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园艺学报 ›› 2015, Vol. 42 ›› Issue (12): 2341-2352.doi: 10.16420/j.issn.0513-353x.2015-0399

• 果树 •    下一篇

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

 1,2,*,谭晓风2,张 2,张靖国3,胡红菊3   

  1. 1湖南工业大学包装与材料工程学院,湖南株洲 4120072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 4100043湖北省农业科学院果树茶叶研究所,武汉 430064
  • 出版日期:2015-12-25 发布日期:2015-12-25
  • 基金资助:

    国家自然科学基金项目(31272124)

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

JIANG Nan1,2,*TAN Xiao-feng2ZHANG Lin2ZHANG Jing-guo3and HU Hong-ju3   

  1. 1School of Packing and Material Engineering,Hunan University of Technology,Zhuzhou,Hunan 412007,China;2Key Laboratory of Cultivation and Protection for Non-wood Forest of Ministry of Education,the Key Laboratory of Non-wood Forest Product of Forestry Ministry,Central-south University of Forestry and Technology,Changsha 410004,China; 3Institute of Fruit and Tea,Hubei Academy of Agricultural Sciences,Wuhan 430064,China
  • Online:2015-12-25 Published:2015-12-25

摘要:

利用东方梨中已鉴定的52S等位基因HVcDNA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55cDNA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

关键词: font-size: 9pt">梨, 自交不亲和, font-size: 9pt">S font-size: 9pt">基因型, font-size: 9pt">cDNAfont-size: 9pt">芯片, S 661.2寡核苷酸芯片

Abstract:

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.

Key words: pear, self-incompatibility, S-genotype, cDNA microarray, oligonucleotide genechip

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