Genetic Characterization and Recombinant History of a Novel Emerging HIV-1 Second-Generation Circulating Recombinant Form (CRF120_0107) Identified Among Sexually Transmitted Infections in Shenzhen, China

Under the background of the main epidemic HIV strains (CRF01_AE and CRF07_BC) co-circulation in China, more HIV second-generation recombinant (SGR) strains with CRF01_AE and CRF07_BC as the backbone are emerging. In this study, we analyzed the characteristics and evolutionary history of a newly emerging HIV-1 CRF120_0107 composed of CRF01_AE and CRF07_BC based on the near full-length genome (NFLG) in Shenzhen, Guangdong Province, China. NFLG phylogenetic analysis revealed that these sequences formed a distinct monophyletic branch with a high bootstrap value (>90%), distantly related to all known HIV-1 genotypes. Recombination analysis showed that CRF120_0107 was composed of the predominant HIV-1 strains in China: CRF01_AE and CRF07_BC. Further subregional phylogenetic analysis was performed that possible parental lineages of CRF07_BC segments (Ⅰ, Ⅲ, and Ⅴ) belonged to the CRF07_BC men who have sex with men cluster (MSM cluster), other CRF01_AE segments also mainly belonged to MSM Cluster (such as CRF01_AE Cluster 5). Bayesian analysis results inferred that CRF120_0107 placed its emergence in Shenzhen approximately between 2009-2011. The appearance of CRF120_0107 further highlights that more and more HIV-1 SGR strains containing CRF01_AE and CRF07_BC will be more generated frequently and will most likely be more conducive to accelerating the spread of HIV in China. This highlighted it is necessary to monitor MSM high-risk individuals with HIV-1 CRF01_AE and CRF07_BC dual infection to prevent the generation of CRF01_AE/CRF07_BC recombinant strains, thus reducing the possibility of HIV-1 genotype resistance and the complexity of treatment in China.

Phylotranscriptomics reveals discordance in the phylogeny of Hawaiian Drosophila and Scaptomyza (Diptera: Drosophilidae)

Island radiations present natural laboratories for studying the evolutionary process. The Hawaiian Drosophilidae are one such radiation, with nearly 600 described species and substantial morphological and ecological diversification. These species are largely divided into a few major clades, but the relationship between these clades remains uncertain. Here we present 12 new assembled transcriptomes from across these clades, and use these transcriptomes to resolve the base of the evolutionary radiation. We recover a new hypothesis for the relationship between clades, and demonstrate its support over previously published hypotheses. We then use the evolutionary radiation to explore dynamics of concordance in phylogenetic support, by analyzing the gene and site concordance factors for every possible topological combination of major groups. We show that high bootstrap values mask low evolutionary concordance, and we demonstrate that the most likely topology is distinct from the topology with the highest support across gene trees and from the topology with highest support across sites. We then combine all previously published genetic data for the group to estimate a time-calibrated tree for over 300 species of drosophilids. Finally, we digitize dozens of published Hawaiian Drosophilidae descriptions, and use this to pinpoint probable evolutionary shifts in reproductive ecology as well as body, wing, and egg size. We show that by examining the entire landscape of tree and trait space, we can gain a more complete understanding of how evolutionary dynamics play out across an island radiation.

Identification of Two Similar Novel HIV-1 Recombinant Forms (CRF01_AE/CRF07_BC) among Intravenous Drug Users in Guangxi, China

New kinds of HIV-1 circulating recombinant forms (CRFs) and unique recombinant forms (URFs) earn a great prevalence in China nowadays. In this study, we identified 2 similar URFs (2016GXNNIDU037 and 2019QZLSIDU253) both isolated from intravenous drug users (IDUs) in Guangxi, China. Phylogenetic analysis of the near full-length genome (NFLG) revealed 2 URFs both clustered with CRF01_AE but setting up a monophyletic branch, supporting a high bootstrap value. Bootscan analysis and subregional recombinant analysis found that the NFLG of 2016GXNNIDU037 and 2019QZLSIDU253 were both composed of CRF01_AE and CRF07_BC, with 3 CRF07_BC mosaic segments inserted into CRF01_AE backbones. The CRF01_AE segments of the 2 URFs clustered with a previously reported cluster 2 lineage of CRF01_AE. The 5 recombinant breakpoints of the 2 URFs were quite similar. Distinct from CRF01_AE/CRF07_BC URFs reported before, 2016GXNNIDU037 and 2019QZLSIDU253 are new evidence of a high genetic variety of HIV-1 in Guangxi, which may pose new challenges to HIV-1 prevention and molecular epidemiological surveillance in China.

Complete Genome Sequence of a Novel Latent Tobamovirus Infecting Hevea Brasiliensis in China

In this work, we describe the complete sequence and genome organization of a novel tobamovirus detected in rubber tree (Hevea brasiliensis) by high-throughput RNA-seq. The infected plants show no identifiable symptoms, therefore this virus is tentatively named rubber tree latent virus 1 (RTLV1). The full genome of RTLV1 is 9,422 nt in length and contains three open reading frames (replicase, movement, and coat proteins) with 157 nt 5' UTR and 316 nt 3' UTR. The replicase encoded by ORF1 shares only 33.29% amino acid sequence identity with that of the closest related Frangipani mosaic virus. Phylogenetic analysis using the ORF1 amino acid sequence clusters RTLV1 with members of genus Tobamovirus (family Virgaviridae) at relative high bootstrap, suggesting that RTLV1 is a novel member of Tobamovirus.

DNA barcoding approach to identification of Cassidula angulifera (Petit, 1841) (Mollusca: Gastropoda)

A large number of gastropod species have similarities in morphology (cryptic) makes misidentification probably happen/occurred. Accurate species identification is needed in studying bioecology of species. This research aims to identify the species of Cassidulla sp. Which was collected from Peyum Beach Merauke with DNA barcoding techniques using COI gene markers. The primers used in this study are forward primers (LCO1490) and reverse primers (HCO2198). The result of identification with DNA barcoding showed that the species analyzed was Cassidula angulifera with a 99.53% similarity level with a DNA sequence length of 650 bp. Phylogenetic reconstruction showing the entire sequence of Cassidula sp. which were analyzed separately based on the type and genetic distance with high bootstrap value. Phylogenetic reconstruction of Cassidula sp. form a monophyletic group, which means that the species come from the same ancestors. DNA barcoding is very good and accurate in identifying species.

Molecular record for the first authentication of Isaria cicadae from Vietnam

The entomopathogenic fungus T011, parasitizing on nymph of Cicada, collected in the coffee garden in Dak Lak Province, Vietnam, was preliminarily morphologically identified as Isaria cicadae, belonged to order Hypocreales and family Clavicipitaceae. To ensure the authenticity of T011, phylogenetic analysis of the concatenated set of multiple genes including ITS, nrLSU, nrSSU, Rpb1, and Tef1 was applied to support the identification. Genomic DNA was isolated from dried sample T011. The PCR assay sequencing was applied to amplify ITS, nrLSU, nrSSU, Rpb1, and Tef1 gene. For phylogenetic analysis, the concatenated data of both target gens were constructed with MEGAX with a 1,000 replicate bootstrap based on the neighbor-joining, maximum likelihood, maximum parsimony method. As the result, the concatenated data containing 62 sequences belonged to order Hypocreales, families Clavicipitaceae, and 2 outgroup sequences belonged to order Hypocreales, genus Verticillium. The phylogenetic analysis results indicated that T011 was accepted at subclade Cordyceps and significantly formed the monophyletic group with referent Cordyceps cicadae (Telemorph of Isaria cicadae) with high bootstrap value. The phylogenetically analyzed result was strongly supported by our morphological analysis described as the Isaria cicadae. In summary, phylogenetic analyses based on the concatenated dataset were successfully applied to strengthen the identification of T011 as Isaria cicadae.

First Phylogenetic Treatment of Apple Cucumber (Family Cucurbitaceae) from Indonesia Utilizing DNA Variation of Internal Transcibed Spacer Region

Cucurbitaceae is one of the largest family in Angiosperm in which the most member of this family is important fruit crops in Indonesia such as Cucumber, Melon, Watermelon, and Apple Cucumber. In particular, Apple Cucumber, currently attracts attention to many researchers due to its phylogeneticand taxonomic problem. In term of its appearance, the fruit looks like an apple but the taste is melon. The purpose of this study was to elucidatephylogenetic relationship between Apple Cucumber and other species of Cucurbitaceae based on variation of DNA sequences derived from internal transcribed spacer (ITS) region. As many as six individuals of Apple Cucumber collected from Karawang, Jember, and Aceh were examined. The ITS sequences of some species of family Cucurbitaceae were retrieved from GenBank, and put them in the analysis. Phylogenetic analysis based on parsimony method with using Begoniaas outgroup reveals that Apple Cucumber are nested in the same clade as Melon (Cucumis melo) with high bootstrap value (100%), suggesting that Apple Cucumber is under the same species as Melon. However, on the basis of morphological characters of fruit, apple cucumber is different with that of Melon. This considerably first phylogenetics treatment provides fundamental knowledge for establishing a subspecies of Melon.

Pantoea ananatis, A New Bacterial Pathogen Affecting Wheat Plants (Triticum L.) in Poland

Wheat (Triticum aestivum) is one of the most economically important crops in the world. During the routine monitoring of wheat pest, the cereal leaf beetle (CLB, Oulema melanopus, Coleoptera, Chrysomelidae), in the Greater Poland region, it was observed that some leaves wounded by CLB also displayed brownish lesions with clear margins and yellow halo, disease symptoms resembling a bacterial infection. The aim of this study was therefore to investigate those symptoms to establish a causal agent of the disease. The identification based on the results of the Biolog’s Gen III system, 16S rRNA, and gyrB genes sequencing, revealed the presence of eight strains of Pantoea ananatis bacteria. Four strains were derived from wheat leaves (Ta024, Ta027, Ta030, Ta046), and four from the CLB’s oral secretion (OUC1, OUD2, OUF2, and OUG1). They shared the nucleotide identity ranging from 99 to 100% to P. ananatis strains deposited in the GenBank database. Additionally, the multi-locus sequence analysis (MLSA) of concatenated sequences of partial atpD, fusA, gyrB, rplB, and rpoB genes was performed. All P. ananatis strains isolated in Poland, grouped into one cluster supported with high bootstrap value. Pathogenicity tests performed on four varieties of wheat plants have identified P. ananatis strains as a causal agent of wheat disease. To our knowledge, this is the first report of P. ananatis affecting wheat plants.

Sequencing and Characterization of Mitochondrial Protein-Coding Genes for Schizothorax niger (Cypriniformes: Cyprinidae) with Phylogenetic Consideration

The present study was conducted to get more information about the genome and locate the taxonomic position of Schizothorax niger in Schizothoracinae through mitochondrial 13 protein-coding genes (PCGs). These PCGs for S. niger were found to be 11409 bps in length ranging from 165 (ATPase 8) to 1824 bps (NADH dehydrogenase subunit 5) and encode 3801 amino acids. In these PCGs, 4 genes overlap on the similar strands, while one shown on the opposite one: ATPase 6+8 and NADH dehydrogenase subunit 4+4L overlap by 7 nucleotides. Similarly, ND5-ND6 overlap by 4 nucleotides, while ATP6 and COIII overlap by 1 nucleotide. Similarly, four commonly used amino acids in S. niger were Leu (15.6 %), Ile (10.12 %), Thr (8.12 %), and Ala (8.7 %). The results presented that COII, COIII, NDI, ND4L, and Cytb had substantial amino acid conservation as compared to the COI gene. Through phylogenetic analysis, it was observed that S. niger is closely linked with S. progastus, S. labiatus, S. plagiostomus, and S. nepalensis with high bootstrap values. The present study provided more genomic data to know the diversity of the mitochondrial genome and its molecular evolution in Schizothoracinae.

Ellipsomyxa ariusi sp. nov. (Myxosporea: Ceratomyxidae), a new myxosporean infecting the gallbladder of threadfin sea catfish Arius arius in India

The present study describes a new species of myxosporean, Ellipsomyxa ariusi sp. nov., infecting the gallbladder of the threadfin sea catfish Arius arius (Hamilton, 1822). E. ariusi sp. nov. is characterized by bivalvular, ellipsoid or elongate-oval myxospores with smooth spore valves and a straight suture, arranged at an angle to the longitudinal spore axis. Mature myxospores measured 10.1 ± 0.8 µm in length, 6.8 ± 0.5 µm in width and 7.7 ± 0.7 µm in thickness. Polar capsules are equal in size and oval to pyriform in shape. They are positioned at an angle to the longitudinal myxospore axis and open in opposite directions. Polar capsules measured 2.8 ± 0.3 µm in length and 2.5 ± 0.4 µm in width; polar filaments formed 4-5 coils, and extended to 32.2 ± 2.1 µm in length. Monosporic and disporic plasmodial stages attached to the wall of gallbladder. Molecular analysis of the type specimen generated a 1703 bp partial SSU rDNA sequence (MN892546), which was identical to the isolates from 3 other locations. In phylogenetic analyses, genus Ellipsomyxa appeared monophyletic and E. ariusi sp. nov. occupied an independent position in maximum likelihood and Bayesian inference trees with high bootstrap values. The overall prevalence of infection was 54.8% and multiway ANOVA revealed that it varied significantly with location, year, season, sex and size of the fish host. Histopathological changes associated with E. ariusi sp. nov. infection included swelling, vacuolation and detachment of epithelial layer, reduced mucus production and altered consistency and colour of bile. Based on the morphologic, morphometric and molecular differences with known species of Ellipsomyxa, and considering differences in host and geographic locations, the present species is treated as new and the name Ellipsomyxa ariusi sp. nov. is proposed.