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Development of microsatellite markers of chloroplastic genome studies in Citrus [S29-10]

Lotfy S., Luro F., Froelicher Y., Risterucci A.M., Filloux D., Ollitrault P.. 2003. In : 7th International Congress of Plant Molecular Biology, ISPMB 2003, Barcelona (Spain), June 23-28. Book of abstracts. Barcelone : ISPMB Office, p. 408-408. International Congress of Plant Molecular Biology. 7, 2003-06-23/2003-06-28, Barcelone (Espagne).

Genetic information of plant chloroplastic DNA presents a great interest for phylogenic studies. Recently, in Citrus, a new interest for cytoplasmic genome analysis risen from the impediment of somatic hybridization. Indeed this technique allows to combine nuclear, chloroplastic and mitochondrial genomes in new patterns that must be characterized. The development of PCR markers appears very suitable for cytoplasmic characterization at an early in vitro step. The application of Cleaved Amplified Polymorphic Sequences method with universal primer has been demonstrated recently to be efficient at the inter-specific level but display very few diversity at the infra-specific one. In the present work, the usefulness of chloroplastic SSR for citrus chloroplast genomes characterization has been studied. Thirty two PCR primers from Oryza sativa and Nicotiana tabacum chloroplast genome have been tested among 8 cultivated Citrus species and 3 related genera. From 26 tobacco couples of primers, 15 have allowed amplification in all citrus species, 10 of them displaying polymorphism. For 6 rice couples of primer, we got successful amplification without any polymorphism. Height couples of primers from tobacco displayed polymorphism have been used for diversity analysis among 50 species of Aurantioideae sub-family. Phylogenic inference were done from a cluster analysis of chloroplastic SSR data. At Citrus genus level, the genetic organization displayed was in agreement with previous hypothesis in cultivated citrus evolution. At the end, we present an example of application of these markers for chloroplastic genome characterization from somatic hybrid. (Texte intégral)

Mots-clés : citrus; adn; chloroplaste; microsatellite; marqueur génétique

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