High-throughput SNP genotyping in the highly heterozygous genome of <it>Eucalyptus</it>: assay success, polymorphism and transferability across species

<p>Abstract</p> <p>Background</p> <p>High-throughput SNP genotyping has become an essential requirement for molecular breeding and population genomics studies in plant species. Large scale SNP developments have been reported for several mainstream crops. A growing inter...

Full description

Bibliographic Details
Main Authors: de Lima Bruno, Kirst Matias, Silva-Junior Orzenil B, Grattapaglia Dario, Faria Danielle A, Pappas Georgios J
Format: Article
Language:English
Published: BMC 2011-04-01
Series:BMC Plant Biology
Online Access:http://www.biomedcentral.com/1471-2229/11/65
_version_ 1828741512005943296
author de Lima Bruno
Kirst Matias
Silva-Junior Orzenil B
Grattapaglia Dario
Faria Danielle A
Pappas Georgios J
author_facet de Lima Bruno
Kirst Matias
Silva-Junior Orzenil B
Grattapaglia Dario
Faria Danielle A
Pappas Georgios J
author_sort de Lima Bruno
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>High-throughput SNP genotyping has become an essential requirement for molecular breeding and population genomics studies in plant species. Large scale SNP developments have been reported for several mainstream crops. A growing interest now exists to expand the speed and resolution of genetic analysis to outbred species with highly heterozygous genomes. When nucleotide diversity is high, a refined diagnosis of the target SNP sequence context is needed to convert queried SNPs into high-quality genotypes using the Golden Gate Genotyping Technology (GGGT). This issue becomes exacerbated when attempting to transfer SNPs across species, a scarcely explored topic in plants, and likely to become significant for population genomics and inter specific breeding applications in less domesticated and less funded plant genera.</p> <p>Results</p> <p>We have successfully developed the first set of 768 SNPs assayed by the GGGT for the highly heterozygous genome of <it>Eucalyptus </it>from a mixed Sanger/454 database with 1,164,695 ESTs and the preliminary 4.5X draft genome sequence for <it>E. grandis</it>. A systematic assessment of <it>in silico </it>SNP filtering requirements showed that stringent constraints on the SNP surrounding sequences have a significant impact on SNP genotyping performance and polymorphism. SNP assay success was high for the 288 SNPs selected with more rigorous <it>in silico </it>constraints; 93% of them provided high quality genotype calls and 71% of them were polymorphic in a diverse panel of 96 individuals of five different species.</p> <p>SNP reliability was high across nine <it>Eucalyptus </it>species belonging to three sections within subgenus Symphomyrtus and still satisfactory across species of two additional subgenera, although polymorphism declined as phylogenetic distance increased.</p> <p>Conclusions</p> <p>This study indicates that the GGGT performs well both within and across species of <it>Eucalyptus </it>notwithstanding its nucleotide diversity ≥2%. The development of a much larger array of informative SNPs across multiple <it>Eucalyptus </it>species is feasible, although strongly dependent on having a representative and sufficiently deep collection of sequences from many individuals of each target species. A higher density SNP platform will be instrumental to undertake genome-wide phylogenetic and population genomics studies and to implement molecular breeding by Genomic Selection in <it>Eucalyptus</it>.</p>
first_indexed 2024-04-13T01:01:22Z
format Article
id doaj.art-b1f577f7c85b4cf8bba05affb366c8cb
institution Directory Open Access Journal
issn 1471-2229
language English
last_indexed 2024-04-13T01:01:22Z
publishDate 2011-04-01
publisher BMC
record_format Article
series BMC Plant Biology
spelling doaj.art-b1f577f7c85b4cf8bba05affb366c8cb2022-12-22T03:09:28ZengBMCBMC Plant Biology1471-22292011-04-011116510.1186/1471-2229-11-65High-throughput SNP genotyping in the highly heterozygous genome of <it>Eucalyptus</it>: assay success, polymorphism and transferability across speciesde Lima BrunoKirst MatiasSilva-Junior Orzenil BGrattapaglia DarioFaria Danielle APappas Georgios J<p>Abstract</p> <p>Background</p> <p>High-throughput SNP genotyping has become an essential requirement for molecular breeding and population genomics studies in plant species. Large scale SNP developments have been reported for several mainstream crops. A growing interest now exists to expand the speed and resolution of genetic analysis to outbred species with highly heterozygous genomes. When nucleotide diversity is high, a refined diagnosis of the target SNP sequence context is needed to convert queried SNPs into high-quality genotypes using the Golden Gate Genotyping Technology (GGGT). This issue becomes exacerbated when attempting to transfer SNPs across species, a scarcely explored topic in plants, and likely to become significant for population genomics and inter specific breeding applications in less domesticated and less funded plant genera.</p> <p>Results</p> <p>We have successfully developed the first set of 768 SNPs assayed by the GGGT for the highly heterozygous genome of <it>Eucalyptus </it>from a mixed Sanger/454 database with 1,164,695 ESTs and the preliminary 4.5X draft genome sequence for <it>E. grandis</it>. A systematic assessment of <it>in silico </it>SNP filtering requirements showed that stringent constraints on the SNP surrounding sequences have a significant impact on SNP genotyping performance and polymorphism. SNP assay success was high for the 288 SNPs selected with more rigorous <it>in silico </it>constraints; 93% of them provided high quality genotype calls and 71% of them were polymorphic in a diverse panel of 96 individuals of five different species.</p> <p>SNP reliability was high across nine <it>Eucalyptus </it>species belonging to three sections within subgenus Symphomyrtus and still satisfactory across species of two additional subgenera, although polymorphism declined as phylogenetic distance increased.</p> <p>Conclusions</p> <p>This study indicates that the GGGT performs well both within and across species of <it>Eucalyptus </it>notwithstanding its nucleotide diversity ≥2%. The development of a much larger array of informative SNPs across multiple <it>Eucalyptus </it>species is feasible, although strongly dependent on having a representative and sufficiently deep collection of sequences from many individuals of each target species. A higher density SNP platform will be instrumental to undertake genome-wide phylogenetic and population genomics studies and to implement molecular breeding by Genomic Selection in <it>Eucalyptus</it>.</p>http://www.biomedcentral.com/1471-2229/11/65
spellingShingle de Lima Bruno
Kirst Matias
Silva-Junior Orzenil B
Grattapaglia Dario
Faria Danielle A
Pappas Georgios J
High-throughput SNP genotyping in the highly heterozygous genome of <it>Eucalyptus</it>: assay success, polymorphism and transferability across species
BMC Plant Biology
title High-throughput SNP genotyping in the highly heterozygous genome of <it>Eucalyptus</it>: assay success, polymorphism and transferability across species
title_full High-throughput SNP genotyping in the highly heterozygous genome of <it>Eucalyptus</it>: assay success, polymorphism and transferability across species
title_fullStr High-throughput SNP genotyping in the highly heterozygous genome of <it>Eucalyptus</it>: assay success, polymorphism and transferability across species
title_full_unstemmed High-throughput SNP genotyping in the highly heterozygous genome of <it>Eucalyptus</it>: assay success, polymorphism and transferability across species
title_short High-throughput SNP genotyping in the highly heterozygous genome of <it>Eucalyptus</it>: assay success, polymorphism and transferability across species
title_sort high throughput snp genotyping in the highly heterozygous genome of it eucalyptus it assay success polymorphism and transferability across species
url http://www.biomedcentral.com/1471-2229/11/65
work_keys_str_mv AT delimabruno highthroughputsnpgenotypinginthehighlyheterozygousgenomeofiteucalyptusitassaysuccesspolymorphismandtransferabilityacrossspecies
AT kirstmatias highthroughputsnpgenotypinginthehighlyheterozygousgenomeofiteucalyptusitassaysuccesspolymorphismandtransferabilityacrossspecies
AT silvajuniororzenilb highthroughputsnpgenotypinginthehighlyheterozygousgenomeofiteucalyptusitassaysuccesspolymorphismandtransferabilityacrossspecies
AT grattapagliadario highthroughputsnpgenotypinginthehighlyheterozygousgenomeofiteucalyptusitassaysuccesspolymorphismandtransferabilityacrossspecies
AT fariadaniellea highthroughputsnpgenotypinginthehighlyheterozygousgenomeofiteucalyptusitassaysuccesspolymorphismandtransferabilityacrossspecies
AT pappasgeorgiosj highthroughputsnpgenotypinginthehighlyheterozygousgenomeofiteucalyptusitassaysuccesspolymorphismandtransferabilityacrossspecies