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根据东方梨中已鉴定的46个S基因序列和S基因的结构特点,设计了86条寡核苷酸探针并制备成S基因寡核苷酸检测芯片,采用Cy3荧光修饰引物标记被检测品种的PCR产物并与芯片杂交,以检测不同品种的S基因型。结果表明:利用芯片与华梨2号、秀玉和德胜香等已知S基因型的品种杂交,杂交信号显示与各品种已知基因型相符合。利用芯片鉴定了丽江黄酸梨等27个未知S基因型的梨品种,获得了各品种的S基因型,随机选取部分品种进行DNA测序和序列分析,结果与芯片杂交结果完全一致,证明利用S基因寡核苷酸芯片鉴定梨品种S基因型结果准确可靠。
According to the identified 46 S gene sequences and the structural features of S gene in oriental pear, 86 oligonucleotide probes were designed and synthesized as S gene oligonucleotide detection chips. Cy3 fluorescence modified primers were used to mark the tested species PCR products and hybridized with the chip to detect different varieties of S genotype. The results showed that the hybridization signals of the hybrids with known S genotypes, such as Huarui 2, Xiuyu and Deshengxiang, were consistent with the known genotypes of each breed. A total of 27 pear cultivars with unknown S genotypes, such as Lihuang yellow pear, were identified by chip. The S genotypes of the cultivars were obtained. Some of them were randomly selected for DNA sequencing and sequence analysis. The results were in good agreement with the results of hybridization. Gene oligonucleotide microarray identification of pear S genotype results accurate and reliable.