Identification and sequencing of the asparagine synthetase genes in Durum Wheat

Project coordinator :

Dr. Gabriella Sonnante
Institute of Plant Genetics (IGV) – CNR
Via Amendola, 165/A
70126 Bari (Italy)

Abstract :
It is well known that many plants rely on the synthesis and degradation of the amides asparagine and glutamine to relocate nitrogen within their tissue. In this context, asparagine plays a prominent role in nitrogen transport and storage in plants.
Glutamine-dependent asparagine synthetase (AsnS) catalyses the transfer of an amide group from glutamine to aspartate forming asparagine in an ATP-dependent reaction. In most plant species, AsnS seems to be encoded by a small gene family. The number of AsnS genes identified in plant species varies between one and four (i.e. one in asparagus, two in bread wheat and barley, three in Arabidopsis and sunflower, four in maize), and in most species only one or two genes are known.
The aim of this work is to identify and characterize AsnS genes in durum wheat. A hybridization of durum wheat macroarrays, representing a durum wheat BAC library accession, was carried out in order to find clones containing asparagine synthetase genes. Probes are developed on full-length AsnS sequences belonging to other close Triticeae taxa already present in NCBI. Positive BAC clones to hybridation will be validated by PCR and Sanger sequencing and entire BAC will be sequenced using 454 technology in order to obtain the different gene family members.

CNRGV involvement :

Responsible: William Marande