Construction and analyses of SSH libraries of cold resistance related genes in Vitis amrensis accession‘Tonghua-3'

【Objective】To clone the related- genes involved in response to cold stress of V. amrensis andanalyse cold stress mechanism of grape.【Methods】With the suppressive subtraction hybridization(SSH)technique,the pros and cons of two differentially expressed c DNA libraries ASSH(V. amurensis‘Tonghua-3') and SSSH(V. vinifera L.‘Red globe') were constructed using V. amurensis accession‘Tonghua-3'with strong cold resistance and V. vinifera L.‘Red Globe'with weak cold resistance as Driver and Tester.【Results】After sequencing the clones expressing differentially and comparing DNAMAN,we selected 70 unigenes from the 657 clones;After blasting their sequences,found some ESTs regulating r RNA gene in-tron-encoded homing endonuclease,aging-related proteins,cytochrome P450,antifreeze proteins,de-fensin- like protein,heat shock proteins and others involved in ASSH library of V. amurensis;In theSSSH c DNA library,we found aging-related proteins in‘Red Globe'cultivar. In addition,we also foundthe fourteen unknown protein of ESTs.【Conclusion】Screened AFPs,HSPs,defensin-like protein,Cyto-chrome P450 monooxygenase of ESTs related to the cold resistance in V. amurensis and the fourteen un-known protein sequences may also be involved in cold metabolism of V. amurensis.