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Gene Mapping and Blast Resistance Analysis of a Lesion Mimic Mutant spl52 in Rice (Oryza sativa) |
LI Yuan-Yuan1, LI Xing1, HUANG Qing-Xiong2, QI Pan1, YIN Wu-Zhong1, ZHANG Jie1, CHEN Hong1, YANG Guo-Tao1, PENG You-Lin2,*, HU Yun-Gao1,* |
1 Rice Research Institute of Southwest University of Science and Technology, Mianyang 621000, China; 2 Yazhou Bay National Laboratory, Sanya 572024, China |
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Abstract Rice (Oryza sativa) blast is one of the factors affecting the stable yield of rice. The lesion mimic mutant is an ideal material for studying the molecular mechanism of crop disease resistance. The analysis of the disease resistance gene of the mutant is the basis for cultivating stable and disease-resistant rice. In this study, a stable genetic lesion mimic mutant spl52 (spotted leaf 52) was obtained by ethylmethylsulfone (EMS) mutagenesis of japonica rice 'Wuyunjing 21'. The effective panicle number, grain number per panicle, 1000-grain weight and seed setting rate of the mutant plants at maturity stage were significantly lower than those of the wild type. From the early tillering stage, the leaf tip of mutant spl52 appeared disease spots. By 3,3'-Diaminobenzidine (DAB) staining, enzyme activity assay and 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) detection results showed that the mutant spl52 lesion leaves showed reactive oxygen species accumulation, and programmed cell death activation was found by TUNEL assay. Transmission electron microscopy analysis of wild-type and mutant leaves showed that the mutant chloroplasts were smaller oval, and the mitochondria were hollowed out. Genetic analysis showed that the phenotype of the mutant was controlled by a pair of recessive nuclear genes. Six possible candidate intervals were found by Mutmap analysis, and the candidate genes were located between RM3.5-RM4.3 on chromosome 6 by map-based cloning. The candidate genes were analyzed, and no reported lesion-like and rice resistance-related genes were found. The leaves of wild-type and mutant plants were inoculated with rice blast fungus, and the results showed that the mutant plants were more sensitive to rice blast fungus (Magnaporthe oryzae). The results of qRT-PCR showed that the expression levels of OsPR1a, OsPR1b, JIOsPR10, OsWRKY6 and OsRSR1 which positively regulated rice resistance in the mutant were significantly lower than those in the wild type. The results showed that the mutant gene was a new gene that had not been reported and was involved in the regulation of rice blast.
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Received: 24 October 2024
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Corresponding Authors:
*ylpeng@swust.edu.cn; huyungao@swust.edu.cn
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[1] Choi C, Hwang S H, Fang I R, et al.2015. Molecular characterization of Oryza sativa WRKY6, which binds to W‐box‐like element 1 of the Oryza sativa pathogenesis‐related PR10a promoter and confers reduced susceptibility to pathogens[J]. New Phytologist, 208: 846-859. [2] Cui Y, Peng Y, Zhang Q, et al.2020. Disruption of EARLY LESION LEAF 1, encoding a cytochrome P450 monooxygenase, induces ROS accumulation and cell death in rice[J]. The Plant Journal, 105: 942-956. [3] Domínguez F, Cejudo F. J2021. Chloroplast dismantling in leaf senescence[J]. Journal of Experimental Botany, 72: 5905-5918. [4] Guan M, Zhang W, Xu P, et al.2022. Mapping and functional analysis of high-copper accumulation mutant oshc1 in rice[J]. Journal of Hazardous Materials, 426: 128063. [5] Hu B, Zhou Y, Zhou Z, et al.2021. Repressed OsMESL expression triggers reactive oxygen species-mediated broad‐spectrum disease resistance in rice[J]. Plant Biotechnology Journal, 19: 1511-1522. [6] Huang Q N, Yang Y, Shi Y F, et al.2010. Spotted-leaf mutants of rice (Oryza sativa)[J]. Rice Science, 17: 247-256. [7] Jiang R, Zhou S, Da X, et al.2023. OsMKK6 regulates disease resistance in rice[J]. International Journal of Molecular Sciences, 24: 12678. [8] Jin B, Zhou X, Jiang B, et al.2015. Transcriptome profiling of the spl5 mutant reveals that SPL5 has a negative role in the biosynthesis of serotonin for rice disease resistance[J]. Rice, 8: 18. [9] Kang S G, Lee K E, Singh M, et al.2021. Rice lesion mimic mutants (LMM): The current understanding of genetic mutations in the failure of ROS scavenging during lesion formation[J]. Plants, 10: 1598. [10] Li B B, Liu Y G, Wu T, et al.2019. OsBGLU19 and OsBGLU23 regulate disease resistance to bacterial leaf streak in rice[J]. Journal of Integrative Agriculture, 18: 1199-1210. [11] Li Z, Ding B, Zhou X, et al.2017. The rice dynamin-related protein OsDRP1E negatively regulates programmed cell death by controlling the release of cytochrome c from mitochondria[J]. PLOS Pathogens, 13(1): 1006157-1006181. [12] Lin A, Wang Y, Tang J, et al.2011. Nitric oxide and protein-nitrosylation are integral to hydrogen peroxide-induced leaf cell death in rice[J]. Plant Physiology, 158: 451-464. [13] Magar N D, Barbadikar K M, Reddy V, et al.2024. Genetic mapping of regions associated with root system architecture in rice using MutMap QTL-seq[J]. Plant Physiology and Biochemistry, 213: 108836. [14] Matin M, Pandeya D, Baek K, et al.2010. Phenotypic and genotypic analysis of rice lesion mimic mutants[J]. Plant Pathology Journal, 26: 159-169. [15] Peng Y, Zou T, Li L, et al.2019. Map-based cloning and functional analysis of YE1 in rice, which is involved in light-dependent chlorophyll biogenesis and photoperiodic flowering pathway[J]. International Journal of Molecular Sciences, 20: 758. [16] Sathe A P, Su X, Chen Z, et al.2019. Identification and characterization of a spotted-leaf mutant spl40 with enhanced bacterial blight resistance in rice[J]. Rice, 12: 68. [17] Tang J, Zhu X, Wang Y, et al.2011. Semi‐dominant mutations in the CC‐NB‐LRR‐type gene, NLS1, lead to constitutive activation of defense responses in rice[J]. The Plant Journal, 66: 996-1007. [18] Tezuka D, Kawamata A, Kato H, et al.2019. The rice ethylene response factor OsERF83 positively regulates disease resistance to Magnaporthe oryzae[J]. Plant Physiology and Biochemistry, 135: 263-271. [19] Ting C, Zheng C, Sathe A P, et al.2019. Characterization of a novel gain-of-function spotted-leaf mutant with enhanced disease resistance in rice[J]. Rice Science, 26: 372-383. [20] Wang A, Shu X, Jing X, et al.2021. Identification of rice (Oryza sativa L.) genes involved in sheath blight resistance via a genome‐wide association study[J]. Plant Biotechnology Journal, 19: 1553-1566. [21] Wang C, Liu W J, Liao X W, et al.2024. The identification and gene mapping of spotted leaf mutant spl43 in rice[J]. International Journal of Molecular Sciences, 25: 6637. [22] Wang H, Zhang Y, Sun L, et al.2018. WB1, a regulator of endosperm development in rice, is identified by a modified mutmap method[M]. International Journal of Molecular Sciences, MDPI AG: 2159. [23] Wang S, Lei C, Wang J, et al.2017. SPL33, encoding an eEF1A-like protein, negatively regulates cell death and defense responses in rice[J]. Journal of Experimental Botany, 68: 899-913. [24] Wang Y, Chen Y, Zhang J, et al.2022. Overexpression of llm1 affects the synthesis of secondary metabolites of Aspergillus cristatus[M]. Microorganisms, MDPI AG: 1707. [25] Wu J, Kim S G, Kang K Y.2016. Overexpression of a pathogenesis-related protein 10 enhances biotic and abiotic stress tolerance in rice[J]. Plant Pathology Journal, 32: 552. [26] Yan J, Fang Y, Xue D.2022. Advances in the genetic basis and molecular mechanism of lesion mimic formation in rice[J]. Plants, 11: 2169. [27] Yan Y, Zhu X, Qi H, et al.2024. Rice seed storability: From molecular mechanisms to agricultural practices[J]. Plant Science, 348: 112215. [28] Yin Z, Chen J, Zeng L, et al.2000. Characterizing rice lesion mimic mutants and identifying a mutant with broad-spectrum resistance to rice blast and bacterial blight[J]. Molecular Plant-Microbe Interactions, 13: 869-876. [29] Yong Y, Lin Q J, Chen X Y, et al.2021. Characterization and proteomic analysis of novel rice lesion mimic mutant with enhanced disease resistance[J]. Rice Science, 28: 466-478. [30] Rao Y C, Ran J, Sheng W, et al.2021. SPL36 encodes a receptor-like protein kinase that regulates programmed cell death and defense responses in rice[J]. Rice, 14: 34. [31] Zeng L R, Qu S, Bordeos A, et al.2004. Spotted leaf11, a negative regulator of plant cell death and defense, encodes a U-Box/Armadillo repeat protein endowed with E3 ubiquitin ligase activity[J]. The Plant Cell, 16: 2795-2808. [32] Zhang A, Jiang H, Chu H, et al.2022. Rice lesion mimic gene cloning and association analysis for disease resistance[M]. Current Issues in Molecular Biology, MDPI AG: 2350-2361. [33] Zhao M, Guo Y, Sun H, et al.2023. Lesion mimic mutant 8 balances disease resistance and growth in rice[J]. Frontiers in Plant Science, 14: 1189926. |
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LUO Xi, FAN Jia-Xing, WEI Yi-Dong, WEI Lin-Yan, ZHU Yong-Sheng, HE Wei, WU Fang-Xi, CAI Qiu-Hua, XIE Hua-An, ZHANG Jian-Fu. The Impact on Resistant-starch Content for Base Mutations Near the 3' Splice Site of the Fourth Intron of the Waxy Gene in Rice (Oryza sativa)[J]. 农业生物技术学报, 2025, 33(9): 1873-1882. |
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