梨S基因芯片的试制及分子杂交条件的优化

 为研究寡核苷酸芯片在确定梨品种S基因型中的应用价值，根据梨自交不亲和基因的结构特点设计了部分S基因的检测探针，并制成寡核苷酸芯片；采用引物Cy3荧光标记法标记检测样品的PCR产物并进行杂交，以检测不同样品的S基因型。结果表明：通过对不同杂交温度、杂交时间等的探索获得了较好的反应条件并用制备的芯片检测了已知S基因型的梨样品，检测结果与各品种的已知基因型相符。证明寡核苷酸芯片检测梨品种的S基因型是一种切实可行的检测方法，若对芯片进一步完善，则今后在梨的自交不亲和性状的机理研究及梨自交不亲和性状的利用等方面将有着广阔的应用前景。

Preparation of Prototype S-RNase Microarray and Molecule Hybridization Condition

To explore the application of oligonucleotide microarray in identification of pears S-genotype, some probes for S-RNase detection were designed according to the structure characteristic of self-incompatibility gene, and oligonucleotide microarray was prepared. The PCR products of S-RNase were amplified by Cy3-labeled primer and hybridized with the microarray in order to detect the S-genotype of pears. Optimized reaction conditions were gained by exploration of hybridization temperature and hybridization time. The microarray results of S-genotype of pear was similar with the RFLP and sequenced results. In conclusion:, the oligonucleotide microarray is an effective method in detecting S-genotype of pear. With the future improvement, it could be widely used in the study of self-incompatibility of pear.