306 For these reasons, any conclusions regarding sexual determinism using ARPA profiles should be carefully made. Use of appro- priate controls such as those used in this study allows accurate conclusions regarding sexual determinism to be correctly drawn. Male plants could now be selected from seedlings issued from seeds. Decreasing prices in molecular biology consumables and technologies would result in affordable sexing services for larger numbers of plants at an early stage. Growing male plants in nurseries instead of a mix of both sexes will optimize the use of nursery surfaces and then contribute to lower the price of such trees, which on the other hand could be more desirable as urban trees once male-certified. Consequently, sexual reproduction and sewing of Ginkgo seeds could benefit this technique and allow the availability of a greater diversity in urban Ginkgo trees. Early sexual determination gained from ARPA allows for ef- fective selection of male Ginkgo biloba trees highly suited by city governments for planting in streets and allows practitioners to easily provide such plants with no need to grow female plants for over 30 years. Acknowledgments. Special thanks go to Georges Wigger (Microsynth, Balgach, Switzerland) for providing helpful advice about ARPA. We are also grateful to Nicolas Hasler (Domaine Nature et Paysage, Départe- ment du Territoire, Republic of Geneva) and Yves Fessler (Service des Espaces Verts of Geneva Town Hall) for authorizing sampling of G. biloba trees. Dominique Verdel, Centre d’Enseignement Professionnel Nature et Paysage, is acknowledged for his stimulating discussions and advice during the conduction of this work. LITERATURE CITED Aoki, T. 1997. Fungal association with Ginkgo biloba, p. 251–258. In: Hori, T., P. Del Tredici, R.W. Ridge, W. Tulecke, J. Tremouillaux- Guiller, and H. Tobe (Eds.). Ginkgo biloba—A Global Treasure. Springer-Verlag, Tokyo, Japan. Carrier, D.J., T.A. van Beek, R. van der Heijden, and R. Verpoorte. 1998. Distribution of ginkgolides and terpenoid biosynthetic activity in Ginkgo biloba. Phytochemistry 48:89–92. Fisher, M.M., and E.W. Triplett. 1999. Automated approach for ribo- somal intergenic spacer analysis of microbial diversity and its appli- cation to freshwater bacterial communities. Applied and Environ- mental Microbiology 65:4630–4636. Handa, M., Y. Lizuka, and N. Fujiwara. 1997. Ginkgo landscapes, p. 259–283. In: Hori, T., P. Del Tredici, R.W. Ridge, W. Tulecke, J. Tremouillaux-Guiller, and H. Tobe (Eds.). Ginkgo biloba—A Global Treasure. Springer-Verlag, Tokyo, Japan. Honda, H. 1997. Ginkgo and insects, p. 259–283. In: Hori, T., P. Del Tredici, R.W. Ridge, W. Tulecke, J. Tremouillaux-Guiller, and H. Tobe (Eds.). Ginkgo biloba—A Global Treasure. Springer-Verlag, Tokyo, Japan. Jiang, L., R.L. You, M.X. Li, and C. Shi. 2003. Identification of a sex associated RAPD marker in Ginkgo biloba. Acta Botanica Sinica 45:742–747. Kim, Y.S., J.K. Lee, and G.C. Chung. 1997. Tolerance and susceptibility of Ginkgo to air pollution, p. 233–242. In: Hori, T., P. Del Tredici, R.W. Ridge, W. Tulecke, J. Tremouillaux-Guiller, and H. Tobe (Eds.). Ginkgo biloba—A Global Treasure. Springer-Verlag, Tokyo, Japan. Kiple, K.F., and K.C. Ornelas. 2000. The Cambridge World History of Food. Cambridge University Press, Cambridge, U.K. 1958 pp. Laurain, D. 2000. Cultivation of Ginkgo biloba on large scale, p. 63–79. In: Van Beek, T.A. (Ed.). Ginkgo biloba. Harwood Academic Pub- lishers, Amsterdam, The Netherlands. Lefort, F., and G.C. Douglas. 1999. An efficient micro-method of DNA isolation from mature leaves of four hardwood tree species Acer, Fraxinus, Prunus and Quercus. Annals of Science 56:259–263. ©2008 International Society of Arboriculture Echenard et al.: Sex Identification in Ginkgo biloba Michel, P.F. 1985. Ginkgo biloba:L’arbre qui a vaincu le temps. Edi- tions du Félin, Paris. 108 pp. Rohr, R. 1989. Maidenhair tree (Ginkgo biloba L.), p. 574–590. In: Bajaj, Y.P.S. (Ed.). Biotechnology in Agriculture and Forestry, Trees II. Springer Verlag, Berlin, Germany. Wada, K., and M. Haga. 1997. Food poisoning by Ginkgo biloba seeds, p. 309–321. In: Hori, T., P. Del Tredici, R.W. Ridge, W. Tulecke, J. Tremouillaux-Guiller, and H. Tobe (Eds.). Ginkgo biloba—A Global Treasure. Springer-Verlag, Tokyo, Japan. Zhong, H.W., Z.H. Yang, G.L. Zhu, and Z.X. Cao. 1982. Peroxidase isozyme pattern as a biochemical test to distinguish the sex of indi- vidual plant in Ginkgo biloba L. Scientia Silvae Sinica 18:1–5. Zhou, Z.Y., and S.L. Zheng. 2003. Palaeobiology: The missing link in Ginkgo evolution—The modern maidenhair tree has barely changed since the days of the dinosaurs. Nature 423:821–822. Vincent Echenard University of Applied Sciences of Western Switzerland Plants and Pathogens Group Research Institute Earth, Nature and Landscape 150 route de Presinge Jussy Geneva 1254, Switzerland François Lefort (corresponding author) University of Applied Sciences of Western Switzerland Plants and Pathogens Group Research Institute Earth, Nature and Landscape 150 route de Presinge Jussy Geneva 1254, Switzerland
[email protected] Gautier Calmin University of Applied Sciences of Western Switzerland Plants and Pathogens Group Research Institute Earth, Nature and Landscape 50 route de Presinge Jussy Geneva 1254, Switzerland Robert Perroulaz University of Applied Sciences of Western Switzerland Agronomy Department Geneva 1254, Switzerland Lassaad Belbahri University of Applied Sciences of Western Switzerland Plants and Pathogens Group Research Institute Earth, Nature and Landscape 50 route de Presinge Jussy Geneva 1254, Switzerland Résumé. Une technique d’amplification polymorphique aléatoire d’ADN du gène S1478 associé au sexe mâle a été utilisée pour amplifier l’ADN de 72 échantillons de feuilles récoltés à partir de Ginkgo biloba dont le sexe était connu dans le canton de Genève en Suisse. Ce mar- queur a été a été découvert comme étant spécifique au mâle et est absent de tous les plants femelles. L’analyse polymorphique aléatoire d’ADN automatisée, une nouvelle technologie d’automatisation développée dans ce but, s’est avérée être hautement effective pour distinguer les plants mâles des plants femelles, et ce à 100% d’efficacité et de succès dans la discrimination mâle versus femelle au sein d’une collection de jeunes semis obtenus à partir de croisements sexuels. Nos découvertes fournissent une preuve sans ambiguïté que l’analyse polymorphique
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