葡萄糖基氟虫腈处理下蓖麻内参基因的稳定性分析

发布时间：2018-03-23 06:01

本文选题：蓖麻　切入点：葡萄糖基氟虫腈　出处：《华南农业大学学报》2016年01期 　论文类型：期刊论文

【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,Sam DC,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ge Norm,Norm Finder,Best Keeper,Delta CT软件及Ref Finder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ge Norm:Actin=ef1aSam DCARCTUA6;Norm Finder:Sam DCARCActinef1aTUA6;Best Keeper:Actinef1aSam DCARCTUA6;Delta CT:Sam DCActinARCef1aTUA6;Ref Finder:ActinSam DCef1aARCTUA6,而单独分析DMSO处理时,稳定性排名则为:ef1aSam DCActinTUA6ARC。【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 the stable internal reference genes of castor Ricinus communis treated with glucosylfluoronitrile and dimethyl sulfoxide (DMSO), and to provide a reference for the study of GTF phloem loader. The gene expression was analyzed by real-time fluorescence quantitative PCR. Five candidate internal reference genes were treated with different time and different concentration of GTF and DMSO in the cotyledons of castor seedlings by using GE Norm Findern Best Keeper Delta CT software and Ref Finder online analysis tool. [results] the stability of the internal reference gene obtained by the software analysis was: ge Norm:Actin=ef1aSam DCARCTUA6, Norm Finder:Sam DCARCActinef1aTUA6, Best Keeper:Actinef1aSam DCARCTUA6, Delta CT:Sam DCDCActinARCef1aTUA6, CT:Sam DCActinARCef1aTUA6, Ref Finder:ActinSam DCCARCTUA6, and DMSO alone, The stability rank is:: ef1aSam DCActinTUA6ARC.ConclusionThe expression of actin in GTF and DMSO treatment is the most stable, and ef1a is the most stable in DMSO treatment.
【作者单位】：华南农业大学农学院/天然农药与化学生物学教育部重点实验室;
【基金】：国家自然科学基金(31171889) 高等学校博士学科点专项科研基金(20114404110020;20134404130003)
【分类号】：S435.656

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