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CRISPR/Cas9-Mediated Gene Editing of the Jasmonate Biosynthesis OsAOC Gene in Rice

  • Trang Hieu Nguyen
  • Huong To Thi Mai
  • Daniel Moukouanga
  • Michel Lebrun
  • Stephane Bellafiore
  • Antony ChampionEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2085)

Abstract

The function of Jasmonate (JA) is well documented in different plant physiological processes as well as in the interactions with their environment. Mutants impaired in JA production and/or signaling are useful materials to study the function of this phytohormone. Genes involved in the JA biosynthesis pathway in rice have been described, but few mutants deficient in JA production and signaling have been identified. Moreover, these mutants are mostly generated through random mutagenesis approaches, such as irradiation, EMS treatment, or T-DNA insertion, and potentially harbor undesired mutations that could affect other biological processes. The CRISPR/Cas9 system is a precise and efficient genome editing tool that creates DNA modification at specific loci and limit undesired mutations.

In this chapter, we describe a procedure to generate new JA-deficient mutant using CRISPR/Cas9 system in rice. The Allene Oxide Cyclase (OsAOC) gene is targeted since it is a single copy gene in the JA biosynthesis pathway in rice. The widely used variety Oryza sativa japonica Kitaake has been chosen due to its short life cycle and its ease of genetic transformation. This protocol describes the selection of the 20-nt target sequence, construction of the binary vector, and strategy for selecting the T-DNA-free mutant.

Key words

Jasmonate Mutant Rice CRISPR/Cas9 

Supplementary material

464998_2_En_15_MOESM1_ESM.docx (13 kb)
Data 1 (DOCX 13 kb)

References

  1. 1.
    Riemann M, Haga K, Shimizu T, Okada K, Ando S, Mochizuki S, Nishizawa Y, Yamanouchi U, Nick P, Yano M et al (2013) Identification of rice Allene oxide Cyclase mutants and the function of jasmonate for defence against Magnaporthe oryzae. Plant J 74:226–238CrossRefGoogle Scholar
  2. 2.
    Cai Q, Yuan Z, Chen M, Yin C, Luo Z, Zhao X, Liang W, Hu J, Zhang D (2014) Jasmonic acid regulates spikelet development in rice. Nat Commun 5:3476CrossRefGoogle Scholar
  3. 3.
    Liao L, Shi CH, Zeng DD, Jin XL, Wu JG (2015) Morphogenesis and molecular basis on the unclosed glumes, a novel mutation related to the floral organ of rice. Plant Mol Biol Rep 33:480–489CrossRefGoogle Scholar
  4. 4.
    Fang C, Zhang H, Wan J, Wu Y, Li K, Jin C, Chen W, Wang S, Wang W, Zhang H et al (2016) Control of leaf senescence by an MeOH-jasmonates cascade that is epigenetically regulated by OsSRT1 in rice. Mol Plant 9:1366–1378CrossRefGoogle Scholar
  5. 5.
    Svyatyna K, Riemann M (2012) Light-dependent regulation of the jasmonate pathway. Protoplasma 249(Suppl 2):S137–S145CrossRefGoogle Scholar
  6. 6.
    Yamada S, Kano A, Tamaoki D, Miyamoto A, Shishido H, Miyoshi S, Taniguchi S, Akimitsu K, Gomi K (2012) Involvement of OsJAZ8 in jasmonate-induced resistance to bacterial blight in rice. Plant Cell Physiol 53:2060–2072CrossRefGoogle Scholar
  7. 7.
    Nahar K, Kyndt T, De Vleesschauwer D, Höfte M, Gheysen G (2011) The jasmonate pathway is a key player in systemically induced defense against root knot nematodes in rice. Plant Physiol 157:305–316CrossRefGoogle Scholar
  8. 8.
    Zhou G, Qi J, Ren N, Cheng J, Erb M, Mao B, Lou Y (2009) Silencing OsHI-LOX makes rice more susceptible to chewing herbivores, but enhances resistance to a phloem feeder. Plant J 60:638–648CrossRefGoogle Scholar
  9. 9.
    Li R, Afsheen S, Xin Z, Han X, Lou Y (2013) OsNPR1 negatively regulates herbivore-induced JA and ethylene signaling and plant resistance to a chewing herbivore in rice. Physiol Plant 147:340–351CrossRefGoogle Scholar
  10. 10.
    Kobayashi T, Itai RN, Senoura T, Oikawa T, Ishimaru Y, Ueda M, Nakanishi H, Nishizawa NK (2016) Jasmonate signaling is activated in the very early stages of iron deficiency responses in rice roots. Plant Mol Biol 91:533–547CrossRefGoogle Scholar
  11. 11.
    Khan GA, Vogiatzaki E, Glauser G, Poirier Y (2016) Phosphate deficiency induces the jasmonate pathway and enhances resistance to insect herbivory. Plant Physiol 171:632–644CrossRefGoogle Scholar
  12. 12.
    Ismail A, Seo M, Takebayashi Y, Kamiya Y, Eiche E, Nick P (2014) Salt adaptation requires efficient fine-tuning of jasmonate signalling. Protoplasma 251:881–898CrossRefGoogle Scholar
  13. 13.
    Fu J, Wu H, Ma S, Xiang D, Liu R, Xiong L (2017) OsJAZ1 attenuates drought resistance by regulating JA and ABA signaling in rice. Front Plant Sci 8:2108CrossRefGoogle Scholar
  14. 14.
    Du H, Liu H, Xiong L (2013) Endogenous auxin and jasmonic acid levels are differentially modulated by abiotic stresses in rice. Front Plant Sci 4:397CrossRefGoogle Scholar
  15. 15.
    Wasternack C, Song S (2017) Jasmonates: biosynthesis, metabolism, and signaling by proteins activating and repressing transcription. J Exp Bot 68:1303–1321Google Scholar
  16. 16.
    Dhakarey R, Kodackattumannil Peethambaran P, Riemann M (2016) Functional analysis of jasmonates in rice through mutant approaches. Plants (Basel) 5Google Scholar
  17. 17.
    Biswas KK, Neumann R, Haga K, Yatoh O, Iino M (2003) Photomorphogenesis of rice seedlings: a mutant impaired in phytochrome-mediated inhibition of coleoptile growth. Plant Cell Physiol 44:242–254CrossRefGoogle Scholar
  18. 18.
    Li X, Wang Y, Duan E, Qi Q, Zhou K, Lin Q, Wang D, Wang Y, Long W, Zhao Z et al (2018) Open Glume1: a key enzyme reducing the precursor of JA, participates in carbohydrate transport of lodicules during anthesis in rice. Plant Cell Rep 37:329–346CrossRefGoogle Scholar
  19. 19.
    Xiao Y, Chen Y, Charnikhova T, Mulder PPJ, Heijmans J, Hoogenboom A, Agalou A, Michel C, Morel J-B, Dreni L et al (2014) OsJAR1 is required for JA-regulated floret opening and anther dehiscence in rice. Plant Mol Biol 86:19–33CrossRefGoogle Scholar
  20. 20.
    Lee S-H, Sakuraba Y, Lee T, Kim K-W, An G, Lee HY, Paek N-C (2015) Mutation of Oryza sativa Coronatine insensitive 1b (OsCOI1b) delays leaf senescence. J Integr Plant Biol 57:562–576CrossRefGoogle Scholar
  21. 21.
    Puchta H (2017) Applying CRISPR/Cas for genome engineering in plants: the best is yet to come. Curr Opin Plant Biol 36:1–8CrossRefGoogle Scholar
  22. 22.
    Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F (2013) Genome engineering using the CRISPR-Cas9 system. Nat Protoc 8:2281–2308CrossRefGoogle Scholar
  23. 23.
    Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337:816–821CrossRefGoogle Scholar
  24. 24.
    Miao J, Guo D, Zhang J, Huang Q, Qin G, Zhang X, Wan J, Gu H, Qu L-J (2013) Targeted mutagenesis in rice using CRISPR-Cas system. Cell Res 23:1233–1236CrossRefGoogle Scholar
  25. 25.
    Shan Q, Wang Y, Li J, Zhang Y, Chen K, Liang Z, Zhang K, Liu J, Xi JJ, Qiu J-L et al (2013) Targeted genome modification of crop plants using a CRISPR-Cas system. Nat Biotechnol 31:686–688CrossRefGoogle Scholar
  26. 26.
    Dehairs J, Talebi A, Cherifi Y, Swinnen JV (2016) CRISP-ID: decoding CRISPR mediated indels by sanger sequencing. Sci Rep 6Google Scholar
  27. 27.
    Hiei Y, Komari T (2008) Agrobacterium-mediated transformation of rice using immature embryos or calli induced from mature seed. Nat Protoc 3:824–834CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Trang Hieu Nguyen
    • 1
  • Huong To Thi Mai
    • 2
  • Daniel Moukouanga
    • 1
  • Michel Lebrun
    • 1
    • 2
    • 3
  • Stephane Bellafiore
    • 4
  • Antony Champion
    • 5
    Email author
  1. 1.Institut de Recherche pour le Developpement (IRD), CiradUniversite fe Montpellier, DIADEMontpellierFrance
  2. 2.Vietnam Academy of Science and Technology (VAST), LMI-RICE2University of Science and Technology of Hanoi (USTH)HanoiVietnam
  3. 3.IRD, CiradUniv Montpellier, LSTMMontpellierFrance
  4. 4.IRD, CiradUniv Montpellier, IPMEMontpellierFrance
  5. 5.UMR DIADE - IRDMontpellierFrance

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