Chinese Journal OF Rice Science >
Osa-miR166i-3 Positively Regulates Resistance to Sheath Blight Through Mediating the Accumulation of Reactive Oxygen Species
#These authors contributed equally to this work
Received date: 2024-01-14
Revised date: 2024-02-28
Online published: 2025-03-19
【Objective】Sheath blight is a major disease in rice, leading to significant yield and quality losses. Osa-miR166i-3p responds to Rhizoctonia solani infection in both the susceptible japonica rice variety Xudao 3 and the resistant indica-japonica hybrid variety YSBR1. This study aimed to clarify the role and molecular mechanism of Osa-miR166i-3p in rice resistance to sheath blight and to explore the potential pathways involving downstream genes. 【Method】Vectors containing Osa-miR166i-3p were constructed, and transgenic plants were generated in the susceptible variety Xudao 3 using Agrobacterium-mediated transformation. The authenticity of the transgenic plants was confirmed by sequencing and detecting the expression level of Osa-miR166i-3p. The plants were inoculated with R. solani in a greenhouse, and lesion lengths were measured. Additionally, the main agronomic traits of the transgenic and control plants grown under field conditions were investigated. Leaf sheath tissues collected at 0 h, 8 h, and 16 h post-inoculation were used for RNA-seq analysis to study the biological function of Osa-miR166i-3p. 【Results】Compared with Xudao 3, the expression level of Osa-miR166i-3p was significantly reduced in knockout plants and significantly increased in overexpressed plants. After inoculation with R. solani, the lesion length of Osa-miR166i-3p knockout plants increased, indicating enhanced susceptibility, while the lesion length of overexpression plants decreased, indicating enhanced resistance. Statistical analysis of agronomic traits, including plant height, panicle length, number of branches per panicle, and 1000-grain weight, showed no significant differences between transgenic and control plants, suggesting that overexpression or knockout of Osa-miR166i-3p did not affect rice agronomic traits. Enrichment analysis revealed that multiple peroxidase genes were induced in Osa-miR166i-3p overexpression plants 8 hours after inoculation with R. solani. 【Conclusion】This study demonstrates that Osa-miR166i-3p positively regulates rice resistance to sheath blight by modulating the expression of class III peroxidase-related genes and influencing reactive oxygen species (ROS) accumulation. These findings provide new insights for improving rice disease resistance.
Key words： rice; sheath blight; Osa-miR166i-3p; biological function; RNA-seq
FENG Tao, ZHANG Zhaoyang, HUANG Xinni, WANG Yue, ZHONG Xuzhi, FENG Zhiming, LIU Xin, ZUO Shimin, OUYANG Shouqiang . Osa-miR166i-3 Positively Regulates Resistance to Sheath Blight Through Mediating the Accumulation of Reactive Oxygen Species[J]. Chinese Journal OF Rice Science, 2025 , 39(2) : 187 -196 . DOI: 10.16819/j.1001-7216.2025.240107
| [1] | Taheri P, Tarighi S. Cytomolecular aspects of rice sheath blight caused by Rhizoctonia solani[J]. European Journal of Plant Pathology, 2011, 129: 511-528. |
| [2] | Zhang C Q, Liu Y H, Ma X Y, Feng Z, Ma Z H. Characterization of sensitivity of Rhizoctonia solani, causing rice sheath blight, to mepronil and boscalid[J]. Crop Protection, 2009, 28(5): 381-386. |
| [3] | Margani R, Hadiwiyono, Widadi S. Utilizing bacillus to inhibit the growth and infection by sheath blight pathogen, Rhizoctonia solani in rice[J]. IOP Conference Series: Earth and Environmental Science, 2018, 142(1): 012070. |
| [4] | Taheri P, Gnanamanickam S, Hofte M. Characterization, genetic structure, and pathogenicity of Rhizoctonia spp. associated with rice sheath diseases in India[J]. Phytopathology, 2007, 97(3): 373-383. |
| [5] | Jasrotia S, Salgotra R K, Sharma M. Efficacy of bioinoculants to control of bacterial and fungal diseases of rice (Oryza sativa L.) in northwestern Himalaya[J]. Brazilian Journal of Microbiology, 2021, 52(2): 687-704. |
| [6] | Ma Y, Wang Y R, He Y H, Ding Y Y, An J X, Zhang Z J, Zhao W B, Hu Y M, Liu Y Q. Drug repurposing strategy part 1: From approved drugs to agri-bactericides leads[J]. The Journal of Antibiotics, 2023, 76(1): 27-51. |
| [7] | Qi P, Wang N, Zhang T, Feng Y, Zhou X, Zeng D, Meng J, Liu L, Jin L, Yang S. Anti-virulence strategy of novel dehydroabietic acid derivatives: Design, synthesis, and antibacterial evaluation[J]. International Journal of Molecular Sciences, 2023, 24(3): 2897-2913. |
| [8] | Ontoy J C, Shrestha B, Karki H S, Barphagha I, Angira B, Famoso A, Ham J H. Genetic characterization of the partial disease resistance of rice to bacterial panicle blight and sheath blight by combined QTL linkage and QTL-seq analyses[J]. Plants, 2023, 12(3): 559-577. |
| [9] | Zuo S M, Zhang L, Wang H, Yin Y J, Zhang Y F, Chen Z X, Ma Y Y, Pan X B. Prospect of the QTL-qSB-9TQ utilized in molecular breeding program of japonica rice against sheath blight[J]. Journal of Genetics and Genomics, 2008, 35(8): 499-505. |
| [10] | Tan C X, Ji X M, Yang Y, Pan X Y, Zuo S M, Zhang Y F, Zou J H, Chen Z X, Zhu L H, Pan X B. Identification and marker-assisted selection of two major quantitative genes controlling rice sheath blight resistance in backcross generations[J]. Journal of Genetics and Genomics, 2005, 32(4): 399-405. |
| [11] | Channamallikarjuna V, Sonah H, Prasad M, Rao G J N, Chand S, Upreti H C, Singh N K, Sharma T R. Identification of major quantitative trait loci qSBR11-1 for sheath blight resistance in rice[J]. Molecular Breeding, 2010, 25(1): 155-166. |
| [12] | 陈燕玲, 岑光莉, 孙婷婷, 尤垂淮, 阙友雄, 苏亚春. 植物几丁质酶和β-1,3-葡聚糖酶及其协同抗病性研究进展[J]. 农业生物技术学报, 2022, 30(7): 1394-1411. |
| Chen Y L, Cen G L, Sun T T, You C H, Que Y X, Su Y C. Progress on plant chitinase and β-1, 3-glucanase and their synergistic function in disease resistance[J]. Journal of Agricultural Biotechnology, 2022, 30(7): 1394-1411. (in Chinese with English abstract) | |
| [13] | Peng X, Hu Y, Tang X, Zhou P, Deng X, Wang H, Guo Z. Constitutive expression of rice WRKY30 gene increases the endogenous jasmonic acid accumulation, PR gene expression and resistance to fungal pathogens in rice[J]. Planta, 2012, 236(5): 1485-1498. |
| [14] | Peng X, Wang H, Jang J C, Xiao T, He H, Jiang D, Tang X. OsWRKY80-OsWRKY4 module as a positive regulatory circuit in rice resistance against Rhizoctonia solani[J]. Rice, 2016, 9(1): 63-77. |
| [15] | Borges F, Martienssen R A. The expanding world of small RNAs in plants[J]. Nature Reviews Molecular Cell Biology, 2015, 16(12): 727-741. |
| [16] | Palatnik J F, Allen E, Wu X, Schommer C, Schwab R, Carrington J C, Weigel D. Control of leaf morphogenesis by microRNAs[J]. Nature, 2003, 425(6955): 257-263. |
| [17] | Weiberg A, Wang M, Lin F M, Zhao H, Zhang Z, Kaloshian I, Huang H D, Jin H. Fungal small RNAs suppress plant immunity by hijacking host RNA interference pathways[J]. Science, 2013, 342(6154): 118-123. |
| [18] | Sunkar R, Chinnusamy V, Zhu J, Zhu J K. Small RNAs as big players in plant abiotic stress responses and nutrient deprivation[J]. Trends in Plant Science, 2007, 12(7): 301-309. |
| [19] | Wang Z, Xia Y, Lin S, Wang Y, Guo B, Song X, Ding S, Zheng L, Feng R, Chen S, Bao Y, Sheng C, Zhang X, Wu J, Niu D, Jin H, Zhao H. Osa-miR164a targets OsNAC60 and negatively regulates rice immunity against the blast fungus Magnaporthe oryzae[J]. The Plant Journal, 2018, 95(4): 584-597. |
| [20] | Feng T, Zhang Z Y, Gao P, Feng Z M, Zuo S M, Ouyang S Q. Suppression of rice Osa-miR444.2 improves the resistance to sheath blight in rice mediating through the phytohormone pathway[J]. International Journal of Molecular Sciences, 2023, 24(4): 3653-3665. |
| [21] | Qiao L, Zheng L, Sheng C, Zhao H, Jin H, Niu D. Rice siR109944 suppresses plant immunity to sheath blight and impacts multiple agronomic traits by affecting auxin homeostasis[J]. The Plant Journal, 2020, 102(5): 948-964. |
| [22] | Cao W L, Cao X X, Zhao J H, Zhang Z Y, Feng Z M, Ouyang S Q, Zuo S M. Comprehensive characteristics of microRNA expression profile conferring to Rhizoctonia solani in rice[J]. Mathematical Research Letters, 2020, 27(2): 101-112. |
| [23] | Ho T T, Zhou N, Huang J, Koirala P, Xu M, Fung R, Wu F, Mo Y Y. Targeting non-coding RNAs with the CRISPR/Cas9 system in human cell lines[J]. Nucleic Acids Research, 2015, 43(3): 17-28. |
| [24] | 贺闽, 尹俊杰, 冯志明, 朱孝波, 赵剑华, 左示敏, 陈学伟. 水稻稻瘟病和纹枯病抗性鉴定方法[J]. 植物学报, 2020, 55(5): 577-587. |
| He M, Yin J J, Feng Z M, Zhu X B, Zhao J H, Zuo S M, Chen X W. Methods for evaluation of rice resistance to blast and sheath blight diseases[J]. Chinese Bulletin of Botany, 2020, 55(5): 577-587. (in Chinese with English abstract) | |
| [25] | Robinson M D, McCarthy D J, Smyth G K. edgeR: A bioconductor package for differential expression analysis of digital gene expression data[J]. Bioinformatics, 2010, 26(1): 139-140. |
| [26] | Li Y, Li T T, He X R, Zhu Y, Feng Q, Yang X M, Zhou X H, Li G B, Ji Y P, Zhao J H, Zhao Z X, Pu M, Zhou S X, Zhang J W, Huang Y Y, Fan J, Wang W M. Blocking Osa- miR1871 enhances rice resistance against Magnaporthe oryzae and yield[J]. Plant Biotechnology Journal, 2022, 20(4): 646-659. |
| [27] | Zhao Y T, Wang M, Wang Z M, Fang R X, Wang X J, Jia Y T. Dynamic and coordinated expression changes of rice small RNAs in response to Xanthomonas oryzae pv. oryzae[J]. Journal of Genetics and Genomics, 2015, 42(11): 625-637. |
| [28] | Zhang J, Zhang H, Srivastava A K, Pan Y, Bai J, Fang J, Shi H, Zhu J K. Knockdown of rice MicroRNA166 confers drought resistance by causing leaf rolling and altering stem xylem development[J]. Plant Physiology, 2018, 176(3): 2082-2094. |
| [29] | Ding Y, Gong S, Wang Y, Wang F, Bao H, Sun J, Cai C, Yi K, Chen Z, Zhu C. MicroRNA166 modulates cadmium tolerance and accumulation in rice[J]. Plant Physiology, 2018, 177(4): 1691-1703. |
| [30] | Tognolli M, Penel C, Greppin H, Simon P. Analysis and expression of the class Ⅲ peroxidase large gene family in Arabidopsis thaliana[J]. Gene, 2002, 288(1): 129-138. |
| [31] | Passardi F, Cosio C, Penel C, Dunand C. Peroxidases have more functions than a Swiss army knife[J]. Plant Cell Reports, 2005, 24(5): 255-265. |
| [32] | Liu X, Zhang Z. A double-edged sword: reactive oxygen species (ROS) during the rice blast fungus and host interaction[J]. The FEBS Journal, 2022, 289(18): 5505-5515. |
| [33] | O'Brien J A, Daudi A, Finch P, Butt V S, Whitelegge J P, Souda P, Ausubel F M, Paul B G. A peroxidase-dependent apoplastic oxidative burst in cultured Arabidopsis cells functions in MAMP-elicited defense[J]. Plant Physiology, 2012, 158(4): 2013-2027. |
| [34] | Zhao L, Phuong L T, Luan M T, Fitrianti A N, Matsui H, Nakagami H, Noutoshi Y, Yamamoto M, Ichinose Y, Shiraishi T, Toyoda K. A class Ⅲ peroxidase PRX34 is a component of disease resistance in Arabidopsis[J]. Journal of General Plant Pathology, 2019, 85(4):1-8. |
| [35] | Wally O, Punja Z K. Enhanced disease resistance in transgenic carrot (Daucus carota L.) plants over- expressing a rice cationic peroxidase[J]. Planta, 2010, 232(5): 1229-1239. |
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