5 days post-inoculation in the control and OsERF93-related transgenic lines during the in vitro stem inoculation assay; C and D respectively display the phenotypic photographs and statistical comparisons of lesion heights at 14 days post-inoculation in the OsERF93-related transgenic lines during the greenhouse adult plant inoculation assay. ** and * indicate statistical significance levels of P< 0.01 and P< 0.05, respectively.

图6 OsERF93超表达和敲除系中部分抗病相关基因在病原菌接种前后的表达变化特征 A和B分别为茉莉酸合成基因OsAOS2、水杨酸途径标志基因OsPAL1在野生型对照及OsERF93相关转基因系接种纹枯病菌后的表达量变化；C和D是乙烯信号相关基因OsACO1和OsACS1在野生型和OsERF93相关转基因系接种纹枯病菌前后的表达量变化；E和F分别为病程相关蛋白基因OsPBZ1和OsPR1a在野生型及OsERF93相关转基因系接种纹枯病菌后的表达量变化。 **和*分别表示P< 0.01和P< 0.05 统计显著性差异水平。

Fig. 6. Expression changes of pathogenesis-related genes in OsERF93 overexpression and knock-out lines after inoculation with sheath blight fungus A and B respectively depict the expression level changes of the jasmonic acid signal related gene OsAOS2 and the salicylic acid pathway marker gene OsPAL1 in the wild-type control and OsERF93-related transgenic lines after inoculation with the sheath blight pathogen; C and D represent the expression changes of ethylene signal related genes OsACO1 and OsACS1 in wild-type and OsERF93-related transgenic lines after inoculation with the sheath blight pathogen; E and F respectively show the expression level changes of the pathogenesis-related protein genes OsPBZ1 and OsPR1a in the wild-type and OsERF93-related transgenic lines after inoculation with the sheath blight pathogen. ** and * denote statistical significance levels of P< 0.01 and P< 0.05, respectively.

图7 OsERF93相关转基因系及其野生型间的部分农艺性状比较 A和B分别为野生型(WT)、OsERF93过表达系（OsERF93OE-1、OsERF93OE-2）及敲除系（oserf93-1、oserf93-1-2）的株型和穗型照片比较；C~K分别为野生型(WT)、OsERF93过表达系及敲除系在株高（C）、分蘖数（D）、剑叶长（E）、剑叶宽（F）、穗长（G）、一次枝梗数（H）、二次枝梗数（I）、穗粒数（J）和千粒重（K）性状上的差异统计比较；**和*分别表示P< 0.01和P< 0.05 统计显著性差异水平。

Fig. 7. Comparison of some agronomic traits among WT, OsERF93 overexpression and knock-out transgenic lines A and B show comparisons of plant architecture and panicle morphology between the wild-type (WT), OsERF93 overexpression lines (OsERF93OE-1, OsERF93OE-2), and knockout lines (oserf93-1, oserf93-2). C-K present statistical comparisons of differences between the wild-type(WT), OsERF93 overexpression lines, and knockout lines for traits including plant height (C), tiller number (D), flag leaf length (E), flag leaf width (F), panicle length (G), primary branch number (H), secondary branch number (I), grain number per panicle (J), and 1000-grain weight (K). ** and * indicate statistically significant differences at P< 0.01 and P< 0.05, respectively.

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OsERF93参与调控水稻纹枯病抗性的研究

Function of OsERF93 in Regulating Resistance to Sheath Blight in Rice

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