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园艺学报 ›› 2014, Vol. 41 ›› Issue (10): 1983-1992 .

• 果树 • 上一篇    下一篇

利用基因芯片鉴定梨品种自交不亲和基因型

江 南1,2,*,谭晓风2,张 琳2,邓 靖1   

  1. 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
  • 收稿日期:2014-06-26 出版日期:2014-10-25 发布日期:2014-10-25
  • 基金资助:

    国家自然科学基金面上项目(31272124);湖南省自然科学基金项目(12JJ6017)

Identifying Pear Cultivars S-genotypes Using Gene Chips

JIANG Nan1,2,*,TAN Xiao-feng2,ZHANG Lin2,and DENG Jing1   

  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
  • Received:2014-06-26 Online:2014-10-25 Published:2014-10-25

摘要: 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。

关键词: 梨, 自交不亲和, S基因型, 寡核苷酸, 芯片

Abstract: Based on the structure characteristic of self-incompatibility gene and the gene group sequences of 46 S genes in Oriental pear cultivars,86 probes for S-RNase detections were designed and oligonucleotide genechip was established. In order to detect the S-genotype of pear cultivars,the PCR products of S-RNase were amplified by Cy3-labeled primers and hybridized with the microarray. The cultivars with known S-genotype such as Huali 2 were hybridized with the genechip and 27 cultivars with unknown S-genotype were S-genotyped using the genechip. The DNA sequenced results of Huali 2,Xiuyu and Deshengxiang pears and randomly selected unknown S-genotype cultivars were consistent with the microarray analyses. In conclusion,the oligonucleotide genechip provided an accurate and reliable method for detecting S-genotype of pear cultivars.

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

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