Abstract
The development of single nucleotide polymorphism (SNP) markers provides the opportunity to improve many areas of plant breeding and population genetics. Unfortunately, for species such as the rubber tree (Hevea brasiliensis), the use of next-generation sequencing for genomic SNP discovery is very difficult because of the large genome size and the abundance of repeated sequences. Access to a set of validated SNP markers is a significant advantage for rubber researchers who wish to apply SNPs in scientific research. Here, we performed genomic sequencing of H. brasiliensis and generated 10,993,648 short reads, which were assembled into 10,071 contigs (N50 = 3078) by a de novo assembly strategy. A total of 2446 contigs presented no hits in the current H. brasiliensis genome assembly and may therefore be considered novel genomic sequences of rubber tree. A total of 143 putative polymorphic positions were selected, gene annotations were available for 58.7 % of the markers, and all of the sequences could be anchored to the released H. brasiliensis genome. These SNPs were validated in eight genotypes of H. brasiliensis and 15 F1 plants from a mapping population, resulting in 30 (20.9 %) positions correctly classified. The analysis revealed key candidate genes responsible for defence mechanisms and provided markers for further genetic improvement of Hevea in breeding programmes.
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References
Bachlava E, Taylor CA, Tang S, Bowers JE, Mandel JR, Burke JM, Knapp SJ (2012) SNP discovery and development of a high-density genotyping array for sunflower. PLoS One 7:e29814. doi:10.1371/journal.pone.0029814
Bentley DR, Balasubramanian S, Swerdlow HP, Smith GP, Milton J, Brown CG, Hall KP, Evers DJ, Barnes CL, Bignell HR, Boutell JM, Bryant J, Carter RJ, Keira Cheetham R, Cox AJ, Ellis DJ, Flatbush MR, Gormley NA, Humphray SJ, Irving LJ (2008) Accurate whole human genome sequencing using reversible terminator chemistry. Nature 456:53–59. doi:10.1038/nature07517
Camacho C, Coulouris G, Avagyan V, Ma N, Papadopoulos J, Bealer K, Madden TL (2009) BLAST+: architecture and applications. BMC Bioinform 10:421. doi:10.1186/1471-2105-10-421
Ching A, Caldwell KS, Jung M, Dolan M, Smith OS, Tingey S, Morgante M, Rafalski AJ (2002) SNP frequency, haplotype structure and linkage disequilibrium in elite maize inbred lines. BMC Genet 3:19–32
Danecek P, Auton A, Abecasis G, Albers CA, Banks E, DePristo MA, Handsaker RE, Lunter G, Marth GT, Sherry ST, McVean G, Durbin R, 1000 Genomes Project Analysis Group (2011) The variant call format and VCFtools. Bioinformatics 27:2156–2158. doi:10.1093/bioinformatics/btr330
DePristo MA, Banks E, Poplin R, Garimella KV, Maguire JR, Hartl C, Philippakis AA, del Angel G, Rivas MA, Hanna M, McKenna A, Fennell TJ, Kernytsky AM, Sivachenko AY, Cibulskis K, Gabriel SB, Altshuler D, Daly MJ (2011) A framework for variation discovery and genotyping using next-generation DNA sequencing data. Nat Genet 43:491–498. doi:10.1038/ng.806
Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15
Flint-Garcia SA, Thuillet AC, Yu J, Pressoir G, Romero SM, Mitchell SE, Doebley J, Kresovich S, Goodman MM, Buckler ES (2005) Maize association population: a high-resolution platform for quantitative trait locus dissection. Plant J 44:1054–1064. doi:10.1111/j.1365-313X.2005.02591.x
Florea L, Hartzell G, Zhang Z, Rubin GM, Miller W (1998) A computer program for aligning a cDNA sequence with a genomic DNA sequence. Genome Res 8:967–974
Garrison E, Marth G (2012) Haplotype-based variant detection from short-read sequencing. Preprint at arXiv:1207.3907[q-bio.GN]
Gonçalves PS, Fontes JRA (2012) Domestication and breeding of the rubber tree. In: Borém A, Lopes MTG, Clement CR, Noda H (eds) Domestication and breeding: Amazon species. UFV, Viçosa, pp 393–420
Goodstein DM, Shu S, Howson R, Neupane R, Hayes RD, Fazo J, Mitros T, Dirks W, Hellsten U, Putnam N, Rokhsar DS (2012) Phytozome: a comparative platform for green plant genomics. Nucleic Acids Res 40:D1178–D1186. doi:10.1093/nar/gkr944
Gupta PK, Roy JK, Prasad M (2001) Single nucleotide polymorphisms: a new paradigm for molecular marker technology and DNA polymorphism detection with emphasis on their use in plants. Curr Sci 80:524–535
Hillier LW, Marth GT, Quinlan AR, Dooling D, Fewell G, Barnett D, Fox P, Glasscock JI, Hickenbotham M, Huang W, Magrini VJ, Richt RJ, Sander SN, Stewart DA, Stromberg M, Tsung EF, Wylie T, Schedl T, Wilson RK, Mardis ER (2008) Whole-genome sequencing and variant discovery in C. elegans. Nat Methods 5:183–188. doi:10.1038/nmeth.1179
Höck J, Meister G (2008) The Argonaute protein family. Genome Biol 9:210. doi:10.1186/gb-2008-9-2-210
Jurka J, Klonowski P, Dagman V, Pelton P (1996) CENSOR—a program for identification and elimination of repetitive elements from DNA sequences. Comput Chem 20:119–121. doi:10.1016/S0097-8485(96)80013-1
Kim D, Pertea G, Trapnell C, Pimentel H, Kelley R, Salzberg SL (2013) TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol 14:R36. doi:10.1186/gb-2013-14-4-r36
Kohany O, Gentles AJ, Hankus L, Jurka J (2006) Annotation, submission and screening of repetitive elements in Repbase: repbaseSubmitter and censor. BMC Bioinform 7:474. doi:10.1186/1471-2105-7-474
Kota R, Rudd S, Facius A, Kolesov G, Thiel T, Zhang H, Stein N, Mayer K, Graner A (2003) Snipping polymorphisms from large EST collections in barley (Hordeum vulgare L.). Mol Genet Genomics 270:24–33. doi:10.1007/s00438-003-0891-6
Langmead B, Salzberg SL (2012) Fast gapped-read alignment with bowtie 2. Nat Methods 9:357–359. doi:10.1038/nmeth.1923
Langmead B, Schatz MC, Lin J, Pop M, Salzberg SL (2009) Searching for SNPs with cloud computing. Genome Biol 10:R134. doi:10.1186/gb-2009-10-11-r134
Le Guen V, Gay C, Xiong TC, Souza LM, Rodier-Goud M, Seguin M (2011) Development and characterization of 296 new polymorphic microsatellite markers for rubber tree (Hevea brasiliensis). Plant Breed 130:294–296. doi:10.1111/j.1439-0523.2010.01774.x
Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N, Marth G, Abecasis G, Durbin R, 1000 Genome Project Data Processing Subgroup (2009) The sequence alignment/map format and SAMtools. Bioinformatics 25:2078–2079. doi:10.1093/bioinformatics/btp352
Li D, Deng Z, Qin B, Liu X, Men Z (2012) De novo assembly and characterization of bark transcriptome using Illumina sequencing and development of EST-SSR markers in rubber tree (Hevea brasiliensis Muell. Arg.). BMC Genomics 13:192. doi:10.1186/1471-2164-13-192
Mantello CC, Suzuki FI, Souza LM, Gonçalves PS, Souza AP (2012) Microsatellite marker development for the rubber tree (hevea brasiliensis): characterization and cross-amplification in wild hevea species. BMC Res Notes 5:329. doi:10.1186/1756-0500-5-329
Mantello CC, Cardoso-Silva CB, da Silva CC, de Souza LM, Scaloppi Junior EJ, de Souza Gonçalves P, Vicentini R, de Souza AP (2014) De novo assembly and transcriptome analysis of the rubber tree (hevea brasiliensis) and SNP markers development for rubber biosynthesis pathways. PLoS One 9:e102665. doi:10.1371/journal.pone.0102665
Pootakham W, Chanprasert J, Jomchai N, Sangsrakru D, Yoocha T, Therawattanasuk K, Tangphatsornruang S (2011) Single nucleotide polymorphism marker development in the rubber tree, hevea brasiliensis (Euphorbiaceae). Am J Bot 98:e337–e338. doi:10.3732/ajb.1100228
Prochnik S, Marri PR, Desany B, Rabinowicz PD, Kodira C, Mohiuddin M, Rodriguez F, Fauquet C, Tohme J, Harkins T, Rokhsar DS, Rounsley S (2012) The cassava genome: current progress, future directions. Trop Plant Biol 5:88–94
Rafalski A (2002) Applications of single nucleotide polymorphisms in crop genetics. Curr Opin Plant Biol 5:94–100. doi:10.1016/S1369-5266(02)00240-6
Rahman AY, Usharraj AO, Misra BB, Thottathil GP, Jayasekaran K, Feng Y, Hou S, Ong SY, Ng FL, Lee LS, Tan HS, Sakaff MK, Teh BS, Khoo BF, Badai SS, Aziz NA, Yuryev A, Knudsen B, Dionne-Laporte A, Mchunu NP (2013) Draft genome sequence of the rubber tree hevea brasiliensis. BMC Genomics 14:75. doi:10.1186/1471-2164-14-75
Salgado LR, Koop DM, Pinheiro DG, Rivallan R, Le Guen V, Nicolás MF, de Almeida LG, Rocha VR, Magalhães M, Gerber AL, Figueira A, Cascardo JC, de Vasconcelos AR, Silva WA, Coutinho LL, Garcia D (2014) De novo transcriptome analysis of Hevea brasiliensis tissues by RNA-seq and screening for molecular markers. BMC Genomics 15:236. doi:10.1186/1471-2164-15-236
Schnable PS, Ware D, Fulton RS, Stein JC, Wei F, Pasternak S, Liang C, Zhang J, Fulton L, Graves TA, Minx P, Reily AD, Courtney L, Kruchowski SS, Tomlinson C, Strong C, Delehaunty K, Fronick C, Courtney B, Rock SM, Belter E, Du F, Kim K, Abbott RM, Cotton M, Levy A, Marchetto P, Ochoa K, Jackson SM, Gillam B (2009) The B73 maize genome: complexity, diversity, and dynamics. Science 326:1112–1115. doi:10.1126/science.1178534
Silva CC, Mantello CC, Campos T, Souza LM, Gonçalves PS, Souza AP (2014) Leaf-, panel- and latex-expressed sequenced tags from the rubber tree (hevea brasiliensis) under cold-stressed and suboptimal growing conditions: the development of gene-targeted functional markers for stress response. Mol Breed 34:1035–1053. doi:10.1007/s11032-014-0095-2
Souza LM, Mantello CC, Santos MO, de Souza Gonçalves P, Souza AP (2009) Microsatellites from rubber tree (Hevea brasiliensis) for genetic diversity analysis and cross-amplification in six hevea wild species. Conserv Genet Resour 1:75–79. doi:10.1007/s12686-009-9018-7
Souza LM, Gazaffi R, Mantello CC, Silva CC, Garcia D et al (2013) QTL mapping of growth-related traits in a full-sib family of rubber tree (Hevea brasiliensis) evaluated in a sub-tropical climate. PLoS One 8:e61238. doi:10.1371/journal.pone.0061238
Thiel T, Michalek W, Varshney RK, Graner A (2003) Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.). Theor Appl Genet 106:411–422. doi:10.1007/s00122-002-1031-0
Trapnell C, Pachter L, Salzberg SL (2009) TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 25:1105–1111. doi:10.1093/bioinformatics/btp120
Triwitayakorn K, Chatkulkawin P, Kanjanawattanawong S, Sraphet S, Yoocha T, Sangsrakru D, Chanprasert J, Ngamphiw C, Jomchai N, Therawattanasuk K, Tangphatsornruang S (2011) Transcriptome sequencing of Hevea brasiliensis for development of microsatellite markers and construction of a genetic linkage map. DNA Res 18:471–482. doi:10.1093/dnares/dsr034
Varshney RK, Beier U, Khlestkina EK, Kota R, Korzun V, Graner A, Börner A (2007) Single nucleotide polymorphisms in rye (Secale cereale L.): discovery, frequency, and applications for genome mapping and diversity studies. Theor Appl Genet 114:1105–1116. doi:10.1007/s00122-007-0504-6
Varshney RK, Nayak SN, May GD, Jackson SA (2009) Next-generation sequencing technologies and their implications for crop genetics and breeding. Trends Biotechnol 27:522–530. doi:10.1016/j.tibtech.2009.05.006
You FM, Huo N, Deal KR, Gu YQ, Luo M-C, McGuire PE, Dvorak J, Anderson OD (2011) Annotation-based genome-wide SNP discovery in the large and complex Aegilops tauschii genome using next-generation sequencing without a reference genome sequence. BMC Genomics 12:59. doi:10.1186/1471-2164-12-59
Acknowledgments
The authors gratefully acknowledge the Fundação de Amparo a Pesquisa do Estado de São Paulo, the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Computational Biology Program and Agropolis Program) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico for financial support and scholarships and a research fellowship.
Authors’ contributions
LMS performed the molecular genetic studies, helped to perform the biocomputational analysis and drafted the manuscript. GTS, CBCS and CCS performed a biocomputational analysis and drafted the manuscript. GTS, ARC, CCM and IAAA assisted in the molecular genetics studies. VLG participated in the evaluations of the molecular data and helped to draft the manuscript. APS conceived the study, participated in its design and coordination and helped to draft the manuscript. All of the authors read and approved the final manuscript.
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de Souza, L.M., Toledo-Silva, G., Cardoso-Silva, C.B. et al. Development of single nucleotide polymorphism markers in the large and complex rubber tree genome using next-generation sequence data. Mol Breeding 36, 115 (2016). https://doi.org/10.1007/s11032-016-0534-3
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DOI: https://doi.org/10.1007/s11032-016-0534-3