梨自交不亲和基因cDNA芯片制备及对部分砂梨品种S基因型的鉴定

利用东方梨中已鉴定的52个S等位基因HV区cDNA序列作为靶基因序列设计探针，制备梨S基因cDNA检测芯片，每张芯片上含有240个位点55个cDNA探针，包含所有序列完善的S基因HV区特异的cDNA序列。以被检测品种雌蕊cDNA为模板，采用Cy3荧光修饰引物经S基因特异PCR扩增标记被检测品种的cDNA序列，并与芯片杂交以检测不同品种的S基因型。结果表明：利用cDNA检测芯片与‘丽江黄酸梨’、‘秀玉’、‘弥渡玉梨’、‘白面梨’和‘德胜香’等已知S基因型品种杂交，杂交结果显示与S基因寡核苷酸芯片检测信号一致，与各品种已知S基因型相符合。利用cDNA芯片和进一步完善的S基因寡核苷酸芯片并行检测鉴定了‘文山红梨’等24个未知S基因型的砂梨品种，获得各品种的S基因型。梨S基因cDNA芯片的构建进一步完善了梨S基因检测平台。

Preparation of S-RNase cDNA Microarray and Its Application in Identifying Pear Cultivars S-genotypes

Using the cDNA sequences from Hyper variable（HV）regions of 52 S-alleles in Oriental pear cultivars，cDNA microarray for S-RNase detections was established. Totally 240 dots and 55 cDNA probes，which includes all the HV specific cDNA sequences of perfected S-RNase gene，were spotted on the chip. By Cy3-labeled primers and cDNA template of tested cultivars pistils，the Cy3-labeled specific cDNA PCR products of S-RNase were amplified and hybridized with the cDNA microarray in order to detect the S genotype of pear cultivars. Cultivars with known S-genotype such as Lijiang Huangsuanli，Xiuyu，Midu Yuli，Baimianli and Deshengxiang were S-genotyped using the cDNA microarray and the results showed that the microarray analyses were consistent with their oligonucleotide genechip analyses and their RFLP and DNA sequenced results. Then the S-RNase cDNA microarrays and the oligonucleotide genechips were used to parallel S-genotyping 24 sand pear cultivars with unknown S-genotype and their S-genotypes were determined. In conclusion，the construction of cDNA microarrays has further improved the pear S-RNase detection platform.