4.6 Article

Selection of reference genes for expression analysis in the potato psyllid, Bactericera cockerelli

Journal

INSECT MOLECULAR BIOLOGY
Volume 25, Issue 3, Pages 227-238

Publisher

WILEY
DOI: 10.1111/imb.12219

Keywords

Psyllids; Bactericera cockerelli; Liberibacter; reference genes; gene expression; polymerase chain reaction

Funding

  1. U.S. Department of Agriculture-National Institute of Food and Agriculture (USDA-NIFA) [2012-67013-19431]
  2. NIFA [2012-67013-19431, 578723] Funding Source: Federal RePORTER

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The selection of reference genes is a crucial step for quantitative real-time PCR analyses and increasingly the use of more than one reference gene for accurate and reliable normalization is being recommended. In this study, a set of six genes was selected and their stability was assessed in different life stages and female organs of Bactericera cockerelli harbouring or not the bacterial pathogen 'Candidatus Liberibacter solanacearum' (Lso) haplotype B. The stability of each gene was determined using the BESTKEEPER, NORMFINDER and GENORM programs. These analyses identified elongation factor-1a, ribosomal protein subunit L5 and ribosomal protein subunit 18 as the most stable genes to analyse gene expression during the insect life stages irrespective of Lso presence; Lso haplotype B only affected their respective ranking. By contrast, a common set of normalizers could not be found amongst the different female organs tested (bacteriomes, alimentary canals and reproductive organs).

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