PMID- 25048176
OWN - NLM
STAT- MEDLINE
DCOM- 20150223
LR  - 20220316
IS  - 1756-0500 (Electronic)
IS  - 1756-0500 (Linking)
VI  - 7
DP  - 2014 Jul 22
TI  - Identification and validation of quantitative real-time reverse transcription PCR 
      reference genes for gene expression analysis in teak (Tectona grandis L.f.).
PG  - 464
LID - 10.1186/1756-0500-7-464 [doi]
AB  - BACKGROUND: Teak (Tectona grandis L.f.) is currently the preferred choice of the 
      timber trade for fabrication of woody products due to its extraordinary qualities 
      and is widely grown around the world. Gene expression studies are essential to 
      explore wood formation of vascular plants, and quantitative real-time reverse 
      transcription PCR (qRT-PCR) is a sensitive technique employed for quantifying 
      gene expression levels. One or more appropriate reference genes are crucial to 
      accurately compare mRNA transcripts through different tissues/organs and 
      experimental conditions. Despite being the focus of some genetic studies, a lack 
      of molecular information has hindered genetic exploration of teak. To date, 
      qRT-PCR reference genes have not been identified and validated for teak. RESULTS: 
      Identification and cloning of nine commonly used qRT-PCR reference genes from 
      teak, including ribosomal protein 60s (rp60s), clathrin adaptor complexes medium 
      subunit family (Cac), actin (Act), histone 3 (His3), sand family (Sand), 
      beta-Tubulin (Beta-Tub), ubiquitin (Ubq), elongation factor 1-alpha (Ef-1alpha), and 
      glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Expression profiles of these 
      genes were evaluated by qRT-PCR in six tissue and organ samples (leaf, flower, 
      seedling, root, stem and branch secondary xylem) of teak. Appropriate gene 
      cloning and sequencing, primer specificity and amplification efficiency was 
      verified for each gene. Their stability as reference genes was validated by 
      NormFinder, BestKeeper, geNorm and Delta Ct programs. Results obtained from all 
      programs showed that TgUbq and TgEf-1alpha are the most stable genes to use as 
      qRT-PCR reference genes and TgAct is the most unstable gene in teak. The relative 
      expression of the teak cinnamyl alcohol dehydrogenase (TgCAD) gene in lignified 
      tissues at different ages was assessed by qRT-PCR, using TgUbq and TgEf-1alpha as 
      internal controls. These analyses exposed a consistent expression pattern with 
      both reference genes. CONCLUSION: This study proposes a first broad collection of 
      teak tissue and organ mRNA expression data for nine selected candidate qRT-PCR 
      reference genes. NormFinder, Bestkeeper, geNorm and Delta Ct analyses suggested 
      that TgUbq and TgEf-1alpha have the highest expression stability and provided similar 
      results when evaluating TgCAD gene expression, while the commonly used Act should 
      be avoided.
FAU - Galeano, Esteban
AU  - Galeano E
FAU - Vasconcelos, Tarcisio Sales
AU  - Vasconcelos TS
FAU - Ramiro, Daniel Alves
AU  - Ramiro DA
FAU - De Martin, Valentina de Fatima
AU  - De Martin Vde F
FAU - Carrer, Helaine
AU  - Carrer H
AD  - Departamento de Ciencias Biologicas, Escola Superior de Agricultura "Luiz de 
      Queiroz", Universidade de Sao Paulo, Av, Padua Dias, 11, Piracicaba, SP 
      13418-900, Brazil. hecarrer@usp.br.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140722
PL  - England
TA  - BMC Res Notes
JT  - BMC research notes
JID - 101462768
RN  - 0 (DNA Primers)
RN  - 0 (Peptide Elongation Factor 1)
RN  - 0 (Ubiquitin)
SB  - IM
MH  - DNA Primers/chemistry
MH  - Flowers/genetics
MH  - Gene Expression Profiling
MH  - *Gene Expression Regulation, Plant
MH  - *Genes, Essential
MH  - *Genes, Plant
MH  - Lamiaceae/*genetics
MH  - Peptide Elongation Factor 1/*genetics
MH  - Plant Leaves/genetics
MH  - Plant Roots/genetics
MH  - Plant Stems/genetics
MH  - Real-Time Polymerase Chain Reaction/standards
MH  - Reverse Transcriptase Polymerase Chain Reaction/standards
MH  - Seedlings/genetics
MH  - Ubiquitin/*genetics
PMC - PMC4114093
EDAT- 2014/07/23 06:00
MHDA- 2015/02/24 06:00
PMCR- 2014/07/22
CRDT- 2014/07/23 06:00
PHST- 2014/02/11 00:00 [received]
PHST- 2014/07/16 00:00 [accepted]
PHST- 2014/07/23 06:00 [entrez]
PHST- 2014/07/23 06:00 [pubmed]
PHST- 2015/02/24 06:00 [medline]
PHST- 2014/07/22 00:00 [pmc-release]
AID - 1756-0500-7-464 [pii]
AID - 10.1186/1756-0500-7-464 [doi]
PST - epublish
SO  - BMC Res Notes. 2014 Jul 22;7:464. doi: 10.1186/1756-0500-7-464.