Reference genes for gene expression studies in wheat flag leaves grown under different farming conditions

被引：41

作者：

Tenea G.N. ^{[1
]}

Peres Bota A. ^{[1
]}

Cordeiro Raposo F. ^{[1
]}

Maquet A. ^{[1
]}

机构：

[1] European Commission, Joint Research Centre (JRC), Institute for Reference Materials and Measurements (IRMM), 2440 Geel

来源：

BMC Research Notes | / 4卷 / 1期

关键词：

Reference Gene; Winter Wheat; Candidate Reference Gene; Flag Leave; Stable Reference Gene;

D O I：

10.1186/1756-0500-4-373

中图分类号：

学科分类号：

摘要：

Background: Internal control genes with highly uniform expression throughout the experimental conditions are required for accurate gene expression analysis as no universal reference genes exists. In this study, the expression stability of 24 candidate genes from Triticum aestivum cv. Cubus flag leaves grown under organic and conventional farming systems was evaluated in two locations in order to select suitable genes that can be used for normalization of real-time quantitative reverse-transcription PCR (RT-qPCR) reactions. The genes were selected among the most common used reference genes as well as genes encoding proteins involved in several metabolic pathways. Findings. Individual genes displayed different expression rates across all samples assayed. Applying geNorm, a set of three potential reference genes were suitable for normalization of RT-qPCR reactions in winter wheat flag leaves cv. Cubus: TaFNRII (ferredoxin-NADP(H) oxidoreductase; AJ457980.1), ACT2 (actin 2; TC234027), and rrn26 (a putative homologue to RNA 26S gene; AL827977.1). In addition of these three genes that were also top-ranked by NormFinder, two extra genes: CYP18-2 (Cyclophilin A, AY456122.1) and TaWIN1 (14-3-3 like protein, AB042193) were most consistently stably expressed. Furthermore, we showed that TaFNRII, ACT2, and CYP18-2 are suitable for gene expression normalization in other two winter wheat varieties (Tommi and Centenaire) grown under three treatments (organic, conventional and no nitrogen) and a different environment than the one tested with cv. Cubus. Conclusions: This study provides a new set of reference genes which should improve the accuracy of gene expression analyses when using wheat flag leaves as those related to the improvement of nitrogen use efficiency for cereal production. © 2010 Maquet et al; licensee BioMed Central Ltd.

引用

共 78 条

[71]

Rogers H.J., Greenland A.J., Hussey P.J., Four members of the maize beta-tubulin gene family are expressed in the male gametophyte, Plant Journal, 4, 5, pp. 875-882, (1993)

[72]

Smidansky E.D., Clancy M., Meyer F.D., Lanning S.P., Blake N.K., Talbert L.E., Giroux M.J., Enhanced ADP-glucose pyrophosphorylase activity in wheat endosperm increases seed yield, Proceedings of the National Academy of Sciences of the United States of America, 99, 3, pp. 1724-1729, (2002)

[73]

Rundle S.J., Zielinski R.E., Alterations in Barley Ribulose-1,5-bisphosphate Carboxylase/ Oxygenase Activase Gene Expression during Development and in Response to Illumination, Journal of Biological Chemistry, 266, 22, pp. 14802-14807, (1991)

[74]

Onda Y., Matsumura T., Kimata-Ariga Y., Sakakibara H., Sugiyama T., Hase T., Differential interaction of maize root ferredoxin:NADP<sup>+</sup> oxidoreductase with photosynthetic and non-photosynthetic ferredoxin isoproteins, Plant Physiol, 123, 3, pp. 1037-1046, (2000)

[75]

Lin C.-M., Koh S., Stacey G., Yu S.-M., Lin T.-Y., Tsay Y.-F., Cloning and functional characterization of a constitutively expressed nitrate transporter gene, OsNRT1, from rice, Plant Physiology, 122, 2, pp. 379-388, (2000)

[76]

Johnson P.E., Patron N.J., Bottrill A.R., Dinges J.R., Fahy B.F., Parker M.L., Waite D.N., Denyer K., A low-starch barley mutant, Risø 16, lacking the cytosolic small subunit of ADP-glucose pyrophosphorylase, reveals the importance of the cytosolic isoform and the identity of the plastidial small subunit, Plant Physiology, 131, 2, pp. 684-696, (2003)

[77]

Wostrikoff K., Stern D., Rubisco large-subunit translation is autoregulated in response to its assembly state in tobacco chloroplasts, Proceedings of the National Academy of Sciences of the United States of America, 104, 15, pp. 6466-6471, (2007)

[78]

Villand P., Aalen R., Olsen O.-A., Luthi E., Lonneborg A., Kleczkowski L.A., PCR amplification and sequences of cDNA clones for the small and large subunits of ADP-glucose pyrophosphorylase from barley tissues, Plant Mol Biol, 19, 3, pp. 381-389, (1992)

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