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Different Reactions of Rice Monogenic Line IRBL9-W Harboring Pi9 Gene to Magnaporthe oryzae Containing AvrPi9 During Seedling and Adult-plant Stages
#These authors contributed equally to this work;
Received date: 2020-07-24
Revised date: 2020-11-24
Online published: 2021-05-10
【Objective】 The Pi9 gene confers broad-spectrum resistance against Magnaporthe oryzae. Rice monogenic line IRBL9-W carrying single blast resistance gene Pi9 is highly resistant to leaf blast during the seedling stage, but it is susceptible to neck blast at adult-plant stage in blast nursery. It is important to analyze the mechanism behind the reversion of resistance to Magnaporthe oryzae, which will provide important information for effective utilization of Pi9 in disease-resistant rice breeding program.【Method】Eight single spore strains of M. oryzae isolated from neck blast samples of diseased IRBL9-W, and one isolate Y363 were artificially inoculated by spraying at seedling stage to pathotype their pathogenicity on 24 monogenic lines including IRBL9-W. Two out of eight strains and control strain Y363 were further selected and inoculated on IRBL9-W at late-booting stage by injecting to evaluate their pathogenicity. The alleles of M. oryzae AvrPi9 gene cognate to rice blast resistance gene Pi9 were amplified and sequenced with AvrPi9-specific primers in nine strains. Total RNA of the rice monogenic line IRBL9-W in seedling and late-booting stage was extracted from young leaf and panicle, respectively. The expression analysis of Pi9 gene was performed by semi-quantitative RT-PCR and real-time qRT-PCR. 【Result】 The monogenic line IRBL9-W was completely resistant to the nine M. oryzae strains at the seedling stage exposed to the artificial spraying. However, IRBL9-W was susceptible to panicle blast when inoculated with both two randomly selected strains (YX2-7-1 and YX2-15-1) isolated from panicle blast lesions of IRBL9-W, and the control strain Y363 by injecting inoculation at the late-booting stage. Compared with AvrPi9, allelic sequence amplified from Y363 was identical to cloned AvrPi9, and the coding region of the other eight strains isolated from IRBL9-W panicle blast was identical to AvrPi9 gene, except for a 16-bp deletion at -264 bp upstream of the coding starting position in the eight strains. Given high resistance of IRBL9-W to these strains at the seedling stage, we contended that this 16-bp deletion did not affect the function of AvrPi9 gene; qRT-PCR results showed that the relative expression level of Pi9 gene in panicle was only 47.3% as compared to that in seedling leaves. 【Conclusion】Compared with the relative expression level of Pi9 gene at the seedling stage in IRBL9-W, the significant decrease of Pi9 gene expression level in panicle of IRBL9-W could be the cause of susceptibility to M. oryzae at late-booting stage.
Key words： Oryza sativa L.; Magnaporthe oryzae; Pi9; seedling stage; adult-plant stage
Shufang LIU, Liying DONG, Xundong LI, Wumin ZHOU, Qinzhong YANG . Different Reactions of Rice Monogenic Line IRBL9-W Harboring Pi9 Gene to Magnaporthe oryzae Containing AvrPi9 During Seedling and Adult-plant Stages[J]. Chinese Journal OF Rice Science, 2021 , 35(3) : 303 -310 . DOI: 10.16819/j.1001-7216.2021.0717
| [1] | Couch B C, Kohn L M.A multilocus gene genealogy concordant with host preference indicates segregation of a new species, Magnaporthe oryzae, from M. grisea[J]. Mycologia, 2002, 94(4): 683-693. |
| [2] | Wilson R A, Talbot N J.Under pressure: Investigating the biology of plant infection by Magnaporthe oryzae[J]. Nature Reviews Microbiology, 2009, 7: 185-195. |
| [3] | Savary S, Willocquet L, Pethybridge S J, Esker P, McRoberts N, Nelson A. The global burden of pathogens and pests on major food crops[J]. Nature Ecology & Evolution, 2019, 3(3): 430-439. |
| [4] | Sesma A, Osbourn A E.The rice leaf blast pathogen undergoes developmental processes typical of root-infecting fungi.Nature, 2004, 431(7008): 582-586. |
| [5] | Skamnioti P, Gurr S J.Against the grain: Safeguarding rice from rice blast disease[J]. Trends in Biotechnology, 2008, 27(3): 141-150. |
| [6] | Dean R, van Kan J A L, Pretorius Z A, Hammond-kosack K E, Pietro A D, Spanu P D, Rudd J J, Dickman M, Kahmann R, Ellis J, Foster G D. The top 10 fungal pathogens in molecular plant pathology[J]. Molecular Plant Pathology, 2012, 13(4): 414-430. |
| [7] | Puri K D, Shreshta S M, Chhetri G B K, Joshi K D. Leaf and neck blast resistance reaction in tropical rice lines under greenhouse condition[J]. Euphytica, 2009, 165(3): 523-532. |
| [8] | Flor H H.Current status of the gene-for-gene concept[J]. Annual Review of Phytopathology, 1971, 9(1): 275-296. |
| [9] | Silué D, Notteghem J L, Tharreau D.Evidence of a gene-for-gene relationship in the Oryza sativa- Magnaporthe grisea pathosystem[J]. Phytopathology, 1992, 82: 577-580. |
| [10] | Jia Y, McAdams S A, Bryan G T, Hershey H P, Valent B. Direct interaction of resistance gene and avirulence gene products confers rice blast resistance[J]. The EMBO Journal, 2000, 19: 4004-4014. PMID: 10921881. |
| [11] | Ebbole D J.Magnaporthe as a model for understanding host-pathogen interactions[J]. Annual Review of Phytopathology, 2007, 45(1): 437-456. |
| [12] | Kalia S, Rathour R.Current status on mapping of genes for resistance to leaf and neck-blast disease in rice[J]. 3 Biotech, 2019, 9: 209. |
| [13] | Zhuang J Y, Ma W B, Wu J L, Chai R Y, Lu J, Fan Y Y, Jin M Z, Leung H, Zheng K L.Mapping of leaf and neck blast resistance genes with resistance gene analog, RAPD and RFLP in rice[J]. Euphytica , 2002, 128: 363-370. |
| [14] | Ma J, Lei C, Xu X, Hao K, Wang J, Cheng Z, Wan J.Pi64, encoding a novel CC-NBS-LRR protein, confers resistance to leaf and neck blast in rice[J]. Molecular Plant-Microbe Interactions, 2015, 28(5): 558-568. |
| [15] | Fujii K, Hayano-Saito Y, Saito K, Sugiura N, Hayashi N, Tsuji T, Izawa T, Iwasaki M.Identification of a RFLP marker tightly linked to the panicle blast resistance gene, Pb1, in rice[J]. Breeding Science, 2000, 50: 183-188. |
| [16] | Chen J, Shi Y F, Liu W Z, Chai R Y, Fu Y P, Zhuang J Y, Wu J L.A Pid3 allele from rice cultivar Gumei 2 confers resistance to Magnaporthe oryzae[J]. Journal of Genetics and Genomics, 2011, 38: 209-216. |
| [17] | Hayashi N, Inoue H, Kato T, Funao T, Shirota M, Shimizu T, Kanamori H, Yamane H, Hayano-Saito Y, Matsumoto T, Yano M, Takatsuji H.Durable panicle blast-resistance gene Pb1 encodes an atypical CC-NBS-LRR protein and was generated by acquiring a promoter through local genome duplication[J]. The Plant Journal, 2010, 64(3): 498-510. |
| [18] | Fang N, Wei X, Shen L, Yu Y, Li M, Yin C, He W, Guan C, Chen H, Zhang H, Bao Y.Fine mapping of a panicle blast resistance gene Pb-bd1 in japonica landrace Bodao and its application in rice breeding[J]. Rice, 2019, 12(1): 18. |
| [19] | Wang R, Fang N, Guan C, He W, Bao Y, Zhang H.Characterization and fine mapping of a blast resistant gene Pi-jnw1 from the japonica rice landrace Jiangnanwan[J]. PLOS ONE,2016, 11(12): e0169417. doi: 10.1371/journal.pone.0169417. |
| [20] | 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. |
| [21] | Liu G, Lu G, Zeng L, Wang G L.Two broad-spectrum blast resistance genes, Pi9(t) and Pi2(t), are physically linked on rice chromosome 6[J]. Molecular Genetic and Genomics, 2002, 267(4): 472-480. |
| [22] | 雷财林, 张国民, 程治军, 马军滔, 王久林, 辛爱华, 陈平, 肖家雷, 张欣, 刘迎雪, 郭秀平, 王洁, 翟虎渠, 万建民. 黑龙江省稻瘟病菌生理小种毒力基因分析与抗病育种策略[J]. 作物学报, 2011, 37(1): 18-27. |
| [22] | Lei C L, Zhang G M, Cheng Z J, Ma J T, Wang J L, Xin A H, Chen P, Xiao J L, Zhang X, Liu Y X, Guo X P, Wang J, Zhai H Q, Wan J M.Pathogenic races and virulence gene structure of Magnaporthe oryzae population and rice breeding strategy for blast resistance in Heilongjiang Province[J]. Acta Agronomica Sinica, 2011, 37(1): 18-27. (in Chinese with English abstract) |
| [23] | 董丽英, 王群, 刘树芳, 郑凤萍, 李迅东, 杨勤忠. 云南省稻瘟病菌群体对稻瘟病抗性单基因系的致病性分析[J]. 西南农业学报, 2012, 25(2): 467-473. |
| [23] | Dong L, Wang Q, Liu S, Zheng F, Li X, Yang Q.Pathogenicity analysis of Magnaporthe oryzae populations of Yunnan on monogenic lines for resistance to rice blast[J]. Southwest China Journal of Agricultural Sciences, 2012, 25(2): 467-473. (in Chinese with English abstract) |
| [24] | 汪文娟, 苏菁, 杨健源, 韦小燕, 陈凯玲, 陈珍, 陈深, 朱小源. 源于广8A 杂交稻组合的稻瘟病菌无毒基因型分析[J]. 中国农业科学, 2018, 51(24): 4633-4646. |
| [24] | Wang W J, Su Q, Yang J Y, Wei X Y, Chen K L, Chen Z, Chen S, Zhu X Y.Analysis of Magnaporthe oryzae avirulent genes in the infected hybrid rice combinations derived from a sterile line of Guang 8A[J]. Scientia Agricultura Sinica, 2018, 51(24): 4633-4646. (in Chinese with English abstract) |
| [25] | Wang J C, Jia Y, Wen J W, Liu W P, Liu X M, Li L, Jiang Z Y, Zhang J H, Guo X L, Ren J P.Identification of rice blast resistance genes using international monogenic differentials[J]. Crop Protection, 2013, 45: 109-116. |
| [26] | 刘水芳, 杨秀荣, 孙淑琴, 刘春艳, 王勇, 张春祥, 顾红艳. 水稻品种抗稻瘟病鉴定技术[J]. 天津农业科学, 2007, 13(4): 55-58. |
| [26] | Liu S, Yang X, Sun S, Liu C, Wang Y, Zhang C, Gu H.Identification technique of rice resistance to Magnaporthe grisea[J]. Tianjin Agricultural Sciences, 2007, 13(4): 55-58. (in Chinese) |
| [27] | Wu J, Kou Y J, Bao J D, Li Y, Tang M Z, Zhu X L, Ponaya A, Xiao G, Li J B, Li C Y, Song M Y, Cumagun C J R, Deng Q Y, Lu G D, Jeon J S, Naqvi N, Zhou B. Comparative genomics identifies the Magnaporthe oryzae avirulence effector Avr-Pi9 that triggers Pi9-mediated blast resistance in rice[J]. New Phytologist, 2015, 206(4): 1463-1475. |
| [28] | Wu Y Y, Yu L, Pan C H, Dai Z Y, Li Y H, Xiao N, Zhang X X, Ji H J, Huang N S, Zhao B H, Zhou C H, Liu G Q, Liu X J, Pan X B, Liang C Z, Li A H.Development of near-isogenic lines with different alleles of Piz locus and analysis of their breeding effect under Yangdao 6 background[J]. Molecular Breeding, 2016, 36: 12. |
| [29] | Wu Y Y, Chen Y, Pan C H, Xiao N, Yu L, Li Y H, Zhang X X, Pan X B, Chen X J, Dai Z Y, Li A H.Development and evaluation of near-isogenic lines with different blast resistance alleles at the Piz locus in japonica rice from the lower region of the Yangtze River, China[J]. Plant Disease, 2017, 101: 1283-1291. |
| [30] | Collard B C Y, Mackill D J. Marker-assisted selection: An approach for precision plant breeding in the twenty-first century.Philosophical Transactions of the Royal Society of London: Series B, 2008, 363: 557-572. |
| [31] | 倪大虎, 易成新, 李莉, 汪秀峰, 王文相, 杨剑波. 利用分子标记辅助选择聚合水稻基因Xa21和Pi9(t)[J]. 分子植物育种, 2005, 3(3): 329-334. |
| [31] | Ni D H, Yi C X, Li L, Wang X F, Wang W X, Yang J B.Pyramiding Xa21 and Pi9(t) in rice by marker-assisted selection[J]. Molecular Plant Breeding, 2005, 3(3): 329-334. (in Chinese with English abstract) |
| [32] | 殷得所, 夏明元, 李进波, 万丙良, 査中萍, 杜雪树, 戚华雄. 抗稻瘟病基因Pi9的STS连锁标记开发及在分子标记辅助育种中的应用[J]. 中国水稻科学, 2011, 25(1): 25-30. |
| [32] | Yin D S, Xia M Y, Li J B, Wan B L, Zha Z P, Du X S, Qi H X.Development of STS marker linked to rice blast resistance gene Pi9 in marker-assisted selection breeding[J]. Chinese Journal of Rice Science, 2011, 25(1): 25-30. (in Chinese with English abstract) |
| [33] | 陈建民, 付志英, 权宝权, 田大刚, 李刚, 王锋. 分子标记辅助培育双抗稻瘟病和白叶枯病杂交稻恢复系[J]. 分子植物育种, 2009, 7(3): 465-470. |
| [33] | Chen J M, Fu Z Y, Quan B Q, Tian D G, Li G, Wang F.Breeding hybrid rice restoring line with double resistance to rice blast and bacterial blight by marker-assisted selection[J]. Molecular Plant Breeding, 2009, 7(3): 465-470. (in Chinese with English abstract) |
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