SSR Markers for Analysing South American Nothofagus Species

Abstract

Summary11 newly discovered microsatellites were used to identify SSR markers for characterising South American Nothofagus species. This was carried out in six species. The sample sizes used were between four and six individuals per species. The cross-genera transferability of 34 Quercus SSRs was also essayed. Out of the 11 new microsatellite markers, three proved to be polymorphic (NnBIO 11, NgBIO 13 and NgBIO 14). The qualitative confirmation of the inheritance of these markers could also be verified. Polymorphism was also observed in five of the cross-genera transferred SSRs (QrBIO7, quru-GA-0A01, quru-GA-0C11, quru-GA-0I01, quru-GA-0M07). The number of alleles per locus found range between 1 and 6 per species. The eight polymorphic SSRs identified in this study will constitute a valuable tool in the gene flow studies that are currently being carried out in natural populations of South American Nothofagus species. The confirmation of crossspecies and cross-genera transferability opens the way for the use of SSRs as bridge markers in genetic mapping.

The effect of volcanism on postglacial migration and seed dispersal. A case study in southern South America

“…Three simple sequence repeats (SSRs) were scored: NnBIO11, NgBIO14 (Azpilicueta et al 2004), and NnBIO111 (Marchelli et al 2007). PCR was performed in a total volume of 20 μl containing about 10 ng of total DNA, 1,5 mM MgCl 2 , 100 μM each dNTP, 0.2 μM each primer, and 0.8 U Taq polymerase (Invitrogen) with the respective 10× PCR buffer, using a Biometra Uno Thermo Block.…”

“…PCR was performed in a total volume of 20 μl containing about 10 ng of total DNA, 1,5 mM MgCl 2 , 100 μM each dNTP, 0.2 μM each primer, and 0.8 U Taq polymerase (Invitrogen) with the respective 10× PCR buffer, using a Biometra Uno Thermo Block. Amplification was done according to Azpilicueta et al (2004) with annealing temperatures of 59°C for NnBIO11 and 55°C for NgBIO14 and NnBIO111. PCR products were denaturized by adding denaturing blue followed by incubation for 10 min at 94°C.…”

Out in the cold: genetic variation of Nothofagus pumilio (Nothofagaceae) provides evidence for latitudinally distinct evolutionary histories in austral South America

“…2008). Although microsatellites for Nothofagus were developed for some species within subgenus Lophozonia (Azpilicueta et al. 2004; Jones et al.…”

Characterisation of seven polymorphic microsatellites forNothofagussubgenusFuscosporafrom New Zealand

“…Isolation Azpilicutea et al 2004). Cross-species amplification of some of these SSR loci has been shown in these studies, but primer pairs appear to work better when transferred to a species from within the same subgenus rather than across subgenera.…”