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Perini P.,Federal University of Rio Grande do Sul | Pasquali G.,Federal University of Rio Grande do Sul | Margis-Pinheiro M.,Federal University of Rio Grande do Sul | de Oliviera P.R.D.,Laboratorio Of Genetica Molecular Vegetal | Revers L.F.,Laboratorio Of Genetica Molecular Vegetal
Molecular Breeding | Year: 2014

Apple (Malus × domestica Borkh.) is the most important deciduous tree fruit crop grown around the world. Comparisons of gene expression profiles from different tissues, conditions or cultivars are valuable scientific tools to better understand the gene expression changes behind important silvicultural and nutritional traits. However, the accuracy of techniques employed to access gene expression is dependent on the evaluation of stable reference genes for data normalization to avoid statistical significance undue or incorrect conclusions. The objective of this work was to select the best genes to be used as references for gene expression studies in apple trees by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Vegetative and reproductive tissues of the apple "Gala" cultivar were evaluated during their seasonal cycle of growth and dormancy. The expression of 23 traditional housekeeping genes or genes suggested as constitutive by microarray data was investigated. Tested combinations of primers allowed the specific amplification and the generation of suitable efficiency curves for gene expression studies by RT-qPCR. Gene stability was determined by two different statistical descriptors, geNorm and NormFinder. The known variable PAL gene expression was used to validate selected normalizers. Results obtained allowed us to conclude that MDH, SAND, THFS, TMp1 and WD40 are the best reference genes to accurately normalize the relative transcript abundances using RT-qPCR in various tissues of apple. © 2014 Springer Science+Business Media Dordrecht.

Cely J.A.B.,Laboratorio Of Genetica Molecular Vegetal | Rodriguez F.E.,Laboratorio Of Genetica Molecular Vegetal | Almario C.G.,Lider Laboratorio Of Microbiologia Molecular | Meneses L.S.B.,Lider Laboratorio Of Genetica Molecular Vegetal
Revista Brasileira de Fruticultura | Year: 2015

Cape gooseberry, Physalis peruviana, is an Andean fruit of great importance for export markets; its main limitation for production in Colombia is the vascular wilt caused by Fusarium oxysporum. The present study proposed the generation of F1 populations between contrasting pathogen response parents and their evaluation at molecular level to support knowledge and use of the species genetics resources. To do this, four genotypes of P. peruviana, and one of the related species P. floridana, were characterized at morphological level using 34 qualitative and 20 quantitative variables and at molecular level using 328 COSII and 154 IRGs markers. The genotypes were used as parents for generation and molecular characterization of F1 populations. Quantitative variables were able to distinguish the species P. floridana and P. peruviana as well as cultivated and wild genotypes within P. peruviana. One hundred percent viability was found in intraspecific F1 crosses and 50% in interspecific crosses, remaining viable only when P. floridana was the receptor of pollen. Molecular characterization did not identify polymorphisms within the P. peruviana but between P. floridana and P. peruviana. An F1 population of 51 individuals generated between the species a total of 127 alleles with an average of 3.18 per locus, a PIC of 0.358 and high values of heterozygosity (Ho: 0.737 and He: 0.449). PCA and cluster analysis allowed discrimination of the F1 population in three groups, more similar to the P. floridana parental. This was reflected by a 75% Mendelian distortion favored by the presence of 63.75% maternal alleles. The study provides insights into Cape gooseberry crossability and the genetic variability of parental genotypes and F1 populations. © 2015, Sociedade Brasileira de Fruticultura. All rights reserved.

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