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Article Dans Une Revue Methods in Molecular Biology Année : 2013

Analyzing small and long RNAs in plant development using non-radioactive in situ hybridization.

Pilar Bustos-Sanmamed
  • Fonction : Auteur
Carole Laffont
  • Fonction : Auteur
  • PersonId : 1205165
Florian Frugier
Christine Lelandais-Brière
  • Fonction : Auteur
  • PersonId : 1355839
  • IdRef : 073427055
Martin Crespi

Résumé

In the past decade, hundreds of non-coding RNAs (small and long RNAs) have been identified as crucial elements in developmental processes and stress response in plants. Among small RNAs, the microRNAs or miRNAs control levels of specific mRNA by inhibiting translation or reducing the stability of their mRNA targets through integration into different ribonucleoproteins (RNP). Spatio-temporal expression of small and long RNAs, using reporter genes or in situ hybridization, is essential to understand their functions. We are interested in understanding the role of various non-coding RNAs (including miRNAs) in the regulation of root and nodule development in legumes, which are agriculturally important crops. Here, we present the protocol we are currently using for detection of small and long RNA in model legume plants and tissues, like nodules and roots. The probe selection, as well as the fixation and permeabilization steps allowing to preserve tissues and cell integrity and to increase accessibility to RNA targets, will be specifically discussed.
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Dates et versions

hal-00856162 , version 1 (30-08-2013)

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Citer

Pilar Bustos-Sanmamed, Carole Laffont, Florian Frugier, Christine Lelandais-Brière, Martin Crespi. Analyzing small and long RNAs in plant development using non-radioactive in situ hybridization.. Methods in Molecular Biology, 2013, 959, pp.303-16. ⟨10.1007/978-1-62703-221-6_20⟩. ⟨hal-00856162⟩
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