Sequence of the Sugar Pine Megagenome

Abstract

Until very recently, complete characterization of the megagenomes of conifers has remained elusive. The diploid genome of sugar pine (Pinus lambertiana Dougl.) has a highly repetitive, 31 billion bp genome. It is the largest genome sequenced and assembled to date, and the first from the subgenus Strobus, or white pines, a group that is notable for having the largest genomes among the pines. The genome represents a unique opportunity to investigate genome "obesity" in conifers and white pines. Comparative analysis of P. lambertiana and P. taeda L. reveals new insights on the conservation, age, and diversity of the highly abundant transposable elements, the primary factor determining genome size. Like most North American white pines, the principal pathogen of P. lambertiana is white pine blister rust (Cronartium ribicola J.C. Fischer ex Raben.). Identification of candidate genes for resistance to this pathogen is of great ecological importance. The genome sequence afforded us the opportunity to make substantial progress on locating the major dominant gene for simple resistance hypersensitive response, Cr1. We describe new markers and gene annotation that are both tightly linked to Cr1 in a mapping population, and associated with Cr1 in unrelated sugar pine individuals sampled throughout the species' range, creating a solid foundation for future mapping. This genomic variation and annotated candidate genes characterized in our study of the Cr1 region are resources for future marker-assisted breeding efforts as well as for investigations of fundamental mechanisms of invasive disease and evolutionary response.KEYWORDS conifer genome; transposable elements; white pine blister rust T HE gymnosperm genus Pinus is diverse and ubiquitous in temperate zones (Critchfield and Little 1966;Farjon and Filer 2013). Pines are often the keystone trees of terrestrial ecosystems (Richardson and Rundel 1998;Keane et al. 2012, and citations therein). Typical of conifers, pines have megagenomes that vary greatly in size among species, yet their karyotype is highly conserved. Pinus is divided into two large, ancient monophyletic subgenera, Strobus and Pinus, "white pines" and "yellow pines," respectively (Critchfield and Little 1966;Gernandt et al. 2005). The first Pinus genome sequence (22 Gbp) was recently reported for Pinus taeda L. ), a yellow pine commonly known as loblolly pine. The genomes of white pines are larger and more variable in size (Tomback 1982). Fossils allied with Strobus are known from the early Tertiary and late Cretaceous (Millar 1998) et al. 2006), the discovery of the underlying genes, and of markers serviceable for genetic improvement in reforestation, may be greatly accelerated by the genome sequence itself. P. lambertiana, commonly known as sugar pine, is a white pine native to western North America that is distributed from northern Oregon to Baja California at a wide span of altitudes. It is currently the tallest pine species, with heights reaching 76 m. The female cones of sugar pine are also gigan...

A genome for gnetophytes and early evolution of seed plants

“…The genome of this species is considered to have retained many features that were likely to have been present in the ancestral angiosperm genome 22 . It is notable that its repeat content 13 and lower abundance of intact LTR-RTs (solo LTR/intact LTR ratio = 2.43/1.0; Supplementary Table 10) is similar to that observed in G. montanum. These observations suggest that neither A. trichopoda nor G. montanum genomes have experienced recent, extensive (retro) transposon activity, although they continue to eliminate repetitive sequences.…”

“…So far, the availability of whole genome sequences for gymnosperms has been limited to conifers (specifically to Pinaceae) [10][11][12][13] and G. biloba 14 , with no whole genome assemblies available for the two remaining major seed plant lineages-cycads and gnetophytes. This deficiency, together with the conflicting phylogenetic evidence for relationships among these groups, is impeding our understanding of genome evolution across all seed plants.…”

“…A BUSCO (Benchmarking Universal Single-Copy Orthologs) analysis to assess the quality of the genome and annotation completeness suggested that 81% of the genes have been recovered (Supplementary Table 6). Unlike conifer genomes, which contain numerous pseudogenes 15 (for example,8,328 in Picea abies,13,550 in Pinus taeda), many fewer were found in the G. montanum genome (3,122, Supplementary Information). The read depth distribution across genic regions ( Supplementary Fig.…”

“…Repetitive sequences have been shown to account for the major component of all gymnosperm genomes that have been sequenced to date [11][12][13][14] , with diverse and ancient transposable elements (TEs), especially LTR retrotransposons (LTR-RTs), being particularly prevalent. Overall, the repetitive element content of G. montanum was also high (85.9%) and dominated by LTR-RTs (especially gypsy-like Table 8 and Supplementary Information).…”

“…Consequently, several possible hypotheses have been put forward that place gnetophytes as sister to (1) Pinaceae ('Gnepine' hypothesis); (2) cupressophytes ('Gnecup' hypothesis); (3) all conifers ('Gnetifer' hypothesis); (4) all other gymnosperms; or (5) all seed plants 9 . Currently, the emerging consensus, based on both older and more recent studies, and recently released data from the 1KP initiative (see https://sites.google.com/a/ualberta.ca/onekp/, and Wickett et al 8 ), indicates that gnetophytes are sister to, or within, the conifers.So far, the availability of whole genome sequences for gymnosperms has been limited to conifers (specifically to Pinaceae) [10][11][12][13] and G. biloba 14 , with no whole genome assemblies available for the two remaining major seed plant lineages-cycads and gnetophytes. This deficiency, together with the conflicting phylogenetic evidence for relationships among these groups, is impeding our understanding of genome evolution across all seed plants.…”