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Approaches to obtain grapefruit-like hybrids; diploid hybrids were recovered from 2x 2x sexual hybridization, and triploid hybrids were recovered from 4x 2x sexual hybridization. We applied diploid and tetraploid “Clemenules” clementines as female monoembryonic and self-incompatible parents (hereafter Clem 2x and Clem 4x, respectively) and “Pink” pummelo as male parent (hereafter Pum 2x). Clem 4x was obtained by shoot tip grafting in vitro combined with colchicine therapy (Aleza et al., 2009). Sensible specifics around the recovery of diploid and triploid hybrids from sexual hybridizations followed by embryo rescue and flow cytometry could be found in Aleza et al. (2010, 2012b), respectively. From these two populations, 116 diploid and 91 triploid hybrids had been recovered and grafted onto “Carrizo” citrange rootstock (C. sinensis P. trifoliata) in 2011 and 2007 respectively, for field evaluation at IVIA plots. Fifteen diploid and 15 triploid hybrids (numbers 15 for diploids and 160 for triploids along the text) have been selected Adenosine A2A receptor (A2AR) Inhibitor MedChemExpress randomly to Adenosine A3 receptor (A3R) Antagonist Source perform this study. Fruit samples had been taken at 3 harvest occasions (December, January, and February) and in three consecutive seasons, 2015/2016, 2016/2017, and 2017/2018. In parallel, 4 diploid grapefruits, “Flame,” “Rio Red,” “Star Ruby,” and “Duncan,” along with their corresponding tetraploid genotypes, were analyzed to study the impact of ploidy level on FC and flavonoid compounds. Tetraploid grapefruits have been obtained from spontaneous chromosome doubling of nucellar cells (Aleza et al., 2009).GenotypingSelected diploid and triploid hybrids with each other with their parents have been analyzed with six uncomplicated sequence repeat (SSR) markers [TAA41 (Kijas et al., 1997), MEST56 (Aleza et al., 2009), mCrCIR07B05 and Ci07C09 (Froelicher et al., 2008), mCrCIR02F12 and mCrCIR01C06 (Cuenca et al., 2011)], displaying heterozygosity and polymorphism among parents (A1 A2 A3 A4 ). The genetic configuration and allele dosage of diploid and triploid hybrids have been inferred straight from electroferograms. Genomic DNA was isolated employing a Plant DNeasy kit from Qiagen Inc. (Valencia, CA, United states of america) following the manufacturer’s protocol. PCR circumstances and data evaluation were as reported by Garcia-Lor et al. (2012). Denaturation and capillary electrophoresis were carried out on a Capillary Gel Electrophoresis CEQTM 8000 Genetic Evaluation Method employing linear polyacrylamide based on the manufacturer’s directions (Beckman Coulter Inc.). Genetic evaluation program computer software (GenomeLabTM GeXP version 10.0) was made use of for dataPhenotypic Information AnalysisA factorial analysis was performed for the phenotypic information using DARwin5 (Dissimilarity Analysis and Representation for Windows) software version 5.0.159 (Perrier and JacquemoudCollet, 2006). For the analysis with the FC and flavonoid contents, the complete dataset of all replicates from the three collection dates on the totally completed season 2015/2016 was used. Single data file format was applied as DARwin input, where every unitFrontiers in Plant Science | www.frontiersin.orgFebruary 2021 | Volume 12 | ArticleGarcia-Lor et al.Breeding Grapefruit-Like Citrus Varietieswas characterized by a single value for each variable. The dissimilarity matrix was calculated working with the usual Euclidean index for continuous information. This process performs on distance matrices, thinking of the higher dimension space defined by the distances among two units. Factorial procedures aim mainly to provide an general representation of.