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  

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

作　　者：江南[1,2] 谭晓风[2] 张琳[2] 张靖国[3] 胡红菊[3] 

机构地区：[1]湖南工业大学包装与材料工程学院,湖南株洲412007 [2]中南林业科技大学经济林培育与保护教育部重点实验室,经济林育种与栽培国家林业局重点实验室,长沙410004 [3]湖北省农业科学院果树茶叶研究所,武汉430064

出　　处：《园艺学报》2015年第12期2341-2352,共12页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31272124)

摘　　要：利用东方梨中已鉴定的52个S等位基因HV区c DNA序列作为靶基因序列设计探针,制备梨S基因c DNA检测芯片,每张芯片上含有240个位点55个c DNA探针,包含所有序列完善的S基因HV区特异的c DNA序列。以被检测品种雌蕊c DNA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c DNA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c DNA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c DNA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c DNA芯片的构建进一步完善了梨S基因检测平台。Using the c DNA sequences from Hyper variable（HV）regions of 52 S-alleles in Oriental pear cultivars,c DNA microarray for S-RNase detections was established. Totally 240 dots and 55 c DNA probes,which includes all the HV specific c DNA sequences of perfected S-RNase gene,were spotted on the chip. By Cy3-labeled primers and c DNA template of tested cultivars pistils,the Cy3-labeled specific c DNA PCR products of S-RNase were amplified and hybridized with the c DNA 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 c DNA 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 c DNA 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 c DNA microarrays has further improved the pear S-RNase detection platform.

关 键 词：梨 自交不亲和 S基因型 c DNA芯片 寡核苷酸芯片 

分 类 号：S661.2[农业科学—果树学]