Full Text:   <2985>

CLC number: Q7

On-line Access: 

Received: 2008-05-05

Revision Accepted: 2008-10-28

Crosschecked: 2008-12-25

Cited: 2

Clicked: 5334

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2009 Vol.10 No.2 P.112-119


Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique

Author(s):  Manuel TORAL IBAÑ,EZ, Margarita CARU, Miguel A. HERRERA, Luis GONZALEZ, Luis M. MARTIN, Jorge MIRANDA, Rafael M. NAVARRO-CERRILLO

Affiliation(s):  Department of Forest Management, Faculty of Forestry; more

Corresponding email(s):   mtoral@uchile.cl

Key Words:  Sequoia sempervirens (D. Don) Endl., Random amplified polymorphic DNAs (PCR-RAPDs), Molecular markers, Clone assays, Chile, Genetic relationships

Manuel TORAL IBAÑEZ, Margarita CARU, Miguel A. HERRERA, Luis GONZALEZ, Luis M. MARTIN, Jorge MIRANDA, Rafael M. NAVARRO-CERRILLO. Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique[J]. Journal of Zhejiang University Science B, 2009, 10(2): 112-119.

@article{title="Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique",
author="Manuel TORAL IBAÑEZ, Margarita CARU, Miguel A. HERRERA, Luis GONZALEZ, Luis M. MARTIN, Jorge MIRANDA, Rafael M. NAVARRO-CERRILLO",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique
%A Margarita CARU
%A Miguel A. HERRERA
%J Journal of Zhejiang University SCIENCE B
%V 10
%N 2
%P 112-119
%@ 1673-1581
%D 2009
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0820162

T1 - Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique
A1 - Manuel TORAL IBAÑ
A1 - EZ
A1 - Margarita CARU
A1 - Miguel A. HERRERA
A1 - Luis M. MARTIN
A1 - Jorge MIRANDA
J0 - Journal of Zhejiang University Science B
VL - 10
IS - 2
SP - 112
EP - 119
%@ 1673-1581
Y1 - 2009
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0820162

A protocol of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPDs) was established to analyse the gene diversity and genotype identification for clones of Sequoia sempervirens (D. Don) Endl. in chile. Ten (out of 34) clones from introduction trial located in Voipir-Villarrica, chile, were studied. The PCR-RAPDs technique and a modified hexadecyltrimethylammonium bromide (CTAB) protocol were used for genomic DNA extraction. The PCR tests were carried out employing 10-mer random primers. The amplification products were detected by electrophoresis in agarose gels. Forty nine polymorphic bands were obtained with the selected primers (BG04, BF07, BF12, BF13, and BF14) and were ordered according to their molecular size. The genetic similarity between samples was calculated by the Jaccard index and a dendrogram was constructed using a cluster analysis of unweighted pair group method using arithmetic averages (UPGMA). Of the primers tested, 5 (out of 60) RAPD primers were selected for their reproducibility and high polymorphism. A total of 49 polymorphic RAPD bands were detected out of 252 bands. The genetic similarity analysis demonstrates an extensive genetic variability between the tested clones and the dendrogram depicts the genetic relationships among the clones, suggesting a geographic relationship. The results indicate that the RAPD markers permitted the identification of the assayed clones, although they are derived from the same geographic origin.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1] Ahuja, M.R., Neale, D.B., 2002. Origins of polyploidy in coast redwood (Sequoia sempervirens (D. Don) Endl.) and relationship of coast redwood to other genera of Taxodiaceae. Silvae Genetica, 51:93-100.

[2] Albert, F.F., 1908. Algunas observaciones sobre las especies forestales en la sección de aguas y bosques. Santiago, Chile, p.52 (in Spanish).

[3] Allnutt, T.R., Newton, A., Premoli, A., Lara, A., 2003. Genetic variation in the threatened South American conifer Pilgerodendron uviferum (Cupressaceae), detected using RAPD markers. Biological Conservation, 114(2): 245-253.

[4] Anekonda, T.S., Criddle, R.S., Libby, W.J., Hansen, L.D., 1993. Spatial and temporal relationships between growth traits and metabolic heat rates in coast redwood. Canadian Journal of Forest Research, 23(9):1793-1798.

[5] Anekonda, T.S., Criddle, R.S., Libby, W.J., 1994. Calorimetric evidence for site-adapted biosynthetic metabolism in coast redwood (Sequoia sempervirens). Canadian Journal of Forest Research, 24(2):380-389.

[6] Bruno, C., Brinegar, C., 2004. Microsatellite markers in coast redwood (Sequoia sempervirens). Molecular Ecology Notes, 4(3):482-484.

[7] Doyle, J.J., Doyle, J.L., 1990. Isolation of plant DNA from fresh tissue. Focus, 12:13-15.

[8] Furman, B.J., Grattapaglia, D., Dvorak, W.S., O′Malley, D.M., 1997. Analysis of genetic relationships of Central American and Mexican pines using RAPD markers that distinguish species. Molecular Ecology, 6(4):321-331.

[9] Grotkopp, E., Rejmanek, M., Sanderson, M., Rost, T., 2004. Evolution of genome size in pines (Pinus) and its life-history correlates: supertree analyses. Evolution, 58(8):1705-1729.

[10] Hall, G.D., Langenheim, J.H., 1987. Geographic variation in leaf monoterpenes of Sequoia sempervirens. Biochemical Systematics and Ecology, 15(1):31-43.

[11] Hansen, O.K., 2008. Mating patterns, genetic composition and diversity levels in two seed orchards with few clones— impact on planting crop. Forest Ecology and Management, 256(5):1167-1177.

[12] Harris, S.A., 1999. RAPDs in Systematicsda Useful Methodology? In: Hollingsworth, P.M., Batesman, R.M., Gornall, R.J. (Eds.), Molecular Systematics and Plant Evolution. Taylor and Francis, London, p.211-228.

[13] Jaccard, P., 1908. Nouvelles recherches sur la distribution florale. Bulletin Société Vaudoise des Sciences Naturelles, 44:223-270 (in French).

[14] Krutovskii, K.V., Vollmer, S., Sorensen, F.C., Adams, W., Knapp, S., Strauss, S., 1998. RAPD genome maps of Douglas-fir. The Journal of Heredity, 89(3):197-205.

[15] Kuser, J.E., 1996. Redwood as an Exotic. Conference on Coast Redwood Forest Ecology and Management. Humbolt State University, Arcata, California, the United States, p.55-59.

[16] Kuser, J., Bailley, A., Franclet, A., Libby, W., Martin, J., Rydelius, J., Schoenike, R., Vagle, N., 1995. Early results of a rangewide provenance test of Sequoia sempervirens. Forest Genetic Resources, 23:21-25.

[17] Manly, B., 1991. Randomization, Bootstrap and Monte Carlo Methods in Biology, 1st Ed. Chapman and Hall, London, p.300.

[18] Millar, C.I., Dunlap, J.M., Walker, N.K., 1985. Analysis of Growth and Specific Gravity in a 20-Year-Old Provenance Test of Sequoia sempervirens. Rep. No. 59. Department of Agriculture, California Agriculture and Experiment Station, Forest Service, Berkeley, CA, USA, Vol. 59, p.1-6.

[19] Moran, G.F., Bell, C., 1987. The origin and genetic diversity of Pinus radiata in Australia. Theoretical and Applied Genetics, 73(4):616-622.

[20] Nkongolo, K.K., Michael, P., Gratton, W.S., 2002. Identification and characterization of RAPD markers inferring genetic relationships among pine species. Genome, 45(1): 51-58.

[21] Noss, R.F. (Ed.), 2000. The Redwood Forest. History, Ecology, and Conservation of the Coast Redwoods. Island Press, p.335.

[22] Osses, A.T., 2004. El estado de las plantaciones de Sequoia sempervirens (D. Don) Endl. y su relación con las características de los suelos para localidades de la VII, IX y IX región. Memoria. Ing. For., Fac. Cs. Forestales. Universidad de Chile, p.215 (in Spanish).

[23] Pavlicek, A., Hrda, S., Flegr, J., 1999. FreeTree—Freeware program for construction of phylogenetic trees on the basis of distance data and bootstrap/jackknife analysis of the tree robustness. Application in the RAPD analysis of the genus Frenkelia. Folia Biologica (Praha), 45:97-99.

[24] Rogers, D.L., 1997. Inheritance of allozymes from seed tissues of the hexaploid gymnosperm, Sequoia sempervirens (D. Don) Endl. (coast redwood). Heredity, 78(2):166-175.

[25] Rogers, D.L., 1999. Allozyme polymorphisms discriminate among coast redwood (Sequoia sempervirens) siblings. Journal of Heredity, 90(3):429-433.

[26] Rogers, D.L., 2000. Genotypic diversity and clone size in old-growth populations of coast redwood (Sequoia sempervirens). Canadian Journal of Botany, 78(11): 1408-1419.

[27] Rogers, D.L., Westfall, R.D., 2007. Spatial Genetic Patterns in Four Old-growth Populations of Coast Redwood. In: Standiford, R.B., Giusti, G.A., Valachovic, Y., Zielinski, W.J., Furniss, M.J. (Eds.), Proceedings of the Redwood Region Forest Science Symposium: What does the Future Hold? Pacific Southwest Research Station, Forest Service, U.S. Department of Agriculture, Albany, CA, p.59-63.

[28] Syring, J., Willyard, A., Cronn, R., Liston, A., 2005. Evolutionary relationships among Pinus (Pinaceae) subsections inferred from multiple low-copy nuclear loci. American Journal of Botany, 92(12):2086-2100.

[29] Toral, M., González, L.Y., Garfias, R., 2005. Secuoya: nueva opción, productos y mercados para el sur de Chile. Universidad de Chile, p.201 (in Spanish).

[30] Tsumina, T., Yoshimaru, K., Tomaru, N., Ohba, K., 1995. Molecular phylogeny of conifer using RFLP analysis of PCR-amplifed specific chloroplast genes. Theoretical and Applied Genetics, 91:1222-1236.

[31] Williams, J.G.K., Kubelik, A.R., Livak, K.J., Rafalski, J.A., Tingey, S.V., 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18(22):6531-6535.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2023 Journal of Zhejiang University-SCIENCE