| Self-incompatibility (SI) in Poaceae plants is a hotspot in the research of plant molecular biology and genetics breeding, which is important in research of cell signal transduction and heteosis. In this study, the molecular mechanism of an ecological self-incompatibility line, HE97, were analysed through differential gene expression between self-incompatible and self-compatible silk. On the basis of studying for several years in succession, the inbred line HE97 was sowed by stages in the farm of Henan agricultural university from April 22, 2004 to July 20,2004. The marterial was self-pollinated and the silk of each plant were sampled in every stage. After maturity, seeding percentage was calculated, and the silk selecting form two stages that represent the self-incompatibility and self-compatibility was used as material for total RNA isolation. There were two cDNA libraries including 1152 clones for forward library and 1056 clones for reverse library constructed by suppression subtractive hybridization (SSH) method.Reverse Northern blot was performed to analysis all of the clons in order to exclude false positives further. And then we selected and sequenced the difference signal intensity positive cDNAs. Forty-nine unique expressed sequence tags(ESTs) in forward library and 20 unique ESTs in reverse library were obtained by clustering and blast analysis.Most of them had homology with to known genes in Oryza sativa or Arabidopsis thaliana.they involved in the process of metabolism, transcription, signal transduction, cell growth/division, transporters, diseases defense and so on.The ESTs were sorted into 12 functional categories. But there are still some unknown function gene and 6 ESTs in forward subtract library were novel sequence, they are expected to be novel genes.The EST blast result showed that the forward clon 1A7 shared homology with the enoyl CoA hydratase gene of Oryza sativa or Arabidopsis thaliana. Enoyl CoA hydratase play an important part in fatty acid catabolism and CoA synthesization. For advanced understanding characteristic of this gene, a novel 1090bp contig was assembled by silicon cloning,it contains a 789bp open reading frame and encodes 262 amino acids. The Predicted protein has the Conserved Domain ECH (Enoyl-CoA hydratase/isomerase) and transmembrane helices between the 105aa and 124aa to the N terminal. Multiple homology sequence alignment and phylogram tree show that the contig1c7 and Enoyl-CoA hydrates gene in Oryza sativa has... |
