Cytogenetic and flow cytometry data expand knowledge of genome evolution in three Coffea species

Abstract

Karyotype and nuclear 2C-value data are considered important in taxonomic and evolutionary approaches in Coffea. Still, new methods are needed to further support such studies, especially to determine the progenitors of Coffea arabica. In this work, new cytogenetic and flow cytometry data were used to compare Coffea arabica, Coffea canephora and Coffea congensis. These data corroborate the hypothesis that C. canephora and C. congensis originated from a single ancestor, whose basic chromosome number was x = 11. In agreement with the observations of other authors, the karyotype and mean 2C-values confirm that C. arabica is a true allotetraploid originating from two diploid Coffea species with similar genomes. Although C. canephora and C. congensis have been considered potential progenitors of C. arabica, karyotype comparison revealed that only one of these species may be parental to C. arabica. These accurate cytogenetic and flow cytometry data contribute to expand our knowledge of the Coffea genome, as well as of possible progenitors of C. arabica.

Regeneration of Allotriploid <i>Coffea</i> Plants from Tissue Culture: Resolving the Propagation Problems Promoted by Irregular Meiosis

“…According to Doležel and Bartoš (2005), histograms with peak CVs lower than 5% have been considered acceptable for FCM assessments in plants. However, as a consequence of the high production of secondary metabolites, obtaining CV values at this level for coffee plants has been considered very difficult (Clarindo et al 2012). Nevertheless, the appropriate FCM procedures adopted in the present work enabled the reduction of secondary metabolite interference with nuclear chromatin staining, providing intact and stoichiometrically stained nuclei as well as low CV values.…”

“…After 24 h, the leaves were surfacewashed by rinsing with liquid detergent, and kept under running tap water for 2 h. Next, in a laminar flow hood, the leaves were disinfected by immersion in 70% ethanol (Merck ® ) for 20 s, followed by 1.5% sodium hypochlorite (Merck ® ) solution (containing 10 drops L 1 of Tween 20, Sigma ® ) for 20 min, and subsequently washed four times with sterile dH 2 O (Clarindo et al 2012).…”

Analysis of Genome Size of Sixteen Coffea arabica Cultivars Using Flow Cytometry

“…Experimental design. The experimental design was adopted following the recommendations of Clarindo et al (2012) for nuclei extraction from multiple leaves, from two standards, soybean (2C = 2.30 pg and C. canephora (2C = 1.29 pg) (values obtained in the present work). FCM parameters, such as gain and channel, were determined for each DNA content measurement, based on FCM assessments of standard and samples (control and target) as described by Clarindo et al (2013).…”

“…Likewise, some variations in nuclear DNA content between species are possibly artifacts caused by the inability of the dye to access DNA, since the fluorescent stains PI and DAPI can yield significantly different values for the same sample (Noirot et al, 2002;2003b). The studies on nuclear DNA content in C. arabica cultivars by FCM have been reported Clarindo and Carvalho, 2009;Clarindo et al, 2012Clarindo et al, , 2013Sanglard et al, 2017;Sattler et al, 2016), and therefore additional studies on more cultivars will be helpful for breeding programs, as well as, from a basic standpoint, for phylogenetic analysis of coffee cultivars.…”

FLOW CYTOMETRIC ANALYSIS OF NUCLEAR DNA BETWEEN OKRA LANDRACES (<i>ABELMOSCHUS ESCULENTUS</i> L.)

“…Genome size of different animals e.g Dairy goat (Fletcher et al, 2013) as well as different, plants has been estimated by flow cytometry like Pinus (O'Brien et al, 1996), Lemon (Iannelli et al, 1998), Arachis hypogeal (Temsch and Greilhuber, 2000), Musa (Roux et al, 2003), Atriplex halimus (Walker et al, 2005), Consolea (Negron-Ortiz, 2007), Vicia faba (Kovarova et al, 2007) Cactus (Lema-Ruminska, 2011), Coffea (Clarindo et al, 2012), Calendula species (Nora et al, 2013), Eryngium (Tavares et al, 2013), Lathyrus (Ochatt et al, 2013) and Phragmites austalis (Nakagawa et al, 2013).…”