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    毛根林, 解云, 赵俊龙, 陈炎, 徐汉虹, 林菲. 葡萄糖基氟虫腈处理下蓖麻内参基因的稳定性分析[J]. 华南农业大学学报, 2016, 37(1): 52-57. DOI: 10.7671/j.issn.1001-411X.2016.01.009
    引用本文: 毛根林, 解云, 赵俊龙, 陈炎, 徐汉虹, 林菲. 葡萄糖基氟虫腈处理下蓖麻内参基因的稳定性分析[J]. 华南农业大学学报, 2016, 37(1): 52-57. DOI: 10.7671/j.issn.1001-411X.2016.01.009
    shu
    MAO Genlin, XIE Yun, ZHAO Junlong, CHEN Yan, XU Hanhong, LIN Fei. Stability analysis of internal reference gene of Ricinus communis treated by glucose-fipronil[J]. Journal of South China Agricultural University, 2016, 37(1): 52-57. DOI: 10.7671/j.issn.1001-411X.2016.01.009
    Citation: MAO Genlin, XIE Yun, ZHAO Junlong, CHEN Yan, XU Hanhong, LIN Fei. Stability analysis of internal reference gene of Ricinus communis treated by glucose-fipronil[J]. Journal of South China Agricultural University, 2016, 37(1): 52-57. DOI: 10.7671/j.issn.1001-411X.2016.01.009
    shu

    葡萄糖基氟虫腈处理下蓖麻内参基因的稳定性分析

    Stability analysis of internal reference gene of Ricinus communis treated by glucose-fipronil

      摘要
    • 摘要:
      目的 筛选葡萄糖基氟虫腈(GTF)及溶剂二甲基亚砜(DMSO)处理下蓖麻Ricinus communis稳定的内参基因，为研究GTF的韧皮部装载机制提供参考。
      方法 选取Actin，ARC，ef1a，SamDC，TUA6为内参基因，通过实时荧光定量PCR分析基因表达量并利用geNorm，NormFinder，BestKeeper，Delta CT软件及RefFinder在线分析工具综合比较不同时间和不同浓度的GTF与DMSO处理后，5个候选内参基因在蓖麻幼苗子叶中表达的稳定性。
      结果 各软件分析得出的内参基因稳定性排名依次为geNorm：Actin=ef1a > SamDC > ARC > TUA6；NormFinder：SamDC > ARC > Actin > ef1a > TUA6; BestKeeper：Actin > ef1a > SamDC > ARC > TUA6；Delta CT：SamDC > Actin > ARC > ef1a > TUA6; RefFinder：Actin > SamDC > ef1a > ARC > TUA6，而单独分析DMSO处理时，稳定性排名则为：ef1a > SamDC > Actin > TUA6 > ARC。
      结论 综合分析GTF和DMSO处理，Actin的表达最稳定；在DMSO处理下，则ef1a最为稳定。

       

      Abstract:
      Objective To screen reliable internal reference genes of castor, Ricinus communis, treated by glucose-fipronil (GTF) and the solvent dimethyl sulfoxide (DMSO), and to provide a basis for studying the uptake mechanism of GTF.
      Method Actin, ARC, ef1a, SamDC and TUA6 genes were selected as candidate reference genes. Specific primers for each gene were designed and real-time quantitative PCR was conducted. Five softwares, including geNorm, NormFinder, BestKeeper, Delta CT and RefFinder, were employed to analyze the gene expression stabilities in cotyledon of castor seedlings treated by GTF and DMSO with different concentration and time.
      Result The stabilities were variant according to different softwares. The rank orders of stability were geNorm: Actin=ef1a >SamDC > ARC > TUA6; NormFinder:SamDC > ARC > Actin > ef1a > TUA6; BestKeeper: Actin > ef1a > SamDC > ARC > TUA6; Delta CT: Actin > ef1a > SamDC > ARC > TUA6; RefFinder: Actin > SamDC > ef1a > ARC > TUA6. With a single analysis of DMSO treatment by RefFinder showed that ef1a > SamDC > Actin > TUA6 > ARC.
      Conclusion Actin is the stablest reference gene under GTF and DMSO treatments, while ef1a is the stablest one under DMSO treatment.

       

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