Snails and freshwater fish were examined from four ponds in the Meinung township in which Clonorchis sinensis was known to be endemic 18 years ago. No metacercariae were found in 478 Tilapia nilotica, whereas of 451 Ctenopharyngodon idellus examined, 16.2%, 3.3% and 0.9% were found to be infected with Haplorchis pumilio, H. taichui and Clonorchis sinensis, respectively. In addition, there were some unidentified metacercariae in 12.0% of Ctenopharyngodon idellus examined. Overall, no positive correlation between infection rates and sizes of infected fish was shown. Six species of snails were collected in this survey and two frequently-occurring snails, Melanoides tuberculata and Thiara granifera were commonly infected with H. pumilio. Reasons for the prevalence of Haplorchis species and the absence of Clonorchis sinensis in fish and snail hosts in a previously reported endemic area for human clonorchiasis are discussed.

A theoretical model describing the interaction between crystalline swelling and cation exchange selectivity is proposed for expanding 2:1 phyllosilicates. The model is based on the assumption that changes in basal spacing of a clay are phase changes, and that each phase of a clay has a different selectivity constant for a particular cation exchange reaction. Energy barriers stabilize the various phases over a limited range of interlayer ionic composition. These energy barriers cause hysteresis in crystalline swelling, which in turn causes hysteresis in cation exchange. Results are presented for an experiment involving Ba-Mg exchange on a synthetic fiuoro-hectorite. The results demonstrate key aspects of the proposed model, including a correlation between measured selectivity coefficients and basal spacings (R2 = 0.85), an abrupt change in basal spacing that corresponds with an abrupt change in selectivity and corresponding hysteresis in crystalline swelling and cation exchange selectivity. The results also demonstrate increased selectivity for the preferred cation (Ba) at high solution mole fraction of the preferred cation. This trend is opposite of that observed for heterogeneous natural smectites but consistent with predictions of the model for a homogeneous smectite.

The U.S. Department of Agriculture–Agricultural Research Service (USDA-ARS) has been a leader in weed science research covering topics ranging from the development and use of integrated weed management (IWM) tactics to basic mechanistic studies, including biotic resistance of desirable plant communities and herbicide resistance. ARS weed scientists have worked in agricultural and natural ecosystems, including agronomic and horticultural crops, pastures, forests, wild lands, aquatic habitats, wetlands, and riparian areas. Through strong partnerships with academia, state agencies, private industry, and numerous federal programs, ARS weed scientists have made contributions to discoveries in the newest fields of robotics and genetics, as well as the traditional and fundamental subjects of weed–crop competition and physiology and integration of weed control tactics and practices. Weed science at ARS is often overshadowed by other research topics; thus, few are aware of the long history of ARS weed science and its important contributions. This review is the result of a symposium held at the Weed Science Society of America’s 62nd Annual Meeting in 2022 that included 10 separate presentations in a virtual Weed Science Webinar Series. The overarching themes of management tactics (IWM, biological control, and automation), basic mechanisms (competition, invasive plant genetics, and herbicide resistance), and ecosystem impacts (invasive plant spread, climate change, conservation, and restoration) represent core ARS weed science research that is dynamic and efficacious and has been a significant component of the agency’s national and international efforts. This review highlights current studies and future directions that exemplify the science and collaborative relationships both within and outside ARS. Given the constraints of weeds and invasive plants on all aspects of food, feed, and fiber systems, there is an acknowledged need to face new challenges, including agriculture and natural resources sustainability, economic resilience and reliability, and societal health and well-being.

Modern high-throughput molecular and analytical tools offer exciting opportunities to gain a mechanistic understanding of unique traits of weeds. During the past decade, tremendous progress has been made within the weed science discipline using genomic techniques to gain deeper insights into weedy traits such as invasiveness, hybridization, and herbicide resistance. Though the adoption of newer “omics” techniques such as proteomics, metabolomics, and physionomics has been slow, applications of these omics platforms to study plants, especially agriculturally important crops and weeds, have been increasing over the years. In weed science, these platforms are now used more frequently to understand mechanisms of herbicide resistance, weed resistance evolution, and crop–weed interactions. Use of these techniques could help weed scientists to further reduce the knowledge gaps in understanding weedy traits. Although these techniques can provide robust insights about the molecular functioning of plants, employing a single omics platform can rarely elucidate the gene-level regulation and the associated real-time expression of weedy traits due to the complex and overlapping nature of biological interactions. Therefore, it is desirable to integrate the different omics technologies to give a better understanding of molecular functioning of biological systems. This multidimensional integrated approach can therefore offer new avenues for better understanding of questions of interest to weed scientists. This review offers a retrospective and prospective examination of omics platforms employed to investigate weed physiology and novel approaches and new technologies that can provide holistic and knowledge-based weed management strategies for future.

Leafy spurge (Euphorbia esula L.) is an invasive perennial weed infesting range and recreational lands of North America. Previous research and omics projects with E. esula have helped develop it as a model for studying many aspects of perennial plant development and response to abiotic stress. However, the lack of an assembled genome for E. esula has limited the power of previous transcriptomics studies to identify functional promoter elements and transcription factor binding sites. An assembled genome for E. esula would enhance our understanding of signaling processes controlling plant development and responses to environmental stress and provide a better understanding of genetic factors impacting weediness traits, evolution, and herbicide resistance. A comprehensive transcriptome database would also assist in analyzing future RNA-seq studies and is needed to annotate and assess genomic sequence assemblies. Here, we assembled and annotated 56,234 unigenes from an assembly of 589,235 RNA-seq-derived contigs and a previously published Sanger-sequenced expressed sequence tag collection. The resulting data indicate that we now have sequence for >90% of the expressed E. esula protein-coding genes. We also assembled the gene space of E. esula by using a limited coverage (18X) genomic sequence database. In this study, the programs Velvet and Trinity produced the best gene-space assemblies based on representation of expressed and conserved eukaryotic genes. The results indicate that E. esula contains as much as 23% repetitive sequences, of which 11% are unique. Our sequence data were also sufficient for assembling a full chloroplast and partial mitochondrial genome. Further, marker analysis identified more than 150,000 high-quality variants in our E. esula L-RNA–scaffolded, whole-genome, Trinity-assembled genome. Based on these results, E. esula appears to have limited heterozygosity. This study provides a blueprint for low-cost genomic assemblies in weed species and new resources for identifying conserved and novel promoter regions among coordinately expressed genes of E. esula.

Microarray analysis was used to follow changes in gene expression coinciding with seasonal changes in the dormancy status of crown buds of field-grown leafy spurge. Known cold-regulated genes were induced, and numerous gibberellic acid–responsive genes were down-regulated during the transition from paradormancy to endodormancy. Genes involved in photomorphogenesis were induced during endodormancy. Also, ethylene signaling responses were observed during ecodormancy rather than endodormancy. These results provide additional insights into the signals regulating expression of several genes previously associated with transition from paradormancy to growth in root buds.

In this review, we examine current techniques and recent advances directed toward understanding cellular mechanisms involved in controlling dormancy in vegetative propagules. Vegetative propagules (including stems, rhizomes, tubers, bulbs, stolons, creeping roots, etc.) contain axillary and adventitious buds capable of producing new stems/branches under permissive environments. Axillary and adventitious buds are distinct in that axillary buds are formed in the axil of leaves and are responsible for production of lateral shoots (branches). Adventitious buds refer to buds that arise on the plant at places (stems, roots, or leaves) other than leaf axils. Both axillary and adventitious buds generally undergo periods of dormancy. Dormancy has been described as a temporary suspension of visible growth of any plant structure containing a meristem (Lang et al. 1987). Dormancy can be subdivided into three categories: (1) ecodormancy-arrest is under the control of external environmental factors; (2) paradormancy-arrest is under the control of external physiological factors within the plant; and (3) endodormancy-arrest is under the control of internal physiological factors. One common feature in all of these processes is prevention of growth under conditions where growth should otherwise continue. There is growing evidence that lack of growth is due to blockage of cell division resulting from interactions between the signaling pathways controlling dormancy and those controlling the cell cycle.

Invasive species such as Canada thistle pose a significant threat to ecosystems. The risk of introducing invasive species has increased with human activities, and the effects of such events have economic and aesthetic impacts. Native to Europe, Canada thistle is now established throughout temperate North America. Although there is documentation of early occurrences to North America, little is known on how it has become established in diverse habitats or how it continues to spread. We examined genetic diversity within and among nearly 1,700 Canada thistle individuals from 85 North American locations with the use of seven microsatellite markers in order to address these questions. PAUP and STRUCTURE programs were used to assess genetic diversity and relationships within and between populations. Populations exhibited greater within-population diversity (> 60%) than expected for a reported clonally reproducing species. Total diversity of sampled locations in North America (0.183) was less than previously reported for European locations (0.715), but the greater mean difference between North American populations (0.264 relative to 0.246 from England) suggests strong founder effects or restriction of gene flow influencing individual populations. Furthermore, analyses identified numerous instances where individuals from geographically distant regions clustered together, indicating long-distance translocation of propogules. However, isolation by distance analysis showed significant correlation between location and population genetic distances (r = 0.1917, P = 0.006). Within populations, nearly 92% of individuals sampled harbored unique multilocus genotypes, strongly suggesting that sexual reproduction is common. Within populations, analysis of genetic structure indicated significant admixture of genotypes throughout the invasive range in North America. The recurrent distribution of seed throughout North America has led to a highly diverse gene pool and increased the adaptive success Canada thistle to a wide variety of habitats. Future technologies developed for control of Canada thistle should consider this diversity.

Signals from both leaves and apical or axillary meristems of leafy spurge are known to inhibit root bud growth. To test the hypothesis that carbohydrates and growth regulators affect root bud growth, decapitated leafy spurge plants were hydroponically treated with glucose, sucrose, gibberellic acid (GA), abscisic acid (ABA), 1-naphthaleneacetic acid (NAA), 6-benzylaminopurine (BA), and a GA biosynthesis inhibitor, paclobutrazol. Both glucose and sucrose caused suppression of root bud growth at concentrations of 30 mM. The inhibitory effect of sucrose was counteracted by GA at 15 μM. In contrast, BA, ABA, NAA, and paclobutrazol inhibited root bud growth at concentrations as low as 1, 2, 1, and 16 μM, respectively. Sugar and starch levels were also determined in root buds at various times after decapitation. Buds of intact plants contained the highest level of sucrose compared with buds harvested 1, 3, and 5 d after decapitation. To determine how seasonal changes affect root bud dormancy, growth from root buds of field-grown plants was monitored for several years. Root buds of field-grown leafy spurge had the highest level of innate dormancy from October to November, which persisted until a prolonged period of freezing occurred in November or early December. Our data support the hypothesis that carbohydrates may be involved in regulating dormancy status in root buds of leafy spurge.

We have isolated both a genomic and near full length cDNA clone for a D-class cyclin gene from the perennial weed leafy spurge. Sequence analysis indicates that this gene has the highest similarity to CYCLIN D3-2 of Arabidopsis. This gene is preferentially expressed in growing shoot apices and is up-regulated in adventitious buds on resumption of growth following loss of correlative inhibition (apical dominance). CYCLIN D3-2 is also induced in nongrowing adventitious buds of plants treated with gibberellic acid or after removal of leaves—treatments known to up-regulate expression of G1 to S phase transition–specific genes such as HISTONE H3 in adventitious buds. CYCLIN D3-2 was not induced on removal of the apical and axillary buds. Expression of CYCLIN D3-2 is down-regulated in adventitious crown buds during initiation of ecodormancy in early winter. Sequence comparisons of CYCLIN D3-2 with its putative orthologue from Arabidopsis identified several conserved motifs in the promoter region and a conserved region capable of forming a stable hairpin loop in the 5′ untranslated region. Conservation of these noncoding sequences across species strongly suggests they have a regulatory function.

Genomics programs in the weed science community have not developed as rapidly as that of other crop, horticultural, forestry, and model plant systems. Development of genomic resources for selected model weeds are expected to enhance our understanding of weed biology, just as they have in other plant systems. In this report, we describe the development, characteristics, and information gained from an expressed sequence tag (EST) database for the perennial weed leafy spurge. ESTs were obtained using a normalized cDNA library prepared from a comprehensive collection of tissues. During the EST characterization process, redundancy was minimized by periodic subtractions of the normalized cDNA library. A sequencing success rate of 88% yielded 45,314 ESTs with an average read length of 671 nucleotides. Using bioinformatic analysis, the leafy spurge EST database was assembled into 23,472 unique sequences representing 19,015 unigenes (10,293 clusters and 8,722 singletons). Blast similarity searches to the GenBank nonredundant protein database identified 18,186 total matches, of which 14,205 were nonredundant. These data indicate that 77.4% of the 23,472 unique sequences and 74.7% of the 19,015 unigenes are similar to other known proteins. Further bioinformatics analysis indicated that 2,950, or 15.5%, of the unigenes have previously not been identified suggesting that some may be novel to leafy spurge. Functional classifications assigned to leafy spurge unique sequences using Munich Information Center for Protein or Gene Ontology were proportional to functional classifications for genes of arabidopsis, with the exception of unclassified or unknowns and transposable elements which were significantly reduced in leafy spurge. Although these EST resources have been developed for the purpose of constructing high-density leafy spurge microarrays, they are already providing valuable information related to sugar metabolism, cell cycle regulation, dormancy, terpenoid secondary metabolism, and flowering.

We developed two leafy spurge bacterial artificial chromosome (BAC) libraries that together represent approximately 5× coverage of the leafy spurge genome. The BAC libraries have an average insert size of approximately 143 kb, and copies of the library and filters for hybridization-based screening are publicly available through the Arizona Genomics Institute. These libraries were used to clone full-length genomic copies of an AP2/ERF transcription factor of the A4 subfamily of DEHYDRATION-RESPONSIVE ELEMENT-BINDING PROTEINS (DREB) known to be differentially expressed in crown buds of leafy spurge during endodormancy, a DORMANCY ASSOCIATED MADS-BOX (DAM) gene, and several FLOWERING LOCUS T (FT) genes. Sequencing of these BAC clones revealed the presence of multiple FT genes in leafy spurge. Sequencing also provided evidence that two different DAM transcripts expressed in crown buds of leafy spurge during endo- and eco-dormancy result from alternate splicing of a single DAM gene. Sequence data from the FT promoters was used to identify several conserved elements previously recognized in Arabidopsis, as well as potential novel transcription factor binding sites that may regulate FT. These leafy spurge BAC libraries represent a new genomics-based tool that complements existing genomics resources for the study of plant growth and development in this model perennial weed. Furthermore, phylogenetic footprinting using genes identified with this resource demonstrate the usefulness of studying weedy species to further our general knowledge of agriculturally important genes.

Bud dormancy is the primary mechanism by which the many perennial weeds escape herbicidal and mechanical control. We developed a 2,654-element Euphorbiaceae cDNA microarray using 1,886 sequenced cDNAs from the model perennial weed leafy spurge, 384 cDNAs from cassava, and 384 control genes from other plant, animal, and bacterial species. This array was used to follow changes in gene expression in root buds of leafy spurge following loss of paradormancy. The differential expression of several genes previously identified as being induced following loss of paradormancy was confirmed by microarray analysis. In addition, genes encoding an asparagine synthase, a phosphate-inducible protein, and a curculin-like (mannose-binding) lectin family protein were found to be rapidly up-regulated upon loss of paradormancy. Several genes involved in flavonoid biosynthesis were found to be rapidly down-regulated upon loss of paradormancy. The potential impact of flavonoid biosynthesis on auxin transport in response to bud growth is discussed.

Leafy spurge seeds are responsive to alternating temperature rather than constant temperature for germination. Transcriptome changes of dry leafy spurge seeds and seeds imbibed for 1 and 3 d at 20 C constant (C) and 20 : 30 C alternating (A) temperature were determined by microarray analysis to examine temperature responsiveness. Principal component analysis revealed differences in the transcriptome of imbibed seeds based on the temperature regime. Computational methods in bioinformatics parsed the data into overrepresented AraCyc pathways and gene regulation subnetworks providing biological context to temperature responses. After 1 d of imbibition, the degradation of starch and sucrose leading to anaerobic respiration were common pathways at both temperature regimes. Several overrepresented pathways unique to 1 d A were associated with generation of energy, reducing power, and carbon substrates; several of these pathways remained overrepresented and up-regulated at 3 d A. At 1 d C, pathways for the phytohormones jasmonic acid and brassinosteroids were uniquely overrepresented. There was little similarity in overrepresented pathways at 1 d C between leafy spurge and arabidopsis seeds, indicating species-specific effects upon imbibition of dry seeds. Overrepresented gene subnetworks at 1 d and 3 d at both temperature regimes related to signaling processes and stress responses. A major overrepresented subnetwork unique to 1 d C related to photomorphogenesis via the E3 ubiquitin ligase COP1. At 1 d A, major overrepresented subnetworks involved circadian rhythm via LHY and TOC1 proteins and expression of stress-related genes such as DREB1A, which is subject to circadian regulation. Collectively, substantial differences were observed in the transcriptome of leafy spurge seeds imbibed under conditions that affect the capacity to germinate.

We report on the enhancement of turbulent convective heat transport due to vapour-bubble nucleation at the bottom plate of a cylindrical Rayleigh–Bénard sample (aspect ratio 1.00, diameter 8.8 cm) filled with liquid. Microcavities acted as nucleation sites, allowing for well-controlled bubble nucleation. Only the central part of the bottom plate with a triangular array of microcavities (etched over an area with diameter of 2.5 cm) was heated. We studied the influence of the cavity density and of the superheat $T_{b}-T_{on}$ ($T_{b}$ is the bottom-plate temperature and $T_{on}$ is the value of $T_{b}$ below which no nucleation occurred). The effective thermal conductivity, as expressed by the Nusselt number $\mathit{Nu}$, was measured as a function of the superheat by varying $T_{b}$ and keeping a fixed difference $T_{b}-T_{t}\simeq 16$  K ($T_{t}$ is the top-plate temperature). Initially $T_{b}$ was much larger than $T_{on}$ (large superheat), and the cavities vigorously nucleated vapour bubbles, resulting in two-phase flow. Reducing $T_{b}$ in steps until it was below $T_{on}$ resulted in cavity deactivation, i.e. in one-phase flow. Once all cavities were inactive, $T_{b}$ was increased again, but they did not reactivate. This led to one-phase flow for positive superheat. The heat transport of both one- and two-phase flow under nominally the same thermal forcing and degree of superheat was measured. The Nusselt number of the two-phase flow was enhanced relative to the one-phase system by an amount that increased with increasing $T_{b}$. Varying the cavity density (69, 32, 3.2, 1.2 and $0.3~\text{mm}^{-2}$) had only a small effect on the global $\mathit{Nu}$ enhancement; it was found that $\mathit{Nu}$ per active site decreased as the cavity density increased. The heat-flux enhancement of an isolated nucleating site was found to be limited by the rate at which the cavity could generate bubbles. Local bulk temperatures of one- and two-phase flows were measured at two positions along the vertical centreline. Bubbles increased the liquid temperature (compared to one-phase flow) as they rose. The increase was correlated with the heat-flux enhancement. The temperature fluctuations, as well as local thermal gradients, were reduced (relative to one-phase flow) by the vapour bubbles. Blocking the large-scale circulation around the nucleating area, as well as increasing the effective buoyancy of the two-phase flow by thermally isolating the liquid column above the heated area, increased the heat-flux enhancement.