Stability and inheritance of methylation states at Pst I sites in Pisum
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O R I GI N A L P A P E R
M. R. Knox á T. H. N. Ellis
Stability and inheritance of methylation states at Pst I sites in Pisum
Received: 25 August 2000 / Accepted: 5 December 2000 / Published online: 22 February 2001 Ó Springer-Verlag 2001
Abstract The nuclear genome of pea is heavily methylated and the stability of this methylation pattern is unknown. In this study we investigated the stability of DNA methylation and its contribution to restriction fragment polymorphism as judged by AFLP fragment dierences. To do this the results of a conventional AFLP analysis were compared with those obtained by a related procedure performed on pre-ampli®ed DNA which provided a 5-methylcytosine-free DNA template (SDAFLP). Genetic mapping in a recombinant inbred (RI) population showed that polymorphisms attributable to dierent methylation states of PstI sites were abundant, and generally appeared to be stably inherited, although occasional failures of the inheritance of methylation states have been found. Assessments of genetic diversity by AFLP and SDAFLP were in general agreement with each other and with the currently accepted phylogeny of Pisum, but within cultivated groups the number of dierences appeared to be exaggerated by AFLP. The data suggest that epigenetic dierences may have played a role in the domestication of pea. Key words Pea á DNA methylation á Mapping á Phylogeny á AFLP
Introduction AFLP is a versatile and widely used molecular marker method, and since its introduction (Zabeau and Vos 1993) it has been exploited in many ways for the analysis of bacterial and eukaryotic genomes. Two of the major uses of AFLP markers in plants have been for mapping ± in barley (Becker et al. 1995; Powell et al. 1997), in Communicated by R. Hagemann M. R. Knox (&) á T. H. N. Ellis John Innes Centre, Colney Lane, Norwich NR4 7UH, UK E-mail: [email protected] Fax: +44-1603-450027
eucalypts (Marques et al. 1998), lentil (Eujayl et al. 1998), melon (Wang et al. 1997), pea (Hall et al. 1997), pine (Travis et al. 1998; Cato et al. 1999; Remington et al. 1999), rice (Mackill et al. 1996; Xiong et al. 1998), sorghum (Boivin et al. 1999), soybean (Keim et al. 1997), sugar beet (Schondelmaier et al. 1996) and tomato (Thomas et al. 1995); and for diversity analysis ± in Arabidopsis thaliana (Breyne et al. 1999), barley malt (Faccioli et al. 1999), cassava (Wong et al. 1999), cocoa (Perry et al. 1998), grapevine (Cervera et al. 1998), lentil (Sharma et al. 1996), olive (Angiolillo et al. 1999), pea (Ellis et al. 1998), poplar (Win®eld et al. 1998), rice (Subudhi et al. 1998; Zhu et al. 1998), soybean (Maughan et al. 1996), sugarcane (Besse et al. 1998), tea (Paul et al. 1997), and wheat (Barrett et al. 1998). AFLP has been compared with RFLP and a range of PCRbased marker systems and has proved to be a robust and highly reproducible marker system, for banana (Crouch et al. 1999), barley (Russell et al. 1997), gooseberry (Lanham and Brennan 1999), maize (Pejic et al. 1998), pea (Lu et al. 1996), potato (Milbourne et al. 1997), and
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