Towards Improved Molecular Identification Tools in Fine Fescue (FestucaL., Poaceae) Turfgrasses: Nuclear Genome Size, Ploidy, and Chloroplast Genome Sequencing

Abstract

Fine fescues (Festuca L., Poaceae) are turfgrass species that perform well in low-12 input environments. Based on morphological characteristics, the most commonly-utilized 13 fine fescues are divided into five taxa: three are subspecies within F. rubra L. and the 14remaining two are treated as species within the F. ovina L. complex. Morphologically, these 15five taxa are very similar, both identification and classification of fine fescues remain 16challenging. In an effort to develop identification methods for fescues, we used flow 17 cytometry to estimate genome size, ploidy level, and sequenced the chloroplast genome of 18 all five taxa. Fine fescue chloroplast genome sizes ranged from 133,331 to 133,841 bp and 19contained 113 to 114 genes. Phylogenetic relationship reconstruction using whole 20 chloroplast genome sequences agreed with previous work based on morphology. 21Comparative genomics suggested unique repeat signatures for each fine fescue taxon that 22could potentially be used for marker development for taxon identification. 23 Keywords: Fine fescue, chloroplast genome, phylogeny, comparative genomics 24 25 485 project, supervised this research, provided suggestions, and comments. All authors contributed to the revision 486 of the manuscript and approved the final version.

Fine fescues: A review of the species, their improvement, production, establishment, and management

“…Villamil, Duell, Fairbrothers, and Sadowski (1982) also successfully distinguished species and subspecies and cultivars within subspecies using esterase banding patterns developed in polyacrylamide gels. Work is ongoing to create genetic markers useful for species identification (Qiu, Hirsch, Yang, & Watkins, 2019).…”

“…Using flow cytometry, Huff and Palazzo (1998) confirmed ploidy of fine fescues previously reported by Hubbard (1984) and Wilkinson and Stace (1991). Although there has been a wide range reported, chromosome analysis revealed that Chewings fescue, slender creeping red fescue, and hard fescue are hexaploid (2 n = 6 x = 42); while strong creeping red fescue is octoploid (2 n = 8 x = 56); and sheep fescue ( Festuca ovina L.) is tetraploid (2 n = 4 x = 28) (Duyvendak & Vos, 1974; Huff & Palazzo, 1998; Qiu et al., 2019; Ruemmele et al., 2003; Wilkinson & Stace, 1991). Subspecies of F. ovina L., which are not commonly found as commercial turf cultivars, can be both diploid ( F. ovina L. ssp.…”

Building a Reference Transcriptome for the Hexaploid Hard Fescue Turfgrass (Festuca brevipila) Using a Combination of PacBio Isoseq and Illumina Sequencing

“…By sequencing the chloroplast genomes, our previous work suggested F. ovina subsp. ovina (2n=2x=14) and F. ovina (2n=4x=28) were both closely related to F. brevipila (Qiu et al, 2019). Here we adopt a phylotranscriptomic approach to investigate the phylogenetic relationship among F. brevipila and close relatives using nuclear loci.…”

“…The species identity of plant materials used in this study were con rmed following guidelines described by Wilkinson and Stace (1991) using vein number, leaf morphology, and ow cytometry (Qiu et al, 2019).…”

“…fallax, 2n=6x=42; Huff and Palazzo, 1998). A previous phylogenetic study using plastid genomes resolved the relationships among these ve taxa (Qiu et al, 2019). Of the ve fescue taxa, F. brevipila is a particularly attractive target for breeding and genetics improvement due to its excellent performance in low-input environments (Ruemmele et al, 1995).…”