AFLP experiment designed to investiga te the role of Gibberellins in Arabidopsis germination

  1. J. G. De Diego 1
  2. F. D. Rodríguez 1
  3. E. Battaner 1
  4. P. Grappin 1
  5. E. Cervantes 1
  1. 1 Dep. Bioquímica & Biología Molecular. Edificio Departamental, lab 125 CampusUnamuno, Universidad de Salamanca,3 7007 Salamanca, Spain. Laboratoire de Biologie des semences. INRA-Versailles. France. IRNA-CSIC. Apartado 257. Salamanca
Workshop on Biochemistry and Molecular Biology of Gibberellins

Publisher: Instituto Juan March

Year of publication: 1990

Pages: 60

Type: Book chapter


An experiment based on the AFLP protocol was designed to isolate cONAfragments induced during Arabidopsis thaliana seed germination. Poly A+RNA from wild type Wassilejska seeds as well as mutant ABC33 (GAI 1) ,24 hours after imbibition, was isolated and reverse transcribed. Sequencesfrom both cDNAs were amplified according to the AFLP protocoldescribed by Bachem el al. ( 1996), and the resulting radiolabelled PCRproducts were ran on acrylamide gels. Bands showing specific expressionon wildtype or mutant seeds were colected and re-amplifíed, cloned and sequenced. The aminoacid sequences derived from the obtained nucleotide sequences showed among the cDNAs severa! putative transcription factors and other proteins that may be involved in the control of Arahidopsis seedgermination. One of the transcription factors, belongs to the b-Zip familyand conesponds toa gene already sequenced, thus making straightforward the analysis of its prometer region. Other isolated cON As encode proteins with diverse degrees of homology with ABA induced proteins, titin, cytochrome oxidase and others. Some data concerning the regul ation of their mRNAs during germination wi ll be presented