Research Paper

Characterization and Proteomic Analysis of Novel Rice Lesion Mimic Mutant with Enhanced Disease Resistance

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  • 1State Key Laboratory for Managing Biotic and Chemical Treats to the Quality and Safety of Agro-Products / Key Laboratory of Biotechnology for Plant Protection, Ministry of Agriculture and Rural Affairs of China / Zhejiang Provincial Key Laboratory of Biotechnology for Plant Protection, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Science, Hangzhou 310021, China
    2Plant Pathogen Laboratory, College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China
    3State Key Laboratory of Subtropical Silviculture, Zhejiang Agriculture and Forest University, Hangzhou 311300, China
    4College of Plant Protection, Yunnan Agricultural University, Kunming 650201, China
    5State Key Laboratory for Managing Biotic and Chemical Treats to the Quality and Safety of Agro-Products / Key Laboratory of Biotechnology for Plant Protection, Ministry of Agriculture and Rural Affairs of China / Zhejiang Provincial Key Laboratory of Biotechnology for Plant Protection, Institute of Plant Virology, Ningbo University, Ningbo 315211, China
    6Institute of Biotechnology, Ningbo Academy of Agricultural Science, Ningbo 315040, China

#These authors contributed equally to this work

Received date: 2020-08-19

  Accepted date: 2021-03-02

  Online published: 2021-09-28

Abstract

Lesion mimic mutants (LMMs) are plants that spontaneously form lesions without pathogen infection or external stimulus and exhibit resistance to pathogens. Here, a rice LMM was created by ethyl methane sulfonate mutagenesis, named as hpil (hydrogen peroxide induced lesion). Diaminobenzidine and trypan blue staining showed that large amounts of H2O2 were produced and cell death was occurred at and around the parts of lesion mimic in the rice leaves. The phenotype of hpil is controlled by a single recessive gene, localized at a 2 Mb interval on chromosome 2. The data suggested that hpil is a novel LMM with enhanced bacterial and fungal disease resistance, and multiple pathogenesis-related proteins (PRs) were up-regulated. The proteomes of leaves at three positions (different degrees of lesion mimic severity) were characterized in hpil compared with its wild type plant. Differentially expressed proteins were detected by two dimensional difference gel electrophoresis and 274 proteins were identified by MALDI TOF/TOFTM. These proteins were related to metabolic process, cellular process and response to stimulus, with mostly down-regulated in hpil leaves. Many of these proteins were related to the Calvin cycle, photosynthetic electron transport chain, glycolysis/gluconeogenesis and phosphonates pathways. Some resistance-related proteins including 14-3-3 proteins, OsPR10 and antioxidases such as peroxidase, superoxide dismutase and ascorbate peroxidase were up-regulated in leaves with lesion mimic. These results provide the foundation for cloning of the target gene and shed light on the mechanism involved in autoimmunity of rice.

Cite this article

Yong Yang, Qiujun Lin, Xinyu Chen, Weifang Liang, Yuwen Fu, Zhengjin Xu, Yuanhua Wu, Xuming Wang, Jie Zhou, Chulang Yu, Chengqi Yan, Qiong Mei, Jianping Chen . Characterization and Proteomic Analysis of Novel Rice Lesion Mimic Mutant with Enhanced Disease Resistance[J]. Rice Science, 2021 , 28(5) : 466 -478 . DOI: 10.1016/j.rsci.2021.07.007

References

[1] Akhkha A, Clarke D D. 2003. Relative tolerances of wild and cultivated barleys to infection by Blumeria graminis f. sp. hordei (Syn. Erysiphe graminis f. sp. hordei): I. The effects of infection on growth and development. Physiol Mol Plant Pathol, 62(4): 237-250.
[2] Anonymous.2002. Find out how the qualities of rice are evaluated and scored in this authoritative sourcebook. In: Standard Evaluation System for Rice. Laguna, the Philippines: International Rice Research Institute: 1518.
[3] Arnon D I. 1949. Copper enzymes in isolated chloroplasts: Polyphenoloxidase in Beta vulagaris. Plant Physiol, 24(1): 1-15.
[4] Beckman K B, Ames B N. 1997. Oxidative decay of DNA. J Biol Chem, 272(32): 19633?19636.
[5] Bowling S A, Clarke J D, Liu Y D, Klessig D F, Dong X. 1997. The cpr5 mutant of Arabidopsis expresses both NPR1-dependent and NPR1-independent resistance. Plant Cell, 9(9): 1573-1584.
[6] Bruggeman Q, Raynaud C, Benhamed M, Delarue M. 2015. To die or not to die? Lessons from lesion mimic mutants. Front Plant Sci, 6: 24.
[7] Chen F, Li Q, Sun L X, He Z H. 2006. The rice 14-3-3 gene family and its involvement in responses to biotic and abiotic stress. DNA Res, 13(2): 53-63.
[8] Chen T, Chen Z, Sathe A P, Zhang Z H, Li L J, Shang H H, Tang S Q, Zhang X B, Wu J L. 2019. Characterization of a novel gain-of-function spotted-leaf mutant with enhanced disease resistance in rice. Rice Sci, 26(6): 372-383.
[9] Cheng Y T, Li Y Z, Huang S, Huang Y, Dong X N, Zhang Y L, Li X. 2011. Stability of plant immune-receptor resistance proteins is controlled by SKP1-CULLIN1-F-box (SCF)-mediated protein degradation. Proc Natl Acad Sci USA, 108(35): 14694-14699.
[10] Cheng Y T, Li X. 2012. Ubiquitination in NB-LRR-mediated immunity. Curr Opin Plant Biol, 15(4): 392-399.
[11] Coll N S, Epple P, Dangl J L. 2011. Programmed cell death in the plant immune system. Cell Death Differ, 18(8): 1247-1256.
[12] Corpas F J, González-Gordo S, Palma J M. 2020. Plant peroxisomes: A factory of reactive species. Front Plant Sci, 11: 853.
[13] Crafts-Brandner S J, van de Loo F J, Salvucci M E. 1997. The two forms of ribulose-1,5-bisphosphate carboxylase/oxygenase activase differ in sensitivity to elevated temperature. Plant Physiol, 114(2): 439-444.
[14] da S Andrade L B, Oliveira A S, Ribeiro J K C, Kiyota S, Vasconcelos I M, de Oliveira J T A, de Sales M P. 2010. Effects of a novel pathogenesis-related class 10 (PR-10) protein from Crotalaria pallida roots with papain inhibitory activity against root-knot nematode Meloidogyne incognita. J Agric Food Chem, 58(7): 4145-4152.
[15] Dangl J L, Dietrich R A, Richberg M H. 1996. Death don’t have no mercy: Cell death programs in plant-icrobe interactions. Plant Cell, 8(10): 1793-1807.
[16] Deng Z P, Zhang X, Tang W Q, Oses-Prieto J A, Suzuki N, Gendron J M, Chen H J, Guan S H, Chalkley R J, Peterman T K, Burlingame A L, Wang Z Y. 2007. A proteomics study of brassinosteroid response in Arabidopsis. Mol Cell Proteomics, 6(12): 2058-2071.
[17] Dong Y, Fang X P, Yang Y, Xue G P, Chen X, Zhang W L, Wang X M, Yu C L, Zhou J, Mei Q, Fang W, Yan C Q, Chen J P. 2017. Comparative proteomic analysis of susceptible and resistant rice plants during early infestation by small brown planthopper. Front Plant Sci, 8: 1744.
[18] Fang X P, Chen W Y, Xin Y, Zhang H M, Yan C Q, Yu H, Liu H, Xiao W F, Wang S Z, Zheng G Z, Liu H B, Jin L, Ma H S, Ruan S L. 2012. Proteomic analysis of strawberry leaves infected with Colletotrichum fragariae. J Proteomics, 75(13): 4074-4090.
[19] Fekih R, Tamiru M, Kanzaki H, Abe A, Yoshida K, Kanzaki E, Saitoh H, Takaqi H, Natsume S, Undan J R, Undan J, Terauchi R. 2015. The rice (Oryza sativa L.) LESION MIMIC RESEMBLING, which encodes an AAA-type ATPase, is implicated in defense response. Mol Genet Genomics, 290(2): 611-622.
[20] Flores T, Alape-Girón A, Flores-Díaz M, Flores H E. 2002. Ocatin: A novel tuber storage protein from the andean tuber crop oca with antibacterial and antifungal activities. Plant Physiol, 128(4): 1291-1302.
[21] Giannopolitis C N, Ries S K. 1977. Superoxide dismutases: I. occurrence in higher plants. Plant Physiol, 59(2): 309-314.
[22] Gordon T R, Duniway J M. 1982. Effects of powdery mildew infection on the efficiency of CO2 fixation and light utilization by sugar beet leaves. Plant Physiol, 69(1): 139-142.
[23] Hashimoto M, Kisseleva L, Sawa S, Furukawa T, Komatsu S, Koshiba T. 2004. A novel rice PR10 protein, RSOsPR10, specifically induced in roots by biotic and abiotic stresses, possibly via the jasmonic acid signaling pathway. Plant Cell Physiol, 45(5): 550-559.
[24] Henkes S, Sonnewald U, Badur R, Flachmann R, Stitt M. 2001. A small decrease of plastid transketolase activity in antisense tobacco transformants has dramatic effects on photosynthesis and phenylpropanoid metabolism. Plant Cell, 13(3): 535-552.
[25] Hiraga S, Yamamoto K, Ito H, Sasaki K, Matsui H, Honma M, Nagamura Y, Sasaki T, Ohashi Y. 2000. Diverse expression profiles of 21 rice peroxidase genes. FEBS Lett, 471: 245?250.
[26] Huang L F, Lin K H, He S L, Chen J L, Jiang J Z, Chen B H, Hou Y S, Chen R S, Hong C Y, Ho S L. 2016. Multiple patterns of regulation and overexpression of a ribonuclease-like pathogenesis- related protein gene,OsPR10a, conferring disease resistance in rice and Arabidopsis. PLoS One, 11(6): e0156414.
[27] Huang Q N, Yang Y, Shi Y F, Chen J, Wu J L. 2010. Spotted-leaf mutants of rice (Oryza sativa). Rice Sci, 17(4): 247-256.
[28] Jones J D G, Dangl J L. 2006. The plant immune system. Nature, 444: 323?329.
[29] Jwa N S, Agrawal G K, Rakwal R, Park C H, Agrawal V P. 2001. Molecular cloning and characterization of a novel jasmonate inducible pathogenesis-related class 10 protein gene, JIOsPR10, from rice (Oryza sativa L.) seedling leaves. Biochem Biophy Res Commun, 286(5): 973-983.
[30] Kang S G, Matin M N, Bae H, Natarajan S. 2007. Proteome analysis and characterization of phenotypes of lesion mimic mutant spotted leaf 6 in rice. Proteomics, 7(14): 2447-2458.
[31] Kauffman H E, Reddy A P K, Hsieh S P Y, Merca S D. 1973. An improved technique for 436 evaluating resistance of rice varieties to Xanthomonas oryzae. Plant Dis Rep, 57: 737?741.
[32] Liu Q E, Ning Y S, Zhang Y X, Yu N, Zhao C D, Zhan X D, Wu W X, Chen D B, Wei X J, Wang G L, Cheng S H, Cao L Y. 2017. OsCUL3a negatively regulates cell death and immunity by degrading OsNPR1 in rice. Plant Cell, 29(2): 345-359.
[33] Lorrain S, Vailleau F, Balagué C, Roby D. 2003. Lesion mimic mutants: Keys for deciphering cell death and defense pathways in plants? Trends Plant Sci, 8(6): 263-271.
[34] Lozano-Durán R, Robatzek S. 2015. 14-3-3 proteins in plant- pathogen interactions. Mol Plant Microbe Interact, 28(5): 511-518.
[35] Manosalva P M, Bruce M, Leach J E. 2011. Rice 14-3-3 protein (GF14e) negatively affects cell death and disease resistance. Plant J, 68(5): 777-787.
[36] Mcgee J D, Hamer J E, Hodges T K. 2001. Characterization of a PR-10 pathogenesis-related gene family induced in rice during infection with Magnaporthe grisea. Mol Plant Microbe Interact, 14(7): 877-886.
[37] Mei Q, Yang Y, Ye S H, Liang W F, Wang X M, Zhou J, Yu C L, Yan C Q, Chen J P. 2019. H2O2 induces association of RCA with the thylakoid membrane to enhance resistance of Oryza meyeriana to Xanthomonas oryzae pv. oryzae. Plants, 8(9): 351.
[38] Nakano Y, Asada K. 1981. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol, 22(5): 867-880.
[39] Navarro L, Zipfel C, Rowland O, Keller I, Robatzek S, Boller T, Jones J D G. 2004. The transcriptional innate immune response to flg22: Interplay and overlap with Avr gene-dependent defense responses and bacterial pathogenesis. Plant Physiol, 135(2): 1113-1128.
[40] Park C J, Kim K J, Shin R, Park J M, Shin Y C, Peak K H. 2004. Pathogenesis-related protein 10 isolated from hot pepper functions as a ribonuclease in an antiviral pathway. Plant J, 37(2): 186-198.
[41] Qin P, Fan S J, Deng L C, Zhong G R, Zhang S W, Li M, Chen W L, Wang G L, Tu B, Wang Y P, Chen X W, Ma B T, Li S G. 2018. LML1, encoding a conserved eukaryotic release factor 1 protein, regulates cell death and pathogen resistance by forming a conserved complex with SPL33 in rice. Plant Cell Physiol, 59(5): 887-902.
[42] Rodriguez E, Ghoul H E, Mundy J, Petersen M. 2015. Making sense of plant autoimmunity and ‘negative regulators’. FEBS J, 283(8): 1385-1391.
[43] Sharkey T D, Badger M R, von Caemmerer S, Andrews T J. 2001. Increased heat sensitivity of photosynthesis in tobacco plants with reduced Rubisco activase. Photosynth Res, 67(1): 147-156.
[44] Shen X L, Liu H B, Yuan B, Li X H, Xu C G, Wang S P. 2011. OsEDR1 negatively regulates rice bacterial resistance via activation of ethylene biosynthesis. Plant Cell Environ, 34(2): 179-191.
[45] Suzuki N, Miller G, Morales J, Shulaev V, Torres M A, Mittler R. 2011. Respiratory burst oxidases: The engines of ROS signaling. Curr Opin Plant Biol, 14(6): 691-699.
[46] Tang J Y, Zhu X D, Wang Y Q, Liu L C, Xu B, Li F, Fang J, Chu C C. 2011. Semi-dominant mutations in the CC-NB-LRR-type R gene, NLS1, lead to constitutive activation of defense responses in rice. Plant J, 66(6): 996-1007.
[47] Thordal-Christensen H, Zhang Z G, Wei Y D, Collinge D B. 1997. Subcellular localization of H2O2 in plants: H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J, 11(6): 1187-1194.
[48] Tsuda K, Katagiri F. 2010. Comparing signaling mechanisms engaged in pattern-triggered and effector-triggered immunity. Curr Opin Plant Biol, 13(4): 459-465.
[49] van Loon L C, van Strien E A. 1999. The families of pathogenesis- related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol Mol Plant Pathol, 55(2): 85-97.
[50] Wang L J, Pei Z Y, Tian Y C, He C Z. 2005. OsLSD1, a rice zinc finger protein, regulates programmed cell death and callus differentiation. Mol Plant Microbe Interact, 18(5): 375-384.
[51] Wang S, Lei C L, Wang J L, Ma J, Tang S, Wang C L, Zhao K J, Tian P, Zhang H, Qi C Y, Cheng Z J, Zhang X, Guo X P, Liu L L, Wu C Y, Wan J M. 2017. SPL33, encoding an eEF1A-like protein, negatively regulates cell death and defense responses in rice. J Exp Bot, 68(5): 899-913.
[52] Wang X C, Wang D Y, Wang D, Wang H Y, Chang L L, Yi X P, Peng M, Guo A P. 2012. Systematic comparison of technical details in CBB methods and development of a sensitive GAP stain for comparative proteomic analysis. Electrophoresis, 33(2): 296-306.
[53] Wang Z H, Wang Y, Xiao H, Hu D H, Liu C X, Yang J, Li Yang, Huang Y Q, Feng Y Q, Gong H Y, Li Y, Fang G, Tang H R, Li Y S. 2015. Functional inactivation of UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1) induces early leaf senescence and defence responses in rice. J Exp Bot, 66(3): 973-987.
[54] Waszczak C, Carmody M, Kangasjärvi J. 2018. Reactive oxygen species in plant signaling. Annu Rev Plant Biol, 69: 209-236.
[55] Wu J N, Kim S G, Kang KY, Kim J G, Park S R, Gupta R, Kim Y H, Wang Y M, Kim S T. 2016. Overexpression of a pathogenesis- related protein 10 enhances biotic and abiotic stress tolerance in rice. Plant Pathol J, 32(6): 552-562.
[56] Xia Z L, Wei Y Y, Sun K L, Wu J Y, Wang Y X, Wu K. 2013. The maize AAA-type protein SKD1 confers enhanced salt and drought stress tolerance in transgenic tobacco by interacting with lyst-interacting protein 5. PLoS One, 8(7): e69787.
[57] Xie Y R, Chen Z Y, Brown R L, Bhatnagar D. 2010. Expression and functional characterization of two pathogenesis-related protein 10 genes from Zea mays. J Plant Physiol, 167(2): 121-130.
[58] Yan S P, Zhang Q Y, Tang Z C, Su W A, Sun W N. 2005. Comparative proteomic analysis provides new insights into chilling stress responses in rice. Mol Cell Proteomics, 5(3): 484-496.
[59] Yin Z, Chen J, Zeng L, Goh M, Leung H, Khush G S, Wang G L. 2000. Characterizing rice lesion mimic mutants and identifying a mutant with broad-spectrum resistance to rice blast and bacterial blight. Mol Plant Microbe Interact, 13(8): 869-876.
[60] Zeng L R, Qu S H, Bordeos A, Yang C W, Baraoidan M, Yan H Y, Xie Q, Nahm B H, Leung H, Wang G L. 2004. Spotted leaf 11, a negative regulator of plant cell death and defense, encodes a U-box/armadillo repeat protein endowed with E3 ubiquitin ligase activity. Plant Cell, 16(10): 2795-2808.
[61] Zhang H F, Tang W, Liu K Y, Huang Q, Zhang X, Yan X, Chen Y, Wang J S, Qi Z Q, Wang Z Y, Zheng X B, Wang P, Zhang Z G. 2011. Eight RGS and RGS-like proteins orchestrate growth, differentiation, and pathogenicity of Magnaporthe oryzae. PLoS Pathog, 7(12): e1002450.
[62] Zhang X B, Feng B H, Wang H M, Xu X, Shi Y F, He Y, Chen Z, Sathe A P, Shi L, Wu J L. 2018. A substitution mutation in OsPELOTA confers bacterial blight resistance by activating the salicylic acid pathway. J Integr Plant Biol, 60(2): 160-172.
[63] Zhao J Y, Liu P C, Li C R, Wang Y Y, Guo L Q, Jiang G H, Zhai W X. 2017. LMM5.1 and LMM5.4, two eukaryotic translation elongation factor 1A-like gene family members, negatively affect cell death and disease resistance in rice. J Genet Genomics, 44(2): 107-118.
[64] Zhou L, Cheung M Y, Li M W, Fu Y P, Sun Z X, Sun S M, Lam M H. 2010. Rice hypersensitive induced reaction protein 1 (OsHIR1) associates with plasma membrane and triggers hypersensitive cell death. BMC Plant Biol, 10(1): 290.
[65] Zhu X B, Mu Z, Mawsheng C, Chen X W, Wang J. 2020. Deciphering rice lesion mimic mutants to understand molecular network governing plant immunity and growth. Rice Sci, 27(4): 278-288.
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