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2025年4月17日 星期四
  2018, Vol. 26 Issue (1): 53-63    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
苹果柱型基因候选基因的筛选
仇占南1,付艳2,杜传慧3,王利敏1,李晨辉1,朱元娣4
1. 中国农业大学
2.
3. 中国农业大学西校区
4. 北京市圆明园西路2号中国农业大学西校区科研楼1235室
Screening Candidate Genes of the Columnar Gene in Apple (Malus×domestica Borkh.)
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摘要 苹果(Malus × domestica Borkh.)柱型生长性状受位于10号染色体上的单显性基因Co(Columnar)控制,Co区域内有多个预测基因。筛选与苹果柱型性状相关的候选基因,为确定Co的身份,提供理论依据。通过NCBI中的序列阅读存档/表达序列标签(sequence read archive/expressed sequence tags, SRA/EST)数据库检索Co区域(18.51~19.1 Mb)的64个预测基因;以柱型苹果‘威赛克’和非柱型苹果‘旭’的茎尖、叶片、芽为试材,分别提取RNA,反转录后等体积混合作为cDNA混合样,用于反转录PCR(reverse transcription PCR, RT-PCR)扩增预测基因的cDNA序列;利用实时荧光定量PCR(quantitative Real-time PCR, qRT-PCR)分析苹果茎尖组织中的基因差异表达。通过SRA/EST数据库检索,筛查到在不同苹果组织或器官中表达的32个预测基因;RT-PCR扩增获得29个预测基因的cDNA序列;qRT-PCR分析获得15个在柱型苹果茎尖中差异表达显著的基因,其中6个基因(6号、41号、33号、38号、28号和32号基因)上调显著,9个基因(30号、9号、37号、6号、44号、10号、27号、11号和4号基因)下调显著。本研究在柱型苹果茎尖中筛选到的15个表达显著的基因,可作为Co候选基因。候选基因的确定旨在阐明Co基因区域的关键基因功能和苹果柱型突变的分子机制,为苹果树型的定向遗传改良服务。
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仇占南
付艳
杜传慧
王利敏
李晨辉
朱元娣
关键词 苹果, 柱型性状, Co区域, 差异表达, 候选基因    
Abstract:Columnar trait of apple (Malus × domestica Borkh.) is controlled by a single dominant Columnar (Co) gene, located in apple genome chromosome 10. Many putative genes are predicted in the Co region. Screening genes related to the columnar trait in apple provides theoretical basis for identification of the Co gene. Sixty-four putative genes in the Co region (18.51~19.1 Mb) were screened by SRA/EST (Sequence Read Archive/Expressed Sequence Tags) database. Samples of shoot tips, leaves and buds from columnar cultivar ‘Wijcik’ and non-columnar cultivar ‘McIntosh’ were used to extract total RNA, respectively, and the cDNA mixture of an equal volume of each sample after reverse transcription, was used as materials to conduct. Reverse transcription PCR (RT- PCR) was to amplify partial cDNA sequences of putative gene. Quantitative Real-time PCR (qRT-PCR) reactions were performed to analyze the differential expression of putative genes in apple shoot tips. 32 putative genes expressed in different apple tissues or organs were detected by searching SRA/EST database. 29 putative genes were obtained by RT-PCR amplification, and 15 genes were with significant differences in apical tips of the columnar apple by qRT-PCR analysis. Among them, 6 genes (No.26, No.41, No.33, No.38, No28 and No.32) were up-regulated significantly (P<0.05), and 9 genes (No.30, No.9, No.37, No.6, No.44, No.10, No.27, No.11 and No.4) were down-regulated significantly in apical tips of the columnar cultivar(P<0.05). In this study, 15 genes with significant differences in the apical tips of the columnar apple could be regarded as candidate genes of the Co. The candidate genes are identified to elucidate the function of critical genes and molecular mechanisms of the apple columnar mutation in the Co gene region, and serve as a directional genetic improvement for apple trees.
Key wordsMalus × domestica Borkh., Columnar trait, The Co locus, Differential expression, Candidate genes
收稿日期: 2017-05-12      出版日期: 2018-01-01
基金资助:苹果柱型相关的基因筛选与鉴定
通讯作者: 朱元娣     E-mail: yuandi7232@sina.com
引用本文:   
仇占南 付艳 杜传慧 王利敏 李晨辉 朱元娣. 苹果柱型基因候选基因的筛选[J]. , 2018, 26(1): 53-63.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2018/V26/I1/53
 
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