Function of OsERF93 in Regulating Resistance to Sheath Blight in Rice

doi:10.16819/j.1001-7216.2026.250202

Abstract

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

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

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.

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.

Figures/Tables 7

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.

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.

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.

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.

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.

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.

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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