Phylogenetic Relationships among Portuguese Rye Based on Isozyme, RAPD and ISSR Markers

Abstract

The phylogenetic relationships of 10 rye landraces and cultivars from the north of Portugal and from Brazil were analysed using 20 isozyme loci, and a total of 511 PCR markers (342 ISSRs and 169 RAPDs). The isozymes were analysed in at least 100 plants of each population/cultivar and, therefore, we have data about intra and inter population/cultivar genetic variability. However, the analyses with ISSRs and RAPDs were obtained using a mix of 25 plants of each population. Therefore, each population/cultivar was reduced to one tube and we have no data about intra genetic variability. As expected in a cross pollinated crop we found genetic diversity and a larger variation within than among the populations using isozymes. Somewhat unexpectedly, however, we found that the breeding cultivars have the same level of heterozygosity as the landraces. The phylogenetic relationships obtained using isozymes among the landraces, synthetic cultivar and the cultivars from breeding programs do not reflect their origin. Moreover, the cultivar from Brazil is not separated from the remaining populations/cultivars studied. However, the data observed using RAPDs and ISSRs are in agreement with their known origin. The populations maintained by the farmers in the north of Portugal are grouped in a cluster in the phenogram and the C902591 (from Brazil), the Alvão (synthetic variety) and Larouco (a hybrid between Montalegre and Brazil) are in a different cluster. The ISSRs and RAPDs provide a rapid method for the production of polymorphic markers, which appear to correspond to known pedigree information.

Molecular diversity and genetic relationships in Secale

“…Molecular data have shown more effectiveness than the morphological and physiological data. Matos et al (2001) and Fernández et al (2002) concluded that for genetic relationships studies, RAPD and ISSR markers were more useful than isozymes. Two RAPD and nine ISSR primers generated more than 80% of polymorphism.…”

“…RAPD and ISSR reactions were performed with minor modifications as described by Matos et al (2001). These reactions were carried out in 25 and 20 μL volume, respectively using T-Professional Thermocycler (Biometra, Göt-tingen, Germany).…”

“…Vavilov 1926;Stutz 1972), cytogenetic (Khush 1962;Cuadrado and Jouve 2002;Zhou et al 2010), chemotaxonomic (Dedio et al 1969), biochemical, like isozymes (Vences et al 1987;Matos et al 2001); and molecular methods such as amplified fragment length polymorphism (AFLP) (Chikmawati et al 2005(Chikmawati et al , 2012, ISSR (Matos et al 2001;Ren et al 2011), random amplified polymorphic DNA (RAPD) (Matos et al 2001;Ma et al 2004), restriction fragment length polymorphism (RFLP) (Loarce et al 1996), simple sequence repeat (SSR) (Shang et al 2006;Jenabi et al 2011) and ribosomal DNA studies (De Bustos and Jouve 2002).…”

Genetic Variation ofZoysiaAs Revealed By Random Amplified Polymorphic Dna (Rapd) and Isozyme Pattern

“…Isozymes and DNA-based markers are frequently used as molecular markers for taxonomic and systematic analyses of plants Bessega et al, 2000;Garkava et al, 2000;Matos et al, 2001;Ochiai et al, 2001). Many researchers Matos et al, 2001;Ochiai et al, 2001) have pointed out that DNA-based markers, such as random amplified polymorphic DNA (RAPD), were superior to isozymes in detecting genetic diversity. The isozymes represents allelic expression of the same locus, but fragments produced by RAPD are independent genetic markers (Ochiai et al, 2001) with a lower proportion of non-neutral markers than the former .…”

Fingerprinting of Vaccinium corymbosum cultivars using DNA of fruits

“…PCR amplifications were performed in a final volume of 20 µl containing 2 μl of DNA template (~20 ng), 2 μl of Taq buffer with (NH 4 ) 2 SO 4 (10×), 1.6 μl of MgCl 2 (25 mM), 2 μl of ISSR primer (5 μM), 0.5 μl of dNTPs mix (10 mM each), 0.4 μl of Taq polymerase (5 U/μl) (Thermo Scientific, Waltham, USA). The amplification programme was described by Matos et al (2001). The PCR products were analysed on 1.7% agarose gels in 1× TBE running at 100 V for 210 min and stained in an ethidium bromide solution.…”