International Network of Plant Abiotic Stress

COST STSM Reference Number: COST-STSM-FA0605-05496

Period: 11/01/2010 to 25/01/2010

STSM Applicant: Anna Rolicka, Warsaw University/Institute of Biochemistry and Biophysics PAS Pawinskiego 5A, Warsaw 02-106 Poland, ( arolicka@ibb.waw.pl )

Purpose of the visit:

    During my COST funded stay I chose to perform my work in Max Planck Institute for Plant Breeding Research under the tutelage of Prof Csaba Koncz and with help of Stefano Torti. Purpose of the visit was to conduct an in situ hybrydization experiment of two major shoot meristem genes, WUSCHEL and SHOOT MERISTEMLESS, in order to reveal characteristic changes in WUS and STM transcription underlying growth abnormalities observed in the atswi3c  Arabidopsis thaliana mutant and of the double atswi3c and flowering time mutants fd, fy and fca. Additionally, as we had more time and samples than we originally expected, we also performed similar analysis for AS2 (ASYMMETRIC LEAVES 2) gene. The tissue was already fixed and embedded in wax.

   The experimental procedure included sectioning of a total 48 meristem samples. We transferred the ribbons of cut meristem tissue onto slides and prepared digoxygenin – labeled riboprobes. Later, the proper hybridization procedure was performed, including tissue pretreatment, an overnight hybridization with probes, and later the washing, antibody staining, second washing and counter staining. Finally, pictures were taken and analyzed.

  My stay in Max Planck Institute was very helpful and I learned a useful and difficult technique that I will be able to perform now in my home lab. The experiment was a success; we obtained signal from WUS and AS2 probes and we were able to observe the differences in expression level and pattern between chosen mutants.

   Our group will continue to collaborate with the host lab especially on the subject of identifying partners and targets of SWI3 proteins, core subunits of Arabidopsis thaliana SWI/SNF remodeling complex.