PMID- 22095481
OWN - NLM
STAT- MEDLINE
DCOM- 20120904
LR  - 20220408
IS  - 1676-5680 (Electronic)
IS  - 1676-5680 (Linking)
VI  - 10
IP  - 4
DP  - 2011 Nov 17
TI  - Establishing references for gene expression analyses by RT-qPCR in Theobroma 
      cacao tissues.
PG  - 3291-305
LID - 10.4238/2011.November.17.4 [doi]
AB  - Lack of continuous progress in Theobroma cacao (Malvaceae) breeding, especially 
      associated with seed quality traits, requires more efficient selection methods 
      based on genomic information. Reverse transcript quantitative PCR (RT-qPCR) has 
      become the method of choice for gene expression analysis, but relative expression 
      analysis requires various reference genes, which must be stable across various 
      biological conditions. We sought suitable reference genes for various tissues of 
      cacao, especially developing seeds. Ten potential reference genes were analyzed 
      for stability at various stages of embryo development, leaves, stems, roots, 
      flowers, and pod epicarp; seven of them were also evaluated in shoot tips treated 
      either with hormones (salicylate; ethefon; methyl-jasmonate) or after inoculation 
      with the fungus Moniliophthora perniciosa (Marasmiaceae sensu lato). For 
      developing embryos, the three most stable genes were actin (ACT), polyubiquitin 
      (PUB), and ribosomal protein L35 (Rpl35). In the analyses of various tissues, the 
      most stable genes were malate dehydrogenase (MDH), glyceraldehyde 3-phosphate 
      dehydrogenase (GAPDH), and acyl-carrier protein B (ACP B). GAPDH, MDH and tubulin 
      (TUB) were the most appropriate for normalization when shoot apexes were treated 
      with hormones, while ACT, TUB and Rpl35 were the most appropriate after 
      inoculation with M. perniciosa. We conclude that for each plant system and 
      biological or ontogenetical condition, there is a need to define suitable 
      reference genes. This is the first report to define reference genes for 
      expression studies in cacao.
FAU - Pinheiro, T T
AU  - Pinheiro TT
AD  - Centro de Energia Nuclear na Agricultura, Universidade de Sao Paulo, Piracicaba, 
      SP, Brasil.
FAU - Litholdo, C G Jr
AU  - Litholdo CG Jr
FAU - Sereno, M L
AU  - Sereno ML
FAU - Leal, G A Jr
AU  - Leal GA Jr
FAU - Albuquerque, P S B
AU  - Albuquerque PS
FAU - Figueira, A
AU  - Figueira A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20111117
PL  - Brazil
TA  - Genet Mol Res
JT  - Genetics and molecular research : GMR
JID - 101169387
SB  - IM
MH  - Cacao/*genetics
MH  - Gene Expression Profiling
MH  - *Gene Expression Regulation, Plant
MH  - Genes, Plant/genetics
MH  - Genetic Association Studies
MH  - Organ Specificity/*genetics
MH  - Reference Standards
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods/*standards
MH  - Software
EDAT- 2011/11/19 06:00
MHDA- 2012/09/05 06:00
CRDT- 2011/11/19 06:00
PHST- 2011/11/19 06:00 [entrez]
PHST- 2011/11/19 06:00 [pubmed]
PHST- 2012/09/05 06:00 [medline]
AID - gmr1337 [pii]
AID - 10.4238/2011.November.17.4 [doi]
PST - epublish
SO  - Genet Mol Res. 2011 Nov 17;10(4):3291-305. doi: 10.4238/2011.November.17.4.