摘要
Chinese cabbage (Brassica rapa) is widely recognized for its economic importance and contribution to human nutrition but abiotic and biotic stresses are main obstacle for its quality, nutritional status and production. In this study, 3,429 Express Sequence Tag (EST) sequences were generated from B. rapa cv. Osome cDNA library and the unique transcripts were classified functionally using a gene ontology (GO) hierarchy, Kyoto encyclopedia of genes and genomes (KEGG). KEGG orthology and the structural domain data were obtained from the biological database for stress related genes (SRG). EST datasets provided a wide outlook of functional characterization of B. rapa cv. Osome. In silico analysis revealed % 83 of ESTs to be well annotated towards reeds one dimensional concept. Clustering of ESTs returned 333 contigs and 2,446 singlets, giving a total of 3,284 putative unigene sequences. This dataset contained 1,017 EST sequences functionally annotated to stress responses and from which expression of randomly selected SRGs were analyzed against cold, salt, drought, ABA, water and PEG stresses. Most of the SRGs showed differentially expression against these stresses. Thus, the EST dataset is very important for discovering the potential genes related to stress resistance in chinese cabbage, and can be of useful resources for genetic engineering of Brassica sp.
Subject(s)
Brassica/drug effects , Brassica/genetics , Brassica/growth & development , Databases, Genetic , Expressed Sequence Tags/metabolism , Gene Expression Profiling , Gene Library , Gene Regulatory Networks , Genes, Plant/genetics , Genome, Plant , Humans , Molecular Sequence Annotation , Oligonucleotide Array Sequence Analysis , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , Sodium Chloride/pharmacology , Stress, Physiological/genetics摘要
BACKGROUND & OBJECTIVES: The spinal expression of the c-Fos immediate early gene in response to formalin pain of the hind paw of rat was used as a marker of neuronal activity. Ketamine, a Nmethyl-D-aspartate (NMDA) receptor antagonist produces analgesic action due to the blockade of glutamate action at the NMDA receptor. Earlier study showed that ketamine acts differently depending on its route of administration. We undertook this study to compare a preemptive suppression of noxious stimulation induced spinal Fos-like immunoreactivity (FLI) after receiving intrathecal ketamine before or after formalin pain. METHODS: Male Sprague-Dawley rats received ketamine 1 mg/kg or saline (control group) intrathecally either 5 min before (pre-treatment group) formalin or 5 min after (post-treatment group) formalin (5%, 50 microl) injection. Animals were killed 2 h after the formalin injection, and the lumbar spinal cord was dissected, and processed by immunoperoxidase staining using an antibody against Fos protein. RESULTS: The FLI was significantly reduced in the pre-treatment group, only laminae I-II of the side ipsilateral to the formalin injection (P < 0.05 vs. control). In laminae V-VI, neither of the ketamine treatment groups showed a significant decrease than the control group. INTERPRETATION & CONCLUSION: The results provide evidence that intrathecal ketamine does not have a preemptive blocking effect of FLI expression in whole spinal laminae area. FLI expression of laminae I-II only might not be a good predictor of the ability of agents to produce preemptive effect. The central patterns of activity generated during central sensitization differ regionally in the spinal dorsal horn.