Reference genes for quantitative reverse transcription-polymerase chain reaction expression studies in wild and cultivated peanut

被引：64

作者：

Morgante C.V. ^{[1
,2
]}

Guimarães P.M. ^{[1
]}

Martins A.C.Q. ^{[1
,3
]}

Araújo A.C.G. ^{[1
]}

Leal-Bertioli S.C.M. ^{[1
]}

Bertioli D.J. ^{[3
,4
]}

Brasileiro A.C.M. ^{[1
]}

机构：

[1] EMBRAPA Recursos Genéticos e Biotecnologia, CP 02372. Final W5 Norte, Brasília, DF, Parque Estação Biolágica

[2] EMBRAPA Semiárido, Petrolina, PE

[3] Universidade de Brasília, Campus i, Brasília, DF

[4] Universidade Catálica de Brasília, Campus II, Brasília, DF

来源：

BMC Research Notes | / 4卷 / 1期

关键词：

Reference Gene; Arachis; Candidate Reference Gene; Internal Control Gene; Pairwise Variation;

D O I：

10.1186/1756-0500-4-339

中图分类号：

学科分类号：

摘要：

Background: Wild peanut species (Arachis spp.) are a rich source of new alleles for peanut improvement. Plant transcriptome analysis under specific experimental conditions helps the understanding of cellular processes related, for instance, to development, stress response, and crop yield. The validation of these studies has been generally accomplished by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) which requires normalization of mRNA levels among samples. This can be achieved by comparing the expression ratio between a gene of interest and a reference gene which is constitutively expressed. Nowadays there is a lack of appropriate reference genes for both wild and cultivated Arachis. The identification of such genes would allow a consistent analysis of qRT-PCR data and speed up candidate gene validation in peanut. Results: A set of ten reference genes were analyzed in four Arachis species (A. magna; A. duranensis; A. stenosperma and A. hypogaea) subjected to biotic (root-knot nematode and leaf spot fungus) and abiotic (drought) stresses, in two distinct plant organs (roots and leaves). By the use of three programs (GeNorm, NormFinder and BestKeeper) and taking into account the entire dataset, five of these ten genes, ACT1 (actin depolymerizing factor-like protein), UBI1 (polyubiquitin), GAPDH (glyceraldehyde-3-phosphate dehydrogenase), 60S (60S ribosomal protein L10) and UBI2 (ubiquitin/ribosomal protein S27a) emerged as top reference genes, with their stability varying in eight subsets. The former three genes were the most stable across all species, organs and treatments studied. Conclusions: This first in-depth study of reference genes validation in wild Arachis species will allow the use of specific combinations of secure and stable reference genes in qRT-PCR assays. The use of these appropriate references characterized here should improve the accuracy and reliability of gene expression analysis in both wild and cultivated Arachis and contribute for the better understanding of gene expression in, for instance, stress tolerance/resistance mechanisms in plants. © 2011Brasileiro et al; licensee BioMed Central Ltd.

引用

共 42 条

[1]

Isleib T.G., Pattee H.E., Giesbrecht F.G., Oil, sugar, and starch characteristics in peanut breeding lines selected for low and high oil content and their combining ability, J Agric Food Chem, 52, 10, pp. 3165-3168, (2004)

[2]

Leal-Bertioli S.C., Jose A.C., Alves-Freitas D.M., Moretzsohn M.C., Guimaraes P.M., Nielen S.A.C., Jensen J.L., Vidigal B.S., Pereira R.W., Pike J., Favero A.P., Identification of candidate genome regions controlling disease resistance in Arachis, BMC Plant Biol, 9, (2009)

[3]

Moretzsohn M.C., Barbosa A.V.G., Alves-Freitas D.M.T., Teixeira C., Leal-Bertioli S.C.M., Guimaraes P.M., Pereira R.W., Lopes C.R., Cavallari M.M., Valls J.F.M., A linkage map for the B-genome of Arachis (Fabaceae) and its synteny to the A-genome, BMC Plant Biol, 9, (2009)

[4]

Moretzsohn M.C., Leoi L., Proite K., Guimaraes P.M., Leal-Bertioli S.C.M., Gimenes M.A., Martins W.S., Valls J.F.M., Grattapaglia D., Bertioli D.J., A microsatellite-based, gene-rich linkage map for the AA genome of Arachis (Fabaceae), Theoretical and Applied Genetics, 111, 6, pp. 1060-1071, (2005)

[5]

Bertioli D.J., Moretzsohn M.C., Madsen L.H., Sandal N., Leal-Bertioli S.C., Guimaraes P.M., Hougaard B.K., Fredslund J., Schauser L., Nielsen A.M., An analysis of synteny of Arachis with Lotus and Medicago sheds new light on the structure, stability and evolution of legume genomes, BMC Genomics, 10, (2009)

[6]

Guimaraes P.M., Garsmeur O., Proite K., Leal-Bertioli S.C.M., Seijo G., Chaine C., Bertioli D.J., D'Hont A., BAC libraries construction from the ancestral diploid genomes of the allotetraploid cultivated peanut, BMC Plant Biology, 8, (2008)

[7]

Proite K., Leal-Bertioli S.C.M., Bertioli D.J., Moretzsohn M.C., Da Silva F.R., Martins N.F., Guimaraes P.M., ESTs from a wild Arachis species for gene discovery and marker development, BMC Plant Biology, 7, (2007)

[8]

Guimaraes P., Brasileiro A., Proite K., De Araujo A., Leal-Bertioli S., Pic-Taylor A., Da Silva F., Morgante C., Ribeiro S., Bertioli D., A study of gene expression in the nematode resistant wild peanut relative, Arachis stenosperma, in response to challenge with Meloidogyne arenaria, Trop Plant Biol, 3, 4, pp. 183-192, (2010)

[9]

Govind G., Thammegowda H.V., Kalaiarasi P.J., Iyer D.R., Muthappa S.K., Nese S., Makarla U.K., Identification and functional validation of a unique set of drought induced genes preferentially expressed in response to gradual water stress in peanut, Mol Genet Genomics, 281, 6, pp. 591-605, (2009)

[10]

Luo M., Dang P., Bausher M.G., Holbrook C.C., Lee R.D., Lynch R.E., Guo B.Z., Identification of transcripts involved in resistance responses to leaf spot disease caused by Cercosporidium personatum in peanut (Arachis hypogaea), Phytopathology, 95, 4, pp. 381-387, (2005)

← 1 2 3 4 5 →