Theoretical and Applied Genetics

, Volume 90, Issue 7–8, pp 933–947 | Cite as

Genetic mapping of QTLs controlling vegetative propagation in Eucalyptus grandis and E. urophylla using a pseudo-testcross strategy and RAPD markers

  • D. Grattapaglia
  • F. L. Bertolucci
  • R. R. Sederoff


We have extended the combined use of the “pseudo-testcross” mapping strategy and RAPD markers to map quantitative trait loci (QTLs) controlling traits related to vegetative propagation in Eucalyptus. QTL analyses were performed using two different interval mapping approaches, MAPMAKER-QTL (maximum likelihood) and QTL-STAT (non-linear least squares). A total of ten QTLs were detected for micropropagation response (measured as fresh weight of shoots, FWS), six for stump sprouting ability (measured as # stump sprout cuttings, #Cutt) and four for rooting ability (measured as % rooting of cuttings, %Root). With the exception of three QTLs, both interval-mapping methods yielded similar results in terms of QTL detection. Discrepancies in the most likely QTL location were observed between the two methods. In 75% of the cases the most likely position was in the same, or in an adjacent, interval. Standardized gene substitution effects for the QTLs detected were typically between 0.46 and 2.1 phenotypic standard deviations (σp), while differences between the family mean and the favorable QTL genotype were between 0.25 and 1.07 (σp). Multipoint estimates of the total genetic variation explained by the QTLs (89.0% for FWS, 67.1 % for#Cutt, 62.7% for %Root) indicate that a large proportion of the variation in these traits is controlled by a relatively small number of major-effect QTLs. In this cross, E. grandis is responsible for most of the inherited variation in the ability to form shoots, while E. urophylla contributes most of the ability in rooting. QTL mapping in the pseudo-testcross configuration relies on withinfamily linkage disequilibrium to establish marker/trait associations. With this approach QTL analysis is possible in any available full-sib family generated from undomesticated and highly heterozygous organisms such as forest trees. QTL mapping on two-generation pedigrees opens the possibility of using already existing families in retrospective QTL analyses to gather the quantitative data necessary for marker-assisted tree breeding.

Key words

RAPD Pseudo-testcross Eucalyptus QTL Vegetative propagation 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Armstrong CL, Romero-Severson J, Hodges TK (1992) Improved tissue culture response of an elite maize inbred through backcross breeding, and identification of chromosomal regions important for regeneration by RFLP analysis. Theor Appl Genet 84: 755–762Google Scholar
  2. Beavis WD, Grant D, Albertsen M, Fincher R (1991) Quantitative trait loci for plant height in four maize populations and their associations with qualitative genetic loci. Theor Appl Genet 83:141–145Google Scholar
  3. Beckmann JS, Soller M (1983) Restriction fragment length polymorphisms in genetic improvement methodologies, mapping and costs. Theor Appl Genet 67:35–43Google Scholar
  4. Blake TJ (1972) Studies on the lignotubers of Eucalyptus obliqua L'herit. III. The effects of seasonal and nutritional factors on dormant bud development. New Phytol 71:327–334Google Scholar
  5. Bradshaw HD Jr, Foster GS (1992) Marker-aided selection and propagation systems in trees: advantages of cloning for studying quantitative inheritance. Can J For Res 22:1044–1049Google Scholar
  6. Brandao LG (1984) The new eucalypt forest. Marcus Wallenberg Symposium, Proc # 1. Falun, Sweden, pp 3–15Google Scholar
  7. Campinhos E, Ikemori Y (1980) Mass production of Eucalyptus s. by rooting cuttings. In: IUFRO Symposium Genetic Improvement and Productivity of Fast-growing Trees. Sao Paulo, Brazil, pp 60–67Google Scholar
  8. Cowen NM, Johnson CD, Armstrong K, Miller M, Woosley A, Pescitelli S, Skokut M, Belmar S, Petolino JF (1992) Mapping genes conditioning in vitro androgenesis in maize using RFLP analysis. Theor Appl Genet 84:720–724Google Scholar
  9. Cremer KW (1973) Ability of Eucalyptus regnans and associated evergreen hardwoods to recover from cutting or complete defoliation in different seasons. Aust For Res 6:9–22Google Scholar
  10. Darvasi A, Weinreb A, Minke V, Weller JI, Soller M (1993) Detecting marker-QTL linkage and estimating QTL gene effect and map location using a saturated genetic map. Genetics 134:943–951Google Scholar
  11. De Fossard RA (1974) Tissue culture of Eucalyptus. Aust For 37:43–54Google Scholar
  12. Delwaulle JC (1985) Plantations clonales d'Eucalyptus hybridas an Congo. Rev Bois Forets Trop 208:37–42Google Scholar
  13. Doebley J, Stec A (1993) Inheritance of the morphological differences between maize and teosinte: comparision of results for two F2 populations. Genetics 134:559–570Google Scholar
  14. Dudley JW (1993) Molecular markers in plant improvement: manipulation of genes affecting quantitative traits. Crop Sci 33:660–668Google Scholar
  15. Eldridge K, Davidson J, Harwood C, Van Wyk G (1993) Eucalypt domestication and breeding. Oxford Science Publications. Oxford University Press, OxfordGoogle Scholar
  16. Falconer DS (1989) Introduction to quantitative genetics. 3rd edn, John Wiley and Sons, New YorkGoogle Scholar
  17. Fazio S (1964) Propagating Eucalyptus from cuttings. Comb Proc Int Plant Propagators Soc 14:288–290Google Scholar
  18. Grattapaglia D, Sederoff R (1994) Genetic linkage maps of Eucalyptus grandis and E. urophylla using a pseudo-testcross mapping strategy and RAPD markers. Genetics 137:1121–1137Google Scholar
  19. Grattapaglia D, Assis TF, Caldas LS (1987) Residual effects of benzyladenine and naphthalenacetic acid on the “in vitro” multiplication and rooting of eight Eucalyptus species. In: Abstracts 2nd National Symp Plant Tissue Culture. Brasilia, DF, Brazil, Abstract # 12Google Scholar
  20. Grattapaglia D, Caldas LS, Machado MA, Assis TF (1990) Large scale micropropagation of Eucalyptus species and hybrids. In: Abstracts 7th Int Congr on Plant Tissue and Cell Culture. Amsterdam, Abstract # A3–123Google Scholar
  21. Gupta PK, Mascarenhas AF (1987) Eucalyptus. In: Bonga JM, Durzan DJ (eds) Cell and tissue culture in forestry vol. 3. Martinus Nijhoff Publishers, Dordrecht, pp 385–399Google Scholar
  22. Hartman HT, Kester D (1983) Plant propagation: principles and practices, 2nd edn. Prentice Hall, Englewood Cliffs, New JerseyGoogle Scholar
  23. Hartney VJ (1980) Vegetative propagation in the eucalypts. Aust For Res 10:191–211Google Scholar
  24. Keats BJS, Sherman SL, Morton NE, Robson EB, Buetow KH, Cann HM, Cartwright PE, Chakravarti A, Francke U, Green PP, Ott J (1991) Guidelines for human linkage maps: an international system for human linkage maps (ISLM, 1990). Genomics 9:557–560Google Scholar
  25. Knapp SJ, Bridges WC (1990) Using molecular markers to estimate quantitative genetic parameters: power and genetic variances for unreplicated and replicated progeny. Genetics 126:769–777Google Scholar
  26. Knapp SJ, Bridges WC, Liu BH (1992) Mapping quantitative trait loci using nonsimultaneous and simultaneous estimators and hypothesis tests. In: Beckmann J, Osborn TC (eds) Plant genomes: methods for genetic and physical mapping. Kluwer Academic Publishers, Dordrecht, pp 209–238Google Scholar
  27. Knott SA, Haley CS (1992) Maximum-likelihood mapping of quantitative trait loci using full-sib families. Genetics 132:1211–1222Google Scholar
  28. Lande R, Thompson R (1990) Efficiency of marker-assisted selection in the improvement of quantitative traits. Genetics 124:743–756Google Scholar
  29. Lander ES, Botstein D (1989) Mapping Mendelian factors underlying quantitative traits using RFLP linkage maps. Genetics 121:185–199Google Scholar
  30. Lander ES, Green P, Abrahamson J, Baarlow A, Daly MJ, Lincoln SE, Newburg L (1987) MapMaker: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174–181Google Scholar
  31. Liu BH, Knapp SJ (1990) GMENDEL: a program for Mendelian segregation and linkage analysis of individual or multiple progeny using log-likelihood ratios. J Hered 81:407Google Scholar
  32. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497Google Scholar
  33. SAS (1988) SAS/STAT User's Guide, version 6.03. SAS Institute, Cary, North CarolinaGoogle Scholar
  34. Soller M (1978) The use of loci associated with quantitative traits in dairy cattle improvement. Anim Prod 27:133–139Google Scholar
  35. Soller M (1991) Mapping quantitative trait loci affecting traits of economic importance in animal populations using molecular markers. In: Schook LB, Lewin HA, Mclaren DG (eds) Gene-mapping techniques and applications. Marcel Dekker Inc., New York, pp 21–50Google Scholar
  36. Strauss SH, Lande R, Namkoong G (1992) Limitations of molecular-marker aided selection in forest tree breeding. Can J For Res 22:1050–1061Google Scholar
  37. Stuber CW (1992) Biochemical and molecular markers in plant breeding. In: Dudley JW, Hallauer AR, Ryder M (eds) Plant breeding reviews vol. 9. John Wiley and Sons, New York, pp 37–61Google Scholar
  38. Stuber CW, Lincoln SE, Wolff DW, Helentjaris T, Lander ES (1992) Identification of genetic factors contributing to heterosis in a hybrid from two elite maize inbred lines using molecular markers. Genetics 132:823–839Google Scholar
  39. Tanksley SD, Hewitt JD (1988) Use of molecular markers in breeding for soluble solids in tomato — a re-examination. Theor Appl Genet 75:811–823Google Scholar
  40. Van Eck HJ, Jacobs JME, Stam P, Ton J, Stiekema WJ, Jacobsen E (1994) Multiple alleles for tuber shape in dipoid potato detected by qualitative and quantitative genetic analysis using RFLPs. Genetics 137:303–309Google Scholar
  41. Van Wyk G (1985) Tree breeding in support of vegetative propagation of Eucalyptus grandis (Hill) Maiden. S Afr For J 12:33–40Google Scholar
  42. Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535Google Scholar
  43. Yu K, Pauls KP (1993) Identification of a RAPD marker associated with somatic embryogenesis in alfalfa. Plant Mol Biol 22: 269–277Google Scholar
  44. Zeng Z (1994) Precision mapping of quantitative trait loci. Genetics 136:1457–1468Google Scholar
  45. Zobel BJ (1993) Clonal forestry in the eucalypts. In: Ahuja MR, Libby WJ (eds) Clonal Forestry II. Conservation and application. Springer Verlag, Berlin, pp 139–148Google Scholar
  46. Zobel BJ, Talbert J (1984) Applied forest tree improvement. John Wiley and Sons, New YorkGoogle Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • D. Grattapaglia
    • 1
  • F. L. Bertolucci
    • 2
  • R. R. Sederoff
    • 1
  1. 1.Forest Biotechnology Group, Departments of Genetics and ForestyNorth Carolina State UniversityRaleighUSA
  2. 2.Gerencia de Tecnologia, Aracruz Celuose S.A.-C.PAracruz, ESBrazil

Personalised recommendations