葡萄糖基氟虫腈处理下蓖麻内参基因的稳定性分析

【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=ef1a〉Sam DC〉ARC...

Full description

Saved in:
Bibliographic Details
Published in华南农业大学学报 Vol. 37; no. 1; pp. 52 - 57
Main Author 毛根林 解云 赵俊龙 陈炎 徐汉虹 林菲
Format Journal Article
LanguageChinese
Published 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,广东广州,510642 2016
Subjects
二甲基亚砜
内参基因
实时荧光定量PCR
稳定性
葡萄糖基氟虫腈
蓖麻
内参基因
internal reference gene
蓖麻
实时荧光定量PCR
DMSO
稳定性
GTF
葡萄糖基氟虫腈
RT-qPCR
二甲基亚砜
stability
Ricinus communis
Online AccessGet full text
ISSN1001-411X
DOI10.7671/j.issn.1001-411X.2016.01.009

Cover

More Information
Summary:【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=ef1a〉Sam DC〉ARC〉TUA6;Norm Finder：Sam DC〉ARC〉Actin〉ef1a〉TUA6;Best Keeper：Actin〉ef1a〉Sam DC〉ARC〉TUA6;Delta CT：Sam DC〉Actin〉ARC〉ef1a〉TUA6;Ref Finder：Actin〉Sam DC〉ef1a〉ARC〉TUA6,而单独分析DMSO处理时,稳定性排名则为：ef1a〉Sam DC〉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。
Bibliography: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,ef1 a,Sam DC 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 ge Norm,Norm Finder,Best Keeper,Delta CT and Ref Finder,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 ge Norm： Actin = ef1 a Sam DC ARC TUA6; NormFinder： Sam DC ARC Actin ef1 a TUA6; Best Keeper： Actin ef1 a Sam DC ARC TUA6;Delta CT： Actin ef1 a Sam DC ARC TUA6; Ref Finder： Actin Sam DC ef1 a ARC TUA6. With a single analysis of DMSO treatment by Ref Finder showed that ef1 a Sam DC
ISSN:1001-411X
DOI:10.7671/j.issn.1001-411X.2016.01.009