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Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]
2016 Jan-Mar;47 (1): 259-65.

Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis.

Chun Chen, Tingna Xie, Sudan Ye, Annette Bruun Jensen, Jørgen Eilenberg
Author Information
  1. Chun Chen: China Jiliang University, Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, Hangzhou 310018, China. Electronic address: aspring@cjlu.edu.cn.
  2. Tingna Xie: China Jiliang University, Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, Hangzhou 310018, China.
  3. Sudan Ye: Zhejiang Economic & Trade Polytechnic, Hangzhou 310018, China.
  4. Annette Bruun Jensen: Department of Plant and Environmental Sciences, University of Copenhagen, DK 1871 Frederiksberg C, Denmark.
  5. Jørgen Eilenberg: Department of Plant and Environmental Sciences, University of Copenhagen, DK 1871 Frederiksberg C, Denmark.
PMID: 26887253 DOI: 10.1016/j.bjm.2015.11.031

Abstract

The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host-pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414) and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis.

Keywords

Pandora neoaphidis Quantitative PCR Reference genes

References

  1. Mycol Res. 2006 Oct;110(Pt 10):1165-71 [PMID: 17010593]
  2. Mol Biol Rep. 2013 Dec;40(12):6691-9 [PMID: 24057254]
  3. Biotechnol Lett. 2003 Nov;25(21):1869-72 [PMID: 14677714]
  4. Appl Environ Microbiol. 2009 Jun;75(11):3787-95 [PMID: 19363067]
  5. Genome Biol. 2002 Jun 18;3(7):RESEARCH0034 [PMID: 12184808]
  6. BMC Mol Biol. 2009;10:94 [PMID: 19818117]
  7. Methods. 2001 Dec;25(4):402-8 [PMID: 11846609]
  8. FEMS Microbiol Lett. 2014 Jul;356(1):97-104 [PMID: 24953133]
  9. Nucleic Acids Res. 2000 Mar 15;28(6):1283-98 [PMID: 10684922]
  10. Biotechniques. 2000 Aug;29(2):332-7 [PMID: 10948434]
  11. Fungal Biol. 2011 Sep;115(9):891-901 [PMID: 21872186]
  12. Appl Environ Microbiol. 2010 Jan;76(1):254-63 [PMID: 19854926]
  13. Mol Plant Pathol. 2010 Nov;11(6):805-16 [PMID: 21029324]
  14. BMC Mol Biol. 2006;7:33 [PMID: 17026756]
  15. Planta. 2012 Nov;236(5):1381-93 [PMID: 22718310]
  16. Biochem Biophys Res Commun. 2011 Oct 7;413(4):537-40 [PMID: 21925148]
  17. Genetics. 2007 Feb;175(2):477-85 [PMID: 17322354]
  18. Biotechnol Lett. 2004 Mar;26(6):509-15 [PMID: 15127793]
  19. J Biotechnol. 1999 Oct 8;75(2-3):291-5 [PMID: 10617337]
  20. Fungal Genet Biol. 2011 Apr;48(4):343-52 [PMID: 21156213]
  21. Braz J Microbiol. 2012 Jan;43(1):325-31 [PMID: 24031835]
  22. J Mol Endocrinol. 2005 Jun;34(3):597-601 [PMID: 15956331]
  23. Anal Biochem. 2005 Sep 1;344(1):141-3 [PMID: 16054107]
  24. Genes Immun. 2005 Jun;6(4):279-84 [PMID: 15815687]
  25. J Exp Bot. 2009;60(2):487-93 [PMID: 19264760]
  26. Exp Hematol. 2002 Jun;30(6):503-12 [PMID: 12063017]
  27. PLoS One. 2012;7(8):e43028 [PMID: 22916200]
  28. BMC Mol Biol. 2010;11:73 [PMID: 20854682]
  29. Appl Environ Microbiol. 2010 Apr;76(7):2262-70 [PMID: 20139313]
  30. Cancer Res. 2004 Aug 1;64(15):5245-50 [PMID: 15289330]
  31. BMC Mol Biol. 2009;10:99 [PMID: 19874630]
  32. J Appl Genet. 2013 Nov;54(4):391-406 [PMID: 24078518]
  33. Environ Microbiol. 2006 Jan;8(1):69-76 [PMID: 16343323]
  34. Gene. 2014 Aug 15;547(1):55-62 [PMID: 24914494]

MeSH Term

Entomophthorales
Gene Expression Profiling
Genes, Fungal
Peptide Elongation Factor 1
RNA, Ribosomal, 18S
RNA, Ribosomal, 28S
Real-Time Polymerase Chain Reaction
Reference Standards

Chemicals

Peptide Elongation Factor 1
RNA, Ribosomal, 18S
RNA, Ribosomal, 28S
Journal Article Research Support, Non-U.S. Gov't

Word Cloud

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