5S rDNA genome regions of Lens species

Authors: Fernández, M; Ruiz, M L; Linares, C; Fominaya, A; Pérez de la Vega, M

Source: Genome, Volume 48, Number 5, October 2005 , pp. 937-942(6)

Publisher: NRC Research Press

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Abstract:

The length variability of the nontranscribed spacer (NTS) of the 5S rDNA repeats was analyzed in species of the genus Lens by means of PCR amplification. The NTS ranged from ~227 to ~952 bp. The polymorphism detected was higher than previous NTS polymorphisms described in this genus. Three NTS length variants from Lens culinaris subsp. culinaris and 2 from Lens culinaris subsp. orientalis were sequenced. The culinaris NTS fragment lengths were 239, 371, and 838 bp, whereas the orientalis ones were 472 bp and 506 bp, respectively. As a result of sequence similarities, 2 families of sequences were distinguished, 1 including the sequences of 838 and 506 bp, and others with the sequences of 239, 371, and 472 bp. The 1st family was characterized by the presence of a repeated sequence designated A, whereas the 2nd family showed a single A sequence and other repeated sequences designated B, C, and D. The presence of an (AT)n microsatellite was also observed in the 2nd family of sequences. The fragments, which included the 239-bp and 838-bp NTS sequences, as well as the intergenic spacer (IGS) of the 18S–5.8S–26S ribosomal DNA also from L. culinaris subsp. culinaris, were used to localize the nucleolar organizer region (NOR) and the 5S rDNA loci in the chromosomes of several species of the genus Lens by means of fluorescence in situ hybridization (FISH). The selective hybridization of the 2 NTS probes allowed us to distinguish between different 5S rDNA chromosomal loci.Key words: Lens, lentil, ribosomal loci, 5S, FISH, NTS polymorphism, NOR.

La variabilité de la longueur de l'espaceur nontranscrit (NTS) au sein de l'ADNr 5S a été examinée par amplification PCR chez diverses espèces du genre Lens. Les NTS variaient entre environ 227 et 952 pb. Le polymorphisme détecté était plus grand que ce qui avait été rapporté antérieurement pour ce genre. Trois variants de la taille du NTS chez le Lens culinaris subsp. culinaris et 2 chez le L. culinaris subsp. orientalis ont été séquencés. Les tailles des NTS chez les culinaris étaient de 239, 371 et 838 bp alors que chez l'orientalis ils étaient de 472 et 506 pb. Sur la base de la similarité des séquences, 2 familles ont été révélées, l'une regroupant les séquences de 838 et 506 pb et l'autre comprenant les séquences de 239, 371 et 472 pb. La première famille était caractérisée par la présence d'une répétition, appelée séquence A, tandis que les NTS de la seconde famille ne contenaient qu'un seul exemplaire de la séquence A, mais des répétitions des séquences B, C et D. Un microsatellite (AT)n a également été observé chez la seconde famille de séquences. Des fragments contenant les séquences NTS de 239 et 838 pb, de même que l'espaceur intergénique (IGS) de l'ADNr 18S–5,8S–26S du L. culinaris subsp. culinaris, ont été employés pour situer l'organisateur nucléolaire (NOR) et les locus d'ADNr 5S sur les chromosomes de plusieurs espèces du genre Lens au moyen de l'hybridation in situ en fluorescence (FISH). L'hybridation sélective avec les 2 sondes NTS a permis de distinguer les différents locus d'ADNr 5S.Mots clés : Lens, lentille, locus ribosomiques, 5S, FISH, polymorphisme des NTS, NOR.

Document Type: Research Article

Publication date: October 1, 2005

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  • From its inception in 1957, this international cytogenetics journal has catered to the research areas of the members of the Genetics Society of Canada; traditionally, these have included agriculture, entomology, genetics/cytogenetics, and evolutionary mechanisms. The contents of the journal have evolved as contributors developed new technologies and interests. A 20-member Editorial Board is composed of scientists from around the world. Reviews and commentary from respected experts are often featured.
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