Proceedings of the XLVI Italian Society of Agricultural Genetics - SIGA Annual Congress

Giardini Naxos, Italy - 18/21 September, 2002

ISBN 88-900622-3-1

Poster Abstract - 3.30

cDNA-AFLP based transcript profiling during tobacco Bright Yellow 2 cell cycle

ATALLAH M., SAVO SARDARO M.L., PORCEDDU E.

Dipartimernto di Agrobiologia e Agrochimica, Universita’ della Tuscia, Via S.C. de Lellis, 01100 Viterbo

Bright Yellow2, Cell cycle, cDNA-AFLP

Genome wide expression analysis is an emerging tool for identifying genes involved in various biological process ranging from basic aspects such as cell division to complex responses to environmental cues.

The precise execution of the molecular process of DNA replication, chromosome segregation and mitosis during the cell cycle is likely to be governed by a precise regulation of gene activity. This makes the study of cell cycle dependent gene expression an attractive strategy to identify genes putatively involved in cell cycle control.

With the aim of identifying genes involved in mitosis exit and G1/S transition, tobacco Bright yellow 2 cell suspension were synchronised by the use of propizamide and twelve points covering the cell cycle were sampled and subjected to cDNA-AFLP protocol. The amplification products of 75 Eco(+2)/Mse(+2) primer combinations were analysed in polyacrilammide gels. 109 bands were eluted from the gel and 74 have been characterised by DNA sequencing.

25 of these AFLP bands were homologous to genes of unknown function, 24 to genes not (yet) considered as directly related to cell cycle whereas 19 to previously characterised cell cycle genes.

Putative cell cycle related AFLP tags are now being characterised further by isolating the corresponding full length cDNA clones and by studying the effect of their overexpression in transgenic tobacco plants.