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 - 4.12

IDENTIFICATION OF CANDIDATE GENES EXPRESSED AT LOW TEMPERATURE IN MAIZE BY USING THE cDNA-AFLP APPROACH

BERTOLI A., MAROCCO A.

Istituto di Botanica e Genetica vegetale, Università Cattolica del Sacro Cuore, Piacenza

cold regulated genes, virescent, Zea mays

The virescent character is a common genetic variant in pigmentation of higher plants for which a large number is known in maize. It is generally accepted that expression of this character is influenced by temperature sensitivity. Availability of temperature-sensitive virescent mutants is a useful tool to identify genes involved in low-temperature resistance. The cDNA-AFLP tecnique has been employed in order to obtain differentially expressed genes. RNA, isolated from wilde type and virescent mutants grown at low temperature, was used to synthetize cDNA. Subsequently thirty-two different primer combinations, each of the primer carrying a selective base at 3’ end, were employed in PCR assay to amplify the sequences generated by restriction reaction with BstYI and MseI. Employment of the optimized protocol for isolating, reamplifying and sequencing transcript-derived framgments, allowed the identification of 534 sequences wich are related to virescent temperature-sensitive events.