Cross-species amplification of microsatellite markers and genetic diversity in the macaw palm (Acrocomia aculeata)

segunda-feira, março 18, 2019

Allelic profiles showing cross-species amplifications of seven SSR loci on Acrocomia aculeata accessions. Loci include Aac04 and Aac12 (Astrocarym aculeatum); sMo00020 and sMo00137 (Elaies oleifera); and Aacu38, Aacu45 and Aacu74 (Acrocomia aculeata). The amplification is on 48 selected A. aculeata accessions of twelve families. BGP99 (Pará); BGP82 (Pernambuco); BGP124 (Paraiba); BGP51, BGP34 and BGP47 (São Paulo); BGP20, BGP68, BGP18, BGP11 and BGP21 (Minas Gerais) and BGP102 (Mato Grosso do Sul). Allele amplicon size (bp) indicated for each locus.

Author: Mengistu, Fekadu Gebretensay

Abstract: Macaw palm (Acrocomia aculeata) is newly emerging oleaginous species in South America with abundant natural distribution in Brazil. It is considered as a great potential palm for production of biodiesel and being under domestication in Brazil. Recently researches show that macaw palm is under the threat of predatory extractivism, climate change and land use policies in the natural population and need to be conserved ex situ for future genetic improvement and sustainable use of its genetic resources. Microsatellites (Simple Sequence Repeats-SSR) are one of the most applicable molecular markers in characterization of germplasm collections in plants and help to conserve genetic variability in germplasm banks.

In the past only few SSR markers were developed for the macaw palm owing to the high development cost and limitation in knowledge about the importance of the palm. Two experiments were set up in this study: (1) to demonstrate the use of cross-species amplification as a cost-effective altarnative to establish SSR markers for A. aculeata; and (2) to study the genetic diversity in A. aculeata ex situ germplasm collections which were originally collected from different provenances in Brasil. In the the first part of the work, a cross-species amplification study was conducted to evaluate the transferability of 34 SSR markers, originally developed for two Arecaceae species (Astrocaryum aculeatum and Elaies oleifera) in A. aculeata using 192 accessions from 41 families collected from six provenances in Brazil. From the total markers evaluated, 15 SSR (44%) successfully amplified the genomic DNA in A. aculeata, of which four SSR (26%) were polymorphic.

The low success of the cross-amplification was accounted for the relatively wider taxonomic distance between the sources (A. aculeatum and E. oleifera) and the target (A. aculeata) species. However, the polymorphic markers identified by this study detected a high average percentage of polymorphic loci (P=79%) per provenance. The markers also revealed heterozygote deficiency in the accessions and this was confirmed by positive inbreeding coefficients obtained in all the loci analyzed. In the second work, genetic diversity study was carried out in the 192 accessions of A. aculeata, based on ten SSR markers (including two polymorphic SSR indentified in the transferability study and the rest eight from sets of SSR previously developed for A. xiii aculeata). The study resulted in different levels of allelic diversity, heterozygosity and polymorphism among the accessions analyzed. Based on genetic distances, three distinct groups of provenances were established using different methods of grouping. However, Mantel test detected a non-significant correlation between the genetic and geographic distances among the provenances, revealing genetic similarities among geographically distant provenances in the country. Analysis of Molecular Variance (AMOVA) revealed the proportions of genetic variation, in which more genetic variation was obtained within family than among families of A. aculeata followed by variation among provenances.

The study proved the efficiency of inter-species transferability of SSR markers between different species in Arecaceae. It also reaffirmed that SSR are useful molecular markers in characterizing A. aculeata germplasm and the information that were generated could be utilized in A. aculeata germplasm conservation and breeding program. The markers could be used to help the breeding program through genotyping each individuals in the germplasm bank that would help to identify genetically distinct groups and also minimize the unnecessary redundancies of entries in the germplasm bank that would potentially maximize conservation efficiencies and reduce its costs.

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