Chromosomal‐level reference genome of the incense tree Aquilaria sinensis

This is the author accepted manuscript. The final version is available on open access from Wiley via the DOI in this recordData Accessibility: The raw genome and RNA sequencing data have been deposited in the SRA under Bioproject numbers SRR10737433 and PRJNA534170. The final chromosome assembly was submitted to NCBI Assembly under accession number VZPZ00000000 in NCBI.Trees in the genus Aquilaria (Thymelaeaceae) are known as lign aloes, and are native to the forests of southeast Asia. Lign aloes produce agarwood as an antimicrobial defence. Agarwood has a long history of cultural and medicinal use, and is of considerable commercial value. However, due to habitat destruction and over collection, lign aloes are threatened in the wild. We present a chromosomal‐level assembly for Aquilaria sinensis, a lign aloe endemic to China known as the incense tree, based on Illumina short‐read, 10X Genomics linked‐read, and Hi‐C sequencing data. Our 783.8Mbp A. sinensis genome assembly is of high physical contiguity, with a scaffold N50 of 87.6Mbp, and high completeness, with a 95.8% BUSCO score for eudicotyledon genes. We include 17 transcriptomes from various plant tissues, providing a total of 35,965 gene models. We reveal the first complete set of genes involved in sesquiterpenoid production, plant defence, and agarwood production for the genus Aquilaria, including genes involved in the biosynthesis of sesquiterpenoids via the mevalonic acid (MVA), 1‐deoxy‐D‐xylulose‐5‐phosphate (DXP), and methylerythritol phosphate (MEP) pathways. We perform a detailed repeat content analysis, revealing that transposable elements account for ~61% of the genome, with major contributions from gypsy‐like and copia‐like LTR retroelements. We also provide a comparative analysis of repeat content across sequenced species in the order Malvales. Our study reveals the first chromosomal‐level genome assembly for a tree in the genus Aquilaria and provides an unprecedented opportunity to address a variety of applied, genomic and evolutionary questions in the Thymelaeaceae more widely.Chinese University of Hong KongAgriculture, Fisheries and Conservation Department, Government of the Hong Kong Special Administrative RegionBiotechnology and Biological Sciences Research Council (BBSRC

Millipede genomes reveal unique adaptations during myriapod evolution

This is the final version. Available from Public Library of Science via the DOI in this record.The final genome assemblies have been deposited to NCBI database with accession numbers JAAFCF000000000 and JAAFCE000000000. The mRNA and sRNA transcriptomic data generated in this study have been deposited to the NCBI database under the following BioProject accessions: PRJNA564202 (Helicorthomorpha holstii) and PRJNA564195 (Trigoniulus corallinus).The Myriapoda, composed of millipedes and centipedes, is a fascinating but poorly understood branch of life, including species with a highly unusual body plan and a range of unique adaptations to their environment. Here, we sequenced and assembled 2 chromosomal-level genomes of the millipedes Helicorthomorpha holstii (assembly size = 182 Mb; shortest scaffold/contig length needed to cover 50% of the genome [N50] = 18.11 Mb mainly on 8 pseudomolecules) and Trigoniulus corallinus (assembly size = 449 Mb, N50 = 26.78 Mb mainly on 17 pseudomolecules). Unique genomic features, patterns of gene regulation, and defence systems in millipedes, not observed in other arthropods, are revealed. Both repeat content and intron size are major contributors to the observed differences in millipede genome size. Tight Hox and the first loose ecdysozoan ParaHox homeobox clusters are identified, and a myriapod-specific genomic rearrangement including Hox3 is also observed. The Argonaute (AGO) proteins for loading small RNAs are duplicated in both millipedes, but unlike in insects, an AGO duplicate has become a pseudogene. Evidence of post-transcriptional modification in small RNAs-including species-specific microRNA arm switching-providing differential gene regulation is also obtained. Millipedes possesses a unique ozadene defensive gland unlike the venomous forcipules found in centipedes. We identify sets of genes associated with the ozadene that play roles in chemical defence as well as antimicrobial activity. Macro-synteny analyses revealed highly conserved genomic blocks between the 2 millipedes and deuterostomes. Collectively, our analyses of millipede genomes reveal that a series of unique adaptations have occurred in this major lineage of arthropod diversity. The 2 high-quality millipede genomes provided here shed new light on the conserved and lineage-specific features of millipedes and centipedes. These findings demonstrate the importance of the consideration of both centipede and millipede genomes-and in particular the reconstruction of the myriapod ancestral situation-for future research to improve understanding of arthropod evolution, and animal evolutionary genomics more widely.Hong Kong Research Grants Council (RGC) General Research FundHong Kong Research Grants Council (RGC) General Research FundThe Chinese University of Hong Kong (CUHK

Analysis of multilocus sequence typing schemes for 35 different bacteria revealed that gene loci of 10 bacteria could be replaced to improve cost-effectiveness

Although multilocus sequence typing (MLST) has been widely used for bacterial typing, the contribution of the gene loci to the discriminatory power of each MLST scheme is unknown. We analyzed the discriminatory powers of 36 MLST schemes using all combinations of the 7 loci and contributions of each locus to the schemes. In 10 schemes, sequencing 6 loci can achieve the discriminatory powers of 7 loci. For the other 26 schemes, the median marginal increase in discriminatory power when 7 instead of 6 loci were used is 0.0004. Sequencing the 7 loci of 50 strains each of Pseudomonas aeruginosa and Acinetobacter baumannii revealed that the discriminatory power for P. aeruginosa was 0.9861 when either 6 (without trp) or 7 loci were used and that for A. baumannii was 0.9363 when 5, 6, or 7 loci were used. Genes that have no additional or minimal contribution to the overall discriminatory powers should be replaced. © 2011 Elsevier Inc.link_to_subscribed_fulltex

Characterization of porcine parainfluenza virus 1, a virus with possible association with respiratory disease, from deceased pigs.

Paper Poster Session IV: Viral infection and disease - Poster presentation no. P0887Objectives: Pigs, being farm animals and an important food source of humans, would also be a source of zoonotic disease pathogens, e.g. Influenenza virus A (HIN1) and Nipah virus, in humans. Therefore, understanding more about porcine pathogens is important in preparing for emerging diseases in humans. Recently, our research group has found a porcine paramyxovirus, named porcine parainfluenza 1 (PPIV-1) and detailed analysis was performed based on its genomic data in this study. Methods: The virus studied, PPIV-1, was found by RT-PCR of RNA extracted from samples of deceased pigs from a slaughter house with conserved primers. Complete genomes of three viruses were then sequenced with primer walking strategy. Potential of mRNA editing of phosphoprotein gene was checked by cloning. Characterisation of genomic features, phylogentic analysis and estimation of divergence time were performed using different bioinformatics software tools. Results: RT-PCR for paramyxovirus gene fragment was positive in nasopharyngeal (3.1%) and rectal (0.7%) swab samples, but negative in other samples, namely, liver, lung and blood samples. Annotation of complete genome sequences showed PPIV-1 contain six major genes similar to other paramyxoviruses. Amino acid identities of PPIV-1 were highest with respiroviruses, especially human parainfluenza virus 1 (HPIV-1) and Sendai virus (SeV). Characteristics of intergenic regions and proteins and phylogenetic analysis also suggested PPIV-1 as a member of the genus Respirovirus, with a closer relationship with SeV and HPIV-1 than other respiroviruses. Estimation of divergence dates suggested the divergence time of all respiroviruses were earlier than the group of viruses comprising HPIV-1, SeV and PPIV-1. The Ka/Ks ratios of most coding regions in PPIV-1 were low, suggesting these genes were under purifying selection. Conclusion: The presence of PPIV-1 in mainly respiratory samples suggested a possible association with respiratory disease. A number of common genome characteristics, the phylogenetic analysis and divergence time estimation showed that PPIV-1 is a respirovirus more closely related to HPIV-1 and SeV than other respiroviruses. This suggested that there should be a subtype of 'group 1' viruses in the respiroviruses. The purifying selection in most genes supported swine as the primary host of PPIV-1. To summarise, PPIV-1 should be classified as a 'group 1' respirovirus with a possible association with respiratory disease in swine, its possibly primary host

Identification and complete genome analysis of three novel paramyxoviruses, Tuhoko virus 1, 2 and 3, in fruit bats from China

Among 489 bats of 11 species in China, three novel paramyxoviruses [Tuhokovirus 1, 2 and 3 (ThkPV-1, ThkPV-2 and ThkPV-3)] were discovered in 15 Leschenault's rousettes. Phylogenetically, the three viruses are most closely related to Menangle and Tioman virus. Genome analysis showed that their 3'-leader sequences are unique by possessing GA instead of AG at the 5th and 6th positions. Unlike Menangle and Tioman virus, key amino acids for neuraminidase activity characteristic of rubulavirus attachment proteins are present. The genome of ThkPV-1 represents the largest rubulavirus genome. Unique features between the three viruses include perfect complementary 5'-trailer and 3'-leader sequence and a unique cysteine pair in attachment protein of ThkPV-1, G at +. 1 position in all predicted mRNA sequences of ThkPV-2, and amino acid substitutions in the conserved N-terminal motif of nucleocapsid of ThkPV-3. Analysis of phosphoprotein gene mRNA products confirmed mRNA editing. Antibodies to the viruses were detected in 48-60% of Leschenault's rousettes. © 2010 Elsevier Inc.link_to_subscribed_fulltex

Feline morbillivirus, a previously undescribed paramyxovirus associated with tubulointerstitial nephritis in domestic cats

We describe the discovery and isolation of a paramyxovirus, feline morbillivirus (FmoPV), from domestic cat (Felis catus). FmoPV RNA was detected in 56 (12.3%) of 457 stray cats (53 urine, four rectal swabs, and one blood sample) by RT-PCR. Complete genome sequencing of three FmoPV strains showed genome sizes of 16,050 bases, the largest among morbilliviruses, because of unusually long 5′ trailer sequences of 400 nt. FmoPV possesses identical gene contents (3′-N-P/V/C-M-F-H-L-5′) and is phylogenetically clustered with other morbilliviruses. IgG against FmoPV N protein was positive in 49 sera (76.7%) of 56 RT-PCR-positive cats, but 78 (19.4%) of 401 RT-PCR-negative cats (P < 0.0001) byWestern blot. FmoPV was isolated from CRFK feline kidney cells, causing cytopathic effects with cell rounding, detachment, lysis, and syncytia formation. FmoPV could also replicate in subsequent passages in primate Vero E6 cells. Infected cell lines exhibited finely granular and diffuse cytoplasmic fluorescence on immunostaining for FmoPV N protein. Electron microscopy showed enveloped virus with typical "herringbone" appearance of helical N in paramyxoviruses. Histological examination of necropsy tissues in two FmoPV-positive cats revealed interstitial inflammatory infiltrate and tubular degeneration/necrosis in kidneys, with decreased cauxin expression in degenerated tubular epithelial cells, compatible with tubulointerstitial nephritis (TIN). Immunohistochemical staining revealed FmoPV N protein-positive renal tubular cells and mononuclear cells in lymph nodes. A case-control study showed the presence of TIN in seven of 12 cats with FmoPV infection, but only two of 15 cats without FmoPV infection (P < 0.05), suggesting an association between FmoPV and TIN.link_to_OA_fulltex