Screening of valid reference genes for real-time RT-PCR data normalization in Hevea brasiliensis and expression validation of a sucrose transporter gene HbSUT3.
Plant Sci
; 181(2): 132-9, 2011 Aug.
Article
in En
| MEDLINE
| ID: mdl-21683878
ABSTRACT
Real-time RT-PCR (RT-qPCR) is a sensitive and precise method of quantifying gene expression, however, suitable reference genes are required. Here, a systematic reference gene screening was performed by RT-qPCR on 22 candidate genes in Hevea brasiliensis. Two ubiquitin-protein ligases (UBC2a and UBC4) were the most stable when all samples were analyzed together. A mitosis protein (YLS8) and a eukaryotic translation initiation factor (eIF1Aa) were the most stable in response to tapping. UBC2b and UBC1 were the most stable among different genotypes. UBC2b and a DEAD box RNA helicase (RH2b) were the most stable across individual trees. YLS8 and RH8 were most stably expressed in hormone-treated samples. Expression of the candidate reference genes varied significantly across different tissues, and at least four genes (RH2b, RH8, UBC2a and eIF2) were needed for expression normalization. In addition, examination of relative expression of a sucrose transporter HbSUT3 in different RNA samples demonstrated the importance of additional reference genes to ensure accurate quantitative expression analysis. Overall, our work serves as a guide for selection of reference genes in RT-qPCR gene expression studies in H. brasiliensis.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Genes, Plant
/
Reverse Transcriptase Polymerase Chain Reaction
/
Hevea
Type of study:
Diagnostic_studies
/
Screening_studies
Country/Region as subject:
America do sul
/
Brasil
Language:
En
Journal:
Plant Sci
Year:
2011
Document type:
Article
Affiliation country:
China