Abstract
Premise of the Study Alectryon ramiflorus (Sapindaceae) is an endangered rainforest tree known from only two populations. In this study, we identified polymorphic microsatellites, in silico, improving the effectiveness and efficiency of microsatellite development of nonmodel species. The development of genetic markers will support future conservation management of the species.Methods and ResultsWe used next‐generation sequencing and bioinformatics to detect polymorphic microsatellites, in silico, reducing both the time and cost of marker development. A panel of 15 microsatellites, 12 of which were polymorphic, were subsequently characterized in 64 adult trees representing the entire species range. Mean observed heterozygosity and expected heterozygosity were 0.471 and 0.425, respectively. The polymorphism information content across loci ranged from 0.152 to 0.875.ConclusionsThe microsatellite markers developed in this study will be useful in gaining an understanding of A. ramiflorus’ genetic diversity, level of inbreeding, and population structure and for guiding future restoration and management efforts.
Highlights
PREMISE OF THE STUDY: Alectryon ramiflorus (Sapindaceae) is an endangered rainforest tree known from only two populations
The microsatellite markers developed in this study will be useful in gaining an understanding of A. ramiflorus’ genetic diversity, level of inbreeding, and population structure and for guiding future restoration and management efforts
The Isis tamarind, Alectryon ramiflorus (Sapindaceae), is a dioecious tree restricted to the dry Araucarian microphyll vine forests near Childers in southern Queensland, the majority of which have been cleared for agriculture over the last century (Threatened Species Scientific Committee, 2016)
Summary
Genomic DNA from a single A. ramiflorus individual from each of the two known populations (Appendix 1) was extracted using the DNeasy Plant Mini Kit (QIAGEN, Hilden, Germany) and used to construct two DNA libraries at the Australian Genome Research Facility (Brisbane, Australia; http://agrf.org.au/). PCR was completed for 14 A. ramiflorus individuals using the same reaction components described above and the following cycling conditions: initial denaturation at 95°C for 5 min; followed by 35 cycles of 94°C for 30 s, 56°C for 90 s, and 72°C for 30 s; with a final extension at 68°C for 10 min. Twelve high-quality polymorphic microsatellite loci (Table 1) were selected and validated in 64 individuals, 60 collected from the only two known populations (plus two individuals each from two roadside patches) located near Childers, Queensland (Appendix 1). Polymorphism information content, observed and expected heterozygosity, and null allele frequencies for each locus were calculated in CERVUS version 3.0.8 (Kalinowski et al, 2007). Six of the 12 loci exhibited an excess of homozygosity and significant deviation from conditions of Hardy–Weinberg equilibrium, which was expected given the small population sizes and extremely low number of individuals remaining in the species.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
