Development of a SCAR Marker for Rapid Identification of New Kentucky Bluegrass Breeding Lines

Abstract

As a commonly used turfgrass, Kentucky bluegrass (Poa pratensis L.) (KBG) has many commercially available cultivars for production. After several years of screening, two new lines were obtained ('KBG03' and 'KBG04'), which have high tolerance to summer. The study showed that the two lines revealed similar morphological characteristics, with light green leaf color, narrow leaf blade, high plant height and light 1,000-grain weight. A total of 400 random amplified polymorphic DNA (RAPD) primers and 256 sequence-related amplified polymorphism (SRAP) primer combinations were screened among the two lines and other 4 imported commercial cultivars. The percentages of polymorphic sites were 65.5% (RAPD) and 22.6% (SRAP) respectively. By cluster analysis of RAPD and SRAP data, the dendrogram at a similarity of 0.29 gave two main clusters, of which one group had 4 commercial cultivars, and the other had the two new breeding lines. Furthermore, one specific band of 'KBG04' was successfully converted into a dominant sequence characterized amplified region marker (SCAR196). Then the SCAR marker was verified by 39 KBG DNA samples, including imported varieties, domestic varieties and self-breeding lines of our laboratory, and it exhibited high consistency with the original RAPD polymorphic amplification. The results showed that the SCAR marker can be used to distinguish the new line 'KBG04' from numerous KBG germplasms, which would be useful for cultivar identification and property rights protection in the future.

Genetic diversity ofPoa pratensisL. depending on geographical origin and compared with genetic markers

“…Molecular analysis, in conjunction with the characteristics of morphological traits, may be used to classify plants and select the initial forms for creating new cultivars or breeding lines. In the case of P. pratensis , this was confirmed by Shortell, Meyer & Bonos (2009) , Tamkoc & Arslan (2010) , Raggi et al (2015) and Yuan et al (2015) . In this study, two markers (RAPD and ISSR) based on DNA amplification reactions were used to assess the genetic diversity of ecotypes and cultivars of P. pratensis .…”

“…In grasses, RAPD and ISSR markers are most commonly used to evaluate genetic variability within and between populations, and for molecular characterization and identification of different species and their hybrids, cultivars and genotypes ( Posselt et al, 2006 ; Pivorienė et al, 2008 ; Al-Humaid, Ibrahim & Motawei, 2011 ; Motawei & Al-Ghumaiz, 2012 ; Madesis et al, 2014 ; Yuan et al, 2014 ). In P. pratensis , genetic analyses employing RAPD and ISSR focus mainly on the identification of sexual and apomictic genotypes ( Huff & Bara, 1993 ; Mazzucato et al, 1995 ; Barcaccia et al, 1997 ; Barcaccia, Veronesi & Falcinelli, 1998 ; Stephens et al, 2006 ); the assessment of genetic variability and identification of genotypes and cultivars ( Lickfeldt, Voigt & Hamblin, 2002 ; Ning et al, 2005 ; Liang et al, 2009 ; Fard et al, 2012 ; Wang et al, 2012 ; Yuan et al, 2015 ); determining the genetic relationships between different genotypes and species belonging to the genus Poa and their hybrids ( Johnson et al, 2002 ; Curley & Jung, 2004 ; Patterson, Larson & Johnson, 2005 ; Goldman, 2008 ; Goldman, 2013 ); and identification of pathogens responsible for fungal diseases in grasses ( Hsiang et al, 2000 ). Both marker systems can generate high resolution band patterns and high levels of polymorphism as high as 90%.…”