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

Verona, Italy - 24/27 September, 2003

ISBN 88-900622-4-X

Poster Abstract - 4.08

GENE EXPRESSION IN BARLEY THROUGH MICROARRAY TECHNOLOGY BY USING NEAR ISOGENIC LINES DERIVED FROM THE “LEAFY LEMMA (LEL)” MUTANT AND THE MALTING QUALITY VARIETY KASKADE

P. Faccioli, F. Rizza, A. Finocchiaro, V. Terzi

Istituto Sperimentale per la Cerealicoltura, S.O.P. Fiorenzuola d’Arda, Via San Protaso 302, 29017 Fiorenzuola d’Arda (PC), Italy

barley, morphological mutant, cDNA microarray, real-time RT-PCR, photosynthetic efficiency

Leafy lemma barley mutant carries recessive alleles of two genes, both of which are necessary to cause the transformation of the lemma into a structure having all characteristics of a vegetative leaf, as shown by SEM analysis. The presence of a sheats, blade, and ligule in the mutant lemma suggests that wild type lemma development is interrupted at a leaf-like stage. Mapping positions of the two lel genes on barley chromosomes 5 and 7 has been determined using F₂ populations derived from leafy lemma x Nudinka crosses and an optimized AFLP- based procedure (Castiglioni et al., 1998; Pozzi et al., 2000).

Starting from the cross leafy lemma x Kaskade, near isogenic lines were developed after a six years breeding program: each pair of lines shows the wild type and mutant version of lemma in the same genetic background. The characterization of the isogenic lines was done at three different level. A first characterization of these isogenic lines was done evaluating a set of agronomic traits in two years trials. A preliminary evaluation of photosynthetic efficiency of the leafy lemma structure in comparison with other photosynthetic tissues of the plant, like awn and flag leaf was done. Finally, gene expression was studied and compared in leafy lemma, awn and flag leaf from pairs of near isogenic lines using microarray technology (Faccioli et al., 2002). EST clones from a barley leaf cDNA library were arrayed and used to test differences in mRNA expression level in the mutant lemma in comparison with other photosynthetic tissues of the plant. To support the microarray-based results, the variation in hybridization signals in some clones was confirmed by real time RT-PCR analysis.

References

Castiglioni P., C. Pozzi, M. Heun, K.J. Muller, V.Terzi et al. 1998. An AFLP-based procedure for the efficient mapping of mutations and DNA probes in barley. Genetics 149: 2039-2056.

Pozzi C., P. Faccioli, V. Terzi, A.M. Stanca, S. Cerioli, et al. 2000. Genetics of mutations affecting the development of a barley floral bract. Genetics 154: 1335-1346.

Faccioli P., M.S. Lagonigro, L. De Cecco, A.M. Stanca, R. Alberici, V. Terzi. 2002. Analysis of differential expression of barley ESTs during cold acclimatization using microarray technology. Plant biol. 4: 630-639.