Author:
Moura Jullyana Cristina Magalhães Silva,Araújo Pedro,Brito Michael dos Santos,Souza Uiara Romero,Viana Julianade Oliveira Fernandes,Mazzafera Paulo
Abstract
Abstract
Background
The genus Eucalyptus consists of approximately 600 species and subspecies and has a physiological plasticity that allows some species to propagate in different regions of the world. Eucalyptus is a major source of cellulose for paper manufacturing, and its cultivation is limited by weather conditions, particularly water stress and low temperatures. Gene expression studies using quantitative reverse transcription polymerase chain reaction (qPCR) require reference genes, which must have stable expression to facilitate the comparison of the results from analyses using different species, tissues, and treatments. Such studies have been limited in eucalyptus.
Results
Eucalyptus globulus Labill, Eucalyptus urograndis (hybrid from Eucalyptus urophylla S.T. Blake X Eucalyptus grandis Hill ex-Maiden) and E. uroglobulus (hybrid from E. urograndis X E. globulus) were subjected to different treatments, including water deficiency and stress recovery, low temperatures, presence or absence of light, and their respective controls. Except for treatment with light, which examined the seedling hypocotyl or apical portion of the stem, the expression analyses were conducted in the apical and basal parts of the stem. To select the best pair of genes, the bioinformatics tools GeNorm and NormFinder were compared. Comprehensive analyses that did not differentiate between species, treatments, or tissue types, showed that IDH (isocitrate dehydrogenase), SAND (SAND protein), ACT (actin), and A-Tub (α-tubulin) genes were the most stable. IDH was the most stable gene in all of the treatments.
Conclusion
Comparing these results with those of other studies on eucalyptus, we concluded that five genes are stable in different species and experimental conditions: IDH, SAND, ACT, A-Tub, and UBQ (ubiquitin). It is usually recommended a minimum of two reference genes is expression analysis; therefore, we propose that IDH and two others genes among the five identified genes in this study should be used as reference genes for a wide range of conditions in eucalyptus.
Publisher
Springer Science and Business Media LLC
Subject
General Biochemistry, Genetics and Molecular Biology,General Medicine
Reference58 articles.
1. Stape JL, Binkley D, Ryan MG: Eucalyptus production and the supply, use and efficiency of use of water, light and nitrogen across a geographic gradient in Brazil. Forest Ecol Manag. 2004, 193: 17-31. 10.1016/j.foreco.2004.01.020.
2. Mora AL, Garcia CH: The Eucalyptus culture in Brazil (A cultura do eucalipto no Brasil). 2000, São Paulo - Brazil: Sociedade Brasileira de Silvicultura-SBS
3. Grattapaglia D, Vaillancourt R, Shepherd M, Thumma B, Foley W, Külheim C, Potts B, Myburg A: Progress in Myrtaceae genetics and genomics: Eucalyptus as the pivotal genus. Tree Genet Genomes. 2012, 8: 463-508. 10.1007/s11295-012-0491-x.
4. Cotterill PP, Brolin A: Proceedings of the IUFRO Conference on Silviculture and Improvement of Eucalypts. Improving Eucalyptus wood, pulp and paper quality by genetic selection. 1997, Salvador, Brazil: EMBRAPA-Brazil, 1-14.
5. Pascoal-Neto C, Evtuguin D, Pinto P: Componentes macromoleculares das madeiras de Eucalyptus e de outras folhosas: estrutura e influencia na aptidão ao cozimento e branqueamento. O Papel - Aveiro. 2005, 1: 17-27.