Mitochondrial genome sequencing and analysis of the invasive Microstegium vimineum: a resource for systematics, invasion history, and management.

Premise of the Research: Plants remain underrepresented among species with sequenced mitochondrial genomes (mitogenomes), due to the difficulty in assembly with short-read technology. Invasive species lag behind crops and other economically important species in this respect, representing a lack of tools for management and land conservation efforts. Methodology: The mitogenome of Microstegium vimineum, one of the most damaging invasive plant species in North America, was sequenced and analyzed using long-read data, providing a resource for biologists and managers. We conducted analyses of genome content, phylogenomic analyses among grasses and relatives based on mitochondrial coding regions, and an analysis of mitochondrial single nucleotide polymorphism in this invasive grass species. Pivotal Results: The assembly is 478,010 bp in length and characterized by two large, inverted repeats, and a large, direct repeat. However, the genome could not be circularized, arguing against a "master circle" structure. Long-read assemblies with data subsets revealed several alternative genomic conformations, predominantly associated with large repeats. Plastid-like sequences comprise 2.4% of the genome, with further evidence of Class I and Class II transposable element-like sequences. Phylogenetic analysis placed M. vimineum with other Microstegium species, excluding M. nudum, but with weak support. Analysis of polymorphic sites across 112 accessions of M. vimineum from the native and invasive ranges revealed a complex invasion history. Conclusions: We present an in-depth analysis of mitogenome structure, content, phylogenetic relationships, and range-wide genomic variation in M. vimineum's invasive US range. The mitogenome of M. vimineum is typical of other andropogonoid grasses, yet mitochondrial sequence variation across the invasive and native ranges is extensive. Our findings suggest multiple introductions to the US over the last century, with subsequent spread, secondary contact, long-distance dispersal, and possibly post-invasion selection on awn phenotypes. Efforts to produce genomic resources for invasive species, including sequenced mitochondrial genomes, will continue to provide tools for their effective management, and to help predict and prevent future invasions.

Digitized collections elucidate invasion history and patterns of awn polymorphism in Microstegium vimineum.

PREMISE: Digitized collections can help illuminate the mechanisms behind the establishment and spread of invasive plants. These databases provide a record of traits in space and time that allows for investigation of abiotic and biotic factors that influence invasive species. METHODS: Over 1100 digitized herbarium records were examined to investigate the invasion history and trait variation of Microstegium vimineum. Presence-absence of awns was investigated to quantify geographic patterns of this polymorphic trait, which serves several functions in grasses, including diaspore burial and dispersal to germination sites. Floret traits were further quantified, and genomic analyses of contemporary samples were conducted to investigate the history of M. vimineum's introduction and spread into North America. RESULTS: Herbarium records revealed similar patterns of awn polymorphism in native and invaded ranges of M. vimineum, with awned forms predominating at higher latitudes and awnless forms at lower latitudes. Herbarium records and genomic data suggested initial introduction and spread of the awnless form in the southeastern United States, followed by a putative secondary invasion and spread of the awned form from eastern Pennsylvania. Awned forms have longer florets, and floret size varies significantly with latitude. There is evidence of a transition zone with short-awned specimens at mid-latitudes. Genomic analyses revealed two distinct clusters corresponding to awnless and awned forms, with evidence of admixture. CONCLUSIONS: Our results demonstrate the power of herbarium data to elucidate the invasion history of a problematic weed in North America and, together with genomic data, reveal a possible key trait in introduction success: presence or absence of an awn.

Chromosome Level Genome Assembly and Annotation of Highly Invasive Japanese Stiltgrass (Microstegium vimineum).

The invasive Japanese stiltgrass (Microstegium vimineum) affects a wide range of ecosystems and threatens biodiversity across the eastern USA. However, the mechanisms underlying rapid adaptation, plasticity, and epigenetics in the invasive range are largely unknown. We present a chromosome-level assembly for M. vimineum to investigate genome dynamics, evolution, adaptation, and the genomics of phenotypic plasticity. We generated a 1.12-Gb genome with scaffold N50 length of 53.44 Mb respectively, taking a de novo assembly approach that combined PacBio and Dovetail Genomics Omni-C sequencing. The assembly contains 23 pseudochromosomes, representing 99.96% of the genome. BUSCO assessment indicated that 80.3% of Poales gene groups are present in the assembly. The genome is predicted to contain 39,604 protein-coding genes, of which 26,288 are functionally annotated. Furthermore, 66.68% of the genome is repetitive, of which unclassified (35.63%) and long-terminal repeat (LTR) retrotransposons (26.90%) are predominant. Similar to other grasses, Gypsy (41.07%) and Copia (32%) are the most abundant LTR-retrotransposon families. The majority of LTR-retrotransposons are derived from a significant expansion in the past 1-2 Myr, suggesting the presence of relatively young LTR-retrotransposon lineages. We find corroborating evidence from Ks plots for a stiltgrass-specific duplication event, distinct from the more ancient grass-specific duplication event. The assembly and annotation of M. vimineum will serve as an essential genomic resource facilitating studies of the invasion process, the history and consequences of polyploidy in grasses, and provides a crucial tool for natural resource managers.

Spatially Targeted Biological Control of Mile-a-Minute Weed Using Rhinoncomimus latipes (Coleoptera: Curculionidae) and an Unmanned Aircraft System.

Rhinoncomimus latipes Korotyaev is a specialist biocontrol agent of mile-a-minute weed, Persicaria perfoliata (L.) H. Gross (Caryophyllales: Polygonaceae). Currently, R. latipes is released by hand where the presence of the weed is readily detected. However, the hand-release method is not applicable to weed patches spread in hard-to-access areas. This study was conducted to develop a spatially targeted biocontrol strategy by using an unmanned aircraft system (UAS, a.k.a. drone) for the detection of P. perfoliata and aerial release of R. latipes. A ground survey was performed to locate P. perfoliata patches and then a rotary-wing UAS was flown at 15 different altitudes to determine the detectability of P. perfoliata patches. We developed an insect-release system including a pod that housed R. latipes for aerial release. The pod was 3D printed with biodegradable polyvinyl alcohol (PVA), and field tests were conducted to determine the ability of R. latipes to escape the pod and assess their post-release mortality and feeding ability. The results of this study showed that P. perfoliata patches were readily detectable on the aerial images taken at ≤15 m above the ground. More than 98% of R. latipes (n = 118) successfully escaped from the pod within 24 h after aerial deployment. There were no significant (P > 0.05) effects of PVA exposure on the mortality and feeding ability of R. latipes. These results indicate that aerial detection of P. perfoliata and deployment of R. latipes for spatially targeted biological control in hard-to-access areas can be accomplished using a rotary-wing UAS.

Housing is positively associated with invasive exotic plant species richness in New England, USA.

Understanding the factors related to invasive exotic species distributions at broad spatial scales has important theoretical and management implications, because biological invasions are detrimental to many ecosystem functions and processes. Housing development facilitates invasions by disturbing land cover, introducing nonnative landscaping plants, and facilitating dispersal of propagules along roads. To evaluate relationships between housing and the distribution of invasive exotic plants, we asked (1) how strongly is housing associated with the spatial distribution of invasive exotic plants compared to other anthropogenic and environmental factors; (2) what type of housing pattern is related to the richness of invasive exotic plants; and (3) do invasive plants represent ecological traits associated with specific housing patterns? Using two types of regression analysis (best subset analysis and hierarchical partitioning analysis), we found that invasive exotic plant richness was equally or more strongly related to housing variables than to other human (e.g., mean income and roads) and environmental (e.g., topography and forest cover) variables at the county level across New England. Richness of invasive exotic plants was positively related to area of wildland-urban interface (WUI), low-density residential areas, change in number of housing units between 1940 and 2000, mean income, plant productivity (NDVI), and altitudinal range and rainfall; it was negatively related to forest area and connectivity. Plant life history traits were not strongly related to housing patterns. We expect the number of invasive exotic plants to increase as a result of future housing growth and suggest that housing development be considered a primary factor in plans to manage and monitor invasive exotic plant species.