Cloning of acyl-ACP thioesterase FatA from Arachis hypogaea L. and its expression in Escherichia coli.

In this study, a full-length cDNA of the acyl-ACP thioesterase, AhFatA, was cloned from developing seeds of Arachis hypogaea L. by 3'-RACE. Sequence analysis showed that the open reading frame encodes a peptide of 372 amino acids and has 50-70% identity with FatA from other plants. Real-time quantitative PCR analysis revealed that AhFatA was expressed in all tissues of A. hypogaea L., but most strongly in the immature seeds harvested at 60 days after pegging. Heterologous expression of AhFatA in Escherichia coli affected bacterial growth and changed the fatty acid profiles of the membrane lipid, resulting in directed accumulation towards palmitoleic acid and oleic acid. These results indicate that AhFatA is at least partially responsible for determining the high palmitoleic acid and oleic acid composition of E. coli.

Proteomic analysis on a high salt tolerance introgression strain of Triticum aestivum/Thinopyrum ponticum.

Soil salinity is a major abiotic constraint to agricultural productivity. We successfully bred a new common wheat (Triticum aestivum L.) introgression variety (Shanrong No. 3) with high salt-tolerance via asymmetric somatic hybridization between common wheat cultivar (Jinan 177) and UV-irradiated Agropyron elongatum (Thinopyrum ponticum Podp). We report here a comparative proteomic analysis to investigate variety-specific and salt-responsive proteins between seedling-roots of Shanrong No. 3 and Jinan 177. In total, 114 spots reproducibly presented differential expression patterns on 2-DE maps. Of them, 34 were variety-specific and 49 were salt-responsive. We identified 110 spots by MALDI-TOF MS and partially confirmed by MALDI-TOF-TOF MS, and functionally classified them into signal transduction, transcription and translation, transporting, chaperones, proteolysis and detoxification, etc. Meanwhile, we also found the alteration of protein expression of Shanrong No. 3 through inhibition of old proteins and production of novel ones, change in abundance and sensitivity of some nonsalt-responsive and salt-responsive proteins, as well as PTMs. Furthermore, comparison between proteome and transcripteome using cDNA microarray showed that there were only 20 proteins with abundances correlative to signal densities of corresponding EST probes. This study gives us a global insight into proteomic difference between Shanrong No. 3 and Jinan 177 in constitute and to salt-response.