Expressed sequence tag-derived microsatellite markers of perennial ryegrass (Lolium perenne L.)

Abstract

An expressed sequence tag (EST) library of the key grassland species perennial ryegrass (Lolium perenne L.) has been exploited as a resource for microsatellite marker development. Out of 955 simple sequence repeat (SSR) containing ESTs, 744 were used for primer design. Primer amplification was tested in eight genotypes of L. perenne and L. multiflorum representing (grand-) parents of four mapping populations and resulted in 464 successfully amplified ESTSSRs. Three hundred and six primer pairs successfully amplified products in the mapping population VrnA derived from two of the eight genotypes included in the original screening and revealed SSR polymorphisms for 143 ESTs. Here, we report on 464 EST-derived SSR primer sequences of perennial ryegrass established in laboratory assays, providing a dedicated tool for marker assisted breeding and comparative mapping within and among forage and turf grasses.Electronic supplementary material The online version of this article

Inheritance patterns of the response to in vitro doubled haploid induction in perennial ryegrass (Lolium perenne L.)

“…Simple sequence repeat (SSR) markers G05_065 and G05_134 (Studer et al 2008) and high resolution melting curve analysis (HRM) marker 05_02833 (Manzanares et al 2016), selected because of their heterozygosity in a large number of anther donor genotypes, were used to characterize the green regenerants. A detailed primer description and the PCR amplification protocol used for the SSR markers can be found in Studer et al (2008). Amplification products were separated, visualised and scored using an ABI 3130 16 capillary electrophoresis system (Applied Biosystems, Foster City, CA, USA) and the GeneMarker software version 1.5 (SoftGenetics, PA, USA).…”

Orchardgrass (Dactylis glomerata L.) EST and SSR marker development, annotation, and transferability

“…As EST-SSRs markers are derived from transcribed regions, they are likely to have a high success of amplification and associated gene annotations (Varshney et al 2005), and are useful for in silico comparisons to the homologous sequences in other species (Bushman et al 2008). EST-SSR markers have been used in genetic and comparative mapping in a number of forage grass species (e.g., Jones et al 2002b;Warnke et al 2004;Saha et al 2005;Alm et al 2003;Studer et al 2008). Additionally, EST libraries with long sequence reads constitute crucial reference sequences for further sequencing and functional genomics studies using ''Next-Generation'' methods.…”

Genetic and Agronomic Analysis of Latvian Fescue (Festuca Spp.), Ryegrass (Lolium Spp.) Accessions and their Hybrids

“…For samples collected from field trials, DNA was extracted from a leaf sample from a single individual, using the previously described method. Genotyping was done using eight simple sequence repeat (SSR) markers -G03_020, G05_033, G07_037, G01_053, G07_065, G05_071, G05_088, and G05_099 (Studer et al, 2008). Each forward primer was labelled with a different fluorophore (6-FAM, HEX or TMR) to facilitate visualisation using capillary electrophoresis.…”