, Volume 128, Issue 4, pp 547–560 | Cite as

DNA transposon invasion and microsatellite accumulation guide W chromosome differentiation in a Neotropical fish genome

  • Michelle Orane Schemberger
  • Viviane Demetrio Nascimento
  • Rafael Coan
  • Érica Ramos
  • Viviane Nogaroto
  • Kaline Ziemniczak
  • Guilherme Targino Valente
  • Orlando Moreira-Filho
  • Cesar Martins
  • Marcelo Ricardo VicariEmail author
Original Article


Sex chromosome differentiation is subject to independent evolutionary processes among different lineages. The accumulation of repetitive DNAs and consequent crossing-over restriction guide the origin of the heteromorphic sex chromosome region. Several Neotropical fish species have emerged as interesting models for understanding evolution and genome diversity, although knowledge of their genomes is scarce. Here, we investigate the content of repetitive DNAs between males and females of Apareiodon sp. based on large-scale genomic data focusing on W sex chromosome differentiation. In Apareiodon, females are the heterogametic sex (ZW) and males are the homogametic sex (ZZ). The genome size estimate for Apareiodon was 1.2 Gb (with ~ 42× and ~ 47× coverage for males and females, respectively). In Apareiodon sp., approximately 36% of the genome was composed of repetitive DNAs and transposable elements (TEs) were the most abundant class. Read coverage analysis revealed different amounts of repetitive DNAs in males and females. The female-enriched clusters were located on the W sex chromosome and were mostly composed of microsatellite expansions and DNA transposons. Landscape analysis of TE contents demonstrated two major waves of invasions of TEs in the Apareiodon genome. Estimation of TE insertion times correlated with in situ locations permitted the inference that helitron, Tc1-mariner, and CMC EnSpm DNA transposons accumulated repeated copies during W chromosome differentiation between 20 and 12 million years ago. DNA transposons and microsatellite expansions appeared to be major players in W chromosome differentiation and to guide modifications in the genome content of the heteromorphic sex chromosomes.


Cytogenomics Read coverage analysis Sex chromosomes Tandem repeats Transposable elements W chromosome region 



The authors are grateful to ICMBio (Instituto Chico Mendes de Conservação da Biodiversidade) (license number 10538-1 to collect specimens). This study was supported by the Fundação Araucária (Fundação Araucária de Apoio ao Desenvolvimento Científico e Tecnológico do Estado do Paraná), FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo, grant number: 2015/16661-1), CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Finance Code 001), and CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico, grant number: 304166/2016-2).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

412_2019_721_MOESM1_ESM.pdf (449 kb)
ESM 1 (PDF 448 kb)
412_2019_721_MOESM2_ESM.pdf (2.7 mb)
ESM 2 (PDF 2809 kb)
412_2019_721_MOESM3_ESM.html (26 kb)
Supplementary figure 2 Repeat landscape of DNA transposons in the male Apareiodon sp. genome. The graphs show, for each element, the sequence divergence from their consensus sequence (x-axis) in relation to their number of copies in the genome (y-axis). Peaks represent waves of insertion (yellow arrows) of specific elements in the genome. Elements with older waves of insertion are presented on the right side of the graph, while recent waves of insertions are depicted on the left side. Different colors indicate the distinct element types described on the right side. (HTML 26 kb)
412_2019_721_MOESM4_ESM.html (26 kb)
Supplementary figure 3 Repeat landscape of DNA transposons in the female Apareiodon sp. genome. The graphs show, for each element, the sequence divergence from their consensus sequence (x-axis) in relation to their number of copies in the genome (y-axis). Peaks represent waves of insertion (yellow arrows) of specific elements in the genome. Elements with older waves of insertion are presented on the right side of the graph, while recent waves of insertions are depicted on the left side. Different colors indicate distinct element types described on the right side. (HTML 25 kb)
412_2019_721_MOESM5_ESM.html (40 kb)
Supplementary figure 4 Repeat landscape of retrotransposons in the male Apareiodon sp. genome. The graphs show, for each element, the sequence divergence from their consensus sequence (x-axis) in relation to their number of copies in the genome (y-axis). Peaks represent waves of insertion (yellow arrows) of specific elements in the genome. Elements with older waves of insertion are presented on the right side of the graph, while recent waves of insertions are depicted on the left side. Different colors indicate distinct element types described on the right. (HTML 40 kb)
412_2019_721_MOESM6_ESM.html (41 kb)
Supplementary figure 5 Repeat landscape of retrotransposons in the male Apareiodon sp. genome. The graphs show, for each element, the sequence divergence from their consensus sequence (x-axis) in relation to their number of copies in the genome (y-axis). Peaks represent waves of insertion (yellow arrows) of specific elements in the genome. Elements with older waves of insertion are presented on the right side of the graph, while recent waves of insertions are depicted on the left side. Different colors indicate distinct element types described on the right side. (HTML 40 kb)


  1. Arkhipova I, Meselson M (2005) Deleterious transposable elements and the extinction of asexuals. Bioessays 27:76–85. CrossRefPubMedGoogle Scholar
  2. Bachtrog D (2013) Y-chromosome evolution: emerging insights into processes of Y-chromosome degeneration. Nat Rev Genet 14:113–124. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Bellafronte E, Schemberger MO, Moreira-Filho O, Almeida MC, Artoni RF, Margarido VP, Vicari MR (2011) Chromosomal markers in Parodontidae: an analysis of new and reviewed data with phylogenetic inferences. Rev Fish Biol Fish 12:559–570. CrossRefGoogle Scholar
  4. Bergero R, Charlesworth D (2009) The evolution of restricted recombination in sex chromosomes. Trends Ecol Evol 24:94–102. CrossRefPubMedGoogle Scholar
  5. Bertollo LAC, Takahashi CS, Moreira-Filho O (1978) Cytotaxonomic considerations on Hoplias lacerdae (Pisces, Erythrinidae). Braz J Genet 1:103–120Google Scholar
  6. Bradnam KR, Fass JN, Alexandrov A, Baranay P, Bechner M, Birol I, Boisvert S, Chapman JA, Chapuis G, Chikhi R, Chitsaz H, Chou WC, Corbeil J, del Fabbro C, Docking TR, Durbin R, Earl D, Emrich S, Fedotov P, Fonseca NA, Ganapathy G, Gibbs RA, Gnerre S, Godzaridis É, Goldstein S, Haimel M, Hall G, Haussler D, Hiatt JB, Ho IY, Howard J, Hunt M, Jackman SD, Jaffe DB, Jarvis ED, Jiang H, Kazakov S, Kersey PJ, Kitzman JO, Knight JR, Koren S, Lam TW, Lavenier D, Laviolette F, Li Y, Li Z, Liu B, Liu Y, Luo R, MacCallum I, MacManes MD, Maillet N, Melnikov S, Naquin D, Ning Z, Otto TD, Paten B, Paulo OS, Phillippy AM, Pina-Martins F, Place M, Przybylski D, Qin X, Qu C, Ribeiro FJ, Richards S, Rokhsar DS, Ruby JG, Scalabrin S, Schatz MC, Schwartz DC, Sergushichev A, Sharpe T, Shaw TI, Shendure J, Shi Y, Simpson JT, Song H, Tsarev F, Vezzi F, Vicedomini R, Vieira BM, Wang J, Worley KC, Yin S, Yiu SM, Yuan J, Zhang G, Zhang H, Zhou S, Korf IF (2013) Assemblathon 2: evaluating de novo methods of genome assembly in three vertebrate species. GigaScience 2(10).
  7. Chalopin D, Volff JN, Galiana D, Anderson JL, Schartl M (2015) Transposable elements and early evolution of sex chromosomes in fish. Chromosom Res 23:545–560. CrossRefGoogle Scholar
  8. Charlesworth B (1991) The evolution of sex chromosomes. Science 251:1030–1033. CrossRefPubMedGoogle Scholar
  9. Charlesworth B (1996) The evolution of chromosomal sex determination and dosage compensation. Curr Biol 6:149–162. CrossRefPubMedGoogle Scholar
  10. Charlesworth B, Sniegowski P, Stephan W (1994) The evolutionary dynamics of repetitive DNA in eukaryotes. Nature 371:215–220. CrossRefPubMedGoogle Scholar
  11. Charlesworth D, Charlesworth B, Marais G (2005) Steps in the evolution of heteromorphic sex chromosomes. Heredity 95:118–128. CrossRefPubMedGoogle Scholar
  12. Chen S, Zhang G, Shao C, Huang Q, Liu G, Zhang P, Song W, An N, Chalopin D, Volff JN, Hong Y, Li Q, Sha Z, Zhou H, Xie M, Yu Q, Liu Y, Xiang H, Wang N, Wu K, Yang C, Zhou Q, Liao X, Yang L, Hu Q, Zhang J, Meng L, Jin L, Tian Y, Lian J, Yang J, Miao G, Liu S, Liang Z, Yan F, Li Y, Sun B, Zhang H, Zhang J, Zhu Y, du M, Zhao Y, Schartl M, Tang Q, Wang J (2014) Whole-genome sequence of a flatfish provides insights into ZW sex chromosome evolution and adaptation to a benthic lifestyle. Nat Genet 46:253–260. CrossRefPubMedGoogle Scholar
  13. Chen X, Mei J, Wu J, Jing J, Ma W, Zhang J, Gui JF (2015) A comprehensive transcriptome provides candidate genes for sex determination/differentiation and SSR/SNP markers in yellow catfish. Mar Biotechnol 17:190–198. CrossRefPubMedGoogle Scholar
  14. Coates BS, Sumerford DV, Hellmich RL, Lewis LC (2010) Helitron-like transposon superfamily from Lepidoptera disrupts (GAAA)n microsatellites and is responsible for flanking sequence similarity within a microsatellite family. J Mol Evol 70:275–288. CrossRefPubMedGoogle Scholar
  15. de Koning AJ, Gu W, Castoe TA, Batzer MA, Pollock DD (2011) Repetitive elements may comprise over two-thirds of the human genome. PLoS Genet 71:e1002384. CrossRefGoogle Scholar
  16. de Oliveira EA, Sember A, Bertollo LAC, Yano CF, Ezaz T, Moreira-Filho O, Hatanaka T, Trifonov V, Liehr T, al-Rikabi ABH, Ráb P, Pains H, Cioffi MB (2018) Tracking the evolutionary pathway of sex chromosomes among fishes: characterizing the unique XX/XY1Y2 system in Hoplias malabaricus (Teleostei, Characiformes). Chromosoma 127:115–128. CrossRefPubMedGoogle Scholar
  17. do Nascimento VD, Coelho KA, Nogaroto V, Almeida RB, Ziemniczak K, Centofante L, Pavanelli CS, Torres RA, Moreira-Filho O, Vicari MR (2018) Do multiple karyomorphs and population genetics of freshwater darter characines (Apareiodon affinis) indicate chromosomal speciation? Zool Anz 272:93–103. CrossRefGoogle Scholar
  18. Fernández-Medina RD, Ribeiro JMC, Carareto CMA, Velasque L, Struchiner CJ (2012) Losing identity: structural diversity of transposable elements belonging to different classes in the genome of Anopheles gambiae. BMC Genomics 13:272. CrossRefPubMedPubMedCentralGoogle Scholar
  19. Feschotte C (2008) Transposable elements and the evolution of regulatory networks. Nat Rev Genet 9:397–405. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Frank-Kamenetskii MD, Mirkin SM (1995) Triplex DNA structures. Annu Rev Biochem 64:65–95. CrossRefPubMedGoogle Scholar
  21. Gazoni T, Haddad CFB, Narimatsu H, Cabral-de-Mello DC, Lyra ML, Parise-Maltempi PP (2018) More sex chromosomes than autosomes in the Amazonian frog Leptodactylus pentadactylus. Chromosoma 127:269–278. CrossRefPubMedGoogle Scholar
  22. Heikkinen E, Launonen V, Muller E, Bachmann L (1995) The pvB370 BamIII satellite DNA family of the Drosophila virilis group and its evolutionary relation to mobile dispersed genetic pDv elements. J Mol Evol 41:604–614. CrossRefPubMedGoogle Scholar
  23. Herpin A, Braasch I, Kraeussling M, Schmidt C, Thoma EC, Nakamura S, Tanaka M, Schartl M (2010) Transcriptional rewiring of the sex determining Dmrt1 gene duplicate by transposable elements. PLoS Genet 6:e1000844. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Kapitonov VV, Holmquist GP, Jurka J (1998) L1 repeat is a basic unit of heterochromatin satellites in Cetaceans. Mol Biol Evol 15:611–612CrossRefGoogle Scholar
  25. Kapusta A, Kronenberg Z, Lynch VJ, Zhuo X, Ramsay L, Bourque G, Yandell M, Feschotte C (2013) Transposable elements are major contributors to the origin, diversification, and regulation of vertebrate long noncoding RNAs. PLoS Genet 9:e1003470. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Keinath MC, Timoshevskaya N, Timoshevskiy VA, Voss SR, Smith JJ (2018) Miniscule differences between the sex chromosomes in the giant genome of a salamander, Ambystoma mexicanum. Sci Rep 8:17882. CrossRefPubMedPubMedCentralGoogle Scholar
  27. Kelkar YD, Eckert KA, Chiaromonte F, Makova KD (2011) A matter of life or death: how microsatellites emerge in and vanish from the human genome. Genome Res 21:2038–2048 CrossRefGoogle Scholar
  28. Kidwell MG, Lisch DR (2001) Perspective: transposable elements, parasitic DNA, and genome evolution. Evolution 55:1–24. CrossRefPubMedGoogle Scholar
  29. Kirkness EF, Bafna V, Halpern AL, Levy S, Remington K, Rusch DB, Delcher AL, Pop M, Wang W, Fraser CM, Venter JC (2003) The dog genome: survey sequencing and comparative analysis. Science 301:1898–1903. CrossRefPubMedGoogle Scholar
  30. Komissarov AS, Galkina SA, Koshel EI, Kulak MM, Dyomin AG, O’Brien SJ, Gaginskaya ER, Saifitdinova AF (2018) New high copy tandem repeat in the content of the chicken W chromosome. Chromosoma 127:73–83. CrossRefPubMedGoogle Scholar
  31. Li SF, Guo YJ, Li JR, Zhang DX, Wang BX, Li N, Deng CL, Gao WJ (2019) The landscape of transposable elements and satellite DNAs in the genome of a dioecious plant spinach (Spinacia oleracea L.). Mob DNA 10(3).
  32. Marino-Ramirez L, Lewis KC, Landsman D, Jordan IK (2005) Transposable elements donate lineage-specific regulatory sequences to host genomes. Cytogenet Genome Res 110:333–341. CrossRefPubMedPubMedCentralGoogle Scholar
  33. Milani D, Cabral-de-Mello DC (2014) Microsatellite organization in the grasshopper Abracris flavolineata (Orthoptera: Acrididae) revealed by FISH mapping: remarkable spreading in the A and B chromosomes. PLoS One 9:e97956. CrossRefPubMedPubMedCentralGoogle Scholar
  34. Mongue AJ, Nguyen P, Volenikova A, Walters JR (2017) Neo-sex chromosomes in the Monarch butterfly, Danaus plexippus. G3 (Bethesda) 7:3281–3294. CrossRefGoogle Scholar
  35. Novak P, Neumann P, Macas J (2010) Graph-based clustering and characterization of repetitive sequences in 592 next-generation sequencing data. BMC Bioinformatics 11:378. CrossRefPubMedPubMedCentralGoogle Scholar
  36. Novak P, Neumann P, Pech J, Steinhaisl J, Macas J (2013) RepeatExplorer: a galaxy-based web server for genome-wide characterization of eukaryotic repetitive elements from next-generation sequence reads. Bioinformatics 29:792–793. CrossRefPubMedGoogle Scholar
  37. Pinkel D, Straume T, Gray JW (1986) Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci U S A 83:2934–2938. CrossRefPubMedPubMedCentralGoogle Scholar
  38. Priyadarshini P, Murthy BS, Nagaraju J, Singh L (2003) GATA-binding protein expressed predominantly in the pupal ovary of the silkworm, Bombyx mori. Insect Biochem Mol Biol 33:185–195. CrossRefPubMedGoogle Scholar
  39. Pucci MB, Barbosa P, Nogaroto V, Almeida MC, Artoni RF, Pansonato-Alves JC, Scacchetti PC, Foresti F, Moreira-Filho O, Vicari MR (2016) Chromosomal spreading of microsatellites and (TTAGGG)n sequences in the Characidium zebra and C. gomesi genomes (Characiformes: Crenuchidae). Cytogenet Genome Res 149:182–190. CrossRefPubMedGoogle Scholar
  40. Raymond CS, Murphy MW, O'Sullivan MG, Bardwell VJ, Zarkower D (2000) Dmrt1, a gene related to worm and fly sexual regulators, is required for mammalian testis differentiation. Genes Dev 14:2587–2595. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Saitoh Y, Saitoh H, Ohtomo K, Mizuno S (1991) Occupancy of the majority of DNA in the chicken W chromosome by bent-repetitive sequences. Chromosoma 101:32–40. CrossRefPubMedGoogle Scholar
  42. Schemberger MO, Bellafronte E, Nogaroto V, Almeida MC, Schühli GS, Artoni RF, Moreira-Filho O, Vicari MR (2011) Differentiation of repetitive DNA sites and sex chromosome systems reveal closely related group in Parodontidae (Actinopterygii: Characiformes). Genetica 139:1499–1508. CrossRefPubMedGoogle Scholar
  43. Schemberger MO, Oliveira JIN, Nogaroto V, Almeida MC, Artoni RF, Cestari MM, Moreira-Filho O, Vicari MR (2014) Construction and characterization of a repetitive DNA library in Parodontidae (Actinopterygii: Characiformes): a genomic and evolutionary approach to the degeneration of the W sex chromosome. Zebrafish 11:518–527. CrossRefPubMedPubMedCentralGoogle Scholar
  44. Schemberger MO, Nogaroto V, Almeida MC, Artoni RF, Valente GT, Martins C, Moreira-Filho O, Cestari MM, Vicari MR (2016) Sequence analyses and chromosomal distribution of the Tc1/Mariner element in Parodontidae fish (Teleostei: Characiformes). Gene 593:308–314. CrossRefPubMedGoogle Scholar
  45. Singh L, Wadhwan R, Naidu S, Nagaraj R, Ganesan M (1994) Sex- and tissue-specific Bkm (GATA)-binding protein in the germ cells of heterogametic sex. J Biol Chem 269:25321–25327PubMedGoogle Scholar
  46. Smit AFA, Hubley R (2008–2015) RepeatModeler Open—1.0 Available from Accessed 26 Nov 2018
  47. Smit AFA, Hubley R, Green P (2013–2015) RepeatMasker Open—4.0. Available at: Accessed 26 Nov 2018
  48. Steinemann S, Steinemann M (2005) Retroelements: tools for sex chromosome evolution. Cytogenet Genome Res 110:134–143. CrossRefPubMedGoogle Scholar
  49. Sumner AT (1972) A simple technique for demonstrating centromeric heterochromatin. Exp Cell Res 75:304–306. CrossRefPubMedGoogle Scholar
  50. Teaniniuraitemoana V, Huvet A, Levy P, Klopp C, Lhuillier E, Gaertner-Mazouni N, Le Moullac G (2014) Gonad transcriptome analysis of pearl oyster Pinctada margaritifera: identification of potential sex differentiation and sex determining genes. BMC Genomics 15:491. CrossRefPubMedPubMedCentralGoogle Scholar
  51. Traldi JB, Vicari MR, Martinez JDF, Blanco DR, Lui RL, Moreira-Filho O (2016) Chromosome analyses of Apareiodon argenteus and Apareiodon davisi (Characiformes, Parodontidae): an extensive chromosomal polymorphism of 45S and 5S ribosomal DNAs. Zebrafish 13:19–25. CrossRefPubMedGoogle Scholar
  52. Treangen TJ, Salzberg SL (2012) Repetitive DNA and next-generation sequencing: computational challenges and solutions. Nat Rev Genet 13:36–46. CrossRefGoogle Scholar
  53. Vicari MR, Moreira-Filho O, Artoni RF, Bertollo LAC (2006) ZZ/ZW sex chromosome system in an undescribed species of the genus Apareiodon Characiformes, Parodontidae. Cytogenet Genome Res 114:163–168. CrossRefPubMedGoogle Scholar
  54. Vicari MR, Nogaroto V, Noleto RB, Cestari MM, Cioffi MB, Almeida MC, Moreira-Filho O, Bertollo LAC, Artoni RF (2010) Satellite DNA and chromosomes in Neotropical fishes: methods, applications and perspectives. J Fish Biol 76:1094–1116. CrossRefPubMedGoogle Scholar
  55. Vicente VE, Bertollo LA, Valentini SR, Moreira-Filho O (2003) Origin and differentiation of sex chromosome system in Parodon hilarii (Pisces, Parodontidae). Satellite DNA, G and C banding. Genetica 119:115–120. CrossRefPubMedGoogle Scholar
  56. Viger RS, Mertineit C, Trasler JM, Nemer M (1998) Transcription factor GATA 4 is expressed in a sexually dimorphic pattern during mouse gonadal development and is a potent activator of the MIS promoter. Development 125:2665–2675PubMedGoogle Scholar
  57. Volff JN, Bouneau L, Ozouf-Costaz C, Fischer C (2003) Diversity of retrotransposable elements in compact pufferfish genomes. Trends Genet 19:674–678. CrossRefPubMedGoogle Scholar
  58. Waterhouse RM, Seppey M, Simão FA, Manni M, Ioannidis P, Klioutchnikov G, Kriventseva EV, Zdobnov EM (2017) BUSCO applications from quality assessments to gene prediction and phylogenomics. Mol Biol Evol 35:543–548. CrossRefPubMedCentralGoogle Scholar
  59. White MA, Kitano J, Peichel CL (2015) Purifying selection maintains dosage-sensitive genes during degeneration of the threespine stickleback Y chromosome. Mol Biol Evol 32:1981–1995. CrossRefPubMedPubMedCentralGoogle Scholar
  60. Wilder J, Hollocher H (2001) Mobile elements and the genesis of microsatellites in dipterans. Mol Biol Evol 18:384–392. CrossRefPubMedGoogle Scholar
  61. Xin H, Zhang T, Han Y, Wu Y, Shi J, Xi M, Jiang J (2018) Chromosome painting and comparative physical mapping of the sex chromosomes in Populus tomentosa and Populus deltoides. Chromosoma 127:313–321. CrossRefPubMedGoogle Scholar
  62. Zerbino D, Birney E (2008) Velvet: algorithms for de novo short read assembly using de Bruijn graphs. Genome Res gr-074492. CrossRefGoogle Scholar
  63. Ziemniczak K, Traldi JB, Nogaroto V, Almeida MC, Artoni RF, Moreira-Filho O, Vicari MR (2014) In situ localization of (GATA)n and (TTAGGG)n repeated DNAs and W sex chromosome differentiation in Parodontidae (Actinopterygii: Characiformes). Cytogenet Genome Res 144:325–332. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Michelle Orane Schemberger
    • 1
  • Viviane Demetrio Nascimento
    • 2
  • Rafael Coan
    • 3
  • Érica Ramos
    • 3
  • Viviane Nogaroto
    • 1
  • Kaline Ziemniczak
    • 4
  • Guilherme Targino Valente
    • 5
  • Orlando Moreira-Filho
    • 4
  • Cesar Martins
    • 3
  • Marcelo Ricardo Vicari
    • 1
    • 2
    Email author
  1. 1.Departamento de Biologia Estrutural, Molecular e GenéticaUniversidade Estadual de Ponta GrossaPonta GrossaBrazil
  2. 2.Programa de Pós-Graduação em Genética, Centro PolitécnicoUniversidade Federal do ParanáCuritibaBrazil
  3. 3.Departamento de Morfologia, Instituto de Biociências de BotucatuUniversidade Estadual PaulistaBotucatuBrazil
  4. 4.Departamento de Genética e EvoluçãoUniversidade Federal de São CarlosSão CarlosBrazil
  5. 5.Departamento de Bioprocessos e Biotecnologia, Fazenda LageadoUniversidade Estadual PaulistaBotucatuBrazil

Personalised recommendations