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Wang, X.-R.

Publications and source records attributed to Wang, X.-R..

8 recordsLinked to original sources

The vitellogenesis pathway of the tick host Ixodes scapularis regulates Rickettsia buchneri transcriptional state and persistence

Obligate endosymbionts relying on transovarial transmission must coordinate with host reproduction, yet the regulatory mechanisms remain poorly understood. Here we show that the vitellogenesis pathway of the tick Ixodes scapularis regulates the transcriptional state of its endosymbiont Rickettsia buchneri (Rb), separately from its abundance. In males, Rb DNA remained detectable, but bacterial transcription was strongly reduced across all examined genes, with markedly lower RNA/DNA ratios. In females, Rb was restricted to ovarian tissues (developing oocytes and interstitial cells), with no detection in salivary glands or midgut by TEM, FISH, or PCR. After blood feeding, Rb density within size-matched early-stage oocytes was significantly reduced. We further characterized two gene families mediating vitellogenesis: vitellogenin synthesis genes (Vgs, n = 20) and vitellogenin receptor genes (Vgr, n = 15). Vgs proteins showed conserved domain organization, whereas Vgr paralogs showed greater structural diversification. RNAi silencing of Vgs20 or Vgr12 altered Rb transcriptional profiles in both tick cells and ticks, although changes in bacterial load were not consistent between the two systems. Antibiotic depletion of Rb increased expression of both host genes. Together, these findings show that tick vitellogenesis pathways contribute to Rb regulation during reproduction and identify the I. scapularis-Rb system as a useful model for studying host control of obligate endosymbionts. IMPORTANCEMaternally inherited bacterial symbionts are commonly characterized by bacterial abundance. This study shows that bacterial abundance alone does not necessarily reflect symbiont functional state. In the blacklegged tick, the inherited symbiont Rickettsia buchneri persists in males but exhibits little transcriptional activity. In females, the host reproductive pathway determines whether the symbiont is active: silencing one component of this pathway changed bacterial gene expression without changing bacterial abundance. Hosts therefore regulate not only the abundance of inherited symbionts, but also their functional state. The same bacterial abundance can correspond to very different functional states. Understanding inherited symbioses therefore requires considering bacterial function alongside bacterial abundance.

microbiology↗

A novel downstream factor in willows replaces the ancestral sex determining gene

Sex chromosome turnover has occurred in many groups, and is mediated by the translocation or duplication of apical sex-determining genes, or the replacement of original sex determination genes by new ones. In dioecious plants, the former frequently occurs, while the latter is rarely reported. Here, we assembled four haplotype-resolved chromosome-level genomes from four lineages of the Salix polyclona complex, including three diploids (E, E-TS, and W1) and one autotetraploid (W2). Our analyses reveal that diploids have a ZW/ZZ system, while autotetraploid has a ZZZW/ZZZZ system on chromosome 15. The apical sex-determining genes of Salicaceae, ARR17-like duplicates on chromosomes 15 and 19, which can indirectly turn-on/off the expression of PISTILLATA (PI) gene and determine sex, appears to have lost the function of sex determination in the S. polyclona complex. We found a novel sex determining factor on the 15W of the complex, namely partial PI-like duplicates, which has taken over the function from the ARR17-like duplicates. In conclusion, a new dominant sex-determining gene was recruited in the S. polyclona complex, replacing the ancestral apical sex-determining ARR17-like duplicates. The newly identified partial PI-like duplicates exhibit a direct influence on the downstream intact PI-like duplications, providing valuable insights into the evolutionary trajectory of the top-down sex determination pathway.

evolutionary biology↗

Low genetic differentiation across an entire continent: the Scots pine paradox

O_LIScots pine is the foundation species of diverse forested ecosystems across Eurasia and displays remarkable ecological breadth, occurring in environments ranging from temperate rainforests to arid tundra margins. Such expansive distributions can be favored by various demographic and adaptive processes and the interactions between them. C_LIO_LITo understand the impact of neutral and selective forces on genetic structure in Scots pine, we conducted range-wide population genetic analyses on 2,321 trees from 202 populations using genotyping-by-sequencing, reconstructed the recent demography of the species, and examined signals of genetic adaptation. C_LIO_LIWe found a high and uniform genetic diversity across the entire range (global FST 0.048), no increased genetic load in expending populations and minor impact of the last glacial maximum on historical population sizes. Genetic-environmental associations identified only a handful of SNPs significantly linked to environmental gradients. C_LIO_LIThe results suggest that extensive gene flow is predominantly responsible for the observed genetic patterns in Scots pine. The apparent missing signal of genetic adaptation is likely attributed to the intricate genetic architecture controlling adaptation to multi-dimensional environments. The panmixia metapopulation of Scots pine offers a good study system for further exploration into how genetic adaptation and plasticity evolve under gene flow and changing environment. C_LI

evolutionary biology↗

Examination of rickettsial host range for shuttle vectors based on dnaA and parA genes from the pRM plasmid of Rickettsia monacensis

The genus Rickettsia encompasses a diverse group of obligate intracellular bacteria that are highly virulent disease agents of mankind as well as symbionts of arthropods. Native plasmids of Rickettsia amblyommatis (AaR/SC) have been used as models to construct shuttle vectors for genetic manipulation of several Rickettsia species. Here we report on the isolation of the complete plasmid (pRM658B) from Rickettsia monacensis (IrR/Munich) mutant Rmona658B and the construction of shuttle vectors based on pRM. To identify regions essential for replication, we made vectors containing the dnaA and parA genes of pRM with varying portions of the region surrounding these genes and a selection-reporter cassette conferring resistance to spectinomycin and expression of green fluorescent protein. Rickettsia amblyommatis (AaR/SC), R. monacensis (IrR/Munich), Rickettsia bellii (RML 369-C), Rickettsia parkeri (Tates Hell), and Rickettsia montanensis (M5/6) were successfully transformed with shuttle vectors containing pRM parA and dnaA. PCR assays targeting pRM regions not included in the vectors revealed that native pRM was retained in R. monacensis transformants. Determination of native pRM copy number using a plasmid-encoded gene (RM_p5) in comparison to chromosomal encoded gltA indicated reduced copy numbers in R. monacensis transformants. In transformed R. monacensis, native pRM and shuttle vectors with homologous parA and dnaA formed native plasmid-shuttle vector complexes. These studies provide insight on the maintenance of plasmids and shuttle vectors in rickettsiae. IMPORTANCEO_LIThis paper describes a new series of plasmid shuttle vectors for the transformation of rickettsiae. C_LIO_LIShuttle vectors based on parA and dnaA sequences of the plasmid pRM can be used to transform diverse rickettsia as well as its native host Rickettsia monacensis. C_LIO_LIOur results provide insight into the maintenance of rickettsial-based shuttle vectors in rickettsiae. C_LI

microbiology↗

The mitogenome of Norway spruce and a reappraisal of mitochondrial recombination in plants

Plant mitogenomes can be difficult to assemble because they are structurally dynamic and prone to intergenomic DNA transfers, leading to the unusual situation where an organelle genome is far outnumbered by its nuclear counterparts. As a result, comparative mitogenome studies are in their infancy and some key aspects of genome evolution are still known mainly from pre-genome, qualitative methods. To help address these limitations, we combined machine learning and in silico enrichment of mitochondrial-like long reads to assemble the bacterial-sized mitogenome of Norway spruce (Pinaceae: Picea abies). We conducted comparative analyses of repeat abundance, intergenomic transfers, substitution and rearrangement rates, and estimated repeat-by-repeat homologous recombination rates. Prompted by our discovery of highly recombinogenic small repeats in P. abies, we assessed the genomic support for the prevailing hypothesis that intramolecular recombination is predominantly driven by repeat length, with larger repeats facilitating DNA exchange more readily. Overall, we found mixed support for this view: recombination dynamics were heterogeneous across vascular plants and highly active small repeats (ca. 200 bp) were present in about a third of studied mitogenomes. As in previous studies, we did not observe any robust relationships among commonly-studied genome attributes, but we identify variation in recombination rates as a underinvestigated source of plant mitogenome diversity.

genomics↗

Universal and naked-eye gene detection platform based on CRISPR/Cas12a/13a system

Colorimetric gene detection based on gold nanoparticles (AuNPs) is an attractive detection format due to its simplicity. Here, we report a new design for a colorimetric gene-sensing platform based on the CRISPR/Cas system that has improved specificity, sensitivity, and universality. CRISPR/Cas12a and CRISPR/Cas13a have two distinct catalytic activities and are used for specific target gene recognition. Programmable recognition of DNA by Cas12a/crRNA and RNA by Cas13a/crRNA with a complementary sequence activates the nonspecific trans-ssDNA or -RNA cleavage, respectively, thus degrading the ssDNA or RNA linkers which are designed as a hybridization template for the AuNP-DNA probe pair. Target-induced trans -ssDNA or RNA cleavage leads to a distance-dependent color change for the AuNP-DNA probe pair. In this platform, naked eye detection of transgenic rice, African swine fever virus (ASFV), and a miRNA can be completed within 1 hour. Our colorimetric gene-sensing method shows superior characteristics, such as probe universality, isothermal reaction conditions, on-site detection capability, and sensitivity that is comparable to that of the fluorescent detection; thus, this method represents a robust next generation gene detection platform.

bioengineering↗

Overexpressed microRNA-141-3p enhance proliferation via targeting PLAG1 in non-diabetic macrosomia

Several studies have shown microRNAs (miRNAs) could regulate the placental development, yet the role and mechanism of miRNAs in the development of non-diabetic macrosomia (NDFMS) remains unclear. The key miRNA that abnormal expressed in NDFMS placentas was screened out by miRNA microarray and verified using qRT-PCR in 91 subjects. The effects of the key miRNA were verified by proliferation assay and invasion assay in HTR-8/SVneo cell, and also in pregnant C57BL/6J mice. miR-141-3p was determined as the key miRNA with the most significant difference, which could promote the proliferation and invasion by regulating the expression of target gene PLAG1. Overexpression of PLAG1 could reverse the effect of cell proliferation and invasion ability caused by miR-141-3p overexpression. Significant difference in fetal birth weight was observed between the control group and treated group with miR-141-3p agomir in late pregnancy, but not in early pregnancy. This study revealed miR-141-3p could increase the proliferation of placenta to participate in the occurrence and development of NDFMS through regulating PLAG1 expression.

developmental biology↗

Interspecific plastome recombination reveals ancient reticulate evolution in Picea (Pinaceae)

Plastid sequences are a cornerstone in plant systematic studies and key aspects of their evolution, such as uniparental inheritance and absent recombination, are often treated as axioms. While exceptions to these assumptions can profoundly influence evolutionary inference, detecting them can require extensive sampling, abundant sequence data, and detailed testing. Using advancements in high-throughput sequencing, we analyzed the whole plastomes of 65 accessions of Picea, a genus of ~35 coniferous forest tree species, to test for deviations from canonical plastome evolution. Using complementary hypothesis and data-driven tests, we found evidence for chimeric plastomes generated by interspecific hybridization and recombination in the clade comprising Norway spruce (P. abies) and ten other species. Support for interspecific recombination remained after controlling for sequence saturation, positive selection, and potential alignment artifacts. These results reconcile previous conflicting plastid-based phylogenies and strengthen the mounting evidence of reticulate evolution in Picea. Given the relatively high frequency of hybridization and biparental plastid inheritance in plants, we suggest interspecific plastome recombination may be more widespread than currently appreciated and could underlie reported cases of discordant plastid phylogenies.

genomics↗