CROSS-SPECIES AMPLIFICATION OF MICROSATELLITE MARKERS IN CASTANEA SPP. AND OTHER RELATED SPECIES

Abstract

Genetic variation and differentiation within and between species can be investigated using microsatellite or simple sequence repeats (SSRs). This can be done using primer sequences identified in one species, which can be amplified across species depending on whether or not the microsatellite flanking regions are conserved between species. This study provides an insight into the distribution of SSRs in related genera of the Fagaceae, Betulaceae, and Nothofagaceae families. Cross-species amplification of 17 SSR loci isolated in Castanea sativa Mill. were evaluated across 16 different species [C. sativa Mill., C. crenata Sieb. et Zucc., C. dentata Borkhausen (Marshall), C. mollissima Blume, C. seguinii Dode, Quercus robur L., Q. cerris L., Q. ilex L., Q. petraea (Mattuschka) Liebl., Q, suber L., Fagus sylvatica L., Betula pendula Roth, B. pubescens Ehrh., Carpinus betulus L., Corylus avellana L., Nothofagus antarctica (Forster) Oerst.] belonging to 7 genera. Moreover, 6 Q. petraea and 3 Q. robur SSR loci were evaluated only in the Betulaceae species. Among the C. sativa SSR loci, 13 of them generated one or more amplification products, in the expected size range, in Castanea species other than C. sativa, and 4 loci failed to amplify C., seguinii, C. mollissima or C. dentata. Surprisingly 5 C. sativa and 2 Q. robur primer pair amplified in the Betulaceae family. The apparent broad cross-species/genus/family transferability are being confirmed by cloning and sequencing the amplified alleles. The results indicate a high level of nucleotide sequence conservation within the primer region of 16 different species and have potential in phylogenetic and genetic variation studies and open up new interesting prospects for comparative genetic.