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Isolation, Purification, and Detection of Micro RNAs in Plant Senescence

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Plant Senescence

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1744))

Abstract

Micro RNAs (miRNAs) are small noncoding RNA molecules that function in transcriptional level to regulate gene expression both in plants and animals. Increasing researches have shown that miRNAs are key regulators in plant development and stress responses, and emerging evidence indicates the potential role of miRNAs on plant senescence. In this chapter we summarize the daily methods used for identification and study of miRNAs in plants, including the isolation of total RNA, the purification of miRNAs, and the methods used to detect miRNAs in plants. The committed steps or modifications of these methods used in plant senescence research are noted.

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References

  1. Lim PO, Kim HJ, Nam HG (2007) Leaf senescence. Annu Rev Plant Biol 58:115–136

    Article  CAS  PubMed  Google Scholar 

  2. Gregersen PL, Culetic A, Boschian L et al (2013) Plant senescence and crop productivity. Plant Mol Biol 82:603–622

    Article  CAS  PubMed  Google Scholar 

  3. Balazadeh S, Riaño-Pachón DM, Mueller-Roeber B (2008) Transcription factors regulating leaf senescence in Arabidopsis thaliana. Plant Biol 10:63–75

    Article  PubMed  Google Scholar 

  4. Humbeck K (2013) Epigenetic and small RNA regulation of senescence. Plant Mol Biol 82:529–537

    Article  CAS  PubMed  Google Scholar 

  5. Carthew RW, Sontheimer EJ (2009) Origins and mechanisms of miRNAs and siRNAs. Cell 136:642–655

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Vionnet O (2009) Origin, biogenesis, and activity of plant microRNAs. Cell 136:669–687

    Article  Google Scholar 

  7. Carrington JC, Ambros V (2003) Role of microRNAs in plant and animal development. Science 301:336–338

    Article  CAS  PubMed  Google Scholar 

  8. Jones-Rhoades MW, Bartel DP, Bartel B (2006) MicroRNAs and their regulatory roles in plants. Annu Rev Plant Biol 57:19–53

    Article  CAS  PubMed  Google Scholar 

  9. Gandikota M, Birkenbihl RP, Höhmann S et al (2007) The miRNA156/157 recognition element in the 3’UTR of the Arabidopsis SBP box gene SPL3 prevents early flowering by translational inhibition in seedlings. Plant J 49:683–693

    Article  CAS  PubMed  Google Scholar 

  10. Palatnik JF, Allen E, Wu X et al (2003) Control of leaf morphogenesis by microRNAs. Nature 425:257–263

    Article  CAS  PubMed  Google Scholar 

  11. Aukerman MJ, Sakai H (2003) Regulation of flowering time and floral organ identity by a microRNA and its APETALA2-like target genes. Plant Cell 15:2730–2741

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Sunkar R, Zhu JK (2004) Novel and stress-regulated microRNAs and other small RNAs from Arabidopsis. Plant Cell 16:2001–2019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Kim JH, Woo HR, Kim J et al (2009) Trifurcate feed-forward regulation of age-dependent cell death involving miR164 in Arabidopsis. Science 323:1053–1057

    Article  CAS  PubMed  Google Scholar 

  14. Schommer C, Palatnik JF, Aggarwal P et al (2008) Control of jasmonate biosynthesis and senescence by miR319 targets. PLoS Biol 6:1991–2001

    Article  CAS  Google Scholar 

  15. Adenot X, Elmayan T, Lauressergues D, Boutet S, Bouche N, Gasciolli V, Vaucheret H (2006) DRB4-dependent TAS3 trans-acting siRNAs control leaf morphology through AGO7. Curr Biol 16:927–932

    Article  CAS  PubMed  Google Scholar 

  16. Fahlgren N, Montgomery TA, Howell MD et al (2006) Regulation of AUXIN RESPONSE FACTOR3 by TAS3 ta-siRNA affects developmental timing and patterning in Arabidopsis. Curr Biol 16:939–944

    Article  CAS  PubMed  Google Scholar 

  17. Chomczynski P, Sacchi N (2006) The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol chloroform extraction: twenty-something years on. Nat Protoc 1:581–585

    Article  CAS  PubMed  Google Scholar 

  18. Rosas-Cardenas FD, Duran-Figueroa N, Vielle-Calzada JP et al (2011) A simple and efficient method for isolating small RNAs from different plant species. Plant Methods 7:4

    Article  CAS  PubMed Central  Google Scholar 

  19. Verwoerd TC, Dekker BM, Hoekema A (1989) A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 17:2362

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. An Z, Li Y, Lili X et al (2013) A rapid and economical method for low molecular weight RNA isolation from a wide variety of plant species. Biosci Biotechnol Biochem 77:1599–1601

    Article  CAS  PubMed  Google Scholar 

  21. Carra A, Gambino G, Schubert A (2007) A cetyltrimethylammonium bromide-based method to extract low-molecular-weight RNA from polysaccharide-rich plant tissues. Anal Biochem 60:318–320

    Article  Google Scholar 

  22. Carra A, Mica E, Gambino G et al (2009) Cloning and characterization of small non-coding RNAs from grape. Plant J 59:750–763

    Article  CAS  PubMed  Google Scholar 

  23. Chen CF, Ridzon DA, Broomer AJ et al (2005) Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res 33:e179

    Article  PubMed  PubMed Central  Google Scholar 

  24. Wu RM, Wood M, Thrush A et al (2007) Real-time PCR quantification of plant miRNAs using universal ProbeLibrary technology. Biochemica 2:12–15

    Google Scholar 

  25. Varkonyi-Gasic E, Wu RM, Wood M et al (2007) Protocols: a highly sensitive RT-PCR method for detection and quantification of microRNAs. Plant Methods 3:12

    Article  PubMed  PubMed Central  Google Scholar 

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Correspondence to Ying Miao .

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Ren, Y., Miao, Y. (2018). Isolation, Purification, and Detection of Micro RNAs in Plant Senescence. In: Guo, Y. (eds) Plant Senescence. Methods in Molecular Biology, vol 1744. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7672-0_20

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  • DOI: https://doi.org/10.1007/978-1-4939-7672-0_20

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7670-6

  • Online ISBN: 978-1-4939-7672-0

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