Development of microsatellite markers for the wind cave-associated shrub Lonicera alpigena subsp. glehnii (Caprifoliaceae).
ABSTRACT: Premise of the Study:Microsatellite markers were developed for the wind cave-associated shrub Lonicera alpigena subsp. glehnii to conduct phylogeographic studies on the species. Methods and Results:Based on the sequence data obtained by 454 sequencing, a total of 81 primer pairs were designed and 18 successfully amplified the microsatellite regions. These markers were highly variable (i.e., average number of alleles per locus = 6.2 [range = 2-15]; average expected heterozygosity per locus = 0.489 [range = 0.149-0.729]). Cross-species amplification of the primers was tested in 10 congeneric taxa (L. caerulea var. emphyllocalyx, L. chamissoi, L. chrysantha, L. gracilipes var. glandulosa, L. japonica, L. kurobushiensis, L. morrowii, L. ramosissima, L. sachalinensis, and L. strophiophora), and six to 11 primers amplified the microsatellite markers. Conclusions:The microsatellite markers developed in this study will be useful for phylogeographic studies and conservation genetics of L. alpigena subsp. glehnii as well as congeneric species.
Project description:PREMISE OF THE STUDY:Nuclear microsatellite markers were developed in Rumex bucephalophorus subsp. canariensis (Polygonaceae) to investigate its genetic diversity and structure. METHODS AND RESULTS:Sixteen polymorphic microsatellite markers were obtained using 454 next-generation sequencing with di-, tri-, and tetranucleotide repeats. The average number of alleles was 5.688 and 3.813 for R. bucephalophorus subsp. canariensis var. canariensis and var. fruticescens, respectively. Slightly higher levels of mean genetic diversity were found in var. canariensis (expected heterozygosity = 0.600) than in var. fruticescens (expected heterozygosity = 0.514). Cross-amplifications in related taxa within R. bucephalophorus showed good amplification and polymorphic patterns. CONCLUSIONS:These 16 novel nuclear microsatellite markers are the first in the genus Rumex and may serve as valuable tools to carry out studies on genetic diversity and structure as well as progeny studies.
Project description:Banana is a nutritionally important crop across tropical and sub-tropical countries in sub-Saharan Africa, Central and South America and Asia. Although cultivars have evolved from diploid, triploid and tetraploid wild Asian species of Musa acuminata (A genome) and Musa balbisiana (B genome), many of today's commercial cultivars are sterile triploids or diploids, with fruit developing via parthenocarpy. As a result of restricted genetic variation, improvement has been limited, resulting in a crop frequently lacking resistance to pests and disease. Considering the importance of molecular tools to facilitate development of disease resistant genotypes, the objectives of this study were to develop polymorphic microsatellite markers from BAC clone sequences for M. acuminata subsp. burmannicoides, var. Calcutta 4. This wild diploid species is used as a donor cultivar in breeding programs as a source of resistance to diverse biotic stresses.Microsatellite sequences were identified from five Calcutta 4 BAC consensi datasets. Specific primers were designed for 41 loci. Isolated di-nucleotide repeat motifs were the most abundant, followed by tri-nucleotides. From 33 tested loci, 20 displayed polymorphism when screened across 21 diploid M. acuminata accessions, contrasting in resistance to Sigatoka diseases. The number of alleles per SSR locus ranged from two to four, with a total of 56. Six repeat classes were identified, with di-nucleotides the most abundant. Expected heterozygosity values for polymorphic markers ranged from 0.31 to 0.75.This is the first report identifying polymorphic microsatellite markers from M. acuminata subsp. burmannicoides, var. Calcutta 4 across accessions contrasting in resistance to Sigatoka diseases. These BAC-derived polymorphic microsatellite markers are a useful resource for banana, applicable for genetic map development, germplasm characterization, evolutionary studies and marker assisted selection for traits.
Project description:Premise of the Study:Microsatellite markers were developed for Smilax rotundifolia (Smilacaceae), an understory vine widely distributed in eastern North America, to investigate genetic diversity and structure. Cross-amplification was tested in three congeneric species: S. china, S. riparia, and S. walteri. Methods and Results:A total of 6153 simple sequence repeat primer pairs were detected from the de novo-assembled transcriptome data (88,3962 contigs) of S. rotundifolia. Thirty-three polymorphic microsatellite loci were selected for further analysis among 96 individuals representing four natural populations of the species. The number of alleles ranged from two to 15, and 87.9% of the developed primer pairs could be cross-amplified in at least one of three congeneric Smilax species. Conclusions:The simple sequence repeat markers developed in this study will facilitate further studies on genetic diversity and phylogeographic patterns of S. rotundifolia and provide additional potential microsatellite resources for other Smilax species.
Project description:Premise of the Study:Microsatellite primers were developed for the first time in the genus Filago (Gnaphalieae: Asteraceae). These markers will facilitate low-scale phylogenetic, phylogeographic, and population genetic studies within the genus Filago. Methods and Results:Ten pairs of polymorphic microsatellite primers (as well as five pairs of monomorphic primers) were identified and optimized on two species of Filago (F. gaditana and F. carpetana) using a microsatellite-enrichment library method and 454 GS-FLX technique. The polymorphic primers amplified tri- to hexanucleotide repeats and showed one to six alleles per locus for both species. Transferability was performed in 29 samples corresponding to nine representative species of Filago. Conclusions:The results indicate the utility of the newly developed markers, which will be useful to delve into the phylogenetic relationships among the taxa within Filago. These microsatellites will enable studies of phylogeographic, reproductive, and genetic variation.
Project description:Premise of the Study:We developed microsatellite primers for Rhododendron shanii (Ericaceae), a narrowly distributed species found in the Dabie Mountains, China, to study the genetic diversity, population structure, and evolutionary history of the species. Methods and Results:Two terminal sequencing modes of the Illumina HiSeq platform were used to mine simple sequence repeat markers from large-scale transcriptional groups. In this study, 24 microsatellite loci were screened. The number of alleles ranged from one to 20, and the levels of observed and expected heterozygosity ranged from 0.000 to 1.000 and 0.000 to 0.918, respectively. Most of these primers were successfully amplified in eight congeneric species (R. annae, R. chihsinianum, R. decorum, R. denudatum, R. fortunei, R. neriiflorum, R. rex, and R. simiarum). Conclusions:These newly developed microsatellite loci will be useful for studying the genetic diversity and population structure of R. shanii and congeneric species.
Project description:PREMISE OF THE STUDY:Polymorphic microsatellite loci were developed and used to genotype individuals of Gentianella praecox subsp. bohemica (Gentianaceae), a highly protected taxon in Europe, to study the genetic structure of the remaining populations. METHODS AND RESULTS:Thirty-eight primer pairs were successfully amplified; of these, 12 polymorphic microsatellite loci were developed using a 454 sequencing approach and used to genotype 180 individuals of G. praecox subsp. bohemica from six populations. Allelic richness ranged between one and nine alleles per locus. We detected a high frequency of polyploid individuals (77.8%). The highest average percentage of heterozygous genotypes was identified for samples from the Hroby population (75.5%). All loci can also be amplified in the congeneric species G. praecox subsp. praecox, G. amarella subsp. amarella, and G. obtusifolia subsp. sturmiana. CONCLUSIONS:These markers will provide knowledge on patterns of gene flow and population genetic structure, which is necessary for current protection actions and for effective conservation of this species in the future.
Project description:PREMISE OF THE STUDY:Although several microsatellite markers of Smilax aspera (Smilacaceae) have been reported in a previous study, due to universality issues in cross-population amplification, we have newly developed microsatellite markers for S. aspera based on transcriptome data to further investigate gene flow and genetic structure of its circum-Mediterranean, East African, and South Asian populations. METHODS AND RESULTS:A total of 4854 simple sequence repeat (SSR) primer pairs were designed from 99,193 contigs acquired from public transcriptome data of S. bona-nox. Forty-six microsatellite loci were selected for further genotyping in 12 S. aspera populations. The number of alleles varied from three to 28, and 93.5% of the developed microsatellite markers could be cross-amplified in least one of three congeneric Smilax species. CONCLUSIONS:The SSR markers developed in this study will facilitate further studies on genetic diversity and phylogeographic patterns of S. aspera in intercontinental geographical scales.
Project description:Premise:The common sowthistle, Sonchus oleraceus (Asteraceae), is a globally invasive weedy species. In order to investigate its genetic diversity, population genetic structure, and evolutionary history, we developed and characterized nuclear simple sequence repeat markers (SSRs or microsatellites). Methods and Results:Seventeen microsatellite primer pairs were developed based on the Illumina sequence data. Ten developed SSR loci were polymorphic in four populations sampled from broad geographical regions. The number of alleles per locus ranged from one to 11, and the levels of observed and expected heterozygosity ranged from 0.000 to 1.000 and from 0.000 to 0.801, respectively. Up to 82% of the newly developed primer pairs were successfully amplified in the congeneric taxa S. asper, S. asper subsp. glaucescens, S. canariensis, and S. palmensis. Conclusions:The SSR markers developed in this study will be useful for future population genetic studies on S. oleraceus and other congeneric species.
Project description:Premise of the Study:Boswellia serrata (Burseraceae) is an economically important aromatic, gum-resin-yielding, non-timber forest tree species. Microsatellite markers were developed for B. serrata for the first time to study genetic diversity and population structure. Methods and Results:A magnetic bead enrichment method was used to develop 16 microsatellite markers, of which 11 were polymorphic. The number of alleles per locus in the 60 individuals studied ranged from three to 10, and the levels of observed and expected heterozygosity ranged from 0.50 to 0.90 and 0.666 to 0.861, respectively. The primers successfully amplified in the congeneric species B. ovalifoliolata. Conclusions:These microsatellite markers can be used to study the genetic variation and population structure of B. serrata and to provide crucial information on population and ecological issues for management and conservation of the species.
Project description:UNLABELLED: PREMISE OF THE STUDY:Pinus wangii is an endemic and endangered species in southwestern China, and microsatellite primers were developed to characterize its genetic diversity and population structure. • METHODS AND RESULTS:Using the Fast Isolation by AFLP of Sequences COntaining repeats (FIASCO) protocol, nine sets of microsatellite primers were developed in P. wangii. One population with 26 individuals of P. wangii, as well as 11 individuals each for two congeneric species, P. taiwanensis and P. squamata, were used to test their polymorphism and transferability. The number of alleles per locus ranged from one to seven with an average of 3.7, and the observed heterozygosity and expected heterozygosity ranged from 0 to 0.91 and 0 to 0.75, respectively. • CONCLUSIONS:We developed nine sets of polymorphic microsatellite loci that are suitable for investigating genetic diversity and population structure of P. wangii, and these markers may be useful for other Pinus species.