Characterization of microsatellite markers for Broussonetia papyrifera (Moraceae)

Abstract

Premise of the study:Broussonetia papyrifera (Moraceae) is native to Asia and is used as a medicinal plant and as a source of fiber for making paper. It was dispersed into the Pacific region as a fiber source for making nonwoven textiles (barkcloth). Microsatellites were developed to trace the human-mediated dispersal of this species into the Pacific region.Methods and Results:A set of 36 microsatellites was isolated and initially assayed on 10 accessions to assess polymorphism. We found that 20 markers were polymorphic, with the number of alleles per marker ranging from four to 35 in 70 accessions genotyped from three Asian populations. Observed and expected heterozygosities ranged from 0.04 to 0.85 and from 0.19 to 0.94, respectively. These markers were tested in four Moraceae species and one Rosaceae species.Conclusions:These markers will be useful for the assessment of genetic diversity in B. papyrifera. They show low transferability to other species tested.

A tale of textiles: Genetic characterization of historical paper mulberry barkcloth from Oceania

“…All DNA replicates obtained from the barkcloth samples were analyzed using ten microsatellite markers, as described in the Methods section. The selected markers are those that proved to be most informative for this species, as reported previously [24,25,28]. DNA from the New Guinea barkcloth sample amplified with all markers, while DNA from the sample from the Fijian textile amplified four to five markers, and the samples from the barkcloth from the Gambier Islands amplified between six and eight markers.…”

“…The presence of three or more microsatellite alleles in a single sample suggests that each barkcloth textile was manufactured from several plants of different genetic backgrounds, as all genetic analyses of extant and herbaria samples presented a single allele (homozygous) or a double signal (heterozygous) for each locus [24,25,28], as expected in a diploid species. The presence of three or more microsatellite alleles in a single sample is consistent with the presence of ITS variants in a single sample as detected in several textiles and discussed above.…”

“…PCR reactions included a negative (blank) control, adding the appropriate amounts of sterile distilled water to the mix in all experiments. We analyzed i) the nuclear ITS-1 region using ITS-A (5'-GGA AGG AGA AGT CGT AAC AAG G-3') and ITS-C (5'-GCA ATT CAC ACC AAG TAT CGC-3') primers as described [20,24] and ii) a set of ten nuclear microsatellite (SSR) markers, as reported previously [28]. DNA from all samples was also assayed with a specific paper mulberry sex marker PLOS ONE [23] and described in the Supporting Text.…”

Human mediated translocation of Pacific paper mulberry [Broussonetia papyrifera (L.) L’Hér. ex Vent. (Moraceae)]: Genetic evidence of dispersal routes in Remote Oceania

“…For degraded DNA, specific primer pairs (300 and 165 bp) were designed for the amplification of a smaller region of the sex marker; iii) the ndhF-rpl32 chloroplast DNA region using the primers and protocol as described in Chang et al [26] with a subset of 35 contemporary samples representative of the islands sampled, and iv) a set of four nuclear microsatellite or Single Sequence Repeat (SSR) markers (Bro 07, Bro 08, Bro 13 and Bro 15) developed by one of us (K-FC, S3 Table) and six nuclear microsatellite markers for B . papyrifera (Bropap 2214, Bropap 2801, Bropap 20558, Bropap 25444, Bropap 26985 and Bropap 30248) as described in Peñailillo et al [33]. Primers used in this study are presented in S3 Table.…”