Chinese Journal OF Rice Science >
Comparative Transcriptome Analysis of Transgenic Rice Line Carrying the Rice Blast Resistance Gene Pi9
Received date: 2019-11-23
Revised date: 2020-03-11
Online published: 2020-05-10
【Objective】The purpose of the study is to analyze the mechanism behind Pi9 gene-mediated rice blast resistance at the transcriptional level, and to lay a theoretical basis for breeding disease-resistant rice cultivars. 【Method】Rice cultivar Nipponbare (NPB) and NPB-transgenic line carrying the Pi9 rice blast resistance gene (NPB/Pi9) were inoculated with Magnaporthe oryzae. Leaf tissues were sampled at 0 h, 12 h, 24 h, and 36 h after inoculation, respectively. Gene-chip transcriptome analysis of 12503 rice genes was performed using the rice samples, and the microarray data were verified by qRT-PCR of a set of differentially expressed genes (DEGs). 【Result】7754 DEGs were identified by comparing the gene expression levels in the resistant NPB/Pi9 line at 12 h, 24 h and 36 h after inoculation, respectively, to that at 0 h after inoculation. Accordingly, 7385 DEGs were identified in the susceptible rice NPB at the same time points. At 36 h after inoculation, the DEG number of NPB/Pi9 was significantly higher than that of NPB. 4065 DEGs were identified by comparing the gene expression levels in NPB/Pi9 and NPB at the same time points; there were significantly more DEGs at 36 h after inoculation than at 0 h, 12 h or 24 h after inoculation. Therefore, NPB/Pi9 exhibited more intensive defense responses to Magnaporthe oryzae. Gene Ontology (GO) and KEGG analyses were carried out on the DEGs between NPB/Pi9 and NPB at the same time points. The GO terms classifications related to extracellular regions, plant response to stimuli, transcriptional regulation, redox activity, ion binding activity, secondary metabolism and plant hormones were significantly enriched for the time points post inoculation. The KEGG pathways of phenylalanine metabolism, flavonoid biosynthesis and plant hormone signaling were enriched significantly at the time points post inoculation. Distinctive gene expression patterns were observed between NPB/Pi9 and NPB at the same time points for the genes of the effector-triggered immunity (ETI) related salicylic acid signaling pathway and chitinase, and the PAMP-triggered immunity (PTI) related extracellular regions, response to stimuli, and lignin biosynthesis. Moreover, distinctive gene expression patterns were observed between NPB/Pi9 and NPB at the same time points for the genes of WRKY transcription factors, MAPK kinases, jasmonate and ethylene signaling pathways that were utilized by both PTI and ETI. Based on all the results, the differential expression patterns between NPB/Pi9 and NPB are related to ETI and PTI, which are involved with each other and play important roles in the Pi9-mediated rice blast rice resistance. 【Conclusion】 Compared with NPB, the resistant genotype NPB/Pi9 showed more intensive defense responses against Magnaporthe oryzae. Transcription factors, kinases, NBS-LRR genes, chitinases, salicylic acid-, jasmonic acid-, and ethylene-signaling pathways, and plant secondary metabolism play important roles in Pi9 gene-mediated rice blast resistance.
Key words： rice blast; transcriptome; resistance genes; plant hormones; secondary metabolism
Zhaomeng XU, Lihua LI, Xiaoqing GAO, Zhengjie YUAN, Xin LI, Xudan TIAN, Lanlan WANG, Shaohong QU . Comparative Transcriptome Analysis of Transgenic Rice Line Carrying the Rice Blast Resistance Gene Pi9[J]. Chinese Journal OF Rice Science, 2020 , 34(3) : 245 -255 . DOI: 10.16819/j.1001-7216.2020.9126
| [1] | Deng Y, Zhai K, Xie Z, Yang D, Zhu X, Liu J, Wang X, Qin P, Yang Y, Zhang G, Li Q, Zhang J, Wu S, Milazzo J, Mao B, Wang E, Xie H, Tharreau D, He Z.Epigenetic regulation of antagonistic receptors confers rice blast resistance with yield balance[J]. Science, 2017, 355(6328): 962-965. |
| [2] | Jain P, Singh P K, Kapoor R, Khanna A, Solanke A U, Krishnan S G, Singh A K, Sharma V, Sharma T R.Understanding host-pathogen interactions with expression profiling of NILs carrying rice-blast resistance Pi9 gene[J]. Frontiers of Plant Sciences, 2017, (8): 93-93. |
| [3] | Liu J, Hu Y, Ning Y, Jiang N, Wu J, Jeon J, Xiao Y, Liu X, Dai N, Wang G.Genetic variation and evolution of the Pi9 blast resistance locus in the AA genome Oryza species[J]. Journal of Plant Biology, 2011, 54(5): 294-302. |
| [4] | Wu J, Kou Y, Bao J, Li Y, Tang M, Zhu X, Ponaya A, Xiao G, Li J, Li C, Song M Y, Cumagun C J, Deng Q, Lu G, Jeon J S, Naqvi N I, Zhou B.Comparative genomics identifies the Magnaporthe oryzae avirulence effector AvrPi9 that triggers Pi9-mediated blast resistance in rice[J]. New Phytologist, 2015, 206(4): 1463-1475. |
| [5] | Jones J D, Dang I J L. The plant immune system[J]. Nature, 2006, 444(7117): 323-329. |
| [6] | Liu W, Liu J, Triplett L, Leach J E, Wang G L.Novel insights into rice innate immunity against bacterial and fungal pathogens[J]. Phytopathology, 2014, 52(11): 213-241. |
| [7] | Kou Y, Wang S.Broad-spectrum and durability: Understanding of quantitative disease resistance[J]. Current Opinion in Plant Biology, 2010, 13: 181-185. |
| [8] | Wei T, Ou B, Li J, Zhao Y, Guo D, Zhu Y, Chen Z, Gu H, Li C, Qin G, Qu L J.Transcriptional profiling of rice early response to Magnaporthe oryzae identified OsWRKYs as important regulators in rice blast resistance[J]. PLoS One, 2013, 8(3): e59720. |
| [9] | Zhang Y, Zhao J, Li Y, Yuan Z, He H, Yang H, Qu H, Ma C, Qu S.Transcriptome analysis highlights defense and signaling pathways mediated by rice pi21 gene with partial resistance to Magnaporthe oryzae[J]. Frontiers of Plant Sciences, 2016, 7: 1834. |
| [10] | Li W, Liu Y, Wang J, He M, Zhou X, Yang C, Yuan C, Wang J, Chern M, Yin J, Chen W, Ma B, Wang Y, Qin P, Li S, Ronald P, Chen X.The durably resistant rice cultivar Digu activates defence gene expression before the full maturation of Magnaporthe oryzae appressorium[J]. Molecular Plant Pathology, 2015, 16: 973-986. |
| [11] | Kitony J K.水稻CO39品种及其5个近等基因系对稻瘟病感染应答的转录组分析[D]. 福州: 福建农林大学, 2016. |
| [11] | Kitony J K.Transcriptome analysis of rice cultivar CO39 and its five Near-Isogenic Lines in response to blast fungus infection[D]. Fuzhou: Fujian University of Agriculture and Forestry, 2016. (in Chinese with English abstract) |
| [12] | 王玉, 杨雪, 杨蕊菁, 王玉霞, 杨飞霞, 夏鹏飞, 赵磊. 调控苯丙烷类生物合成的MYB类转录因子研究进展[J]. 安徽农业大学学报, 2019, 46(5): 859-864. |
| [12] | Wang Y, Yang X, Yang R Q, Wang Y X, Yang F X, Xia P F, Zhao L.Advances in research of MYB transcription factors in regulating phenylpropane biosynthesis[J]. Journal of Anhui Agricultural University, 2019, 46(5): 859-864. (in Chinese with English abstract) |
| [13] | 崔慧萍, 周薇, 郭长虹. 植物过氧化物酶体在活性氧信号网络中的作用[J]. 中国生物化学与分子生物学报, 2017, 33(3): 220-226. (in Chinese with English abstract) |
| [13] | Cui H P, Zhou W, Guo C H.The role of plant peroxisomes in ROS signaling network[J]. Chinese Journal of Biochemistry and Molecular Biology, 2017, 33(3): 220-226. (in Chinese with English abstract) |
| [14] | 杜欣谊. 苯丙氨酸解氨酶的研究进展[J]. 现代化农业, 2016(7): 24-26. |
| [14] | Du X Y.Research advances on phenylalanine ammonia-lyase gene[J]. Modernizing Agriculture, 2016(7): 24-26. (in Chinese with English abstract) |
| [15] | 李魏, 谭晓风, 陈鸿鹏. 植物肉桂酰辅酶A还原酶基因的结构功能及应用潜力[J]. 经济林研究, 2009, 27(1): 7-12. |
| [15] | Li W, Tan X F, Chen H P.Structural function and application potential of cinnamoyl-CoA reductase gene in plant[J]. Non-wood Forest Research, 2009, 27(1): 7-12. (in Chinese with English abstract) |
| [16] | Kasprzewska A.Plant chitinases-regulation and function[J]. Cellular & Molecular Biology Letters, 2003, 8(3): 809-824. |
| [17] | 许明辉, 李成云, 李进斌, 谭学林, 田颖川, 陈正华, 唐祚舜, 田文忠. 转几丁质酶-葡聚糖酶基因水稻稻瘟病抗谱分析[J]. 中国水稻科学, 2003, 17(4): 307-310. |
| [17] | Xu M H, Li C Y, Li J B, Tan X L, Tian Y C, Chen Z H, Tang Z S, Tian W Z.Analysis on resistant spectrum to rice blast in transgenic rice lines with chitinase gene and β-1, 3-glucanase gene[J]. Chinese Journal of Rice Science, 2003, 17(4): 307-310. (in Chinese with English abstract) |
| [18] | Qu S, Liu G, Zhou B, Bellizzi M, Zeng L, Dai L, Han B, Wang G L.The broad-spectrum blast resistance gene Pi9 encodes a nucleotide-binding site-leucine-rich repeat protein and is a member of a multigene family in rice[J]. Genetics, 2006, 172(3): 1901-1914. |
| [19] | Liu W, Liu J, Ning Y, Ding B, Wang X, Wang Z, Wang G.Recent progress in understanding PAMP- and effector-triggered immunity against the rice blast fungus Magnaporthe oryzae[J]. Molecular Plant, 2013, 6(3): 605-620. |
| [20] | Zipfel C.Pattern-recognition receptors in plant innate immunity[J]. Current Opinion Immunology, 2008, 20(1): 10-16. |
| [21] | Ramamoorthy R, Jiang S Y, Kumar N, Venkatesh P N, Ramachandran S.A comprehensive transcriptional profiling of the WRKY gene family in rice under various abiotic and phytohormone treatment[J]. Plant Cell Physiology, 2008, 49(6): 865-879. |
| [22] | 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. |
| [23] | Pitzschke A, Schikora A, Hirt H.MAPK cascade signalling networks in plant defence[J]. Current Opinion in Plant Biology, 2009, 12(4): 421-426. |
/
| 〈 |
|
〉 |