The Protein Disulfide Isomerase gene family in bread wheat (T. aestivum L.).

BACKGROUND: The Protein Disulfide Isomerase (PDI) gene family encodes several PDI and PDI-like proteins containing thioredoxin domains and controlling diversified metabolic functions, including disulfide bond formation and isomerisation during protein folding. Genomic, cDNA and promoter sequences of the three homologous wheat genes encoding the "typical" PDI had been cloned and characterized in a previous work. The purpose of present research was the cloning and characterization of the complete set of genes encoding PDI and PDI like proteins in bread wheat (Triticum aestivum cv Chinese Spring) and the comparison of their sequence, structure and expression with homologous genes from other plant species. RESULTS: Eight new non-homologous wheat genes were cloned and characterized. The nine PDI and PDI-like sequences of wheat were located in chromosome regions syntenic to those in rice and assigned to eight plant phylogenetic groups. The nine wheat genes differed in their sequences, genomic organization as well as in the domain composition and architecture of their deduced proteins; conversely each of them showed high structural conservation with genes from other plant species in the same phylogenetic group. The extensive quantitative RT-PCR analysis of the nine genes in a set of 23 wheat samples, including tissues and developmental stages, showed their constitutive, even though highly variable expression. CONCLUSIONS: The nine wheat genes showed high diversity, while the members of each phylogenetic group were highly conserved even between taxonomically distant plant species like the moss Physcomitrella patens. Although constitutively expressed the nine wheat genes were characterized by different expression profiles reflecting their different genomic organization, protein domain architecture and probably promoter sequences; the high conservation among species indicated the ancient origin and diversification of the still evolving gene family. The comprehensive structural and expression characterization of the complete set of PDI and PDI-like wheat genes represents a basis for the functional characterization of this gene family in the hexaploid context of bread wheat.

Identification and characterization of gene sequences expressed in wheat spikelets at the heading stage.

Through differential analysis of transcripts (SDDM), 85 cDNA sequences specifically or preferentially expressed in wheat spikelets at heading time were identified and cloned; 54 of them had significant homology with genomic, cDNA and protein and 16 with EST sequences. Among these 54 clones, 44 matched genes with known functions, whereas 10 detected homology with putative genes encoding proteins whose functions have been deduced on the basis of bioinformatic comparisons. Seventeen clones corresponded to genes that had never been cloned in cereals, 5 were related to wheat genes with known functions, and the remaining 32 to genes cloned in other cereals. On the basis of their presumed functions, the 54 clones were assigned to seven groups. The first four of them contained 40 sequences likely involved in floral organ morphogenesis and gametogenesis, and precisely (i) sequences involved in the morphogenesis of floral organs; (ii) sequences expressed in pollen and/or anther tissues; (iii) sequences encoding transcription factors; (iv) sequences involved in signal perception and transduction (kinases and LRR proteins). The expression patterns of these 40 sequences have been studied by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of transcripts from different tissues and spike organs of wheat.