Chromosome-scale genome assembly of Poa trivialis and population genomics reveal widespread gene flow in a cool-season grass seed production system.

Poa trivialis (L.) is a cool-season grass species found in various environments worldwide. In addition to being a desired turfgrass species, it is a common weed of agricultural systems and natural areas. As a weed, it is an important contaminant of commercial cool-season grass seed lots, resulting in widespread gene flow facilitated by human activities and causing significant economic losses to farmers. To better understand and manage infestations, we assembled and annotated a haploid genome of P. trivialis and studied troublesome field populations from Oregon, the largest cool-season grass seed producing region in the United States. The genome assembly resulted in 1.35 Gb of DNA sequence distributed among seven chromosome-scale scaffolds, revealing a high content of transposable elements, conserved synteny with Poa annua, and a close relationship with other C3 grasses. A reduced-representation sequencing analysis of field populations revealed limited genetic diversity and suggested potential gene flow and human-assisted dispersal in the region. The genetic resources and insights into P. trivialis provided by this study will improve weed management strategies and enable the development of molecular detection tests for contaminated seed lots to limit seed-mediated gene flow. These resources should also be beneficial for turfgrass breeders seeking to improve desirable traits of commercial P. trivialis varieties and help to guide breeding efforts in other crops to enhance the resiliency of agricultural ecosystems under climate change. Significance Statement: The chromosome-scale assembly of Poa trivialis and population genomic analyses provide crucial insights into the gene flow of weedy populations in agricultural systems and contribute a valuable genomic resource for the plant science community.

The genome of Salmacisia buchloëana, the parasitic puppet master pulling strings of sexual phenotypic monstrosities in buffalograss.

To complete its parasitic lifecycle, Salmacisia buchloëana, a biotrophic fungus, manipulates reproductive organ development, meristem determinacy, and resource allocation in its dioecious plant host, buffalograss (Bouteloua dactyloides; Poaceae). To gain insight into S. buchloëana's ability to manipulate its host, we sequenced and assembled the 20.1 Mb genome of S. buchloëana into 22 chromosome-level pseudomolecules. Phylogenetic analysis suggests that S. buchloëana is nested within the genus Tilletia and diverged from Tilletia caries and Tilletia walkeri ∼40 MYA. We find that S. buchloëana contains a novel chromosome arm with no syntenic relationship to other publicly available Tilletia genomes, and that genes on the novel arm are upregulated upon infection, suggesting that this unique chromosomal segment may have played a critical role in S. buchloëana's evolution and host specificity. Salmacisia buchloëana has one of the largest fractions of serine peptidases (1.53% of the proteome) and one of the highest GC contents (62.3%) in all classified fungi. Analysis of codon base composition indicated that GC content is controlled more by selective constraints than directional mutation, and that S. buchloëana has a unique bias for the serine codon UCG. Finally, we identify 3 inteins within the S. buchloëana genome, 2 of which are located in a gene often used in fungal taxonomy. The genomic and transcriptomic resources generated here will aid plant pathologists and breeders by providing insight into the extracellular components contributing to sex determination in dioecious grasses.

Homoeologous evolution of the allotetraploid genome of Poa annua L.

BACKGROUND: Poa annua (annual bluegrass) is an allotetraploid turfgrass, an agronomically significant weed, and one of the most widely dispersed plant species on earth. Here, we report the chromosome-scale genome assemblies of P. annua's diploid progenitors, P. infirma and P. supina, and use multi-omic analyses spanning all three species to better understand P. annua's evolutionary novelty. RESULTS: We find that the diploids diverged from their common ancestor 5.5 - 6.3 million years ago and hybridized to form P. annua ≤ 50,000 years ago. The diploid genomes are similar in chromosome structure and most notably distinguished by the divergent evolutionary histories of their transposable elements, leading to a 1.7 × difference in genome size. In allotetraploid P. annua, we find biased movement of retrotransposons from the larger (A) subgenome to the smaller (B) subgenome. We show that P. annua's B subgenome is preferentially accumulating genes and that its genes are more highly expressed. Whole-genome resequencing of several additional P. annua accessions revealed large-scale chromosomal rearrangements characterized by extensive TE-downsizing and evidence to support the Genome Balance Hypothesis. CONCLUSIONS: The divergent evolutions of the diploid progenitors played a central role in conferring onto P. annua its remarkable phenotypic plasticity. We find that plant genes (guided by selection and drift) and transposable elements (mostly guided by host immunity) each respond to polyploidy in unique ways and that P. annua uses whole-genome duplication to purge highly parasitized heterochromatic sequences. The findings and genomic resources presented here will enable the development of homoeolog-specific markers for accelerated weed science and turfgrass breeding.

Chromosome-Scale Genome Assembly and Annotation of Allotetraploid Annual Bluegrass (Poa annua L.).

Poa annua L. is a globally distributed grass with economic and horticultural significance as a weed and as a turfgrass. This dual significance, and its phenotypic plasticity and ecological adaptation, have made P. annua an intriguing plant for genetic and evolutionary studies. Because of the lack of genomic resources and its allotetraploid (2n = 4x = 28) nature, a reference genome sequence would be a valuable asset to better understand the significance and polyploid origin of P. annua. Here we report a genome assembly with scaffolds representing the 14 haploid chromosomes that are 1.78 Gb in length with an N50 of 112 Mb and 96.7% of BUSCO orthologs. Seventy percent of the genome was identified as repetitive elements, 91.0% of which were Copia- or Gypsy-like long-terminal repeats. The genome was annotated with 76,420 genes spanning 13.3% of the 14 chromosomes. The two subgenomes originating from Poa infirma (Knuth) and Poa supina (Schrad) were sufficiently divergent to be distinguishable but syntenic in sequence and annotation with repetitive elements contributing to the expansion of the P. infirma subgenome.

Draft Genome Sequence of Salmacisia buchloëana (Basidiomycota), Which Induces Hermaphroditism in Dioecious Buffalograss.

Here, we report the draft genome of Salmacisia buchloëana type strain OK1, a monotypic species of fungi that induces ovary development in genetic male plants and increases reproductive biomass allocation in its host buffalograss. This research will enhance our understanding of fungal manipulation of host development at the genomic level.

Pistil Smut Infection Increases Ovary Production, Seed Yield Components, and Pseudosexual Reproductive Allocation in Buffalograss.

Sex expression of dioecious buffalograss [Bouteloua dactyloides Columbus (syn. Buchloë dactyloides (Nutt.) Engelm.)] is known to be environmentally stable with approximate 1:1, male to female, sex ratios. Here we show that infection by the pistil smut fungus [Salmacisia buchloëana Huff & Chandra (syn. Tilletia buchloëana Kellerman and Swingle)] shifts sex ratios of buffalograss to be nearly 100% phenotypically hermaphroditic. In addition, pistil smut infection decreased vegetative reproductive allocation, increased most seed yield components, and increased pseudosexual reproductive allocation in both sex forms compared to uninfected clones. In female sex forms, pistil smut infection resulted in a 26 fold increase in ovary production and a 35 fold increase in potential harvest index. In male sex forms, pistil smut infection resulted in 2.37 fold increase in floret number and over 95% of these florets contained a well-developed pistil. Although all ovaries of infected plants are filled with fungal teliospores and hence reproductively sterile, an average male-female pair of infected plants exhibited an 87 fold increase in potential harvest index compared to their uninfected clones. Acquiring an ability to mimic the effects of pistil smut infection would enhance our understanding of the flowering process in grasses and our efforts to increase seed yield of buffalograss and perhaps other grasses.

Genetic variability in populations of the southern chinch bug, Blissus insularis, assessed using AFLP analysis.

Southern chinch bug, Blissus insularis Barber (Heteroptera: Blissidae), is the most destructive insect pest of St. Augustine grass, Stenotaphrum secundatum Waltz (Kuntze), in the southern United States. The present study is focused on assessing genetic variability in five populations of B. insularis collected from Texas and Florida where St. Augustine grass is widely grown. The amplified fragment length polymorphism technique was used to DNA fingerprint individuals from each population (a total of 46 individuals) using five primer combinations (EcoRI/MSeI). Analysis of molecular variance results show no evidence to support significant genetic variability among Texas and Florida populations of B. insularis. Nearly all genetic variation was found to reside within populations (95%), with only approximately 3% residing among populations between regions. Low G(ST) values obtained from POPGENE and low F(ST) values obtained from the analysis of molecular variance both support the conclusion for high levels of gene flow resulting from interbreeding and/or migratory events among the populations. A Mantel test of Euclidean squared distances showed no correlation between the genetic distance and geographic distance matrices of tested populations of B. insularis. The results of the present study suggests that gene flow is occurring among populations of B. insularis and, therefore, breeders need to be aware that this resistance will most likely not remain localized, and it has the potential to spread as a result of migratory events.

A fungal parasite regulates a putative female-suppressor gene homologous to maize tasselseed2 and causes induced hermaphroditism in male buffalograss.

Parasitically induced hermaphroditism is a fascinating illustration of floral sex organ modification; however, knowledge of how parasites induce hermaphroditism in plants is limited. Here, we show the fungal parasite pistil smut induces development of female sex organs (pistils) in flowers of male buffalograss, potentially by downregulating a putative female-suppressor gene, BdTs2, homologous to maize Tasselseed2 (ZmTs2). Full-length BdTs2, isolated using rapid amplification of cDNA ends, exhibits 89% nucleotide sequence similarity with ZmTs2 and 85% amino acid sequence homology with ZmTs2 protein. Scanning electron micrographs demonstrate that unisexual buffalograss flowers develop through a process of selective abortion of opposite sex organs within hermaphroditic floral primordia. Quantitative real-time polymerase chain reaction showed that high expression levels of BdTs2 within male inflorescences correlate with the selective abortion of gynoecium, leading to the development of unisexual male flowers. RNA in situ hybridization confirmed the expression of BdTs2 precisely within vestigial gynoeciums of male flowers and not in other floral organs of the inflorescence. Furthermore, we show that BdTs2 expression is downregulated by pistil smut infection, which corresponds to the presence of pistils in flowers otherwise destined to become unisexual male. This study provides a potential molecular basis for pistil smut-induced hermaphroditism in male buffalograss.

Salmacisia, a new genus of Tilletiales: reclassification of Tilletia buchloëana causing induced hermaphroditism in buffalograss.

For 119y the fungal parasite Tilletia buchloëana has been known to induce female sex organs (pistils) in male plants of buffalograss, making them hermaphrodite. Here we report the life cycle characteristics and phylogeny for T. buchloëana based on morphology and large subunit region of nuclear ribosomal RNA gene (nLSU-rDNA) to accurately place pistil smut within order Tilletiales. We found that T. buchloëana induces hermaphroditism in not only male sex forms of buffalograss by inducing the development of otherwise vestigial pistils but also in female sex forms by inducing hypertrophy of otherwise vestigial stamens (male sex organs). The fungus also induces the development of additional pistillate flowers in both infected male and female plants. Due to its pistil inducing effects we refer to T. buchloëana as pistil smut. Pistil smut exhibits a combination of morphological and life cycle characteristics that are unique among species of order Tilletiales. Phylogenetic analyses of nLSU-rDNA sequences using maximum parsimony, maximum likelihood and genetic distance-based methods show that pistil smut exhibits an elevated rate of nucleotide substitution and is as, or more, distant from Tilletia species than the basal group Erratomyces patelli. As such pistil smut occupies a phylogenetic position outside the current taxonomic circumscription of genus Tilletia. Therefore a new Latin binomial combination Salmacisia buchloëana is proposed as a sister taxon of Tilletia to accurately describe the phylogenetic position of pistil smut.

Endophyte-Mediated Suppression of Dollar Spot Disease in Fine Fescues.

In 1989, a close association was found between single-plant progenies of strong creeping red fescue infected with the endophyte Epichloë festucae and enhanced suppression of dollar spot, a widespread foliar disease of turfgrass caused by Sclerotinia homoeocarpa. From this limited observation, extensive field evaluations were conducted on a wide range of fine fescue germplasm obtained throughout the United States and Europe to determine the frequency and magnitude of this association. In five field trials established between 1985 and 1991, endophyte-infected Chewings, hard, blue, and strong creeping red fescue cultivars, selections, and crosses consistently exhibited endophyte-mediated suppression of dollar spot, when compared with closely related endophyte-free entries. Endophyte-infected Chewings and hard fescue cultivars and selections also had greater turf density and supported less foliar mycelium of S. homoeocarpa than endophyte-free entries.