Functional genomics analysis reveals the evolutionary adaptation and demographic history of pygmy lorises.

Lorises are a group of globally threatened strepsirrhine primates that exhibit many unusual physiological and behavioral features, including a low metabolic rate, slow movement, and hibernation. Here, we assembled a chromosome-level genome sequence of the pygmy loris (Xanthonycticebus pygmaeus) and resequenced whole genomes from 50 pygmy lorises and 6 Bengal slow lorises (Nycticebus bengalensis). We found that many gene families involved in detoxification have been specifically expanded in the pygmy loris, including the GSTA gene family, with many newly derived copies functioning specifically in the liver. We detected many genes displaying evolutionary convergence between pygmy loris and koala, including PITRM1. Significant decreases in PITRM1 enzymatic activity in these two species may have contributed to their characteristic low rate of metabolism. We also detected many evolutionarily convergent genes and positively selected genes in the pygmy loris that are involved in muscle development. Functional assays demonstrated the decreased ability of one positively selected gene, MYOF, to up-regulate the fast-type muscle fiber, consistent with the lower proportion of fast-twitch muscle fibers in the pygmy loris. The protein product of another positively selected gene in the pygmy loris, PER2, exhibited weaker binding to the key circadian core protein CRY, a finding that may be related to this species' unusual circadian rhythm. Finally, population genomics analysis revealed that these two extant loris species, which coexist in the same habitat, have exhibited an inverse relationship in terms of their demography over the past 1 million years, implying strong interspecies competition after speciation.

Genomic and transcriptomic investigations of the evolutionary transition from oviparity to viviparity.

Viviparous (live-bearing) vertebrates have evolved repeatedly within otherwise oviparous (egg-laying) clades. Over two-thirds of these changes in vertebrate reproductive parity mode happened in squamate reptiles, where the transition has happened between 98 and 129 times. The transition from oviparity to viviparity requires numerous physiological, morphological, and immunological changes to the female reproductive tract, including eggshell reduction, delayed oviposition, placental development for supply of water and nutrition to the embryo by the mother, enhanced gas exchange, and suppression of maternal immune rejection of the embryo. We performed genomic and transcriptomic analyses of a closely related oviparous-viviparous pair of lizards (Phrynocephalus przewalskii and Phrynocephalus vlangalii) to examine these transitions. Expression patterns of maternal oviduct through reproductive development of the egg and embryo differ markedly between the two species. We found changes in expression patterns of appropriate genes that account for each of the major aspects of the oviparity to viviparity transition. In addition, we compared the gene sequences in transcriptomes of four oviparous-viviparous pairs of lizards in different genera (Phrynocephalus, Eremias, Scincella, and Sphenomorphus) to look for possible gene convergence at the sequence level. We discovered low levels of convergence in both amino acid replacement and evolutionary rate shift. This suggests that most of the changes that produce the oviparity-viviparity transition are changes in gene expression, so occasional reversals to oviparity from viviparity may not be as difficult to achieve as has been previously suggested.

Whole-genome sequence of the Tibetan frog Nanorana parkeri and the comparative evolution of tetrapod genomes.

The development of efficient sequencing techniques has resulted in large numbers of genomes being available for evolutionary studies. However, only one genome is available for all amphibians, that of Xenopus tropicalis, which is distantly related from the majority of frogs. More than 96% of frogs belong to the Neobatrachia, and no genome exists for this group. This dearth of amphibian genomes greatly restricts genomic studies of amphibians and, more generally, our understanding of tetrapod genome evolution. To fill this gap, we provide the de novo genome of a Tibetan Plateau frog, Nanorana parkeri, and compare it to that of X. tropicalis and other vertebrates. This genome encodes more than 20,000 protein-coding genes, a number similar to that of Xenopus. Although the genome size of Nanorana is considerably larger than that of Xenopus (2.3 vs. 1.5 Gb), most of the difference is due to the respective number of transposable elements in the two genomes. The two frogs exhibit considerable conserved whole-genome synteny despite having diverged approximately 266 Ma, indicating a slow rate of DNA structural evolution in anurans. Multigenome synteny blocks further show that amphibians have fewer interchromosomal rearrangements than mammals but have a comparable rate of intrachromosomal rearrangements. Our analysis also identifies 11 Mb of anuran-specific highly conserved elements that will be useful for comparative genomic analyses of frogs. The Nanorana genome offers an improved understanding of evolution of tetrapod genomes and also provides a genomic reference for other evolutionary studies.

Stackebrandtia endophytica sp. nov., an actinobacterium isolated from Tripterygium wilfordii.

A novel endophytic actinobacterium, designated strain YIM 64602T, was isolated from healthy stems of Tripterygium wilfordii. It grew at 15-40 °C, pH 6.0-9.0 and in the presence of 0-3 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain YIM 64602T belongs to the genus Stackebrandtia. Whole-cell hydrolysates of strain YIM 64602T contained the amino acid meso-diaminopimelic acid with the sugars mannose, rhamnose and glucose, and a trace of ribose. The major polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine and phosphatidylethanolamine. MK-10(H6), MK-10(H4) and MK-11(H4) were the predominant components in the quinone system. The fatty-acid pattern was mainly composed of the saturated branched-chain acids iso-C16 : 0, anteiso-C17 : 0, iso-C15 : 0 and iso-C17 : 0. The DNA G+C content was 72.4 mol%. 16S rRNA gene sequence analysis showed the highest pairwise sequence identity (96.0-98.5 %) with the members of the genus Stackebrandtia. Strain YIM 64602T displayed a DNA-DNA relatedness of 43.9±0.4 % with the type strain Stackebrandtia albiflava YIM 45751T. Based on evidence from this polyphasic study, strain YIM 64602T ( = BCRC 16954T = DSM 45928T) is considered to represent a novel species of the genus Stackebrandtia, for which the name Stackebrandtia endophytica is proposed.

Mariniluteicoccus flavus gen. nov., sp. nov., a new member of the family Propionibacteriaceae, isolated from a deep-sea sediment.

A Gram-staining-positive, aerobic, non-motile, irregular coccus, designated strain YIM M13146(T), was isolated from a sediment sample collected from the South China Sea at a depth of 2439 m, and its taxonomic position was determined by a polyphasic approach. Optimal growth of the strain was observed at 30 °C (range 5-40 °C), pH 7.0 (pH 6.0-9.0) and 0-1% NaCl (0-6%, w/v) on/in tryptic soy agar/broth. Strain YIM M13146(T) had the major cellular fatty acid anteiso-C15:0, the predominant respiratory menaquinone MK-9(H4), peptidoglycan type A3γ (ll-DAP-Gly) containing alanine, glycine, glutamic acid and ll-diaminopimelic acid (ll-DAP) and the polar lipids phosphatidylcholine, diphosphatidylglycerol, one unknown phospholipid and several glycolipids. The G+C content of the DNA was 67.2 mol%. Phenotypic and chemotaxonomic characteristics together with 16S rRNA gene sequence analyses showed that strain YIM M13146(T) was distinct from its close phylogenetic relatives in the genera Propioniferax and Granulicoccus of the family Propionibacteriaceae. Hence, a new genus and species, Mariniluteicoccus flavus gen. nov., sp. nov., is proposed. The type strain of Mariniluteicoccus flavus is YIM M13146(T) ( = DSM 25892(T) = CCTCC AB 2012055(T)).

Isolation and biological activity of six new polyketide and terpenoid derivatives from Neopestalotiopsis Clavispora AL01.

A fungus strain, Neopestalotiopsis clavispora AL01, was isolated from the leaf spot of the plant Phoenix dactylifera. Further chemical investigation of the fermentation extract of this strain afforded six new secondary metabolites (1-6), along with 11 known compounds (7-17) which included a new natural compound (7). Their structures were determined by extensive spectroscopic analysis including one-and two-dimensional (1D and 2D) NMR spectroscopy, high-resolution electrospray ionization mass spectrometry (HRESIMS), and ECD and NMR calculations. All compounds were evaluated for their phytotoxic activities. Among them, compounds 10, 12 and 13 exhibited phytotoxic activities against Nicotiana tabacum. Compound 3 exhibited weak antibacterial activity against methicillin-resistant Staphylococcus aureus, Micrococcus luteus and Vibrio harveyi. Taken collectively, these findings establish a solid research foundation for future investigations on bioactive natural products derived from phytopathogenic fungi.

Rothia endophytica sp. nov., an actinobacterium isolated from Dysophylla stellata (Lour.) Benth.

A novel endophytic actinobacterium, designated strain YIM 67072(T), was isolated from healthy roots of Dysophylla stellata (Lour.) Benth. Cells of this aerobic, cream-yellow-coloured strain occurred singly, in pairs or in tetrads, were Gram-stain-positive and ovoid- to spherical-shaped. Strain YIM 67072(T) grew at 4-45 °C, pH 5.0-10.0 and in the presence of 0-7 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 67072(T) belonged to the genus Rothia. The isolate contained MK-7 as the major component of the quinone system. The peptidoglycan type was A3α. The polar lipid profile consisted predominantly of diphosphatidylglycerol and glycolipids. The major fatty acids were anteiso-C15 : 0, iso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The DNA G+C content was 53.2 mol%. However, strain YIM 67072(T) differed from its closest relatives Rothia nasimurium CCUG 35957(T) (98.5 % 16S rRNA gene sequence similarity), Rothia amarae JCM 11375(T) (97.6 %) and Rothia terrae L-143(T) (97.3 %) in many phenotypic characteristics. Moreover, the levels of DNA-DNA relatedness between the novel isolate and the three above-mentioned type strains were 28.7±1.3 %, 36.5±1.2 %, 46.8±1.5 %, respectively. Based on comparative analysis of physiological and chemotaxonomic data, strain YIM 67072(T) represents a novel species of the genus Rothia, for which the name Rothia endophytica sp. nov. is proposed. The type strain is YIM 67072(T) ( = DSM 26247(T) = JCM 18541(T)).

Blastococcus endophyticus sp. nov., an actinobacterium isolated from Camptotheca acuminata.

A novel endophytic actinobacterium, designated strain YIM 68236(T), was isolated from healthy leaves of Camptotheca acuminata. and characterized by using a polyphasic approach. Cells of this strain occurred singly, in pairs or in tetrads. It grew at 10-45 °C, at pH 5.0-8.0 (optimum pH 7.0) and in the presence of 0-3% (w/v) NaCl. The DNA G+C content was 71.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 68236(T) belongs to the genus Blastococcus. However, it differed from its closest relatives, Blastococcus aggregatus DSM 4725(T), Blastococcus saxobsidens DSM 44509(T) and Blastococcus jejuensis DSM 19597(T) in many phenotypic characteristics. Moreover, the DNA-DNA relatedness values between the novel isolate and the three above-mentioned type strains were 49.0 ± 1.6%, 46.1 ± 3.2% and 39.8 ± 1.5%, respectively. Based on comparative analysis of physiological and chemotaxonomic data, strain YIM 68236(T) represents a novel species of the genus Blastococcus, for which the name Blastococcus endophyticus sp. nov. is proposed. The type strain is YIM 68236(T) ( =CCTCC AA 209045(T) =DSM 45413(T) =KCTC 19998(T)).

Geomicrobium sediminis sp. nov., a novel bacterium isolated from a sediment sample collected from the South China Sea, and emended description of the genus Geomicrobium.

A novel bacterium, designated YIM M13075(T), was isolated from a sediment sample collected from the South China Sea. Growth occurred from 4 to 45 °C (optimum 28 °C), pH 6.0-11.0 (optimum pH 8.0). The strain formed yellow-cream colonies after 5 days incubation on TSA modified with 5 % NaCl medium at 28 °C. Cells were Gram-positive, short rods and motile. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain YIM M13075(T) was affiliated with the genus Geomicrobium (93.5 %). The strain YIM M13075(T) contained meso-diaminopimelic acid in the cell wall. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The major fatty acids were iso-C15:0 and anteiso-C15:0. The predominant menaquinones were MK-7 and MK-6. The genomic DNA G+C content was 42.7 mol%. On the basis of the morphological and chemotaxonomic characteristics as well as genotypic data, strain YIM M13075(T) represents a novel species in the genus Geomicrobium, for which the name Geomicrobium sediminis sp. nov. is proposed. The type strain is YIM M13075(T) (=DSM 25540(T) =JCM 18144(T) =CCTCC AB 2013245(T)). An emended description of the genus Geomicrobium is also proposed in the light of the new data.

Saccharomonospora oceani sp. nov. isolated from marine sediments in Little Andaman, India.

Two actinomycete strains, designated YIM M11168(T) and YIM M11177, were isolated from marine sediment samples from Little Andaman, Indian Ocean, and their taxonomic position was determined by a polyphasic approach. The two Gram-positive, aerobic strains were observed to produce branched substrate mycelium and aerial hyphae but did not fragment, and no diffusible pigment was produced on the media tested. At maturity, spores were formed singly or in pairs on aerial hyphae and substrate mycelium, and occasionally the single ones were borne on long sporophores. The optimum growth was determined to occur at 28 °C, 0-4 % (w/v) NaCl and pH 7.0-8.0. Whole-cell hydrolysates of both strains contained meso-diaminopimelic acid and the diagnostic sugars were determined to be galactose, glucose and arabinose. Their predominant menaquinone was found to be MK-9(H4). The polar lipids detected in the two strains were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylinositol, phosphatidylmethylethanolamine, phosphatidylethanolamine and two unknown phosphoglycolipids. The major fatty acids (>10 %) identified were iso-C16:0, iso-C16:1 H, iso-C16:0, C17:1 ω6c for strain YIM M11168(T), iso-C16:0 and Summed Feature 3 for strain YIM M11177. The G + C contents of the genomic DNAs of both strains were determined to be 71.4 %. DNA-DNA hybridization relatedness values (78.4 ± 3.7 %) of these two isolates supported the conclusion that they belong to the same species. Based on phylogenetic analysis, phenotypic and genotypic data, it is concluded that the two isolates belong to a novel species of the genus Saccharomonospora of the family Pseudonocardiaceae. The name Saccharomonospora oceani sp. nov. (Type strain YIM M11168(T) = DSM 45700(T) = JCM 18128(T)) is proposed for the novel species.

Nagimycins A and B, Antibacterial Ansamycin-Related Macrolactams from Streptomyces sp. NA07423.

Chemical investigation of Streptomyces sp. NA07423 led to the discovery of two unreported macrolactams, nagimycins A (1) and B (2). Their structures were elucidated by NMR, HRESIMS, X-ray crystallography, and comparison of experimental and theoretical ECD spectra. The nagimycins have a unique butenolide moiety rarely found in ansamycin antibiotics. Genome analysis revealed the putative biosynthetic gene cluster for nagimycins, and a likely biosynthetic pathway was proposed. Notably, compounds 1 and 2 exhibited potent antibacterial activity against two pathogenic Xanthomonas bacteria.

Draft genome of the gayal, Bos frontalis.

Gayal (Bos frontalis), also known as mithan or mithun, is a large endangered semi-domesticated bovine that has a limited geographical distribution in the hill-forests of China, Northeast India, Bangladesh, Myanmar, and Bhutan. Many questions about the gayal such as its origin, population history, and genetic basis of local adaptation remain largely unresolved. De novo sequencing and assembly of the whole gayal genome provides an opportunity to address these issues. We report a high-depth sequencing, de novo assembly, and annotation of a female Chinese gayal genome. Based on the Illumina genomic sequencing platform, we have generated 350.38 Gb of raw data from 16 different insert-size libraries. A total of 276.86 Gb of clean data is retained after quality control. The assembled genome is about 2.85 Gb with scaffold and contig N50 sizes of 2.74 Mb and 14.41 kb, respectively. Repetitive elements account for 48.13% of the genome. Gene annotation has yielded 26 667 protein-coding genes, of which 97.18% have been functionally annotated. BUSCO assessment shows that our assembly captures 93% (3183 of 4104) of the core eukaryotic genes and 83.1% of vertebrate universal single-copy orthologs. We provide the first comprehensive de novo genome of the gayal. This genetic resource is integral for investigating the origin of the gayal and performing comparative genomic studies to improve understanding of the speciation and divergence of bovine species. The assembled genome could be used as reference in future population genetic studies of gayal.