RAPD markers encoding retrotransposable elements are linked to the male sex in Cannabis sativa L.

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Abstract:

Male-associated DNA sequences were analyzed in Cannabis sativa L. (hemp), a dioecious plant with heteromorphic sex chromosomes. DNA was isolated from male and female plants and subjected to random amplified polymorphic DNA analysis. Of 120 primers, 17 yielded 400 to 1500-bp fragments detectable in male, but not female, plants. These fragments were cloned and used as probes in gel-blot analysis of genomic DNA. When male and female DNA was hybridized with 2 of these male-specific fragments, MADC(male-associated DNA sequences in C. sativa)3 and MADC4, particularly intense bands specific to male plants were detected in addition to bands common to both sexes. The MADC3 and MADC4 sequences were shown to encode gag/pol polyproteins of copia-like retrotransposons. Fluorescence in situ hybridization with MADC3 and MADC4 as probes revealed a number of intense signals on the Y chromosome as well as dispersed signals on all chromosomes. The gel-blot analysis and fluorescence in situ hybridization results presented here support the hypothesis that accumulation of retrotransposable elements on the Y chromosome might be 1 cause of heteromorphism of sex chromosomes.Key words: Cannabis sativa, FISH, RAPD, retrotransposon, sex chromosome.

Des séquences d'ADN associées au sexe mâle ont été analysées chez le Cannabis sativa L. (chanvre), une plante dioïque avec des chromosomes sexuels hétéromorphes. L'ADN a été extrait de plantes mâles ou femelles et assujetti à une analyse RAPD (ADN polymorphe amplifié au hasard). Des 120 amorces employées, 17 ont produit des amplicons de 400 à 1500 pb présents chez les mâles mais absents chez les femelles. Ces amplicons ont été clonés et employés comme sondes lors d'hybridations Southern. Lorsque l'ADN mâle ou femelle a été hybridé à 2 de ces sondes mâle-spécifiques, MADC(« male-associated DNA sequences in C. sativa »)3 et MADC4, des bandes particulièrement intenses et mâle-spécifiques ont été détectées en plus de bandes communes aux 2 sexes. Les séquences MADC3 et MADC4 codent pour des polyprotéines gag/pol de rétrotransposons de type copia. Des hybridations in situ en fluorescence avec MADC3 et MADC4 ont révélé plusieurs signaux intenses sur le chromosome Y de même que des signaux dispersés sur tous les chromosomes. Les résultats de l'analyse Southern et de l'hybridation in situ supportent l'hypothèse voulant que l'accumulation de rétrotransposons sur le chromosome Y serait une cause de l'hétéromorphisme des chromosomes sexuels.Mots clés : Cannabis sativa, FISH, RAPD, rétrotransposon, chromosomes sexuels.[Traduit par la Rédaction]

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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