Phylogeny Tree Research Articles

Genetic Identification of Endemic Fish Nomorhamphus liemi Vogt, 1978 from Sigi, Central Sulawesi, Indonesia

Sulawesi Island is geographically included in the Wallacea region, a transitional area between the Oriental and Australian zones, which makes Sulawesi’s fresh waters have a high level of fish species diversity. Some fish species found in Sulawesi freshwater are endemic. Research on Sulawesi endemic fishes in rivers is not as much as research on endemic fishes in lakes. Nomorhamphus liemi or locally known as angculung is one of Sulawesi’s endemic fishes. This species belongs to the sub-family Zenarchopterinae. The majority of species from the Zenarchopterinae sub-family are sexually dimorphic. This study aimed to molecularly identify angculung fish species using COI markers in the region. A total of nine angculung fish were collected and preserved using 96% alcohol. In the laboratory, tissue taken from the back of the dorsal fin was isolated and extracted, then amplified by PCR before sequencing. Sequencing results were validated on BLASTn, showing that nine samples belonged to Nomorhamphus liemi with 98-99% sequence similarity. The nucleotide base sequences were then analyzed using MEGA 7.0 software. The genetic distance of mackerel in these waters showed a close relationship. Phylogeny tree results showed population mixing so that mackerel in these waters can be managed in one management unit, especially in Sulawesi.

Fall Armyworm Spodoptera frugiperda J.E. Smith (Lepidoptera: Noctuidae) in East Nusa Tenggara Province of Indonesia: Genetic Characterization and Strain Detection

<em>Spodoptera frugiperda</em> J.E. Smith (Lepidoptera: Noctuidae) is an invasive pest of corn plants spreading throughout the world, including East Nusa Tenggara (NTT), Indonesia. Despite the wide distribution, there is a lack of information on the strain or genetic diversity of the pest in NTT. Therefore, this study aimed to determine the strain of <em>S. frugiperda</em> from several areas in NTT with a molecular method using <em>cytochrome oxidase subunit 1</em> (<em>COI</em>)<em> </em>and <em>triose phosphate isomerase</em> (<em>Tpi</em>)<em> </em>gene markers. The<em> </em>samples were collected from 3 islands: Timor, Flores, and Sumba. Amplification of the marker genes was carried out using 3 specific primers to identify the strain obtained from samples. Subsequently, polymerase chain reaction (PCR) products were sequenced and the DNA sequences were analyzed using the BioEdit and BLAST programs. Phylogeny analyses were carried out using the MEGA 11 program to verify the strain group of samples with reference isolates from other countries found in GenBank. The PCR results showed product amplicon size 811 bp for the <em>CO1A</em> marker. Based on phylogeny tree analysis using <em>COI </em>marker, <em>S. frugiperda </em>from NTT showed 2 clades, namely corn and rice strains. The characterization results showed that <em>S. frugiperda</em> in NTT comprised 63.6% corn and 36.4% rice strains. <em>COIB </em>classified <em>S. frugiperda</em> from NTT into the h4 haplotype subgroup, while <em>Tpi </em>gene marker was in the corn strain. This study provided valuable information regarding the strain of <em>S. frugiperda</em> in Indonesia to determine the appropriate control strategy.

Phenotypic and transcriptomics characterization uncovers genes underlying tuber yield traits and gene expression marker development in potato under aeroponics.

Transcriptome analysis in potato varieties revealed genes associated with tuber yield-related traits and developed gene expression markers. This study aimed to identify genes involved in high tuber yield and its component traits in test potato varieties (Kufri Frysona, Kufri Khyati, and Kufri Mohan) compared to control (Kufri Sutlej). The aeroponic evaluation showed significant differences in yield-related traits in the varieties. Total RNA sequencing was performed using tuber and leaf tissues on the Illumina platform. The high-quality reads (QV > 25) mapping with the reference potato genomes revealed statistically significant (P < 0.05) differentially expressed genes (DEGs) into two categories: up-regulated (> 2 Log2 fold change) and down-regulated (< -2 Log2 fold change). DEGs were characterized by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Collectively, we identified genes participating in sugar metabolism, stress response, transcription factors, phytohormones, kinase proteins, and other genes greatly affecting tuber yield and its related traits. A few selected genes were UDP-glucose glucosyltransferase, glutathion S-transferase, GDSL esterase/lipase, transcription factors (MYB, WRKY, bHLH63, and BURP), phytohormones (auxin-induced protein X10A, and GA20 oxidase), kinase proteins (Kunitz-type tuber invertase inhibitor, BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1) and laccase. Based on the selected 17 peptide sequences representing 13 genes, a phylogeny tree and motifs were analyzed. Real time-quantitative polymerase chain reaction (RT-qPCR) analysis was used to validate the RNA-seq results. RT-qPCR based gene expression markers were developed for the genes such as 101kDa heat shock protein, catechol oxidase B chloroplastic, cysteine protease inhibitor 1, Kunitz-type tuber invertase inhibitor, and laccase to identify high yielding potato genotypes. Thus, our study paved the path for potential genes associated with tuber yield traits in potato under aeroponics.

The complete chloroplast genome sequence of Isoetes baodongii (Isoetaceae)

Isoetes baodongii is a diploid species of Isoetaceae distributed in low altitude area, its megaspore ornamentation is similar to tetraploid species I. sinensis. We collected leaf material of I. baodongii and sequenced it for low depth whole genome sequence, then, a complete chloroplast genome of I. baodongii was assembled and annotated. This chloroplast genome has a circular structure of 145,494 bp in length with a GC content of 38.0%, comprising a large single copy (LSC) region of 91,860 bp, a pair of inverted repeat (IR) regions of 13,207 bp each, and a small single copy (SSC) region of 27,220 bp. 136 genes were annotated, including 84 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. A maximum likelihood phylogeny tree was reconstructed after the sequences alignment, the result showed that I. baodongii formed a sister clade to the one clustered by I. sinensis, I. taiwanensis and I. orientalis. Although the chloroplast genome structure of I. baodongii is extremely similar to other species distributed in China, a well-supported phylogenetic relationship was reconstructed here, these results may provide new messages for further studies on phylogeny and evolution of vascular plant on the earth.

Functional ecology and evolution of terrestrial and epiphytic species of Rhododendron section Schistanthe (Ericaceae)

Epiphytes are often characterized by small size, specifically adapted to a relatively dry and nutrient-poor habitat. However, this epiphyte syndrome is derived from studies on species-rich and habitat-dominating groups such as orchids and bromeliads. To better understand general trait syndromes of epiphytes, it is important to analyze plant groups that are more transitional in the evolution of epiphytism e.g., the Australasian vireyas (Rhododendron sect. Schistanthe; Ericaceae). Here, we studied the morphology, ecophysiology, and anatomy of 44 species of Rhododendron section Schistanthe from living collections and obtained data for 288 species related to their "life form" (holoepiphytes [= obligatory epiphytes], facultative epiphytes and terrestrial congeners) from the literature. For phylogeny and time-calibrated tree, we used the nuclear ITS (Internal Transcribed Spacers) region. Using different statistics and phylogenetic comparative methods, our results support the hypothesis that holoepiphytic vireyas are smaller in size than the terrestrial ones. Terrestrial vireyas are found at higher minimum and maximum elevation than either facultative or holoepiphytic congeners. We demonstrate that the diversification of tropical Rhododendron is not related to the evolution of the epiphytic life form. Regarding the evolution of traits that are important for water economy, holoepiphytic vireyas do not differ much from their terrestrial relatives indicating that evolutionary steps for specialization towards an holoepiphytic habit are in their initial stages in Rhododendron. Given the large species diversity of the group conclusions based on the subset used here must be considered preliminary. However, our results provide the basis for more detailed future studies.

The influence of tree genus, phylogeny, and richness on the specificity, rarity, and diversity of ectomycorrhizal fungi.

Partner specificity is a well-documented phenomenon in biotic interactions, yet the factors that determine specificity in plant-fungal associations remain largely unknown. By utilizing composite soil samples, we identified the predictors that drive partner specificity in both plants and fungi, with a particular focus on ectomycorrhizal associations. Fungal guilds exhibited significant differences in overall partner preference and avoidance, richness, and specificity to specific tree genera. The highest level of specificity was observed in root endophytic and ectomycorrhizal associations, while the lowest was found in arbuscular mycorrhizal associations. The majority of ectomycorrhizal fungal species showed a preference for one of their partner trees, primarily at the plant genus level. Specialist ectomycorrhizal fungi were dominant in belowground communities in terms of species richness and relative abundance. Moreover, all tree genera (and occasionally species) demonstrated a preference for certain fungal groups. Partner specificity was not related to the rarity of fungi or plants or environmental conditions, except for soil pH. Depending on the partner tree genus, specific fungi became more prevalent and relatively more abundant with increasing stand age, tree dominance, and soil pH conditions optimal for the partner tree genus. The richness of partner tree species and increased evenness of ectomycorrhizal fungi in multi-host communities enhanced the species richness of ectomycorrhizal fungi. However, it was primarily the partner-generalist fungi that contributed to the high diversity of ectomycorrhizal fungi in mixed forests.

Abstract 6924: Structural variant dynamics in melanoma: Unraveling tumor heterogeneity and evolution

Abstract Melanoma is characterized by significant intratumoral heterogeneity and complex evolutionary dynamics. This diversity in genomic alterations leads to the emergence of various subclonal populations within a single tumor. In our research, we established a new model system composed of 24 single-cell-derived clonal sublines (C1-C24), originating from the M4 melanoma model. This model was developed using a genetically engineered hepatocyte growth factor (HGF)-transgenic mouse. We employed Trisicell (Triple-toolkit for single-cell intratumor heterogeneity inference), a cutting-edge computational tool for scalable analysis of intratumor heterogeneity and evaluation based on single-cell RNA mutations. This enabled us to construct a phylogeny tree, revealing melanoma’s intricate branching evolutionary patterns. These patterns show ancestral clones evolving into genetically distinct subclones, which demonstrate varied phenotypic traits such as drug sensitivity or resistance, cellular plasticity, and immunogenicity. In our study, we conducted long-read sequencing on these clonal sublines in the phylogeny and identified structural variants (SVs) using Severus, a tool optimized for phasing in long-read sequencing. The types of SVs we discovered include deletions, insertions, amplifications, translocations, and inversions. We explored their roles in subclonal evolution, particularly focusing on how they disrupt genes and accumulate during melanoma progression. Our initial data from eleven sublines indicated a higher prevalence of ancestral SVs, shared by all sublines, compared to subline-specific SVs, representing later events. Notably, the individual sublines showed a higher rate of gene disruption by SVs, hinting at potential functional selection. Our analysis further revealed that SVs common to all sublines are linked with genes in key cell growth pathways, such as Rap1, Hippo, and calcium signaling pathways. In contrast, subline-specific SVs primarily affect genes involved in neurophysiological pathways, such as glutamatergic synapse and morphine addiction pathways. These findings suggest that different genes, associated with various pathways, are disrupted at distinct stages of melanoma progression, providing insights into the genetic factors that may predispose individual melanocytes to melanomagenesis. This methodology presents a comprehensive tool for characterizing tumor genomes and understanding their relationship with disease progression and therapy resistance. We are continuing our analysis to map SVs across the entire mutation-based phylogeny of all sublines, which will further our understanding of melanoma’s genomic landscape. Citation Format: Xiwen Cui, Ayse G. Keskus, Farid R. Mehrabadi, Salem Malikic, Mikhail Kolmogorov, Chi-Ping Day, Glenn Merlino, S. Cenk Sahinalp. Structural variant dynamics in melanoma: Unraveling tumor heterogeneity and evolution [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6924.

Abstract 6243: Rapid autopsy reveals the interplay of tumor heterogeneity at genomic, transcriptomic, and spatial dimensions

Abstract Incomplete understanding of tumor heterogeneity across primary and metastatic sites often lead to ineffective cancer treatments. To comprehensively interrogate tumor heterogeneity at genomic, transcriptomic, and spatial levels, we established a novel rapid autopsy protocol to obtain multiregional tri-omic data consisted of whole exome sequencing, mRNA-sequencing, and multiplexed immunohistochemistry on primary and metastatic tumors and associated microenvironments (TMEs). We applied this protocol to six patients with various primary tumor types: non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), urothelial cancer, and one mixed case with both NSCLC and SCLC. A total of 50 tumor samples with paired tri-omic data were obtained across 16 different primary and metastatic organs to interrogate heterogeneities underlying tumor evolution and treatment resistance mechanisms. We constructed mutation-based phylogeny trees to reveal structural differences in tumor evolution trajectories across patients and infer heterogeneous tumor antigen landscape. We uncovered highly entropic phylogenetic structures having unique variants gained at each metastasis as well as low entropic structures with high sharing across sites. The mixed case had a clonal sweep structure with two metastases developed prior to the rest of the metastasis. In this case, older clones were shown to have strong association with APOBEC mutation signatures, enhanced subclonal mutation burden and loss of human leukocyte antigen heterozygosity as mechanisms of immune evasion. Integrating transcriptome and spatial profiles, we revealed that tumor intrinsic expressions and extrinsic immune infiltration levels covary with tissue types rather than with mutation-based phylogenetic clusters. Across patients, samples that had high similarities in mutational landscape varied in their pro-tumor expression programs and the cellular composition of TMEs. In particular, lung samples across patients had the highest effector T cell population and major histocompatibility complex (MHC) class I, II presentations compared to other tissue types. Contrasting small cell carcinoma (SCC) and adenocarcinoma (Adeno) samples from lung tissues further revealed that SCC had more immunosuppressive TMEs with downregulation of MHC class II. Liver metastasis samples were further characterized to have the most aggressive phenotypes enriched in genomic instability measured by aneuploidy compared to other metastatic sites. In sum, we present a novel technology and computational framework that enables a three-dimensional profiling of tumor heterogeneity. This work suggests that single-site biopsies are insufficient to capture tumor heterogeneity and calls for multi-modal integrations to design treatments that can overcome tumor-specific resistance mechanisms. Citation Format: Ning Yao, Rami Vanguri, Juber Patel, Jayon Lihm, Andrew Fisher, Irene Jarchum, Shigeaki Umeda, Hao Li, Scott Ely, Travis Hollmann, Christine Iacobuzio-Donahue, Benjamin Greenbaum. Rapid autopsy reveals the interplay of tumor heterogeneity at genomic, transcriptomic, and spatial dimensions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6243.

Polyphyletic origin of saxitoxin biosynthesis genes in the marine dinoflagellate Alexandrium revealed by comparative transcriptomics

The marine dinoflagellate Alexandrium is known to form harmful algal blooms, and at least 14 species within the genus can produce saxitoxins (STXs). STX biosynthesis genes (sxt) are individually revealed in toxic dinoflagellates; however, the evolutionary history remains controversial. Herein, we determined the transcriptome sequences of toxic Alexandrium (A. catenella and A. pacificum) and non-toxic Alexandrium (A. fraterculus and A. fragae) and characterized their sxt by focusing on evolutionary events and STX production. Comparative transcriptome analysis revealed higher homology of the sxt in toxic Alexandrium than in non-toxic species. Notably, non-toxic Alexandrium spp. were found to have lost two sxt core genes, namely sxtA4 and sxtG. Expression levels of 28 transcripts related to eight sxt core genes showed that sxtA, sxtG, and sxtI were relatively high (>1.5) in the toxic group compared to the non-toxic group. In contrast, the non-toxic group showed high expression levels in sxtU (1.9) and sxtD (1.7). Phylogenetic tree comparisons revealed distinct evolutionary patterns between 28S rDNA and sxtA, sxtB, sxtI, sxtD, and sxtU. However, similar topology was observed between 28S rDNA, sxtS, and sxtH/T. In the sxtB and sxtI phylogeny trees, toxic Alexandrium and cyanobacteria were clustered together, separating from non-toxic species. These suggest that Alexandrium may acquire sxt genes independently via horizontal gene transfer from toxic cyanobacteria and other multiple sources, demonstrating monocistronic transcripts of sxt in dinoflagellates.

The roles of environmental filtering and competitive exclusion in the plant community assembly at Mt. Huangshan are forest-type-dependent

Integrating trait and phylogenetic data are increasingly being used to enhance our understanding of the underlying community assembly processes and optimize the interpretation of unmeasured, phylogenically conserved traits. The biodiversity of Huangshan, China, was higher, but it was suffered from some damage, such as biological invasion and human disturbance. We sampled 176 species from three different forests (EBF: evergreen broadleaf forest; DBF: deciduous broadleaf forest; MNBF: mixed needleleaf and broadleaf forest) which are the representative communities of Mt. Huangshan. Then, we measured six functional traits (leaf thickness, leaf area, specific leaf area, leaf carbon concentration, leaf nitrogen concentration and leaf phosphorus concentration) and constructed a molecular phylogeny tree to assess community structure using species trait-phylogenetic distances. Results showed that six traits are more phylogenetically convergent than predicted, which was not closely related to the evolutionary history of species. The contribution of phylogenetic distance and functional distance to community construction was equal in total layer and more importance was given to functional distances for tree and shrub layer, indicating that closely related species tended to be phenotypically different. The functional trait-phylogenetic of EBF and MNBF was clustering, while DBF exhibited functional trait-phylogenetic over-dispersion. In other words, environmental filtering appears to be the dominant force in EBF and MNBF, while competitive exclusion appears to become more important in DBF. Our study also indicates that spatial factors resulted in a higher explanatory power than environmental factors. In according to better protect forest biodiversity, species coexistence mechanisms should be incorporated into the management of forest communities and to predict their presentation in future environmental scenarios.

DNA Barcoding Supports "Color-Pattern''-Based Species of Stictochironomus from China (Diptera: Chironomidae).

The genus Stictochironomus (Diptera: Chironomidae) has an almost worldwide distribution, with more than 30 species. However, species delimitation and identification based on the markings on the wings and legs are controversial and uncertain. In this study, we focused on color patterns to review the adults of the genus from China, and two new species (S. trifuscipes sp. nov. and S. quadrimaculatus sp. nov.) are described and figured. DNA barcodes can accurately separate the two new species with specific color patterns. However, heterospecific individuals form a monophyletic cluster in the phylogeny tree. For example, S. maculipennis (Meigen) and S. pictulus (Meigen), which have a lower interspecific genetic divergence, form a single clade. Sequences with the same species name but with high intraspecific distance form more than one phylogenetic clade, such as S. sticticus (Fabricius) of three clades, S. pictulus of four clades, S. akizukii (Tokunaga) and S. juncaii Qi, Shi, and Wang of two clades, might have potential cryptic species diversity. Species delimitation analysis using ASAP, PTP, and GMYC clearly delineated them as separate species. Consequently, color patterns are a good diagnostic characteristic for species delimitation in Stictochironomus. The distance-based analysis shows that a threshold of 4.5-7.7% is appropriate for species delimitation in Stictochironomus. Additionally, an updated key including color pattern variation for male adults of known Stictochironomus species from China is provided.

Investigation of CaMV-host co-evolution through synonymous codon pattern.

Cauliflower mosaic virus (CaMV) has a double-stranded DNA genome and is globally distributed. The phylogeny tree of 121 CaMV isolates was categorized into two primary groups, with Iranian isolates showing the greatest genetic variations. Nucleotide A demonstrated the highest percentage (36.95%) in the CaMV genome and the dinucleotide odds ratio analysis revealed that TC dinucleotide (1.34 ≥ 1.23) and CG dinucleotide (0.63 ≤ 0.78) are overrepresented and underrepresented, respectively. Relative synonymous codon usage (RSCU) analysis confirmed codon usage bias in CaMV and its hosts. Brassica oleracea and Brassica rapa, among the susceptible hosts of CaMV, showed a codon adaptation index (CAI) value above 0.8. Additionally, relative codon deoptimization index (RCDI) results exhibited the highest degree of deoptimization in Raphanus sativus. These findings suggest that the genes of CaMV underwent codon adaptation with its hosts. Among the CaMV open reading frames (ORFs), genes that produce reverse transcriptase and virus coat proteins showed the highest CAI value of 0.83. These genes are crucial for the creation of new virion particles. The results confirm that CaMV co-evolved with its host to ensure the optimal expression of its genes in the hosts, allowing for easy infection and effective spread. To detect the force behind codon usage bias, an effective number of codons (ENC)-plot and neutrality plot were conducted. The results indicated that natural selection is the primary factor influencing CaMV codon usage bias.

Transcriptome profiling and characterization of genes associated with tuberization under high temperature in aeroponics in potato cv. Kufri Anand.

High temperature stress is an important abiotic factor, which affects tuberization and ultimately causes heavy yield reduction in potato. Identification and characterization of genes associated with tuberization under high temperature stress is essential for future management through biotechnology. Two contrasting potato varieties Kufri Anand (profuse tuber-bearing) versus Kufri Frysona (very less/scanty tuber-bearing, control) were cultivated in aeroponics under high temperature stress, and transcriptomes were analyzed. Potato cv. Kufri Anand was found superior over control (Kufri Frysona) for tuber yield and its component traits along with root morphology under aeroponics. Transcriptomes of tuber and leaf tissues were analyzed. Statistically significant (p < 0.05) differentially expressed genes (DEGs) were categorised into up-regulated (> 2 log2 fold change, FC) and down-regulated (< -2 log2 FC) genes. DEGs were annotated by gene ontology and KEGG pathways. A few selected up-regulated genes of both tissues were identified, and phylogeny tree and motif analysis were analysed based on 36 peptide sequences representing 15 selected DEGs in this study. Further, gene expression markers were developed and validated by real time qPCR analysis for the identification of high temperature tolerant genotypes. A few key genes associated in tuberization under high temperature conditions were heat shock proteins (e.g. 18.5kDa class I heat shock protein), sugar metabolism (e.g. glucosyltransferase), transcription factor (e.g. WRKY), and phytohormones (e.g. auxin-induced beta-glucosidase). Our study provides an overview of key genes involved in tuberization under high temperature stress in potato cv. Kufri Anand under aeroponics.

Molecular Identification of Lactic Acid Bacteria from Broiler Chicken Meat

Lactic acid bacteria are bacteria used to ferment food. Molecular identification of isolates of lactic acid bacteria has a high degree of accuracy compared to morphological identification. This study aimed to molecularly identify isolates of lactic acid bacteria from broiler chickens. Molecular identification is carried out in 3 stages, namely: rRNA gene amplification stage, squencing and phylogeny tree. PCR electrophoresis results obtained amplicone results measuring about 1,500 bp. The results of BLAST analysis showed that the gen 16S rRNA sequence of BR 17 lactic acid bacteria isolate had a 97-98% similarity with the genome sequence of the Lactobacillus fermentum strain. Development of phylogenetic tree isolates of single clusters of lactic acid bacteria BR 17 with strains of Lactobacillus fermentum CAU2036, Lactobacillus fermentum JCM 7772, and Lactobacillus fermentum 10-18. The identification results of lactic acid bacteria isolate BR 17 showed Lactobacillus fermentum BR 17.

Identification and Characterization of Entamoeba histolytica Choline Kinase.

Entamoeba histolytica is one of the death-causing parasites in the world. Study on its lipid composition revealed that it is predominated by phosphatidylcholine and phosphatidylethanolamine. Further study revealed that its phosphorylated metabolites might be produced by the Kennedy pathway. Here, we would like to report on the characterizations of enzymes from this pathway that would provide information for the design of novel inhibitors against these enzymes in future. E. histolytica HM-1:IMSS genomic DNA was isolated and two putative choline/ethanolamine kinase genes (EhCK1 and EhCK2) were cloned and expressed from Escherichia coli BL21 strain. Enzymatic characterizations were further carried out on the purified enzymes. EhCK1 and EhCK2 were identified from E. histolytica genome. The deduced amino acid sequences were more identical to its homologues in human (35-48%) than other organisms. The proteins were clustered as ethanolamine kinase in the constructed phylogeny tree. Sequence analysis showed that they possessed all the conserved motifs in choline kinase family: ATP-binding loop, Brenner's phosphotransferase motif, and choline kinase motif. Here, the open reading frames were cloned, expressed, and purified to apparent homogeneity. EhCK1 showed activity with choline but not ethanolamine. The biochemical characterization showed that it had a Vmax of 1.9 ± 0.1µmol/min/mg. Its Km for choline and ATP was 203 ± 26µM and 3.1 ± 0.4mM, respectively. In contrast, EhCK2 enzymatic activity was only detected when Mn2+ was used as the co-factor instead of Mg2+ like other choline/ethanolamine kinases. Highly sensitive and specific antibody against EhCK1 was developed and used to confirm the endogenous EhCK1 expression using immunoblotting. With the understanding of EhC/EK importance in phospholipid metabolism and their unique characteristic, EhC/EK could be a potential target for future anti-amoebiasis study.

Comparison of genetic diversity of LEP gene between Indonesia domestic goats: Etawa Cross and Senduro Goats

Abstract. Amrullah MF, Utomo B, Utama S, Lestari TD, Suprayogi TW, Restiadi TI, Belgania RH, Pakpahan S, Khairullah AR, Kurniawan SC, Silaen OSM, Hasib A. 2023. Comparison of genetic diversity of LEP gene between Indonesia domestic goats: Etawa Cross and Senduro Goats. Biodiversitas 24: 6567-6573. The leptin (LEP) gene is a gene that functions to produce the hormone leptin secreted by fat tissue that can increase livestock productivity. This study aimed to identify the profile of the LEP gene between Etawah Cross goats and Senduro goats. Gene profiling includes single nucleotide polymorphisms (SNPs) and phylogeny tree reconstruction. Furthermore, 10 whole blood samples from 5 Etawah Cross goats and 5 Senduro goats were collected from goat farms in Lumajang District, Lumajang Regency, Indonesia. The Research procedure involved collecting blood samples, extracting DNA, amplification using the polymerase chain reaction (PCR) method using specific primers, and nucleotide sequencing method. The edited DNA sequences were aligned with some DNA sequences (n=32) of the Capra hircus group published in Genbank. The results of this study obtained the alignment of the LEP gene sequences of Etawah Cross goats and Senduro goats, including T723A, G729A, G758A, A763T, G774C, T1100C, and G1454A. Based on the LEP gene phylogeny tree, Etawah Cross goats and Senduro goats belong to the same clade and have a close kinship with the Bligon goats. The profile of the LEP gene between Etawah Cross goats and Senduro goats varies so that further studies can be carried out to look for the relationship between SNPs and the productivity traits of the LEP gene.

Species discrimination power of ITS DNA barcoding on Tristaniopsis species on Bangka Island, Indonesia

The rDNA ITS barcoding marker has been widely used for plant identification, along with other primers for the chloroplast sequence region. This study applied the ITS primer to discriminate Tristaniopsis spp. on Bangka Island. Twelve leaf samples were collected from five community forests in Bangka district. The pairwise distance between sequences calculated using Kimura’s two parameters in MEGA11 revealed that pelawan air is different from other pelawan specimens (1.58–2.1%), while pelawan orange and pelawan merah specimens showed low variation (0–0.78%). There is a barcoding gap in the application of the ITS markers (full-length ITS including 5.8s, ITS1, and ITS2) for Tristaniopsis, which is shown by the higher value of minimum interspecific divergence than the value of maximum interspecific divergence for all three regions. Phylogeny trees constructed from the nucleotide sequence of 12 pelawan and other related taxa clustered pelawan merah and pelawan orange together and separated them from pelawan air.

Comparison of chloroplast genomes and phylogenetic analysis of four species in Quercus section Cyclobalanopsis

The identification in Quercus L. species was considered to be difficult all the time. The fundamental phylogenies of Quercus have already been discussed by morphological and molecular means. However, the morphological characteristics of some Quercus groups may not be consistent with the molecular results (such as the group Helferiana), which may lead to blurring of species relationships and prevent further evolutionary researches. To understand the interspecific relationships and phylogenetic positions, we sequenced and assembled the CPGs (160,715 bp-160842 bp) of four Quercus section Cyclobalanopsis species by Illumina pair-end sequencing. The genomic structure, GC content, and IR/SC boundaries exhibited significant conservatism. Six highly variable hotspots were detected in comparison analysis, among which rpoC1, clpP and ycf1 could be used as molecular markers. Besides, two genes (petA, ycf2) were detected to be under positive selection pressure. The phylogenetic analysis showed: Trigonobalanus genus and Fagus genus located at the base of the phylogeny tree; The Quercus genus species were distincted to two clades, including five sections. All Compound Trichome Base species clustered into a single branch, which was in accordance with the results of the morphological studies. But neither of group Gilva nor group Helferiana had formed a monophyly. Six Compound Trichome Base species gathered together in pairs to form three branch respectively (Quercus kerrii and Quercus chungii; Quercus austrocochinchinensis with Quercus gilva; Quercus helferiana and Quercus rex). Due to a low support rate (0.338) in the phylogeny tree, the interspecies relationship between the two branches differentiated by this node remained unclear. We believe that Q. helferiana and Q. kerrii can exist as independent species due to their distance in the phylogeny tree. Our study provided genetic information in Quercus genus, which could be applied to further studies in taxonomy and phylogenetics.

Novel Mutations in the Non-Structure Protein 2 of SARS-CoV-2.

Mutation in the genome of SARS-CoV-2 may play a role in immune evasion, pathogenicity, and speed of its transmission. Our investigation aimed to evaluate the mutations that exist in the NSP2. RNA was extracted from nasopharyngeal swabs from 100 COVID-19 patients. RT-PCR was performed on all samples using NSP2-specific primers. Following gel electrophoresis, the bands were cut, purified, and sequenced using the Sanger method. After sequencing, 90 sequences could be used for further analysis. Bioinformatics analysis was conducted to investigate the effect of mutations on protein structure, stability, prediction of homology models, and phylogeny tree. The patients' mean age was 51.08. The results revealed that 8 of the 17 NSP2 mutations (R207C, T224I, G262V, T265I, K337D, N348S, G392D, and I431M) were missense. One deletion was also found in NSP2. Among NSP2 missense mutations studied, K337D and G392D increased structural stability while the others decreased it. The homology-designed models demonstrated that the homologies were comparable to the sequences of the Wuhan-HU-1 virus. Our study suggested that the mutations K337D and G392D modulate the stability of NSP2, and tracking viral evolution should be implemented and vaccine development updated.

The maternal ancestry of the Kavaratti islanders and the last glacial maximum aftermath.

The prehistoric human settlement of the Lakshadweep islands remains a mystery for various reasons. Uncertainty about the existence of indigenous tribes in these islands and the lack of folklore records present major obstacles to the reconstruction of Lakshadweep ancestry. However, with extant population data, we seek to understand the maternal ancestry of theKavaratti islanders. Mitochondrial control region variation analysis of 80 individuals from this island shows maternal links with the populations in the northwestern region of the South Asian mainland. The founder clade R30b2, observed in the Kavaratti islanders, is so far present only in the Scheduled Castes from the Punjab region, Jat Sikhs and Nairs. All other mainland populations carry basal R30 or R30a subclades. The presence of a specific Uralic U4 lineage in our samples, in addition to the Indo-European affinity observed in the phylogeny tree, substantiates a northwestern maternal ancestry of the Kavaratti islanders and implies an ancestral admixture with early humans in the Near East at the time of the last glacial maximum (LGM). Based on our Bayesian analysis, we furthermore propose that a group bearing mostly R30b2 during the LGM recovery, moved eastward and southward, where they received Indian-specific M haplogroups. Hence, the maternal ancestry of the Kavaratti islanders is evidently a consequence of the demographic changes in the northwestern region of the Indian subcontinent caused by the Last Glacial Maximum. The haplogroup distribution pattern and nucleotide sequence data produced in this study will enrich the forensic database of the Lakshadweep islands.