科研之友
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摘要
Polyamines confer abiotic stress tolerance through mechanisms that are not yet fully understood. Previous studies on a transgenic line of European pear (Pyrus communis L., line #32), over-expressing an apple gene for spermidine synthase (MdSPDS1), provided greater insight into the mechanisms of stress tolerance, leading to agronomic benefits, as well as being an elite line to be used to further elucidate the molecular mechanisms underlying stress tolerance that are exerted by spermidine (Spd). In this study, a new and accurate reverse transcription-polymerase chain reaction (RT-PCR) method, involving an annealing control primer (ACP)-based PCR (GeneFishing) system, was employed to identify genes that may be implicated in abiotic stress tolerance. A total of 42 transcripts that were differentially expressed between line #32 and wild-type (WT) pear were identified. After cloning, 24 of these were found to exhibit significant sequence similarities with known genes from other species, and the remaining 18 were annotated as "unknown". To verify the reliability of the ACP-based differential display method, ten clones (five up-regulated and five down-regulated in line #32 compared to WT) were subjected to real-time RT-PCR analysis. These clones were further analysed to determine the relative abundance of the target sequences under conditions of salinity or cadmium stress. The results indicated that GeneFishing is a useful method to identify differentially-expressed genes (DEGs) in an MdSPDS1-overexpressing transgenic line of pear. The possible involvement of these DEGs in salinity and cadmium stress tolerance is also discussed based on an analysis of their levels of expression.
