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Bph6 encodes an exocyst-localized protein and confers broad resistance to planthoppers in rice

Nature Genetics volume 50, pages 297–306 (2018)Cite this article

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Abstract

The brown planthopper (BPH) and white-backed planthopper (WBPH) are the most destructive insect pests of rice, and they pose serious threats to rice production throughout Asia. Thus, there are urgent needs to identify resistance-conferring genes and to breed planthopper-resistant rice varieties. Here we report the map-based cloning and functional analysis of Bph6, a gene that confers resistance to planthoppers in rice. Bph6 encodes a previously uncharacterized protein that localizes to exocysts and interacts with the exocyst subunit OsEXO70E1. Bph6 expression increases exocytosis and participates in cell wall maintenance and reinforcement. A coordinated cytokinin, salicylic acid and jasmonic acid signaling pathway is activated in Bph6-carrying plants, which display broad resistance to all tested BPH biotypes and to WBPH without sacrificing yield, as these plants were found to maintain a high level of performance in a field that was heavily infested with BPH. Our results suggest that a superior resistance gene that evolved long ago in a region where planthoppers are found year round could be very valuable for controlling agricultural insect pests.

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Fig. 1: Map-based cloning of Bph6.
Fig. 2: Molecular characterization of Bph6.
Fig. 3: Demonstration that BPH6 interacts with exocyst subunit OsEXO70E1.
Fig. 4: Characterization of exocyst gene expression and cell walls in 9311-Bph6-NIL and 9311 plants.
Fig. 5: Analyses of phytohormones in Bph6-carrying plants.
Fig. 6: Characterization of Bph6-mediated resistance.
Fig. 7: Evolution of Bph6 alleles in rice.

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References

  1. Grist, D. H. & Lever, R. J. Pests of Rice (Longmans, Green and Co, London, 1969).

    Google Scholar 

  2. Cheng, X., Zhu, L. & He, G. Towards understanding of molecular interactions between rice and the brown planthopper. Mol. Plant 6, 621–634 (2013).

    Article  CAS  Google Scholar 

  3. Sogawa, K. The rice brown planthopper: feeding physiology and host plant interactions. Annu. Rev. Entomol. 27, 49–73 (1982).

    Article  CAS  Google Scholar 

  4. Sezer, M. & Butlin, R. K. The genetic basis of host plant adaptation in the brown planthopper (Nilaparvata lugens). Heredity 80, 499–508 (1998).

    Article  Google Scholar 

  5. Dyck, V. A. & Thomas, B. in Brown Planthopper: Threat to Rice Production in Asia 3–20 (International Rice Research Institute, Manila, Philippines, 1979).

  6. Catindig, J. L. A. et al. in Planthoppers: New Threats to the Sustainability of Intensive Rice Production Systems in Asia 191–220 (International Rice Research Institute, Manila, Philippines, 2009).

  7. Matsumura, M. et al. in Planthoppers: New Threats to the Sustainability of Intensive Rice Production Systems in Asia 233–244 (International Rice Research Institute, Manila, Philippines, 2009).

  8. Pathak, M. D., Cheng, C. H. & Fortuno, M. E. Resistance to Nephotettix impicticeps and Nilaparvata lugens in varieties of rice. Nature 223, 502–504 (1969).

    Article  Google Scholar 

  9. Athwal, D. S., Pathak, M. D., Bacalangco, E. & Pura, C. D. Genetics of resistance to brown planthoppers and green leaf hoppers in Oryza sativa L. Crop Sci. 11, 747–750 (1971).

    Article  Google Scholar 

  10. Ling, Y. & Weilin, Z. Genetic and biochemical mechanisms of rice resistance to planthopper. Plant Cell Rep. 35, 1559–1572 (2016).

    Article  Google Scholar 

  11. Du, B. et al. Identification and characterization of Bph14, a gene conferring resistance to brown planthopper in rice. Proc. Natl. Acad. Sci. USA 106, 22163–22168 (2009).

    Article  CAS  Google Scholar 

  12. Tamura, Y. et al. Map-based cloning and characterization of a brown planthopper resistance gene BPH26 from Oryza sativa L. ssp. indica cultivar ADR52. Sci. Rep. 4, 5872 (2014).

    Article  Google Scholar 

  13. Cheng, X. et al. A rice lectin receptor-like kinase that is involved in innate immune responses also contributes to seed germination. Plant J. 76, 687–698 (2013).

    Article  CAS  Google Scholar 

  14. Liu, Y. et al. A gene cluster encoding lectin receptor kinases confers broad-spectrum and durable insect resistance in rice. Nat. Biotechnol. 33, 301–305 (2015).

    Article  CAS  Google Scholar 

  15. Wang, Y. et al. Map-based cloning and characterization of BPH29, a B3-domain-containing recessive gene conferring brown planthopper resistance in rice. J. Exp. Bot. 66, 6035–6045 (2015).

    Article  CAS  Google Scholar 

  16. Zhao, Y. et al. Allelic diversity in an NLR gene BPH9 enables rice to combat planthopper variation. Proc. Natl. Acad. Sci. USA 113, 12850–12855 (2016).

    Article  CAS  Google Scholar 

  17. Ren, J. et al. Bph32, a novel gene encoding an unknown SCR domain-containing protein, confers resistance against the brown planthopper in rice. Sci. Rep. 6, 37645 (2016).

    Article  CAS  Google Scholar 

  18. Kabir, M. A. & Khush, G. S. Genetic analysis of resistance to brown planthopper resistance gene in rice. Euphytica 107, 23–28 (1988).

    Google Scholar 

  19. Qiu, Y., Guo, J., Jing, S., Zhu, L. & He, G. High-resolution mapping of the brown planthopper resistance gene Bph6 in rice and characterizing its resistance in the 9311 and Nipponbare near-isogenic backgrounds. Theor. Appl. Genet. 121, 1601–1611 (2010).

    Article  Google Scholar 

  20. Wang, J. et al. EXPO, an exocyst-positive organelle distinct from multivesicular endosomes and autophagosomes, mediates cytosol-to-cell-wall exocytosis in Arabidopsis and tobacco cells. Plant Cell 22, 4009–4030 (2010).

    Article  CAS  Google Scholar 

  21. Zárský, V., Kulich, I., Fendrych, M. & Pečenková, T. Exocyst complexes multiple functions in plant cells secretory pathways. Curr. Opin. Plant Biol. 16, 726–733 (2013).

    Article  Google Scholar 

  22. Fujisaki, K. et al. Rice EXO70 interacts with a fungal effector, AVR-Pii, and is required for AVR-Pii-triggered immunity. Plant J. 83, 875–887 (2015).

    Article  CAS  Google Scholar 

  23. Zhao, T. et al. A truncated NLR protein, TIR-NBS2, is required for activated defense responses in the exo70B1 mutant. PLoS Genet. 11, e1004945 (2015).

    Article  Google Scholar 

  24. Kim, S. J. & Brandizzi, F. The plant secretory pathway: an essential factory for building the plant cell wall. Plant Cell Physiol. 55, 687–693 (2014).

    Article  CAS  Google Scholar 

  25. Hao, P. et al. Herbivore-induced callose deposition on the sieve plates of rice: an important mechanism for host resistance. Plant Physiol. 146, 1810–1820 (2008).

    Article  CAS  Google Scholar 

  26. Pieterse, C. M. J., Van der Does, D., Zamioudis, C., Leon-Reyes, A. & Van Wees, S. C. M. Hormonal modulation of plant immunity. Annu. Rev. Cell Dev. Biol. 28, 489–521 (2012).

    Article  CAS  Google Scholar 

  27. McConn, M., Creelman, R. A., Bell, E., Mullet, J. E. & Browse, J. Jasmonate is essential for insect defense in Arabidopsis. Proc. Natl. Acad. Sci. USA 94, 5473–5477 (1997).

    Article  CAS  Google Scholar 

  28. Van der Does, D. et al. Salicylic acid suppresses jasmonic acid signaling downstream of SCFCOI1-JAZ by targeting GCC promoter motifs via transcription factor ORA59. Plant Cell 25, 744–761 (2013).

    Article  Google Scholar 

  29. Zhou, G. et al. Silencing OsHI-LOX makes rice more susceptible to chewing herbivores but enhances resistance to a phloem feeder. Plant J. 60, 638–648 (2009).

    Article  CAS  Google Scholar 

  30. Guo, H. M., Li, H. C., Zhou, S. R., Xue, H. W. & Miao, X. X. Cis-12-oxo-phytodienoic acid stimulates rice defense response to a piercing–sucking insect. Mol. Plant 7, 1683–1692 (2014).

    Article  CAS  Google Scholar 

  31. Howe, G. A. & Jander, G. Plant immunity to insect herbivores. Annu. Rev. Plant Biol. 59, 41–66 (2008).

    Article  CAS  Google Scholar 

  32. Naseem, M., Wölfling, M. & Dandekar, T. Cytokinins for immunity beyond growth, galls and green islands. Trends Plant Sci. 19, 481–484 (2014).

    Article  CAS  Google Scholar 

  33. Grosskinsky, D. K. et al. Cytokinins mediate resistance against Pseudomonas syringae in tobacco through increased antimicrobial phytoalexin synthesis independent of salicylic acid signaling. Plant Physiol. 157, 815–830 (2011).

    Article  CAS  Google Scholar 

  34. Jiang, C. J. et al. Cytokinins act synergistically with salicylic acid to activate defense gene expression in rice. Mol. Plant Microbe Interact. 26, 287–296 (2013).

    Article  CAS  Google Scholar 

  35. Argueso, C. T. et al. Two-component elements mediate interactions between cytokinin and salicylic acid in plant immunity. PLoS Genet. 8, e1002448 (2012).

    Article  CAS  Google Scholar 

  36. Naseem, M., Kaltdorf, M. & Dandekar, T. The nexus between growth and defense signaling: auxin and cytokinin modulate plant immune response pathways. J. Exp. Bot. 66, 4885–4896 (2015).

    Article  CAS  Google Scholar 

  37. Hart, S. V., Kogan, M. & Paxton, J. D. Effect of soybean phytoalexins on the herbivorous insects mexican bean beetle and soybean looper. J. Chem. Ecol. 9, 657–672 (1983).

    Article  CAS  Google Scholar 

  38. Yamane, H. Biosynthesis of phytoalexins and regulatory mechanisms of it in rice. Biosci. Biotechnol. Biochem. 77, 1141–1148 (2013).

    Article  CAS  Google Scholar 

  39. Ahuja, I., Kissen, R. & Bones, A. M. Phytoalexins in defense against pathogens. Trends Plant Sci. 17, 73–90 (2012).

    Article  CAS  Google Scholar 

  40. Miyamoto, K. et al. Overexpression of the bZIP transcription factor OsbZIP79 suppresses the production of diterpenoid phytoalexin in rice cells. J. Plant Physiol. 173, 19–27 (2015).

    Article  CAS  Google Scholar 

  41. Painter, R. H. in Insect Resistance in Crop Plants 23–83 (Macmillan, New York, 1951).

  42. Sang, T. & Ge, S. Understanding rice domestication and implications for cultivar improvement. Curr. Opin. Plant Biol. 16, 139–146 (2013).

    Article  Google Scholar 

  43. Fisher, M. C. et al. Emerging fungal threats to animal, plant and ecosystem health. Nature 484, 186–194 (2012).

    Article  CAS  Google Scholar 

  44. Huang, X. et al. A map of rice genome variation reveals the origin of cultivated rice. Nature 490, 497–501 (2012).

    Article  CAS  Google Scholar 

  45. Xu, X. et al. Resequencing 50 accessions of cultivated and wild rice yields markers for identifying agronomically important genes. Nat. Biotechnol. 30, 105–111 (2011).

    Article  CAS  Google Scholar 

  46. Agrawal, A. A., Hastings, A. P., Johnson, M. T., Maron, J. L. & Salminen, J. P. Insect herbivores drive real-time ecological and evolutionary change in plant populations. Science 338, 113–116 (2012).

    Article  CAS  Google Scholar 

  47. Yan, W. et al. Natural variation in Ghd7.1 plays an important role in grain yield and adaptation in rice. Cell Res. 23, 969–971 (2013).

    Article  CAS  Google Scholar 

  48. Qiu, Y. F. et al. Identification of antibiosis and tolerance in rice varieties carrying brown planthopper resistance genes. Entomol. Exp. Appl. 141, 224–231 (2011).

    Article  CAS  Google Scholar 

  49. Jing, S. et al. Development and use of EST-SSR markers for assessing genetic diversity in the brown planthopper (Nilaparvata lugens Stål). Bull. Entomol. Res. 102, 113–122 (2012).

    Article  CAS  Google Scholar 

  50. Huang, Z., He, G. C., Shu, L. H., Li, X. H. & Zhang, Q. F. Identification and mapping of two brown planthopper resistance genes in rice. Theor. Appl. Genet. 102, 929–934 (2001).

    Article  CAS  Google Scholar 

  51. Heinrichs, E., Medrano, F. & Rapusas, H. in Genetic Evaluation of Insect Resistance in Rice (International Rice Research Institute, Manila, Philippines, 1985).

  52. Yang, Z., Chen, H., Tang, W., Hua, H. & Lin, Y. Development and characterzation of transgenic rice expressing two Bacillus thuringiensis genes. Pest Manag. Sci. 67, 414–422 (2011).

    Article  CAS  Google Scholar 

  53. Walter, M. et al. Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation. Plant J. 40, 428–438 (2004).

    Article  CAS  Google Scholar 

  54. Chen, S., Songkumarn, P., Liu, J. & Wang, G. L. A versatile zero background T-vector system for gene cloning and functional genomics. Plant Physiol. 150, 1111–1121 (2009).

    Article  CAS  Google Scholar 

  55. Zhang, Y. et al. A highly efficient rice green tissue protoplast system for transient gene expression and studying light/- and chloroplast-related processes. Plant Methods 7, 30 (2011).

    Article  CAS  Google Scholar 

  56. Nelson, B. K., Cai, X. & Nebenführ, A. A multicolored set of in vivo organelle markers for colocalization studies in Arabidopsis and other plants. Plant J. 51, 1126–1136 (2007).

    Article  CAS  Google Scholar 

  57. Weng, Q. M., Huang, Z., Wang, X. L., Zhu, L. L. & He, G. C. In situ localization of proteinase inhibitor mRNA in rice plant challenged with brown planthopper. Chin. Sci. Bull. 48, 979–982 (2003).

    Article  CAS  Google Scholar 

  58. Pettolino, F. A., Walsh, C., Fincher, G. B. & Bacic, A. Determining the polysaccharide composition of plant cell walls. Nat. Protoc. 7, 1590–1607 (2012).

    Article  CAS  Google Scholar 

  59. Dubois, M., Gilles, K. A., Hamilton, J. K., Robers, P. A. & Smith, F. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28, 350–356 (1956).

    Article  CAS  Google Scholar 

  60. Updegraff, D. M. Semimicro determination of cellulose in biological materials. Anal. Biochem. 32, 420–424 (1969).

    Article  CAS  Google Scholar 

  61. Smyth, G. K. Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. Stat. Appl. Genet. Mol. Biol. 3, e3 (2004).

    Article  Google Scholar 

  62. Liu, C. et al. Revealing different systems responses to brown planthopper infestation for pest-susceptible and resistant rice plants with the combined metabonomic and gene expression analysis. J. Proteome Res. 9, 6774–6785 (2010).

    Article  CAS  Google Scholar 

  63. Liu, H., Li, X., Xiao, J. & Wang, S. A convenient method for simultaneous quantification of multiple phytohormones and metabolites: application in study of rice–bacterium interaction. Plant Methods 8, 2 (2012).

    Article  CAS  Google Scholar 

  64. Yuan, H. M., Liu, W. C. & Lu, Y. T. CATALASE2 coordinates SA-mediated repression of both auxin accumulation and JA biosynthesis in plant defenses. Cell Host Microbe 21, 143–155 (2017).

    Article  CAS  Google Scholar 

  65. Librado, P. & Rozas, J. DnaSPv5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25, 1451–1452 (2009).

    Article  CAS  Google Scholar 

  66. Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol. 30, 2725–2729 (2013).

    Article  CAS  Google Scholar 

  67. You, A. et al. Identification of quantitative trait loci across recombinant inbred lines and testcross populations for traits of agronomic importance in rice. Genetics 172, 1287–1300 (2006).

    Article  Google Scholar 

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Acknowledgements

We thank Q. Qian (China National Rice Research Institute), L. Han (Chinese Academy of Agricultural Sciences), L. Yan (Jiangxi Academy of Agricultural Sciences) and D. Pan (Guangdong Academy of Agricultural Sciences) for kindly providing rice germplasm, S. Wang (Huazhong Agricultural University) for kindly providing the rice disease pathogen PXO145, Y. Lin (Huazhong Agricultural University) for kindly providing the striped stem borer insects, D. Zeng (China National Rice Research Institute) for kindly providing the WBPH insects, Y. Liu for suggestions for the experiments, and Q. Zhang, R. He and J. Blackwell for edits and suggestions. This work was supported by grants from the National Natural Science Foundation of China (31230060 and 31630063, both to G.H.), the National Program on Research and Development of Transgenic Plants (2016ZX08009-003-001 to G.H.) and the National Key Research and Development Program (2016YFD0100600 and 2016YFD0100900, both to G.H.).

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  1. Jianping Guo, Chunxue Xu and Di Wu contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Hybrid Rice, College of Life Sciences, Wuhan University, Wuhan, China

    Jianping Guo, Chunxue Xu, Di Wu, Yan Zhao, Yongfu Qiu, Xiaoxiao Wang, Shengli Jing, Xinxin Shangguan, Huiying Wang, Yinhua Ma, Liang Hu, Yan Wu, Shaojie Shi, Wenliang Wang, Lili Zhu, Rongzhi Chen, Bo Du & Guangcun He

  2. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China

    Yidan Ouyang

  3. Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, China

    Baodong Cai & Yuqi Feng

  4. BGI-Shenzhen, Shenzhen, China

    Xin Liu & Xun Xu

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  1. Jianping Guo
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Contributions

G.H. conceived and supervised the project; G.H. and J.G. designed the experiments; J.G. performed most of the experiments; C.X., Y.Z., D.W., B.D., X.W., Y.O., X.L., W.W., Y.Q., S.J., B.C., X.S., H.W., Y.M., Y.W., L.H., S.S., L.Z., X.X., R.C. and Y.F. performed some of the experiments; and J.G., C.X., D.W., B.D., Y.Z. and G.H. analyzed data and wrote the manuscript.

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Correspondence to Bo Du or Guangcun He.

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Supplementary Text and Figures

Supplementary Figures 1–27 and Supplementary Tables 1, 2, 4, 5, 7, 10 and 11

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Supplementary Table 3

GeneChip data for comparing Nip-Bph6-NIL and Bph6-RNAi plants

Supplementary Table 6

Information on accessions of wild rice and cultivated varieties that were sequenced in the Bph6 coding region

Supplementary Table 8

List of SNPs between Swarnalata or Nipponbare and the 80 haplotypes

Supplementary Table 9

List of indels between Swarnalata or Nipponbare and the 80 haplotypes

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Guo, J., Xu, C., Wu, D. et al. Bph6 encodes an exocyst-localized protein and confers broad resistance to planthoppers in rice. Nat Genet 50, 297–306 (2018). https://doi.org/10.1038/s41588-018-0039-6

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