Abstract
Seeds of the genus Lolium are difficult to identify based on morphology for morphological likeness and some physical deformation such as friction and flattening during storage and transport. DNA barcoding, a newly-established method, has been used to discriminate a variety of agricultural crops with its own advantages. In present study, DNA barcodes for the genus Lolium were investigated for the first time. DNA sequences of psbA-trnH, rbcL, atpF-atpH, and the ITS2 region were evaluated for their ability to differentiate Lolium from the related genus Festuca. As confirmed by inter-intraspecific divergence and Kimura 2 parameter analysis, the greatest divergence existed in ITS2, followed by psbA-trnH. On the contrary, rbcL and atpF-atpH possessed poor genetic variation of 0-0.0115, and was relatively difficult in discrimination of genus Lolium. For ITS2 sequence, no inter-intraspecific distance overlaps were observed and each species has a distinct barcoding gap. ITS2 could effectively discriminate all species based on a neighbor-joining tree. Thus, the ITS2 region is a candidate for DNA barcoding of Lolium.
Highlights
Lolium, a genus of Poaceae, consists of six species and one variety: Lolium perenne, L. persicum, L. remotum, L.multiflorum, L. rigidum, L. temulentum, and L. temulentum var. arvense (Raven and Zhang, 2013)
The objective of this study was to evaluate four potential DNA barcodes suggested by the CBOL Plant Working Group (2009) and other authors (Kress et al, 2005; Lahaye et al, 2008a) and applied them to Lolium and Festuca species for phylogenetic reconstructions
The results showed that psbA-trnH and ITS2 had the highest inter-specific variability, followed by rbcL and atpF-atpH
Summary
A genus of Poaceae, consists of six species and one variety: Lolium perenne, L. persicum, L. remotum, L.multiflorum, L. rigidum, L. temulentum, and L. temulentum var. arvense (Raven and Zhang, 2013). A genus of Poaceae, consists of six species and one variety: Lolium perenne, L. persicum, L. remotum, L.multiflorum, L. rigidum, L. temulentum, and L. temulentum var. Lolium perenne and L. multiflorum are widely used as cool-season forage and have high economic value. Lolium temulentum and its variety are morphologically similar to L. perenne, L. multiflorum, and L. rigidum and cannot be distinguished routinely. The term “DNA barcode” for global species identification was first proposed by Hebert et al (2003). Compared with the conventional morphology, DNA barcodes are not affected by the part of the plant tested and individual developmental stages. The key to identification is to find an appropriate DNA barcode. RbcL, atpF-atpH, psbA-trnH, and ITS2 have been commonly used as DNA barcodes for plants (CBOL Plant Working Group, 2009; Chen et al, 2010)
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