Diverse Host Research Articles

Identification of a novel mycovirus belonging to the “flexivirus”-related family with icosahedral virion

Abstract The order Tymovirales currently comprises five viral families with positive-sense RNA ((+)RNA) genomes that infect plants, fungi, and insects. Virion morphologies within the order Tymovirales differ between families, with icosahedral virions in the Tymoviridae and filamentous virions in the other “flexi”viridae families. Despite their different morphologies, these viruses are placed in the same order based on phylogenetic analyses of replicase-associated polyproteins. However, one of the families in the Tymovirales, Deltaflexiviridae, is considered to be capsidless because there have been no published reports of virion isolation. Here we report that a new “flexivirus”-related (+)RNA virus, prospectively named Fusarium oxysporum icosahedral virus 1 (FoIV1), is icosahedral, and that most deltaflexiviruses may have icosahedral virions. Phylogenetic analyses based on replicase-associated polyproteins indicated that FoIV1 forms a distinct group in the Tymovirales with some viruses originally assigned to the Deltaflexiviridae. Electron microscopy, protein analysis, and protein structure predictions indicate that FoIV1 ORF4 encodes a single jelly-roll (SJR)-like coat protein (CP) that constitutes the icosahedral virions. Results of clustering analyses based on amino acid sequences and predicted CP structures suggested that most of the deltaflexiviruses have icosahedral virions composed of SJR-like CPs as in FoIV1, rather than having filamentous virions or capsidless. These results challenge the conventional understanding of viruses in the order Tymovirales, with important implications for revising its taxonomic framework and providing insights into the evolutionary relationships within this diverse and broad host range group of (+)RNA viruses.

Near complete assembly of Pyricularia penniseti infecting Cenchrus grass identified its eight core chromosomes

Fungi from the Pyricularia genus cause blast disease in many economically important crops and grasses, such as wheat, rice, and Cenchrus grass JUJUNCAO. Structure variation associated with the gain and loss of effectors contributes largely to the adaptive evolution of this fungus towards diverse host plants. A telomere-to-telomere genome assembly would facilitate the identification of genome-wide structural variations through comparative genomics. Here, we report a telomere-to-telomere, near-complete genome assembly of a Pyricularia penniseti isolate JC-1 infecting JUJUNCAO. The assembly consists of eight core chromosomes and two supernumerary chromosomes, named mini1 and mini2, spanning 42.1 Mb. We annotated 12,156 protein-coding genes and identified 4.54% of the genome as repetitive sequences. The two supernumerary chromosomes contained fewer genes and more repetitive sequences than the core chromosomes. Our genome and results provide valuable resources for the future study in genome evolution, structure variation and host adaptation of the Pyricularia fungus.

PIPdb: a comprehensive plasmid sequence resource for tracking the horizontal transfer of pathogenic factors and antimicrobial resistance genes.

Plasmids, as independent genetic elements, carrying resistance or virulence genes and transfer them among different pathogens, posing a significant threat to human health. Under the 'One Health' approach, it is crucial to control the spread of plasmids carrying such genes. To achieve this, a comprehensive characterization of plasmids in pathogens is essential. Here we present the Plasmids in Pathogens Database (PIPdb), a pioneering resource that includes 792964 plasmid segment clusters (PSCs) derived from 1 009571 assembled genomes across 450 pathogenic species from 110 genera. To our knowledge, PIPdb is the first database specifically dedicated to plasmids in pathogenic bacteria, offering detailed multi-dimensional metadata such as collection date, geographical origin, ecosystem, host taxonomy, and habitat. PIPdb also provides extensive functional annotations, including plasmid type, insertion sequences, integron, oriT, relaxase, T4CP, virulence factors genes, heavy metal resistance genesand antibiotic resistance genes. The database features a user-friendly interface that facilitates studies on plasmids across diverse host taxa, habitats, and ecosystems, with a focus on those carrying antimicrobial resistance genes (ARGs). We have integrated online tools for plasmid identification and annotation from assembled genomes. Additionally, PIPdb includes a risk-scoring system for identifying potentially high-risk plasmids. The PIPdb web interface is accessible at https://nmdc.cn/pipdb.

Bat-associated ticks as a potential link for vector-borne pathogen transmission between bats and other animals.

Potentially zoonotic pathogens have been previously detected in bat-associated ticks, but their role in disease transmission and their frequency of feeding on non-bat hosts is poorly known. We used molecular blood meal analysis to reveal feeding patterns of the bat-associated tick species Ixodes ariadnae, I. simplex, and I. vespertilionis collected from cave and mine walls in Central and Southeastern Europe. Vertebrate DNA, predominantly from bats, was detected in 43.5% of the samples (70 of 161 ticks) but in these ticks we also detected the DNA of non-chiropteran hosts, such as dog, Canis lupus familiaris, wild boar, Sus scrofa, and horse, Equus caballus, suggesting that bat-associated ticks may exhibit a much broader host range than previously thought, including domestic and wild mammals. Furthermore, we detected the zoonotic bacteria Neoehrlichia mikurensis in bat ticks for the first time, and other bacteria, such as Bartonella and Wolbachia. In the light of these findings, the role of bat ticks as disease vectors should be urgently re-evaluated in more diverse host systems, as they may contribute to pathogen transmission between bats and non-chiropteran hosts.

Parasitic plants show striking convergence in host preference across angiosperm lineages.

The host specificity of a parasite underpins its ecology, distribution, invasive potential and adaptability, yet for most parasitic plants host ranges are poorly understood. We examine host-parasite relationships across lineages to infer how host specificity may have influenced the evolution of parasitism in plants. Host preference data for all plant holoparasite species were manually collected from literature and herbarium specimens, then analysed to investigate and visualise host diversity and specificity. We reveal a disproportionality in host preference across host lineages: the Asteraceae contains 10% of angiosperm diversity but is infected by 31% of parasite species; meanwhile Monocots comprise 23% but are infected by just 3.2%. Furthermore, we observe striking convergence in host preference: Asteraceae, Euphorbiaceae and Fabaceae are infected by six, five and four independent parasite lineages, respectively. We also demonstrate considerable variation in the degree of host specificity among closely related parasite species; a result that does not reflect the expectation of holoparasites - especially endoparasites - as host specialists. The marked pattern of convergence in preference across disparate lineages points to a common pathway in the evolution of parasitism of eudicots in preference to monocots, which may have in turn have been driven by a divergence in host root and vascular architecture. The unexpected variation in host specificity among closely related species suggests that even apparent generalists may comprise cryptic host-specific taxa. This highlights the value of host preference as an additional consideration in parasitic plant taxonomy. Together, our data point to a complex interplay between ecological and physiological factors driving the evolution of host-parasite interactions. Moreover, they emphasize how little is known about the ecology of most holoparasitic plants, a group of organisms that are especially vulnerable at a time of unprecedented biodiversity loss and extinction.

Exploring the Genetic and Morphological Diversity of Argulus Ectoparasite Infecting the Aquaculture and Ornamental Fish in Tripura

Background: The present study screened for isolation of Argulus spp. infecting fishes based on the report of causing substantial economic losses both in aquaculture ponds and ornamental shops across several locations in Tripura, India. Methods: Identification of Argulus spp. was performed based on morphological, and molecular techniques. The molecular identification of Argulus spp. was performed using universal eukaryotic 18S rRNA genetic markers. Prevalence analysis was performed to determine the infestation rate of Argulus spp. in different hosts. Result: Seven A. japonicus, one A. coregoni and one A. foliaceus isolates were identified based on molecular and morphological features. In the present study it was found that A. japonicus infected a wide range of host species, including Goldfish, Milky Koi carp, Mrigal, Rohu, Catla and Subunkin. It was the first instance of A. japonicus infecting milky koi carp both in India and globally. On the other hand, A. foliaceus and A. coregoni infected only tiger oscar and goldfish respectively, indicating a rather narrow range of host diversity.

Genetic Variation in the Invaded Population of the Fall Armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), in India

The fall armyworm, Spodoptera frugiperda, an invasive, polyphagous pest, causes significant damage to corn. The majority of insects rely on a broad range of digestive enzymes and an intricate detoxifying mechanism to consume chemically diverse host plants. The genetic variation in S. frugiperda was analyzed using cytochrome oxidase subunit I (COI) and triose phosphate isomerase (Tpi). In addition, a new attempt was made to determine the variation with respect to two detoxifying genes, viz., carboxylesterase and glutathione S-transferase. The highest genetic variation (3.03%) was found between the S. frugiperda populations of Tirupati and Delhi with respect to the Tpi gene and between the populations of Punjab and Hosur regarding COI (3.30%). The results for various genes revealed that populations of the fall armyworm were homogeneous, showing low genetic distance using COI, ranging from 0.40 to 3.30%, and, using Tpi, ranging from 0.43 to 3.03%. The variation in carboxylesterase and glutathione S-transferase ranged from 0.04 to 0.15% and from 0.01 to 0.02, respectively. Amino acid sequences were also produced using DNA sequences from several fall armyworm populations. Populations in Tirupati, Solapur, and Hyderabad shared 98.7% of their sequence with that in Delhi. Fall armyworm amino acid sequences showed 79.7 to 82.0% identity with S. exigua and 69.6 to 73.0% identity with S. litura. Our study provides vital information for understanding the genetic variation in the fall armyworm following its invasion of India.

Molecular Detection of Anaplasma marginale in Capybara (Hydrochoerus hydrochaeris) from Corrientes, Argentina.

Monitoring wildlife health is essential for understanding global disease patterns, particularly as vector-borne infections extend the geographic ranges and thereby hosts due to environmental shifts. Anaplasma marginale, primarily impacting cattle, has economic implications and has been found in diverse hosts, yet its presence in capybaras (Hydrochoerus hydrochaeris), influential in tick-borne pathogen spread, lacks comprehensive understanding. From 2015 to 2022, 14 capybaras were surveyed across two different areas of northeastern Argentina. In 1 of 14 (7%) capybaras, the presence of A. marginale was confirmed through the amplification of specific genes, msp5 and msp1β. In addition, A. marginale DNA was detected in the capybara's blood sample through quantitative PCR, with a cycle threshold value of 30.81 (800 copies per reaction). Amplification of a fragment of the msp1α gene revealed PCR products of three different sizes, suggesting the presence of at least three coinfecting A. marginale variants in the capybara host. This study suggests that capybaras are wild hosts for A. marginale in the Ibera Wetlands in Argentina, potentially influencing the infection dynamics of both domestic and wild species. This finding highlights the necessity for thorough studies on the role of capybaras in disease dynamics, crucial for understanding wildlife health and the spread of disease.

The ageing virus hypothesis: Epigenetic ageing beyond the Tree of Life.

A recent thought-provoking theory argues that complex organisms using epigenetic information for their normal development and functioning must irreversibly age as a result of epigenetic signal loss. Importantly, the scope of this theory could be considerably expanded, with scientific benefits, by analyzing epigenetic ageing beyond the borders of the Tree of Life. Viruses that use epigenetic signals for their normal functioning may also age, that is, present an increasing risk of failing to complete their individual life cycle and to disappear with time. As viruses are ancient, abundant, and infect a considerable diversity of hosts, the ageing virus hypothesis, if verified, would have important consequences for many fields of the Life sciences. Uncovering ageing viruses would integrate the most abundant and biologically central entities on Earth into theories of ageing, enhance virology, gerontology, evolutionary biology, molecular ecology, genomics, and possibly medicine through the development of new therapies manipulating viral ageing.

Systematic, high-throughput characterization of bacteriophage gene essentiality on diverse hosts.

Understanding core and conditional gene essentiality is crucial for decoding genotype-phenotype relationships in organisms. We present PhageMaP, a high-throughput method to create genome-scale phage knockout libraries for systematically assessing gene essentiality in bacteriophages. Using PhageMaP, we generate gene essentiality maps across hundreds of genes in the model phage T7 and the non-model phage Bas63, on diverse hosts. These maps provide fundamental insights into genome organization, gene function, and host-specific conditional essentiality. By applying PhageMaP to a collection of anti-phage defense systems, we uncover phage genes that either inhibit or activate eight defenses and offer novel mechanistic hypotheses. Furthermore, we engineer synthetic phages with enhanced infectivity by modular transfer of a PhageMaP-discovered defense inhibitor from Bas63 to T7. PhageMaP is generalizable, as it leverages homologous recombination, a universal cellular process, for locus-specific barcoding. This versatile tool advances bacteriophage functional genomics and accelerates rational phage design for therapy.

Pathotype determination of sorghum anthracnose (Colletotrichum sublineola) isolates from Ethiopia using sorghum differentials.

Sorghum anthracnose, caused by Colletotrichum sublineola, is the most destructive disease of sorghum, which causes up to 80% grain yield loss in susceptible varieties. The use of resistance varieties is an effective, durable, and eco-friendly strategy for anthracnose control. Knowledge of the phenotypic and genetic variation in C. sublineola is vital for designing appropriate anthracnose management strategies. The present study examined the morphology and virulence of 25 C. sublineola isolates recovered from various sorghum-producing regions of Ethiopia against 18 known sorghum anthracnose differentials, 6 Ethiopian sorghum landraces, and a variety of Bonsa. Analysis of variance (ANOVA) revealed significant differences among sorghum genotypes, C. sublineola isolates, and their interactions. There was a significant difference between the isolates in virulence, with each isolate exhibiting virulence in 8-72% of the sorghum genotypes tested. Among the 25 tested isolates, the top four most virulent isolates were from Pawe, suggesting that this area is suitable for pathogen diversity studies and host plant resistance screening. The sorghum genotypes IS_18760, Brandes, and Bonsa showed resistance to all tested isolates. Consequently, they may provide potential sources of resistance genes for sorghum breeding programs to develop cultivars resistant to different C. sublineola pathotypes. However, the resistant check SC748-5 was susceptible to isolates NK73_F37, while another resistant check SC112-14 was susceptible to isolates PW123_F47 and PW122_F47. Cluster analysis grouped 22 isolates into seven clusters based on their morphological characters, whereas 24 pathotypes were identified among 25 isolates that were tested on 25 sorghum genotypes. Hence, this study revealed high variation in C. sublineola in Ethiopia suggesting the need for broad-spectrum resistance to control the disease. Sorghum genotypes resistant to various C. sublineola isolates were identified in this study, which can be used in sorghum breeding programs aiming to develop resistant cultivars to anthracnose. Highly virulent C. sublineola isolates were also identified which could be used in sorghum germplasm resistance screening. The report is the first to show the existence of C. sublineola pathotypes in Ethiopia.

Diversity and Host Specificity of Avian Haemosporidians in an Afrotropical Conservation Region.

Afrotropical regions have high bird diversity, yet few studies have attempted to unravel the prevalence of avian haemosporidia in conservation areas. The diversity and host specificity of parasites in biodiversity hotspots is crucial to understanding parasite distribution and potential disease emergence. We test the hypothesis that biodiverse regions are associated with highly diverse parasites. By targeting the cytochrome b (Cytb) gene, we molecularly screened 1035 blood samples from 55 bird species for avian haemosporidia infections to determine its prevalence and diversity on sites inside and adjacent to the Kruger National Park. Overall infection prevalence was 28.41%. Haemoproteus, Leucocytozoon, and Plasmodium presented prevalences of 17.39%, 9.24%, and 4.64%, respectively. One hundred distinct parasite lineages were detected, of which 56 were new lineages. Haemoproteus also presented the highest diversity compared to Leucocytozoon and Plasmodium with varying levels of specificity. Haemoproteus lineages were found to be specialists while Plasmodium and Leucocytozoon lineages were generalists. We also found a positive relationship between avian host diversity and parasite diversity, supporting an amplification effect. These findings provide insight data for host-parasite and co-evolutionary relationship models.

Global meta‐analysis reveals the drivers of gut microbiome variation across vertebrates

AbstractShifts in gut microbial diversity and structure are one route by which vertebrate hosts adapt to local environmental conditions. However, recent studies have mostly been limited to a single species, small sample sizes, or restricted geographic ranges. Therefore, drawing a global picture of vertebrate gut microbiome diversity, community structure, and determinants for their adaptive shifts remains to be elucidated. We here collected 6508 samples from 113 vertebrate species covering diverse classes, feeding behaviors, and host habitats based on 16S rRNA gene sequencing. The results showed that host diet pattern had a significant impact on gut microbiome variation, which might drive taxonomic and functional contents of gut microbiome across vertebrates. Of note, the phylum Fusobacteria were enriched in carnivorous vertebrate gut while herbivorous vertebrate gut selectively increased the abundance of Verrucomicrobia. Also, climate factors were strongly associated with gut microbiome variation across vertebrates. Interestingly, we found that the abundance of microbiota belonging to Bacteroidetes increased gradually while the members from Proteobacteria showed a decreasing trend from high‐ to low‐latitude zones, potentially contributing to vertebrate adaptation to local climate condition. Additionally, we comprehensively deciphered the common antibiotic resistomes and their potential mobility between terrestrial vertebrate gut microbiome (n = 487) and their sympatric soil biological environment samples (n = 203) by integrating metagenomic sequencing datasets. Particularly, potential horizontal antibiotic resistance genes (e.g., bacA) transfers were detected between vertebrates gut microbiome and their sympatric soil biological environment. Together, our findings provide new evidence of how external environmental factors affect vertebrate gut microbiome variation.

Host diversity of Aedes albopictus in relation to invasion history: a meta-analysis of blood-feeding studies

BackgroundThe invasive mosquito Aedes albopictus is a major concern for human and animal health given its high potential to spread over large geographical distances, adapt to various habitats and food sources, and act as a vector for pathogens. It is crucial to understand how this species establishes ecological relationships at different locations, as it determines its role in transmission of diseases.MethodsBased on published blood meal surveys, a meta-analysis was performed to investigate how host diversity changes along the process of invasion at a large scale. For 48 independent localities, the Shannon diversity index was calculated and was then assessed against several moderator variables describing invasion status, habitat type, methodology, survey year and the year of introduction for invasive populations.ResultsDiet diversity was higher in the invasive than in the native populations when the strong habitat effects were held constant. Furthermore, the year of introduction also had a significant role, as invasive populations that had been established earlier had wider diet diversity than more recent populations.ConclusionsInvasive Ae. albopictus has considerable ecological flexibility. The species’ ability to adapt to various food sources goes hand in hand with its successful worldwide dispersion, which has strong implications for its role in pathogen transmission.Graphical

Population genetic structure of Montipora digitata coral-algal symbiosis in the South China Sea

Species genetic diversity can reflect their adaptability to environmental changes. Coral reefs worldwide are in rapid decline due to climate change and human activities, highlighting the need for conservation intervention. The South China Sea (SCS) is an important biodiversity hotspot, particularly in the Xisha Islands (XS) that contain the majority of coral species in the SCS. However, few studies exist on coral genetic diversity in the SCS. Montipora digitata is the dominant reef-building coral species in the XS. In this study, two mitochondrial DNA fragments and 11 microsatellite markers were employed to investigate the genetic diversity of coral host of M. digitata, and ITS amplicon sequencing was employed to investigate the Symbiodiniaceae community structure. We sampled five wild populations of M. digitata in Hainan Island and the Xisha Islands. Montipora digitata showed low genetic diversity across populations in the SCS. We found significant genetic differentiation between the Sanya (SY) and XS populations but no significant genetic differentiation within XS populations. We identified four Symbiodiniaceae genera, with Cladocopium and Durusdinium being the most common, as well as Gerakladium and Fugacium. The Symbiodiniaceae types of SY and XS are significantly different, C15 is abundant in all populations, while D1a is also abundant in SY. We analyzed the correlation between 16 environmental factors and the genetic diversity of symbionts, genetic diversity of coral host and Symbiodiniaceae community was significantly correlated with sea surface temperature (SST) and other environmental factors. Our study provided a detailed paradigm from the perspective of genetic diversity of how a dominant coral species can be indicative of poor environmental adaptation potential.

First Report of Haemaphysalis longicornis (Neumann) in Oklahoma, USA.

Haemaphysalis longicornis (Neumann), the Asian longhorned tick, is a species native to East Asia, but invasive to Australia, New Zealand, and most recently, the United States. It has spread rapidly across the eastern United States after being established in New Jersey in 2017. Aiding this rapid expansion is the ability of this tick to reproduce parthenogenically and feed on diverse host species. In cattle, this tick can cause heavy burdens and act as a vector for the pathogenic hemoprotozoan parasite Theileria orientalis, genotype Ikeda, creating economic losses that impact the cattle industry. Here, we report Asian longhorned ticks, collected from cattle, a dog, and pastures and morphologically identified at the Oklahoma Animal Disease Diagnostic Laboratory as H. longicornis before molecular confirmation through PCR amplification of the cox1 gene. Blood samples from infested cattle were collected and assessed molecularly for the presence of T. orientalis, with no pathogenic DNA detected. This report describes the first record of H. longicornis in Oklahoma and the farthest westward detection of this tick in the United States to date.

Reassessing the Diversity of the Arthropod-Pathogenic Genus Pandora Batko (Entomophthoromycotina; Erynioideae)

The fungal order Entomophthorales occurs worldwide, with most species infecting arthropods as pathogens. Species in this order can cause epizootics and change the behavior of infected hosts. Molecular data are available only for 20% of the known species, and distributions of species are seldom summarized. Significant diversity of hosts, poor molecular data availability, and poor resolution of the phylogenetic relationships within this fungal order suggest that the diversity of these fungi is not sufficiently described. The subfamily Erynioideae includes 111 arthropod pathogens, divided among six genera, with the genus Pandora being one of the most diverse genera. Sequences of 18S, 28S, and ITS for two species are used to place these Pandora species in a phylogenic tree of the subfamily; this tree also supports our synonymy of the genus Furia with Pandora. Among the two species specifically covered in this paper, Pandora gloeospora was observed during epizootics occurring in mushroom flies (Diptera: Sciaridae) on Agaricus bisporus cultures in Pennsylvania, Delaware, and Maryland (US) mushroom farms and also in Florida on Pleurotus sp. Outside the US, P. gloeospora was found infecting several Nematocera (Diptera) in Europe (France) and Asia (China). Pandora sylvestris n. sp. was collected during epizootics occurring in larvae of hickory tussock moths, Lophocampa caryae (Lepidoptera: Erebidae), in hardwood forests in Michigan and Vermont.

Cultural Reproduction and Religious Traditions Among Banyumas Migrant Communities Within the NGAPAK Population in The Provinces of Lampung and East Kalimantan

Understanding the perpetuation of cultural and religious traditions among Banyumasan migrant communities presents a multifaceted challenge, necessitating a profound comprehension of Banyumasan cultural and religious norms, the migratory journey to regions such as Lampung and Kalimantan, and the intricate dynamics of cultural transmission. Meticulous research yields the following conclusions: Firstly, Banyumasan culture encompasses communities in Banjarnegara, Purbalingga, Banyumas, Cilacap, Kebumen, Tegal, and Brebes, characterized by the prevalent Ngapak dialect of Javanese. Rooted in language, values, traditions, and rituals, Banyumasan culture reflects egalitarianism, directness, and harmony, emblematic of the wong cilik community. Secondly, a significant number of Banyumasan individuals historically migrated to various parts of Indonesia, notably Lampung and Kalimantan, initially for colonial-era labor needs and later to alleviate population pressures and enhance economic prospects during the New Order era. Thirdly, migration fosters extensive interactions between Banyumasan migrants and host communities, involving a nuanced negotiation process wherein individuals balance preserving their cultural heritage and adapting to new surroundings. The outcome depends on effective communication strategies to convey Banyumasan cultural identity amidst diverse host cultures.

Pathogen contingency loci and the evolution of host specificity: Simple sequence repeats mediate Bartonella adaptation to a wild rodent host.

Parasites, including pathogens, can adapt to better exploit their hosts on many scales, ranging from within an infection of a single individual to series of infections spanning multiple host species. However, little is known about how the genomes of parasites in natural communities evolve when they face diverse hosts. We investigated how Bartonella bacteria that circulate in rodent communities in the dunes of the Negev Desert in Israel adapt to different species of rodent hosts. We propagated 15 Bartonella populations through infections of either a single host species (Gerbillus andersoni or Gerbillus pyramidum) or alternating between the two. After 20 rodent passages, strains with de novo mutations replaced the ancestor in most populations. Mutations in two mononucleotide simple sequence repeats (SSRs) that caused frameshifts in the same adhesin gene dominated the evolutionary dynamics. They appeared exclusively in populations that encountered G. andersoni and altered the dynamics of infections of this host. Similar SSRs in other genes are conserved and exhibit ON/OFF variation in Bartonella isolates from the Negev Desert dunes. Our results suggest that SSR-based contingency loci could be important not only for rapidly and reversibly generating antigenic variation to escape immune responses but that they may also mediate the evolution of host specificity.

Directed evolution of an orthogonal transcription engine for programmable gene expression in eukaryotes.

T7 RNA polymerase has enabled orthogonal control of gene expression and recombinant protein production across diverse prokaryotic host chassis organisms for decades. However, the absence of 5' methyl guanosine caps on T7 RNAP derived transcripts has severely limited its utility and widespread adoption in eukaryotic systems. To address this shortcoming, we evolved a fusion enzyme combining T7 RNAP with the single subunit capping enzyme from African swine fever virus using Saccharomyces cerevisiae. We isolated highly active variants of this fusion enzyme, which exhibited roughly two orders of magnitude higher protein expression compared to the wild-type enzyme. We demonstrate the programmable control of gene expression using T7 RNAP-based genetic circuits in yeast and validate enhanced performance of these engineered variants in mammalian cells. This study presents a robust, orthogonal gene regulatory system applicable across diverse eukaryotic hosts, enhancing the versatility and efficiency of synthetic biology applications.