| 郭清云,胡福初,吴凤芝,王祥和,范鸿雁,冯学杰,陈哲.榴莲蜜实时荧光定量PCR内参基因的克隆与筛选[J].中国南方果树,2022,51(5): |
| 榴莲蜜实时荧光定量PCR内参基因的克隆与筛选 |
| Cloning%20and%20Selection%20of%20Reference%20Genes%20for%20Real-Time%20qPCR%20Analysis%20in%20Cempedak |
| 投稿时间：2021-10-09 修订日期：2021-11-06 |
| DOI： |
| 中文关键词: 榴莲蜜 内参基因 实时荧光定量PCR 不同组织 |
| 英文关键词:cempedak,%20reference gene,%20RT-qPCR,%20different tissues |
| 基金项目:2021年农业种质资源保护项目（菠萝蜜种质资源保护），国家科技资源共享服务平台：国家热带植物种质资源库（National Tropical Plants Germplasm Resource Center）、海南省农业科学院“动植物资源收集与保存”专项项目（ZYBC-2020-01） |
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| 中文摘要: |
| 为筛选榴莲蜜实时荧光定量PCR的稳定内参基因,进一步提高榴莲蜜基因表达的稳定性和准确性。根据课题组测序获得的榴莲蜜转录组数据,筛选出10个候选内参基因；以“多异1号”榴莲蜜花序、茎、叶片以及不同发育时期的果苞为实验材料,通过基因克隆、实时荧光定量PCR分析,并结合4种统计分析软件评价内参基因稳定性。结果表明,克隆获得了5个榴莲蜜内参基因,分别为：Actin1、α-TUB1、β-TUB1、β-TUB2和GAPDH1；这5个内参基因的转录水平在榴莲蜜的不同组织类型及果苞发育过程中存在差异；稳定性综合排名结果表明,在榴莲蜜花序、茎和叶片中表达稳定性最好的是β-TUB2和α-TUB1,在榴莲蜜果苞发育阶段中表达水平最稳定的是β-TUB1和α-TUB1；通过2个成花相关基因和4个蔗糖合成酶基因的表达模式验证了α-TUB1、α-TUB1+β-TUB1及α-TUB1+β-TUB2作为内参基因的可靠性。本研究结果为榴莲蜜的功能基因表达分析提供了理论基础。 |
| 英文摘要: |
| In%20order to screen%20the%20stable%20reference%20genes%20for%20qRT-PCR%20analysis%20of%20cempedak,%20“duoyi%201”%20as%20material,%20five%20candidate%20reference%20genes%20including%20Actin1,%20α-TUB1,%20β-TUB1,%20β-TUB2%20and%20GAPDH1%20were%20screened%20and%20cloned%20based%20on%20the%20RNA-seq%20data%20of%20cempedak.%20The%20expression%20of%20fivet%20candidate%20reference%20genes%20were%20assessed%20by%20qRT-PCR%20in%20different%20tissues%20and%20periods%20of%20bud%20development,%20and%20the%20stability%20of%20them%20were%20analyzed%20by%20four%20programs.The%20results%20showed%20that%20the%20transcription%20level%20and%20stability%20of%20five%20reference%20genes%20were%20different%20in%20the%20tissue%20type%20and%20bud%20development%20stage%20of%20cempedak.%20β-TUB2%20and%20α-TUB1%20were%20the%20most%20stable%20genes%20in%20different%20tissues,%20such%20as%20flowering%20inflorescence,%20stems%20and%20leaves,%20while%20β-TUB1%20and%20α-TUB1%20were%20the%20most%20stable%20genes%20in%20bud%20of%20cempedak.%20The%20expression%20patterns%20of%202%20flower%20formation-related%20genes%20and%204%20sucrose%20synthase%20genes%20verified%20the%20reliability%20of%20α-TUB1,%20α-TUB1+β-TUB1%20and%20α-TUB1+β-TUB2%20as%20internal%20reference%20genes.%20The%20results%20of%20this%20study%20provided%20a%20theoretical%20basis%20for%20the%20functional%20gene%20expression%20analysis%20of%20cempedak. |
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