Strategies to obtain grapefruit-like hybrids; diploid hybrids had been recovered from 2x 2x sexual hybridization, and triploid hybrids have been recovered from 4x 2x sexual hybridization. We made use of 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 treatment (Aleza et al., 2009). Practical details around the recovery of diploid and triploid hybrids from sexual hybridizations followed by embryo rescue and flow cytometry can be discovered in Aleza et al. (2010, 2012b), respectively. From these two populations, 116 diploid and 91 triploid hybrids have 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) were selected randomly to execute this study. Fruit samples had been taken at 3 harvest occasions (December, January, and February) and in 3 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 effect of ploidy level on FC and flavonoid compounds. Tetraploid grapefruits were obtained from spontaneous chromosome doubling of nucellar cells (Aleza et al., 2009).GenotypingSelected diploid and triploid hybrids together with their parents were analyzed with six basic 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 amongst 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 applying a Plant DNeasy kit from Qiagen Inc. (Valencia, CA, United PARP14 Purity & Documentation states) following the manufacturer’s protocol. PCR conditions and information analysis had been as reported by Garcia-Lor et al. (2012). Denaturation and capillary electrophoresis have been carried out on a Capillary Gel Electrophoresis CEQTM 8000 Genetic Analysis Program working with linear polyacrylamide according to the manufacturer’s directions (Beckman Coulter Inc.). Genetic evaluation method software (GenomeLabTM GeXP version ten.0) was used for dataPhenotypic Data RGS19 Formulation AnalysisA factorial evaluation was performed for the phenotypic data employing DARwin5 (Dissimilarity Analysis and Representation for Windows) software program version five.0.159 (Perrier and JacquemoudCollet, 2006). For the analysis on the FC and flavonoid contents, the full dataset of all replicates in the 3 collection dates from the completely completed season 2015/2016 was utilized. Single data file format was made use of 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 worth for each variable. The dissimilarity matrix was calculated applying the usual Euclidean index for continuous data. This strategy functions on distance matrices, considering the high dimension space defined by the distances amongst two units. Factorial methods aim mainly to provide an overall representation of.