Advertisement

Multiprobe In-Situ Hybridization to Whole Mount Arabidopsis Seedlings

  • Leonardo Bruno
  • Fabrizio Araniti
  • Olimpia Gagliardi
Chapter

Abstract

In the multicellular organisms to know the temporal and spatial expression of genes gives specific information about the putative sites of activity of the corresponding proteins and also their function related to processes of growth and development as well as in response to biotic and abiotic stress conditions. In-situ RNA-RNA hybridization (ISH) is a powerful technique that enables the localization of gene transcripts at the cellular level by using labeled probes complementary to nucleic acid of interest, followed by probe specific detection. Original methods for ISH have been developed in the 1980s. Since that time, the methodologies associated with it have undergone a continuous evolution, which is also reviewed in the present chapter.

References

  1. Araniti F, Graña E, Krasuska U, Bogatek R, Reigosa MJ, Abenavoli MR, Sánchez-Moreiras AM (2016) Loss of gravitropism in farnesene-treated arabidopsis is due to microtubule malformations related to hormonal and ROS unbalance. PLoS One 11(8):e0160202CrossRefPubMedPubMedCentralGoogle Scholar
  2. Araniti F, Bruno L, Sunseri F, Pacenza M, Forgione I, Bitonti MB, Abenavoli MR (2017) The allelochemical farnesene affects Arabidopsis thaliana root meristem altering auxin distribution. Plant Physiol Biochem 121:14–20CrossRefPubMedGoogle Scholar
  3. Birnbaum K, Jung JW, Wang JY, Lambert GM, Hirst JA, Galbraith DW, Benfey PN (2005) Cell type–specific expression profiling in plants via cell sorting of protoplasts from fluorescent reporter lines. Nat Methods 2(8):615–619CrossRefPubMedGoogle Scholar
  4. Bruno L, Chiappetta A, Muzzalupo I, Gagliardi C, Iaria D, Bruno A, Greco M, Giannino D, Perri E, Bitonti MB (2009) Role of geranylgeranyl reductase gene in organ development and stress response in olive (Olea europaea) plants. Funct Plant Biol 36(4):370–381CrossRefGoogle Scholar
  5. Bruno L, Muto A, Spadafora ND, Iaria D, Chiappetta A, Van Lijsebettens M, Bitonti MB (2011) Multi-probe in situ hybridization to whole mount Arabidopsis seedlings. Int J Dev Biol 55(2):197–203CrossRefPubMedGoogle Scholar
  6. Bruno L, Ronchini M, Gagliardi O, Corinti T, Chiappetta A, Gerola P, Bitonti MB (2015) Analysis of AtGUS1 and AtGUS2 in Arabidopsis root apex by a highly sensitive TSA-MISH method. Int J Dev Biol 59(4-5-6):221–228CrossRefPubMedGoogle Scholar
  7. Christine T, Bernard T (2008) High-resolution in situ hybridization to whole-mount zebrafish embryo. Nat Protoc 3:59–69CrossRefGoogle Scholar
  8. Chuaqui RF, Bonner RF, Best CJ, Gillespie JW, Flaig MJ, Hewitt SM, Phillips JL, Krizman DB, Tangrea MA, Ahram M, Linehan WM, Knezevic V, Linehan WM (2002) Post-analysis follow-up and validation of microarray experiments. Nat Genet 32:509–514CrossRefPubMedGoogle Scholar
  9. Drews GN, Beals TP, Bui AQ, Goldberg RB (1992) Regional and cell specific gene expression patterns during peta1 development. Plant Cell 4:1383–1404CrossRefPubMedPubMedCentralGoogle Scholar
  10. Giannino D, Condello E, Bruno L, Testone G, Tartarini A, Cozza R, Innocenti AM, Bitonti MB, Mariotti D (2004) The gene geranylgeranyl reductase of peach (Prunus persica [L.] Batsch) is regulated during leaf development and responds differentially to distinct stress factors. J Exp Bot 55(405):2063–2073CrossRefPubMedGoogle Scholar
  11. Harrison PR, Conkie D, Affara N, Paul J (1974) In situ localization of globin messenger RNA formation. J Cell Biol 63(2):402–413CrossRefPubMedPubMedCentralGoogle Scholar
  12. Kim JS, Jung HJ, Lee HJ, Kim K, Goh CH, Woo Y, Seung HO, Yeon SH, Kang H (2008) Glycine-rich RNA-binding protein7 affects abiotic stress responses by regulating stomata opening and closing in Arabidopsis thaliana. Plant J 55(3):455–466CrossRefPubMedGoogle Scholar
  13. Larkin JC, Oppenheimer DG, Pollock S, Marks MD (1993) Arabidopsis GLABROUS l gene requires downstream sequences for function. Plant Cell 5:1739–1748CrossRefPubMedPubMedCentralGoogle Scholar
  14. Lefebvre V, North H, Frey A, Sotta B, Seo M, Okamoto M, Nambara E, Marion-Poll A (2006) Functional analysis of Arabidopsis NCED6 and NCED9 genes indicates that ABA synthesized in the endosperm is involved in the induction of seed dormancy. Plant J 45(3):309–319CrossRefPubMedGoogle Scholar
  15. Mizukami Y, Huang H, Tudor M, Hu Y, Ma H (1996) Functional domains of the floral regulator AGAMOUS: characterization of the DNA binding domain and analysis of dominant negative mutations. Plant Cell 8:831–845CrossRefPubMedPubMedCentralGoogle Scholar
  16. Taylor C (1997) Promoter fusion analysis: an insufficient measure of gene expression. Plant Cell 9(3):273–275CrossRefPubMedCentralGoogle Scholar
  17. Wellmer F, Riechmann JL, Alves-Ferreira M, Meyerowitz EM (2004) Genome-wide analysis of spatial gene expression in Arabidopsis flowers. Plant Cell 16(5):1314–1326CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  • Leonardo Bruno
    • 1
  • Fabrizio Araniti
    • 2
  • Olimpia Gagliardi
    • 1
  1. 1.Department of Biology, Ecology and Earth Science, Laboratory of Plant BiologyUniversity of CalabriaArcavacata di RendeItaly
  2. 2.Department AgrariaMediterranea UniversityReggio CalabriaItaly

Personalised recommendations