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2014 | 36 | 03 |

Tytuł artykułu

Selection of reference genes for quantitative real-time PCR during flower bud development in CMS7311 of heading Chinese cabbage (Brassica rapa L. ssp. pekinensis)

Autorzy

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
The accuracy of quantitative real-time PCR (qRT-PCR) depends on the stability of the reference gene used for normalization. In heading Chinese cabbage (Brassica rapa L. ssp. pekinensis), the most stable reference genes for qRT-PCR during flower bud development have not been elucidated. In this study, the statistical software geNorm was used to test eight candidate reference genes during flower bud development in male sterile (Ms) and fertile (Mf) plants. The result revealed that the stability order was Tub/GAPDH > Cyp > EF1a > U34559 > BrTip41 > Apr > 18S rRNA, Tub and GAPDH were the most stable genes [average expression stability (M) 0.614], and the combined use of six reference genes [pairwise variation (V) 0.15] was suggested to be the optimal reference gene for qRT-PCR during flower bud development. Furthermore, the expressions of BcPME31 during flower bud development normalized with the combined use of six reference genes and with GAPDH or Tub alone were compared; the various results also suggested that selection of the optimal reference gene was necessary for gene expression analysis.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

36

Numer

03

Opis fizyczny

p.809-814,fig.,ref.

Twórcy

autor
  • College of Horticulture, State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A and F University, Yangling, 712100, Shaanxi, People’s Republic of China
  • College of Horticulture, Shanxi Agriculture University, Taigu, 030801, Shanxi, People’s Republic of China
autor
  • College of Horticulture, State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China
autor
  • College of Horticulture, State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A and F University, Yangling, 712100, Shaanxi, People’s Republic of China
autor
  • College of Horticulture, State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A and F University, Yangling, 712100, Shaanxi, People’s Republic of China
autor
  • College of Horticulture, State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A and F University, Yangling, 712100, Shaanxi, People’s Republic of China
  • Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

Bibliografia

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  • Andersen CL, Jensen JL, Orntoft TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64(15):5245–5250. doi:10.1158/0008-5472.can-04-0496
  • Chang E, Shi S, Liu J, Cheng T, Xue L, Yang X, Yang W, Lan Q, Jiang Z (2012) Selection of reference genes for quantitative gene expression studies in Platycladus orientalis (Cupressaceae) using real-time PCR. PLoS One 7(3):e33278. doi:10.1371/journal.pone.0033278
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  • Czechowski T, Stitt M, Altmann T, Udvardi MK, Scheible W-R (2005) Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol 139(1):5–17. doi:10.1104/pp.105.063743
  • de Oliveira LA, Breton MC, Bastolla FM, Camargo Sda S, Margis R, Frazzon J, Pasquali G (2012) Reference genes for the normalization of gene expression in eucalyptus species. Plant Cell Physiol 53(2):405–422. doi:10.1093/pcp/pcr187
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  • Han LQ, Yang GY, Zhu HS, Wang YY, Wang LF, Guo YJ, Lu WF, Li HJ, Wang YL (2010) Selection and use of reference genes in mouse mammary glands. Genet Mol Res 9(1):449–456
  • Liu LC, Yu XL, Ye WZ, Xiang X, Cao JS (2006) Functional confirmation of BcMF3 gene related to male sterility of Chinese cabbage by antisense RNA. J Zhejiang Univ (Agric Life Sci) 32(05):473–478 (in Chinese)
  • Mallona I, Lischewski S, Weiss J, Hause B, Egea-Cortines M (2010) Validation of reference genes for quantitative real-time PCR during leaf and flower development in Petunia hybrida. BMC Plant Biol 10:4. doi:10.1186/1471-2229-10-4
  • Pfaffl MW, Tichopad A, Prgomet C, Neuvians TP (2004) Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper—Excel-based tool using pair-wise correlations. Biotechnol Lett 26(6):509–515. doi:10.1023/b:bile.0000019559.84305.47
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  • Ramakers C, Ruijter JM, Deprez RH, Moorman AF (2003) Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data. Neurosci Lett 339(1):62–66
  • Silveira ED, Alves-Ferreira M, Guimaraes LA, da Silva FR, Carneiro VT (2009) Selection of reference genes for quantitative real-time PCR expression studies in the apomictic and sexual grass Brachiaria brizantha. BMC Plant Biol 9(84):1471–2229
  • Tian GW, Chen MH, Zaltsman A, Citovsky V (2006) Pollen-specific pectin methylesterase involved in pollen tube growth. Dev Biol 294(1):83–91. doi:10.1016/j.ydbio.2006.02.026
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  • Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3(7):research0034.1–research0034.11. doi:10.1186/gb-2002-3-7-research0034
  • Wan H, Zhao Z, Qian C, Sui Y, Malik AA, Chen J (2010) Selection of appropriate reference genes for gene expression studies by quantitative real-time polymerase chain reaction in cucumber. Anal Biochem 399(2):257–261. doi:10.1016/j.ab.2009.12.008
  • Wang Y, Yu K, Poysa V, Shi C, Zhou Y (2012) Selection of reference genes for normalization of qRT-PCR analysis of differentially expressed genes in soybean exposed to cadmium. Mol Biol Rep 39(2):1585–1594
  • Xu X, Sun X, Zhang J, Huang W, Zhang L, Fang Z (2013) Identification of candidate genes associated with male sterility in CMS7311 of heading Chinese cabbage (Brassica campestris L. ssp. pekinensis). Acta Physiologiae Plantarum 35(11):3265–3270. doi:10.1007/s11738-013-1346-2
  • Zhang JY, Li Y, Shi GJ, Chen XF, Wang JJ, Hou XL (2009) Characterization of alpha-tubulin gene distinctively presented in a cytoplasmic male sterile and its maintainer line of non-heading Chinese cabbage. J Sci Food Agric 89(2):274–280. doi:10.1002/Jsfa.3438
  • Zhang CW, Qi L, Hou XL, Shi GJ, Zhang JY (2010) Differential gene expression analysis of a new Ogura CMS line and its maintainer in non-heading Chinese cabbage by cDNA-AFLP. Acta Physiologiae Plantarum 32(4):781–787. doi:10.1007/s11738-010-0463-4

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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