Genetic diversity of Metrodorea nigra (Rutaceae) from a small forest remnant in Brazil assessed with microsatellite markers

Abstract

ABSTRACT. Metrodorea nigra (Rutaceae) is an endemic Brazilian tree of great ecological importance, frequently found in the submontane regions of ombrophilous dense and semideciduous forests. This tree is useful for reforesting degraded areas and the wood can be employed in construction. We developed 12 microsatellite markers from a genomic library enriched for GA/CA repeats, for this species. Polymorphisms were assessed in 40 trees of a highly fragmented population found in Cravinhos, State of São Paulo, in southeastern Brazil. Among the 12 loci, 8 were polymorphic and only one had fixed alleles in this population. The number of alleles per locus and expected heterozygosity ranged from 2 to 11 and from 0.190 to 0.889, respectively. These results revealed mod- Microsatellite markers for Metrodorea nigra erate levels of genetic variation in M. nigra population when compared to other tropical species. Additionally, transferability of the 12 primers was tested in seven other Brazilian Rutaceae tree species (endemics: M. stipularis, Galipea jasminiflora, Esenbeckia leiocarpa and non-endemics: E. febrifuga, E. grandiflora, Balfourodendron riedelianum, Zanthoxylum riedelianum). Transferability ranged among species, but at least 8 loci (~67%) amplified in M. stipularis, demonstrating a high potential for transferring microsatellite markers between species of the same genus in the Rutaceae family.

Fragmentation effects and genetic diversity of the key semidecidual forest species Metrodorea nigra in Southwestern Brazil

“…N A = number of alleles; N E = effective number of alleles; H O = observed heterozygosity; H E = expected heterozygosity; F = fixation index; P.Ex1 = probability of exclusion for the first parental individual; P.Ex2 = probability of exclusion for the second parental individual; NAF = null alleles frequency. *Primers from Guidugli et al (2012). **Unpublished.…”

“…Five microsatellite (SSR) primer pairs (Mtn 1, Mtn 3, Mtn 13, Mtn 16, and Mtn 19) previously developed by Guidugli et al (2012) were used in this study. To increase the number of SSR primers available for analysis, a new set of primers was designed according to the protocol described by Guidugli et al (2012), and 2 newly developed and highly polymorphic SSR primer pairs were obtained (Mtn 87 and Mtn 95,unpublished).…”

“…To increase the number of SSR primers available for analysis, a new set of primers was designed according to the protocol described by Guidugli et al (2012), and 2 newly developed and highly polymorphic SSR primer pairs were obtained (Mtn 87 and Mtn 95,unpublished). All microsatellite loci were amplified by PCR in a final volume of 15 µL using the GoTaq Kit (Promega) consisting of 6 µL H 2 O, 6 µL master mix (400 nM of each dNTPs and 3 mM MgCl 2 ), 1 mM of each primer, and 20 ng DNA.…”

“…Simple sequence repeats (SSRs) are highly polymorphic multiallelic codominant markers that are particularly useful for analyzing genetic diversity within and between populations (Nybom, 2004) and frequently used to resolve the genetic relatedness of tree species (FerreiraRamos et al, 2008;Feres et al, 2009;Guidugli et al, 2012). The polymerase chain reaction (PCR) amplification of inter-simple sequence repeats (ISSRs) can be used to rapidly differentiate between closely related individuals, as well as generate a large number of highly dominant polymorphic loci (Zietkiewicz et al, 1994).…”

Molecular phylogeny and biogeography of the SouthAmerican genus Metrodorea (Rutaceae)

“…For example, Müller et al (1995) isolated several compounds from its stems and leaves; Souza et al (2004) studied its flower and fruit anatomy; Pombal and Morellato (2000) investigated the correlation between its floral traits (color and odor) and main pollinators, as compared to M. stipularis; and recent works by Guidugli et al (2012) and Schwarcz et al (2010) have been focused on its genetic diversity and structure, and its mating system. In addition, many studies have reported M. nigra as a frequent species in floristic surveys (Durigan et al, 2000;Toniato and OliveiraFilho, 2004;Santos et al, 2007b), playing an important ecological role in tropical deciduous forests (Metzger, 2000), and also as a key species for forest restoration purposes in this biome (Chagas et al, 2004).…”