Species-specific microsatellite markers to monitor gene flow between exotic poplars and their natural relatives in eastern North America (original) (raw)

Microsatellite DNA fingerprinting, differentiation, and genetic relationships of clones, cultivars, and varieties of six poplar species from three sections of the genus Populus

Genome, 2002

Accurate identification of Populus clones and cultivars is essential for effective selection, breeding, and genetic resource management programs. The unit of cultivation and breeding in poplars is a clone, and individual cultivars are normally represented by a single clone. Microsatellite DNA markers of 10 simple sequence repeat loci were used for genetic fingerprinting and differentiation of 96 clones/cultivars and varieties belonging to six Populus species (P. deltoides, P. nigra, P. balsamifera, P. trichocarpa, P. grandidentata, and P. maximowiczii) from three sections of the genus. All 96 clones/cultivars could be uniquely fingerprinted based on their single- or multilocus microsatellite genotypes. The five P. grandidentata clones could be differentiated based on their single-locus genotypes, while six clones of P. trichocarpa and 11 clones of P. maximowiczii could be identified by their two-locus genotypes. Twenty clones of P. deltoides and 25 clones of P. nigra could be differ...

Trinucleotide repeat microsatellite markers for black poplar (Populus nigra L.)

Molecular Ecology Notes, 2001

Using an enrichment procedure, we have cloned microsatellite repeats from black poplar ( Populus nigra L.) and developed primers for microsatellite marker analysis. Ten primer pairs, mostly for trinucleotide repeats, produced polymorphic fragments in P. nigra . Some of them also showed amplification in other poplar species. ( P. deltoides , P. tricocarpa , P. tremula , P. tremuloides , P. candicans , P. lasiocarpa ). The best six loci were tested on 23 P. nigra genotypes collected across Europe. The microsatellites produced up to 12 alleles per locus in this set, with observed heterozygosity between 0.32 and 0.91.

Complex patterns of hybridization between exotic and native North American poplar species

American Journal of Botany, 2010

We thank K. Plante, A. Claveau, P. Sylvestre, and S. Senneville for help with the DNA extraction or management of the samples, as well as G. Geneau and Y. Renaud of Genome Quebec for performing the SNPstream assays. The sampling would not have been possible without the help of G. Guigou; P. Gagn é from Ligniculture Qu é bec; and S. Morin, H. Gagnon, F. Caron, and A. Fauchon from the Minist è re des Ressources naturelles et de la Faune du Qu é bec. We are also grateful to M. Neiman, S. Meirmans, and three anonymous reviewers for comments on the manuscript. This work was fi nancially supported by the Canadian Regulatory System for Biotechnology and the FQRNT Quebec Research Fund.

A fast and simple genetic survey reveals the spread of poplar hybrids at a natural Elbe river site

Conservation Genetics, 2008

It is known that various poplar species and cultivated poplar hybrids have the potential to interbreed and produce fertile offspring. Conservation strategies for the genetic resources of the endangered Eurasian black poplar (Populus nigra L.) thus rely on a monitoring which enables the identification and verification of the pure species status. At the same time, the risk of hybrid dispersal and introgressive gene flow has to be estimated. In the present study a combination of two molecular markers, one from chloroplast DNA and the other from nuclear DNA, was applied to evaluate a large P. nigra population on the Elbe River. Hybrid clones of P. × canadensis are scattered within this population and also occur as plantations in the surrounding landscape. By means of the DNA markers the taxonomic status of 208 adult trees in the population and 140 young poplars along the riverbank was monitored. From the analysed young poplars, almost 20 percent were found to exhibit at least one of the two P. deltoides or P. × canadensis diagnostic alleles or genotypes, respectively. Possible vegetative spreads of F1 hybrids and precedent mating scenarios are discussed. Most interestingly we found clear evidence for a small number of backcross hybrids where P. × canadensis acted as pollen donor. This case had long been debated and thought to be less probable, so far.

When exotic poplars and native Populus balsamifera L. meet on the Canadian Prairies: Spontaneous hybridization and establishment of interspecific hybrids

As the interest in plantations of fast-growing tree species increases, concerns regarding the possibility of gene flow between plantations of exotic trees and their wild relatives are being raised. In Canada, the Prairie ecozone provides a unique opportunity to examine spontaneous hybridization and the spread of exotic genetic material because of the historical introduction of large numbers of exotic poplar species and their hybrids. In this region, poplar shelterbelts bearing exotic components such as Populus laurifolia Ledeb. and Populus nigra L. have been in contact with the native populations of Populus balsamifera L. since the 19th century. Given the ability for poplar species to hybridize, the objective of this study was to estimate the rates of spontaneous hybridization between one common poplar cultivar planted in shelterbelts, #'Assiniboine' ($'Walker' [$Populus deltoides Marsh.  #(P. laurifolia  P. nigra)]  #P. del-toides), and its neighbouring native congener, $P. balsamifera, on two study sites. To distinguish the genetic contributions of the species under study, a set of 26 diagnostic single nucleotide polymorphisms (SNPs) representative of 23 genes was used. We genotyped 2116 seeds sampled from P. balsamifera and identified paternal alleles specific to P. laurifolia, P. nigra and P. deltoides, which are typical genetic components of poplar shelterbelts. Surprisingly, the highest hybridization rate (2.3%) was observed at the site where 100-year-old Russian poplar shelterbelts (P. laurifolia  P. nigra) were found. A preliminary study of the P. balsamifera stands established in an 8 km radius around the study site confirmed introgression of P. laurifolia and P. nigra alleles at a rate of 4.6%. These results indicate that spontaneous hybridization between shelterbelts of exotic cultivars and native P. balsamifera does occur in the Canadian Prairies and that interspecific hybrids can establish and survive in this landscape. This study suggests that biological factors such as the genetic composition of the native population and the gender of the exotic cultivars should be taken into consideration in the management strategies of this ecozone. Crown

Allelic and population variation of microsatellite loci in aspen (Populus tremuloides)

New Phytologist, 2005

To develop a robust basis for inferences about population genetics and evolution, this work assayed 192 aspens (Populus tremuloides) from 11 sites in Wisconsin, USA, for allelic and population variation at 16 microsatellite loci distributed across the Populus genome. Frequency distributions of fluorochrome-labeled alleles resolved by capillary electrophoresis were analyzed for relationships to repeat size and number. Population-level statistics were compared with those of other studies, especially in Populus. All loci were polymorphic, varying widely in the number of alleles per locus (mean = 8.25, range 2-20). Expected and observed heterozygosities were high (0.45 and 0.41, respectively), with little differentiation among populations (F(ST) = 0.006-0.045) and a moderate level of inbreeding (F(IS) = 0.09), intermediate among levels reported in studies based on isozymes. Contrary to several other reports, allele frequencies clustered tightly around the modal frequency, and the genetic diversity (measured as alleles per locus or as expected heterozygosity) was not related to either the repeat unit size or to the number of repeats.

Development of Multiplexed Marker Sets to Identify the Most Relevant Poplar Species for Breeding

Forests

Within the genus Populus, about 30 species are classified into six sections, of which some are cross-compatible. Besides naturally occurring hybrids, huge breeding programs have led to a high number of artificially produced hybrids, for which the determination of genetically involved species by morphological characteristics is often difficult. This necessitates the use of molecular markers for the identification of both maternal as well as paternal species, and in the case of complex hybrids, the genealogy. For this reason, we developed new chloroplast and nuclear markers for the differentiation of up to 19 poplar species, with one to 32 individuals per species regularly used in breeding programs based on already known barcoding, other chloroplast regions, and nuclear genes of interest. We developed methods to identify species by either species-specific nucleotide variations or, when no initial information for the species was given, by using a set of markers either in a procedure of...

Development of two microsatellite multiplex PCR systems for high throughput genotyping in Populus euphratica

Journal of Forestry Research, 2009

Eighteen microsatellite primer pairs previously developed at Oak Ridge National Laboratory for Populus tremuloides Michx. and Populus trichocarpa Torr. & Gray were screened for amplification in Euphrates poplar, Populus euphratica Oliv. Thirteen loci were found to express polymorphisms ranging from two to 17 alleles. The eight most variable loci were selected to set up and optimize two multiplex polymerase chain reaction (PCR) assays. Three populations containing altogether 436 trees were used to characterize the selected loci and ascertain their applicability for parentage analysis and genotyping studies. Through cross-checking of clonal identity against sex of the genotyped trees we estimated the maximum error rate for merging genotypes to be less than 0.045.

Genetic relationships and clonal identity in a collection of commercially relevant poplar cultivars assessed by AFLP and SSR

Tree Genetics & Genomes, 2005

A collection of 66 poplar commercial clones widely cultivated in Italy, China and in other countries of southern Europe and belonging to various poplar species and hybrids, have been fingerprinted using both amplified fragment length polymorphism (AFLP) and simple sequence repeats (SSR) techniques. Three AFLP primer combinations and six SSRs unambiguously genotyped the analysed poplar collection, with the exception of three groups of six, four and two individuals, which turned out to be indistinguishable even if they met the standards currently applied for distinctness, uniformity and stability (DUS) testing when registered. High levels of variation were detected with both molecular techniques; a total of 201 AFLP bands were amplified of which 96% turned out to be polymorphic and up to 15 SSR alleles were identified at a single locus, with a mean of 9.3 alleles per locus in the case of Populus × canadensis. The probability of matching fortuitously any two genotypes at all the SSR loci in the case of P. × canadensis was less then 7.5×10 −9 . The AFLP-derived dendrogram and principal coordinate analysis (PCOOR-DA) clustered the clones with respect to their taxonomic classification, and allowed their genetic interrelationships to be established. Correct identification of poplar varieties is essential for ensuring the effective correspondence between the real and the declared identity of a clone, to avoid commercial frauds, and to establish breeding programmes. Molecular markers may play a major role to satisfy all these needs.

Application of Microsatellite Loci for Molecular Identification of Elite Genotypes, Analysis of Clonality, and Genetic Diversity in Aspen Populus tremula L. (Salicaceae)

International Journal of Plant Genomics, 2015

Testing systems for molecular identification of micropropagated elite aspen (Populus tremula L.) genotypes were developed on the base on microsatellite (SSR) loci. Out of 33 tested microsatellite loci, 14 were selected due to sustainable PCR amplification and substantial variability in elite clones of aspen aimed for establishment of fast-rotated forest plantations. All eight tested clones had different multilocus genotypes. Among 114 trees from three reference native stands located near the established plantations, 80 haplotypes were identified while some repeated genotypes were attributed to natural clones which appeared as a result of sprouting. The selected set of SSR markers showed reliable individual identification with low probability of appearance of identical aspen genotypes (a minimum of 4.8·10-10 and 1 × 10−4 for unrelated and related individuals, resp.). Case studies demonstrating practical applications of the test system are described including analysis of clonal struct...