Tuesday, 29 November 2016

Verification of a phenotypic discrimination method for hybrid larch seedlings using DNA markers

Published Date
Volume 19, Issue 5pp 461–468

Original Article
DOI: 10.1007/s10310-014-0

Cite this article as: 
Kita, K., Uchiyama, K., Ichimura, Y. et al. J For Res (2014) 19: 461. doi:10.1007/s10310-014-0438-1

  • Kazuhito Kita
  • Kazuko Uchiyama
  • Yasuhiro Ichimura
  • Yoshinari Moriguchi
  • Yoshihiko Tsumura
  • Makoto Kuromaru

The hybrid between Kuril larch (Larix gmelinii var. japonica) and Japanese larch (Lkaempferi) is an important afforestation tree species in Hokkaido, Japan, because of its rapid juvenile growth, straight stem, and resistance to bark gnawing by voles. To produce desirable hybrid seedlings, precise seedling discrimination is essential. However, continuous variations in morphological and phenological traits occur across L. gmelinii var. japonica × L. kaempferi and L. gmelinii var. japonica × L. gmelinii var. japonica seedlings. Therefore, we used DNA markers to verify and improve the morphological and phenological discrimination method. We collected seeds from an interspecific seed orchard and, using chloroplast DNA analysis, we showed that the hybridization rates of 1-year-old seedlings were different between years (2004, 23.2 %; 2005, 53.6 %) and between mother trees (2004, 5.8–39.4 %; 2005, 20.0–81.0 %). Discriminant analyses revealed that the root collar diameter of 2-year-old seedlings, number of sylleptic branches of 2-year-old seedlings, and day of terminal bud set in 1- and 2-year-old seedlings are traits that aid in discriminating hybrid seedlings. The proportions of correctly discriminated individuals were found to be 81.7–88.2 % when using improved phenotypic discrimination methods. These methods adopted traits with high discrimination abilities during discriminant analyses and determined the selection intensity according to the hybridization rate estimated from the DNA markers. In contrast, the proportions of correctly discriminated individuals obtained using the current method were lower, 72.7–78.5 %, because this method was based on seedling height and adopted a constant selection intensity.


  1. Acheré V, Faivre-Rampant P, Pâques LE, Prat D (2004) Chloroplast and mitochondrial molecular tests identify European × Japanese larch hybrids. Theor Appl Genet 108:1643–1649PubMedCrossRefGoogle Scholar
  2. Arcade A, Faivre-Rampant P, Le Guerroué B, Pâques LE, Prat D (1996) Heterozygosity and hybrid performance in larch. Theor Appl Genet 93:1274–1281PubMedCrossRefGoogle Scholar
  3. Baltunis BS, Greenwood MS, Eysteinsson T (1998) Hybrid vigor in Larix: growth of intra- and interspecific hybrids of Larix deciduaL. laricina, and L. kaempferi after 5 years. Silvae Genet 47:288–293Google Scholar
  4. Bergmann F, Ruetz W (1987) Identifizierung von hybridlärchensaatgut aus samenplantagen mit hilfe eines isoenzym-markers. Silvae Genet 36:102–105Google Scholar
  5. Ennos RA, Qian T (1994) Monitoring the output of a hybrid larch seed orchard using isozyme markers. Forestry 67:63–74CrossRefGoogle Scholar
  6. Gros-Louis MC, Bousquet J, Pâques LE, Isabel N (2005) Species-diagnostic markers in Larixspp. based on RAPDs and nuclear, cpDNA, and mtDNA gene sequences, and their phylogenetic implications. Tree Genet Genomes 1:50–63CrossRefGoogle Scholar
  7. Häcker M, Bergmann F (1991) The proportion of hybrids in seed from a seed orchard composed of two species (L. europaea and L. leptolepis). Ann For Sci 48:631–640CrossRefGoogle Scholar
  8. Hokkaido Forest Nursery Corporation (2010) Price table for silvicultural plantlets in 2010. Hokkaido Forest Nursery Corporation, Sapporo (in Japanese)Google Scholar
  9. Howe GT, Saruul P, Davis J, Chen THH (2000) Quantitative genetics of bud phenology, frost damage, and winter survival in an F2 family of hybrid poplars. Theor Appl Genet 101:632–642CrossRefGoogle Scholar
  10. Howe GT, Aitken SN, Neale DB, Jermstad KD, Wheeler NC, Chen THH (2003) From genotype to phenotype: unraveling the complexities of cold adaptation in forest trees. Can J Bot 81:1247–1266CrossRefGoogle Scholar
  11. Karhu A, Hurme P, Karjalainen M, Karvonen P, Kärkkäinen K, Neale D, Savolainen O (1996) Do molecular markers reflect patterns of differentiation in adaptive traits of conifers? Theor Appl Genet 93:215–221PubMedCrossRefGoogle Scholar
  12. Kurahashi A, Sasaki C, Hamaya T (1966) Flowering time and accumulated temperature. Forest Tree Breed Hokkaido 9(2):20–27 (in Japanese)
  13. Kuromaru M, Uchiyama K, Kita K (2009) Prospective reform of Larix clonal seed orchards in the present Hokkaido. Forest Tree Breed Hokkaido 52(2):1–4 (in Japanese)Google Scholar
  14. Lewandowski A, Nikkanen T, Burczyk J (1994) Production of hybrid seed in a seed orchard of two larch species, Larix sibirica and Larix decidua. Scand J For Res 9:214–217CrossRefGoogle Scholar
  15. Li B, Wyckoff GW (1994) Breeding strategies for Larix deciduaL. leptolepis and their hybrids in the United States. For Genet 1:65–72Google Scholar
  16. Li B, Howe GT, Wu R (1998) Developmental factors responsible for heterosis in aspen hybrids (Populus tremuloides × P. tremula). Tree Physiol 18:29–36PubMedCrossRefGoogle Scholar
  17. Luquez V, Hall D, Albrectsen BR, Karlssoon J, Ingvarsson P, Jansson S (2008) Natural phenological variation in aspen (Populus tremula): the SwAsp collection. Tree Genet Genomes 4:279–292CrossRefGoogle Scholar
  18. Matyssek R, Schulze ED (1987) Heterosis in hybrid larch (Larix decidua × leptolepis). II. Growth characteristics. Trees 1:225–231CrossRefGoogle Scholar
  19. Miyaki Y (1990) Superior characters of the F1 hybrid Larix gmelinii × L. kaempferi and the growth gain in full-sib family selection. Forest Tree Breed Hokkaido 33(1):7–12 (in Japanese)Google Scholar
  20. Moriguchi Y, Kita K, Uchiyama K, Kuromaru M, Tsumura Y (2008) Enhanced hybridization rates in a Larix gmelinii var. japonica × L. kaempferi interspecific seed orchard with a single maternal clone revealed by cytoplasmic DNA markers. Tree Genet Genomes 4:637–645CrossRefGoogle Scholar
  21. Murray M, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325PubMedCentralPubMedCrossRefGoogle Scholar
  22. Oshima T, Nishigori M (1994) Variation of stem crook in hybrid larch (Larix gmelinii var. japonica × L. leptolepis). Trans Mtg Jpn For Soc Hokkaido Branch 42:37–39 (in Japanese)Google Scholar
  23. Pâques LE (1989) A critical review of larch hybridization and its incidence on breeding strategies. Ann For Sci 46:141–153CrossRefGoogle Scholar
  24. Pâques LE (1992) Performance of vegetatively propagated Larix decidua L. laricina hybrids. Ann For Sci 49:63–74CrossRefGoogle Scholar
  25. Pâques LE, Philippe G, Prat D (2006) Identification of European and Japanese larch and their interspecific hybrid with morphological markers: application to young seedlings. Silvae Genet 55:123–134Google Scholar
  26. Perron M (2008) A strategy for the second breeding cycle of Larix × marschlinsii in Québec, Canada including experiments to guide interspecific tree breeding programme. Silvae Genet 57:282–290Google Scholar
  27. Sato M, Seki K, Kita K, Moriguchi Y, Yunoki K, Kofujita H, Ohnishi M (2008) Prominent differences in leaf fatty acid composition in the F1 hybrid compared with parent trees Larix gmelinii var. japonica and L. kaempferi. Biosci Biotechnol Biochem 72:2895–2902PubMedCrossRefGoogle Scholar
  28. Sato M, Seki K, Kita K, Moriguchi Y, Hashimoto M, Yunoki K, Ohnishi M (2009) Comparative analysis of diterpene composition in the bark of the hybrid larch F1, Larix gmelinii var. japonica × L. kaempferi and their parent trees. J Wood Sci 55:32–40CrossRefGoogle Scholar
  29. Scheepers D, Eloy MC, Briquet M (2000) Identification of larch species (Larix deciduaLarix kaempferi and Larix × eurolepis) and estimation of hybrid fraction in seed lots by RAPD fingerprints. Theor Appl Genet 100:71–74CrossRefGoogle Scholar
  30. Semerikov V, Lascoux M (2003) Nuclear and cytoplasmic variation within and between Eurasian Larix (Pinaceae) species. Am J Bot 90:1113–1123PubMedCrossRefGoogle Scholar
  31. Takahashi K (1968) The characteristics of hybrid larch. In: Takahashi K, Yanagisawa T, Kubota Y (eds) The production and application of hybrid larch. Hokkaido Forest Tree Breeding Association, Sapporo, pp 58–108 (in Japanese)Google Scholar

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