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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Establishing references for gene expression analyses by RT-qPCR in Theobroma cacao tissues

Full text
Author(s):
Pinheiro, T. T. [1] ; Litholdo, Jr., C. G. [1] ; Sereno, M. L. [1] ; Leal, Jr., G. A. [1] ; Albuquerque, P. S. B. [2] ; Figueira, A. [1]
Total Authors: 6
Affiliation:
[1] Univ Sao Paulo, Ctr Energia Nucl Agr, Piracicaba, SP - Brazil
[2] Comissao Execut Plano Lavoura Cacaueira, ERJOH, Marituba, PA - Brazil
Total Affiliations: 2
Document type: Journal article
Source: Genetics and Molecular Research; v. 10, n. 4, p. 3291-3305, 2011.
Web of Science Citations: 19
Abstract

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. (AU)

FAPESP's process: 07/07175-0 - Genetic analysis of Theobroma cacao defense response against Moniliophthora perniciosa, the causal agent of witches broom
Grantee:Antonio Vargas de Oliveira Figueira
Support Opportunities: Research Projects - Thematic Grants