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Morphological diversity and genomic DNA fingerprinting of the African rice gall midge Orseolia oryzivora (Diptera: Cecidomyiidae) and of two other species of African Orseolia

Published online by Cambridge University Press:  01 December 2006

Francis E. Nwilene*
Africa Rice Center, PO Box 5320, Ibadan, Nigeria
Keith M. Harris
Africa Rice Center, 81 Linden Way, Ripley, Woking, Surrey, GU23 6LP, UK
Olusegun Okhidievbie
Africa Rice Center, PO Box 5320, Ibadan, Nigeria
Amos Onasanya
Africa Rice Center, 01 BP 2031, Cotonou, Benin Republic
Yacouba Sere
Africa Rice Center, 01 BP 2031, Cotonou, Benin Republic
Ivan Ingelbrecht
International Institute of Tropical Agriculture, PO Box 5320, Ibadan, Nigeria
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A morphological diversity study was conducted on African rice gall midge Orseolia oryzivora Harris & Gagné and two other African species, Orseolia bonzii Harris and Orseolia nwanzei Harris & Nwilene, sp. n. Morphological differences between adult and immature stages of the three species are slight and mainly evident in pupal characters. Due to limited differences in morphological characters, genomic DNA fingerprinting of these three insect species was carried out using random amplified polymorphic DNA (RAPD) and sequence-characterized amplified region (SCAR) methods. Out of 90 operon primers, 15 showed polymorphism among the three species tested, generating 86 bands, 60 (70%) of which were polymorphic. Primer OPV6 produced three RAPD markers that distinguished the three Orseolia species. These markers were cloned and sequenced. Their sequence was then used to design six SCAR primer pairs. Each SCAR primer pair amplified and distinguished the three Orseolia species at genomic DNA level. Both SCAR and RAPD genomic DNA fingerprinting revealed that O. oryzivora and O. bonzii are most closely related and O. nwanzei is distinct. Each of the six SCAR primer pairs produced a specific DNA fragment size specific for O. nwanzei, O. oryzivora and O. bonzii. The DNA fingerprints will be useful for entomological survey, identification of new species and differentiating aggressive from non-aggressive species, aimed at the effective development of rice cultivars with durable resistance to African rice gall midge.

Une étude de la diversité morphologique a été réalisée sur la cécidomyie de la galle du riz africain, Orseolia oryzivora Harris & Gagné et sur deux autres espèces africaines, O. bonzii Harris et O. nwanzei Harris & Nwilene, sp. n. Les différences morphologiques entre les stades adultes et immatures des trois espèces sont minimes et plus marquées au niveau des caractères nymphaux. A cause des caractères morphologiques limités, la cartographie peptidique de l'ADN génomique de ces trois espèces d'insectes a été effectuée à l'aide de la méthode d'amplification aléatoire d'ADN polymorphe (RAPD) et de la méthode SCAR. Quinze des 90 amorces d'opérons ont montré un polymorphisme chez les trois espèces testées, créant 86 bandes, dont 60 (70%) étaient polymorphes. L'amorce OPV6 a produit trois marqueurs RAPD, qui ont permis de différencier les trois espèces d'Orseolia. Ces marqueurs ont été clonés et séquencés. Leur séquence a été ensuite utilisée pour concevoir six paires d'amorces SCAR. Les paires des amorces de chaque marqueur SCAR ont permis d'amplifier et de différencier les trois espèces d'Orseolia au niveau de l'ADN génomique. La cartographie peptidique de l'ADN génomique des marqueurs SCAR et RAPD a révélé que les espèces O. oryzivora et O. bonzii sont plus étroitement liées et que O. nwanzei est distincte. Chacune des six paires d'amorces SCAR a produit un segment d'ADN de taille spécifique pour O. nwanzei, O oryzivora et O, bonzii; ces paires d'amorces pourraient donc être utiles pour des diagnostiques de terrain. La cartographie peptidique de l'ADN définie pour chaque espèce d'Orseolia sera utile pour les études entomologiques, l'identification de nouvelles espèces et la différentiation des espèces agressives des espèces non agressives, en vue de développer de façon efficace des cultivars de riz dotés d'une résistance durable à la cécidomyie de la galle du riz africain.

Research Paper
Copyright © ICIPE 2006

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