CRISPR/Cas9-Targeted Knockout of Rice Susceptibility Genes OsDjA2 and OsERF104 Reveals Alternative Sources of Resistance to Pyricularia oryzae
Received date: 2022-01-07
Accepted date: 2022-04-24
Online published: 2022-09-02
Rice genes OsDjA2 and OsERF104, encoding a chaperone protein and an APETELA2/ ethylene-responsive factor, respectively, are strongly induced in a compatible interaction with blast fungus, and also have function in plant susceptibility validated through gene silencing. Here, we reported the CRISPR/Cas9 knockout of OsDjA2 and OsERF104 genes resulting in considerable improvement of blast resistance. A total of 15 OsDjA2 (62.5%) and 17 OsERF104 (70.8%) T0 transformed lines were identified from 24 regenerated plants for each target and used in downstream experiments. Phenotyping of homozygous T1 mutant lines revealed not only a significant decrease in the number of blast lesions but also a reduction in the percentage of diseased leaf area, compared with the infected control plants. Our results supported CRISPR/Cas9-mediated target mutation in rice susceptibility genes as a potential and alternative breeding strategy for building resistance to blast disease.
Fabiano T. P. K. Távora, Anne Cécile Meunier, Aurore Vernet, Murielle Portefaix, Joëlle Milazzo, Henri Adreit, Didier Tharreau, Octávio L. Franco, Angela Mehta . CRISPR/Cas9-Targeted Knockout of Rice Susceptibility Genes OsDjA2 and OsERF104 Reveals Alternative Sources of Resistance to Pyricularia oryzae[J]. Rice Science, 2022 , 29(6) : 535 -544 . DOI: 10.1016/j.rsci.2022.04.001
| [1] | Abiri R, Shaharuddin N A, Maziah M, Yusof Z N B, Atabaki N, Sahebi M, Valdiani A, Kalhori N, Azizi P, Hanafi M M. 2017. Role of ethylene and the APETALA 2/ethylene response factor superfamily in rice under various abiotic and biotic stress conditions. Environ Exp Bot, 134: 33-44. |
| [2] | Ahn S W, Seshu D V. 1991. Blast reaction of durably resistance rice cultivar in multiplication trials. Phytopathology, 81(10): 1150. |
| [3] | Bes M, Herbert L, Mounier T, Meunier A C, Durandet F, Guiderdoni E, Périn C. 2021. Efficient genome editing in rice protoplasts using CRISPR/CAS9 construct. Methods Mol Biol, 2238: 173-191. |
| [4] | Bevitori R, Sircar S, de Mello R N, Togawa R C, Côrtes M V C B, Oliveira T S, Grossi-de-Sá M F, Parekh N. 2020. Identification of co-expression gene networks controlling rice blast disease during an incompatible reaction. Genet Mol Res, 19(3): gmr18579. |
| [5] | Bonman J M, Khush G S, Nelson R J. 1992. Breeding rice for resistance to pests. Annu Rev Phytopathol, 30: 507-528. |
| [6] | Boston R S, Viitanen P V, Vierling E. 1996. Molecular chaperones and protein folding in plants. Plant Mol Biol, 32(1/2): 191-222. |
| [7] | Büschges R, Hollricher K, Panstruga R, Simons G, Wolter M, Frijters A, van Daelen R, van der Lee T, Diergaarde P, Groenendijk J, Töpsch S, Vos P, Salamini F, Schulze-Lefert P. 1997. The barley Mlo gene: A novel control element of plant pathogen resistance. Cell, 88(5): 695-705. |
| [8] | Choi Y, Sims G E, Murphy S, Miller J R, Chan A P. 2012. Predicting the functional effect of amino acid substitutions and indels. PLoS One, 7(10): e46688. |
| [9] | Concordet J P, Haeussler M. 2018. CRISPOR: Intuitive guide selection for CRISPR/Cas9 genome editing experiments and screens. Nucleic Acids Res, 46(W1): W242-W245. |
| [10] | Dehairs J, Talebi A, Cherifi Y, Swinnen J V. 2016. CRISP-ID: Decoding CRISPR mediated indels by Sanger sequencing. Sci Rep, 6: 28973. |
| [11] | do Vale F X R, Fernandes Filho E I, Liberato J R. 2003. A Software for Plant Disease Severity Assessment. Christchurch, New Zealand: 8th International Congress of Plant Pathology: 105. |
| [12] | Feng K, Hou X L, Xing G M, Liu J X, Duan A Q, Xu Z S, Li M Y, Zhuang J, Xiong A S. 2020. Advances in AP2/ERF super-family transcription factors in plant. Crit Rev Biotechnol, 40(6): 750-776. |
| [13] | Fukagawa N K, Ziska L H. 2019. Rice: Importance for global nutrition. J Nutr Sci Vitaminol, 65: S2-S3. |
| [14] | Gasteiger E, Gattiker A, Hooland C, Ivanyi I, Appel R D, Bairoch A. 2003. ExPASy: The proteomics server for in-death protein knowledge and analysis. Nucleic Acids Res, 31(13): 3784-3788. |
| [15] | Hiei Y, Ohta S, Komari T, Kumashiro T. 1994. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J, 6(2): 271-282. |
| [16] | Hong Y B, Liu Q N, Cao Y R, Zhang Y, Chen D B, Lou X Y, Cheng S H, Cao L Y. 2019. The OsMPK15 negatively regulates Magnaporthe oryzae and Xoo disease resistance via SA and JA signaling pathway in rice. Front Plant Sci, 10: 752. |
| [17] | Jain P, Singh P K, Kapoor R, Khanna A, Solanke A U, Krishnan S G, Singh A K, Sharma V, Sharma T R. 2017. Understanding host-pathogen interactions with expression profiling of NILs carrying rice-blast resistance Pi9 gene. Front Plant Sci, 8: 93. |
| [18] | Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna J A, Charpentier E. 2012. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science, 337: 816-821. |
| [19] | Kampinga H H, Andreasson C, Barducci A, Cheetham M E, Cyr D, Emanuelsson C, Genevaux P, Gestwicki J E, Goloubinoff P, Huerta-Cepas J, Kirstein J, Liberek K, Mayer M P, Nagata K, Nillegoda N B, Pulido P, Ramos C, de Los Rios P, Rospert S, Rosenzweig R, Sahi C, Taipale M, Tomiczek B, Ushioda R, Young J C, Zimmermann R, Zylicz A, Zylicz M, Craig E A, Marszalek J. 2019. Function, evolution, and structure of J-domain proteins. Cell Stress Chaperones, 24(1): 7-15. |
| [20] | Ko S H, Huang L M, Tarn W Y. 2019. The host heat shock protein MRJ/DNAJB6 modulates virus infection. Front Microbiol, 10: 2885. |
| [21] | Kusch S, Panstruga R. 2017. mlo-based resistance: An apparently universal “weapon” to defeat powdery mildew disease. Mol Plant Microbe Interact, 30(3): 179-189. |
| [22] | Li W T, Chern M, Yin J J, Wang J, Chen X W. 2019. Recent advances in broad-spectrum resistance to the rice blast disease. Curr Opin Plant Biol, 50: 114-120. |
| [23] | Luo Y, Fang B H, Wang W P, Yang Y, Rao L Q, Zhang C. 2019. Genome-wide analysis of the rice J-protein family: Identification, genomic organization, and expression profiles under multiple stresses. 3 Biotech, 9(10): 358. |
| [24] | Miao J, Guo D S, Zhang J Z, Huang Q P, Qin G J, Zhang X, Wan J M, Gu H Y, Qu L J. 2013. Targeted mutagenesis in rice using CRISPR-Cas system. Cell Res, 23(10): 1233-1236. |
| [25] | Nakano T, Suzuki K, Fujimura T, Shinshi H. 2006. Genome-wide analysis of the ERF gene family in Arabidopsis and rice. Plant Physiol, 140(2): 411-432. |
| [26] | Oliva R, Ji C H, Atienza-Grande G, Huguet-Tapia J C, Perez-Quintero A, Li T, Eom J S, Li C H, Nguyen H, Liu B, Auguy F, Sciallano C, Luu V T, Dossa G S, Cunnac S, Schmidt S M, Slamet-Loedin I H, Vera Cruz C, Szurek B, Frommer W B, White F F, Yang B. 2019. Broad-spectrum resistance to bacterial blight in rice using genome editing. Nat Biotechnol, 37(11): 1344-1350. |
| [27] | Parisi C, Tillie P, Rodríguez-Cerezo E. 2016. The global pipeline of GM crops out to 2020. Nat Biotechnol, 34(1): 31-36. |
| [28] | Park C J, Seo Y S. 2015. Heat shock proteins: A review of the molecular chaperones for plant immunity. Plant Pathol J, 31(4): 323-333. |
| [29] | Phukan U J, Jeena G S, Tripathi V, Shukla R K. 2017. Regulation of Apetala2/ethylene response factors in plants. Front Plant Sci, 8: 150. |
| [30] | Rashid M, He G Y, Yang G X, Hussain J, Yan X. 2012. AP2/ERF transcription factor in rice: Genome-wide canvas and syntenic relationships between monocots and eudicots. Evol Bioinform Online, 8: 321-355. |
| [31] | Ribot C, Hirsch J, Balzergue S, Tharreau D, Nottéghem J L, Lebrun M H, Morel J B. 2008. Susceptibility of rice to the blast fungus, Magnaporthe grisea. J Plant Physiol, 165(1): 114-124. |
| [32] | Sallaud C, Lorieux M, Roumen E, Tharreau D, Berruyer R, Svestasrani P, Garsmeur O, Ghesquiere A, Notteghem J L. 2003. Identification of five new blast resistance genes in the highly blast-resistant rice variety IR64 using a QTL mapping strategy. Theor Appl Genet, 106(5): 794-803. |
| [33] | Sarkar N K, Thapar U, Kundnani P, Panwar P, Grover A. 2013. Functional relevance of J-protein family of rice (Oryza sativa). Cell Stress Chaperones, 18(3): 321-331. |
| [34] | Srivastava R, Kumar R. 2018. The expanding roles of APETALA2/Ethylene Responsive Factors and their potential applications in crop improvement. Brief Funct Genomics, 18(4): 240-254. |
| [35] | Stam R, McDonald B A. 2018. When resistance gene pyramids are not durable: The role of pathogen diversity. Mol Plant Pathol, 19(3): 521-524. |
| [36] | Streubel J, Pesce C, Hutin M, Koebnik R, Boch J, Szurek B. 2013. Five phylogenetically close rice SWEET genes confer TAL effector-mediated susceptibility to Xanthomonas oryzae pv. oryzae. New Phytol, 200(3): 808-819. |
| [37] | Távora F T P K, Bevitori R, Mello R N, Cintra M M D F, Oliveira-Neto O B, Fontes W, Castro M S, Sousa M V, Franco O L, Mehta A. 2021. Shotgun proteomics coupled to transient- inducible gene silencing reveal rice susceptibility genes as new sources for blast disease resistance. J Proteomics, 241: 104223. |
| [38] | van Schie C C N, Takken F L W. 2014. Susceptibility genes 101: How to be a good host. Annu Rev Phytopathol, 52: 551-581. |
| [39] | Verma A K, Tamadaddi C, Tak Y, Lal S S, Cole S J, Hines J K, Sahi C. 2019. The expanding world of plant J-domain proteins. Crit Rev Plant Sci, 38: 382-400. |
| [40] | Win J, Chaparro-Garcia A, Belhaj K, Saunders D G O, Yoshida K, Dong S, Schornack S, Zipfel C, Robatzek S, Hogenhout S A, Kamoun S. 2012. Effector biology of plant-associated organisms: Concepts and perspectives. Cold Spring Harb Symp Quant Biol, 77: 235-247. |
| [41] | Wen C K. 2015. Ethylene in Plants. Berlin: Springer: 286. |
| [42] | Yang L T, Ding J Y, Zhang C M, Jia J W, Weng H B, Liu W X, Zhang D B. 2005. Estimating the copy number of transgenes in transformed rice by real-time quantitative PCR. Plant Cell Rep, 23(10/11): 759-763. |
| [43] | Zaidi S S E A, Mukhtar M S, Mansoor S. 2018. Genome editing: Targeting susceptibility genes for plant disease resistance. Trends Biotechnol, 36(9): 898-906. |
/
| 〈 |
|
〉 |