Genetic regulatory and biological implications of the 10q24.32 schizophrenia risk locus.

Genome-wide association studies have identified 10q24.32 as a robust schizophrenia risk locus. Here we identify a regulatory variant (rs10786700) that disrupts binding of transcription factors at 10q24.32. We independently confirmed the association between rs10786700 and schizophrenia in a large Chinese cohort (n = 11 547) and uncovered the biological mechanism underlying this association. We found that rs10786700 resides in a super-enhancer element that exhibits dynamic activity change during the development process and that the risk allele (C) of rs10786700 conferred significant lower enhancer activity through enhancing binding affinity to repressor element-1 silencing transcription factor (REST). CRISPR-Cas9-mediated genome editing identified SUFU as a potential target gene by which rs10786700 might exert its risk effect on schizophrenia, as deletion of rs10786700 downregulated SUFU expression. We further investigated the role of Sufu in neurodevelopment and found that Sufu knockdown inhibited proliferation of neural stem cells and neurogenesis, affected molecular pathways (including neurodevelopment-related pathways, PI3K-Akt and ECM-receptor interaction signalling pathways) associated with schizophrenia and altered the density of dendritic spines. These results reveal that the functional risk single nucleotide polymorphism rs10786700 at 10q24.32 interacts with REST synergistically to regulate expression of SUFU, a novel schizophrenia risk gene which is involved in schizophrenia pathogenesis by affecting neurodevelopment and spine morphogenesis.

Streptomyces longhuiensis sp. nov, a novel species isolated from soil of Lilium brownii in Hunan Province, PR China.

An actinobacterium strain, designated BH-MK-02T, was isolated from the soil of Lilium brownii. The taxonomic position was determined using a polyphasic approach. Strain BH-MK-02T grew well on International Streptomyces Project series media and formed well-developed, branched substrate hyphae and aerial mycelium that differentiated into straight spore chains with a wrinkled surface. The diagnostic diamino acid was ll-diaminopimelic acid. The major menaquinones were MK-9(H4), MK-9(H6) and MK-9(H8). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannosides, phosphatidylglycerol and unidentified lipid spots. The predominant fatty acids were anteiso-C15 : 0, iso-C16 : 0, C16 : 0 and C16 : 1 ω7c/C16 : 1 ω6c. The phenotypic characteristics of strain BH-MK-02T indicated that it belonged to the genus Streptomyces. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain BH-MK-02T was most closely related to Streptomyces aureus CGMCC 4.1833T (99.7 %). However, the average nucleotide identity and digital DNA-DNA hybridization values between the whole-genome sequences of strain BH-MK-02T and S. aureus CGMCC 4.1833T were 78.1 and 23.2 %, respectively, below the 96.7 and 70 % cut-off points respectively recommended for delineating Streptomyces species. Furthermore, the novel isolate could be distinguished from S. aureus CGMCC 4.1833T by morphological, physiological and biochemical characteristics. Based on all these data, strain BH-MK-02T (=MCCC 1K06237T=JCM 34789T) clearly represents a novel species within the genus Streptomyces, for which the name Streptomyces longhuiensis sp. nov. is proposed.

Functional variant rs2270363 on 16p13.3 confers schizophrenia risk by regulating NMRAL1.

Recent genome-wide association studies have reported multiple schizophrenia risk loci, yet the functional variants and their roles in schizophrenia remain to be characterized. Here we identify a functional single nucleotide polymorphism (rs2270363: G>A) at the schizophrenia risk locus 16p13.3. rs2270363 lies in the E-box element of the promoter of NMRAL1 and disrupts binding of the basic helix-loop-helix leucine zipper family proteins, including USF1, MAX and MXI1. We validated the regulatory effects of rs2270363 using reporter gene assays and electrophoretic mobility shift assay. Besides, expression quantitative trait loci analysis showed that the risk allele (A) of rs2270363 was significantly associated with elevated NMRAL1 expression in the human brain. Transcription factors knockdown and CRISPR-Cas9-mediated editing further confirmed the regulatory effects of the genomic region containing rs2270363 on NMRAL1. Intriguingly, NMRAL1 was significantly downregulated in the brain of schizophrenia patients compared with healthy subjects, and knockdown of Nmral1 expression affected proliferation and differentiation of mouse neural stem cells, as well as genes and pathways associated with brain development and synaptic transmission. Of note, Nmral1 knockdown resulted in significant decrease of dendritic spine density, revealing the potential pathophysiological mechanisms of NMRAL1 in schizophrenia. Finally, we independently confirmed the association between rs2270363 and schizophrenia in the Chinese population and found that the risk allele of rs2270363 was the same in European and Chinese populations. These lines of evidence suggest that rs2270363 may confer schizophrenia risk by regulating NMRAL1, a gene whose expression dysregulation might be involved in the pathogenesis of schizophrenia by affecting neurodevelopment and synaptic plasticity.

A missense variant in NDUFA6 confers schizophrenia risk by affecting YY1 binding and NAGA expression.

Genome-wide association studies (GWASs) have revealed that genetic variants at the 22q13.2 risk locus were robustly associated with schizophrenia. However, the causal variants at this risk locus and their roles in schizophrenia remain elusive. Here we identify the risk missense variant rs1801311 (located in the 1st exon of NDUFA6 gene) as likely causal for schizophrenia at 22q13.2 by disrupting binding of YY1, TAF1, and POLR2A. We systematically elucidated the regulatory mechanisms of rs1801311 and validated the regulatory effect of this missense variant. Intriguingly, rs1801311 physically interacted with NAGA (encodes the alpha-N-acetylgalactosaminidase, which is mainly involved in regulating metabolisms of glycoproteins and glycolipids in lysosome) and showed the most significant association with NAGA expression in the human brain, with the risk allele (G) associated with higher NAGA expression. Consistent with eQTL analysis, expression analysis showed that NAGA was significantly upregulated in brains of schizophrenia cases compared with controls, further supporting that rs1801311 may confer schizophrenia risk by regulating NAGA expression. Of note, we found that NAGA regulates important neurodevelopmental processes, including proliferation and differentiation of neural stem cells. Transcriptome analysis corroborated that NAGA regulates pathways associated with neuronal differentiation. Finally, we independently confirmed the association between rs1801311 and schizophrenia in a large Chinese cohort. Our study elucidates the regulatory mechanisms of the missense schizophrenia risk variant rs1801311 and provides mechanistic links between risk variant and schizophrenia etiology. In addition, this study also revealed the novel role of coding variants in gene regulation and schizophrenia risk, i.e., genetic variant in coding region of a specific gene may confer disease risk through regulating distal genes (act as regulatory variant for distal genes).

Streptomyces liliifuscus sp. nov and an anti-ginger plague agent Streptomyces liliiviolaceus sp. nov, two novel species isolated from soil of Lilium lancifolium.

Two novel strains of actinobacteria, ZYC-3T and BH-SS-21T, were isolated from Hunan Province, PR China. The fermentation broth of BH-SS-21T inhibited the rapid spread of ginger blast, unlike that of ZYC-3T. The taxonomic characteristics of ZYC-3T and BH-SS-21T were defined using a polyphasic approach. The analysis of the full-length 16S rRNA gene sequence revealed that ZYC-3T and BH-SS-21T represented members of the genus Streptomyces. ZYC-3T had less than 98.7% sequence similarities to all species of the genus Streptomyces, while BH-SS-21T exhibited 99.97, 98.95, 98.83, 98.82, 98.75 and less than 98.7% sequence similarities to 'Streptomyces dioscori' A217, Streptomyces ederensis JCM 4958T, Streptomyces glomeroaurantiacus NBRC 15418T, Streptomyces aurantiacus NBRC 13017T, Streptomyces umbrinus JCM 4521T and other species with validly published names in the genus Streptomyces. However, the digital DNA-DNA relatedness and average nucleotide identity values between ZYC-3T, BH-SS-21T, and their closely related strains were significantly lower than the recommended threshold values. Also, phenotypic, chemotaxonomic and genetic features distinguished ZYC-3T and BH-SS-21T from their reference strains. On the basis of their genotypic and phenotypic characteristics, strains ZYC-3T and BH-SS-21T were classified as representing novel species of the genus Streptomyces under the names Streptomyces liliifuscus sp. nov. ZYC-3T (=CICC 25040T=JCM 34560T=MCCC 1K04978T) and Streptomyces liliiviolaceus sp. nov. BH-SS-21T (=MCCC 1K06236T=JCM 34767T), respectively.

Streptomyces akebiae sp. nov., a novel actinomycete isolated from rhizosphere soil of Akebia trifoliate.

A novel actinomycete strain, designated as MG28T, was isolated from rhizosphere soil of Akebia trifoliate. The taxonomic position of the strain was investigated by using a polyphasic approach. BLAST search of the full-length 16S rRNA gene sequence of strain MG28T indicated it represented a member of the genus Streptomyces, and displayed 99.03%, 98.90%, 98.90%, 98.89%, 98.83% and less than 98.70% sequence similarities with S. phaeolivaceus GY16T, S. deccanensis KCTC 19241T, S. europaeiscabiei KACC 20186T, S. fructofermentans CGMCC 4.1593T, S. scabiei NRRL B-16523T and other species of the genus Streptomyces with validly published names, respectively. Phylogenomic analysis indicated that strain MG28T was closely related to Streptomyces deccanensis KCTC 19241T. However, the average nucleotide identity values and the digital DNA-DNA hybridization values between them indicated that strain MG28T represented a distinct species. Furthermore, strain MG28T was also distinctly differentiated from strain KCTC 19241T by morphological, physiological and biochemical characteristics. Therefore, strain MG28T (= MCCC 1K06895T = JCM 34922T) represents a novel species of the genus Streptomyces, for which the name Streptomyces akebiae sp. nov. is proposed.

Proposal of Streptomyces aureorectus (ex Taig et al. 1969) Taig and Solovieva 1986 as a later heterotypic synonym of Streptomyces calvus Backus et al. 1957 (Approved Lists 1980) on the basis of a polyphasic taxonomic approach.

As two separate genomic species, Streptomyces calvus and Streptomyces aureorectus were approved in 1980 and 1986, respectively. However, recently, it has been found that the average nucleotide identity and digital DNA-DNA hybridization values between S. calvus JCM 4326T and S. aureorectus DSM 41692T were 99.19 and 92.70 %, respectively, much higher than 95-96 and 70  % cut-off points proposed and the generally accepted species boundaries. These data indicated that they should be classified as the same genomic species. Furthermore, this result was also supported by a comprehensive comparison of phenotypic, chemotaxonomic and physio-biochemical characteristics between the two type strains. All these data indicated that S. calvus and S. aureorectus had the same taxonomic position. In accordance with the principle of priority, it is proposed that S. aureorectus is a later heterotypic synonyms of S. calvus.

Streptomyces genisteinicus sp. nov., a novel genistein-producing actinomycete isolated from a Chinese medicinal plant and proposal of Streptomyces michiganensis Corbaz et al. 1957 as a later heterotypic synonym of Streptomyces xanthochromogenes Arishima et al. 1956.

A novel genistein-producing actinobacterial strain, designated strain CRPJ-33T, was isolated from the healthy leaves of a medicinal plant Xanthium sibiricum collected from Hunan Province, PR China. 16S rRNA gene sequence analysis indicated strain CRPJ-33T belonged to the genus Streptomyces and had 99.7, 99.0, 98.9, 98.9, 98.8 and 98.7% sequence similarities to Streptomyces zhihengii YIM T102T, Streptomyces eurocidicus NRRL B-1676T, Streptomyces xanthochromogenes NRRL B-5410T, Streptomyces michiganensis NBRC 12797T, Streptomyces mauvecolor LMG 20100T and Streptomyces lavendofoliae NBRC 12882T, respectively. Phylogenetic analysis of 16S rRNA gene sequences showed that strain CRPJ-33T was most closely related to S. zhihengii YIM T102T. However, digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between them were much less than the recommended threshold values. Furthermore, differential comparisons of the phenotypic characteristics were enough to distinguish strain CRPJ-33T from S. zhihengii YIM T102T. Meanwhile, the ANI and dDDH values or MLSA distances between strain CRPJ-33T and other type strains, which exhibited ≥98.7 % 16S rRNA gene sequence similarities to strain CRPJ-33T, were far away from the recommended threshold values. Based on these results, it is thought that strain CRPJ-33T should represent a novel species of the genus Streptomyces, for which the name Streptomyces genisteinicus sp. nov. is proposed. The type strain is CRPJ-33T (=MCCC 1K04965T=JCM 34526T). In addition, the phenotypic, chemotaxonomic and genotypic characteristics, as well as phylogenetic information revealed that the type strains of S. xanthochromogenes and S. michiganensis should belong to same genomic species. Consequently, it is proposed that S. michiganensis is a heterotypic synonym of S. xanthochromogenes for which an emended description is given.

Proposal of Bacillus ciccensis Liu et al. as a Later Heterotypic Synonym of Cytobacillus solani (Liu et al.) Patel and Gupta Using Comparative Genomic Analysis.

Bacillus ciccensis and Cytobacillus solani were reported to belong to separate genomic species based on a polyphasic taxonomic approach. However, we found that type strains of B. ciccensis and C. solani should belong to the same genomic species based on the clustering patterns resulting from genome phylogenetic analysis. The average nucleotide identity and digital DNA-DNA hybridization values between the genomes of two type strains were 99.6% and 96.9%, respectively, much higher than 95 ~ 96% and 70% cut-off point proposed and generally accepted species boundary. This result was also supported by a comprehensive comparison of phenotypic characteristics between the two type strains. These results suggested that B. ciccensis and C. solani had the same taxonomic position. Consequently, we propose that B. ciccensis is a later heterotypic synonym of C. solani.

Streptomyces paludis sp. nov., isolated from an alpine wetland soil.

A Gram-stain-positive actinobacterium, designated strain GSSD-12T, was isolated from an alpine wetland soil. blast search of the full-length 16S rRNA gene sequence of GSSD-12T indicated it represented a member of the genus Streptomyces, and displayed highest similarity with Streptomyces scopuliridis NRRL B-24574T (99.2 %) and less than 98.6 % similarity with other species of the genus Streptomyces with validly published names. Phylogenetic analysis based on 16S rRNA gene sequences indicated that GSSD-12T was closely related to S. scopuliridis NRRL B-24574T, Streptomyces odonnellii NRRL B-24891T and Streptomyces lushanensis NRRL B-24994T. However, the digital DNA-DNA hybridization value, the average nucleotide identity value and the multilocus sequence analysis evolutionary distance between this strain and its closest relatives indicated that it represented a distinct species. Furthermore, GSSD-12T was also distinctly differentiated from them by morphological, physiological and biochemical characteristics. Therefore, strain GSSD-12T(=CICC 11051T=JCM 33019T) represents a novel species of the genus Streptomyces, for which the name Streptomyces paludis sp. nov. is proposed.

Steptomyces fagopyri sp. nov., a novel actinomycete isolated from rhizospheric soil of Fagopyrum dibotrys.

A novel actinomycete, designated strain QMT-28T, was isolated from rhizosphere soil of Fagopyrum dibotrys collected from Shuangfeng, Hunan Province, PR China. Strain QMT-28T grew well on International Streptomyces Project series media and formed well-developed, branched substrate hyphae and aerial mycelium that differentiated into loose spiral spore chains consisting of cylindrical spores with smooth surfaces. The diagnostic diamino acid was ll-diaminopimelic acid and the whole-cell sugars were galactose and glucose. The predominant fatty acids were C18 : 1 cis9, summed feature 6 (C18 : 2 cis 9,12/C18 : 0 a) and C16 : 0. The polar lipids included diphosphatidylglycerol, hydroxy phospatidylethanolamine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides, phospholipids of unknown structure containing glucosamine and several unidentified phospholipids. The major menaquinones were MK-9, MK-9(H2), MK-9(H4), MK-9(H6) and MK-9(H8). The genome size of strain QMT-28T was about 8.7 Mbp with a G+C content of 71.2 mol%. Phylogenetic analysis showed that the novel strain was closely related to Streptomyces olivochromogenes DSM 40451T (99.5 % similarity), Streptomyces mirabilis NBRC 13450T (98.9 %), Streptomyces kanamyceticus NBRC 13414T (98.9 %), Streptomyces kaempferi I37T (98.9 %) and Streptomyces arcticus ZLN234T (98.8 %). However, the average nucleotide identity values, the digital DNA-DNA hybridization values and the multilocus sequence analysis evolutionary distances between this strain and closely related strains showed that it belonged to a distinct species. In addition, these results were also supported by differences in the phenotypic characteristics between QMT-28T and five closely related type strains. Consequently, strain QMT-28T should represent a novel species of the genus Streptomyces, with the suggested name Streptomyces fagopyri sp. nov. The type strain is QMT-28T (=CICC 24808T=JCM 33796T).

Streptomyces aquilus sp. nov., a novel actinomycete isolated from a Chinese medicinal plant.

The taxonomic position of a novel actinomycete isolate, designated strain GGCR-6T, isolated from the healthy leaves of Xanthium sibiricum collected from the botanic garden of Hunan University of Science and Technology in Hunan province, PR China, was determined by a polyphasic approach. GGCR-6T grew well on ISP series media and formed well-developed, branched substrate hyphae and aerial mycelium that differentiated into straight spore chains consisting of cylindrical spores with smooth surfaces. The diagnostic diamino acid was ll-diaminopimelic acid. The major menaquinones were MK-9(H8), MK-9(H2), MK-9 and MK-9(H6). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphotidylinositol and phosphatidylinositol mannosides. The predominant fatty acids were C16 : 1ω9c, iso-C16 : 0 and C16 : 0. The phenotypic characteristics of GGCR-6T indicated that it represented a member of the genus Streptomyces. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that GGCR-6T was most closely related to Streptomyces cyaneus NRRL B2296T and Streptomyces griseoruber NRRL B1818T. However, the digital DNA-DNA hybridization, the average nucleotide identity and the multi locus sequence analysis evolutionary distance clearly separate GGCR-6T from the phylogenetically closely related species. Furthermore, the novel isolate was distinctly differentiated from S. cyaneus NRRL B2296T and S. griseoruber NRRL B1818T by morphological, physiological and biochemical characteristics. Based on these data, strain GGCR-6T should be designated as a representative of a novel species of the genus Streptomyces, for which the name Streptomyces aquilus sp. nov. is proposed. The type strain is strain GGCR-6T (=CICC 11055T=JCM 33584T).

Streptomyces cadmiisoli sp. nov., a novel actinomycete isolated from cadmium-contaminated soil.

A novel Streptomyces strain, ZFG47T, isolated from a cadmium-contaminated soil sample, was taxonomically studied in detail. Strain ZFG47T formed long, flexuous spiral spore chains consisting of elliptoid spores with spiny surfaces. The cell-wall hydrolysates contained ll-diaminopimelic acid as the diagnostic diamino acid. The major menaquinones consisted of MK-9(H2), MK-9(H4) and MK-9(H8). The major polar lipids contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol mannosides. The predominant cellular fatty acids were iso-C16 : 0, C16 : 0 and anteiso-C15 : 0. The 16S rRNA gene sequence-based phylogenetic analysis indicated that this strain belongs to the genus Streptomyces, showing the highest sequence similarity to Streptomyceskoyangensis VK-A60T (98.7 %). However, the digital DNA-DNA hybridization value, the average nucleotide identity value and the MLSA evolutionary distance between this strain and S. koyangensis VK-A60T showed that it belonged to a distinct species. Furthermore, the novel isolate could be distinctly differentiated from S. koyangensis VK-A60T by morphological, physiological and biochemical characteristics. On the basis of the evidence from this polyphasic study, it is concluded that strain ZFG47T represents a novel species of the genus Streptomyces, for which the name Streptomyces cadmiisoli sp. nov. is proposed, with strain ZFG47T (CICC 11050T=JCM 32897T) as the type strain.

Streptomyces cyaneochromogenes sp. nov., a blue pigment-producing actinomycete from manganese-contaminated soil.

A novel actinomycete isolate, designated strain MK-45T, was isolated from soil sampled at a manganese-contaminated area in Xiangtan, China. The isolate formed extensively branched substrate mycelia and aerial mycelia that differentiated into tightly coiled or spiral spore chains with smooth-surfaced spores. The cell-wall hydrolysates contained ll-diaminopimelic acid and traces of meso-diaminopimelic acid. The major menaquinones consisted of MK-9(H4), MK-9(H6) and MK-9(H8). The major polar lipids contained diphosphatidylgycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphotidylinositol and phosphotidylinositol mannosides. The predominant cellular fatty acids were iso-C16 : 0, C16 : 0 and summed feature 3. Phylogenetic analysis based on the 16S rRNA gene sequences and concatenated partial sequences of the five protein-coding genes indicated that strain MK-45T was a member of the genus Streptomyces and most closely related to Streptomyces caeruleatus GIMN4.002T (99.5 % similarity), Streptomyces curacoi CGMCC 4.1680T (99.5 %), Streptomyces shaanxiensis CCNWHQ 0031T (99.4 %) and Streptomyces cinnabarinus NRRL B12382T (98.9 %), respectively. However, the digital DNA-DNA hybridization values, the average nucleotide identity values and MLSA evolutionary distances between strain MK-45T and them showed that it belonged to a distinct species. Furthermore, the results of morphological, physiological and biochemical tests allowed further phenotypic differentiation of strain MK-45T from the four closely related type strains mentioned above and other species of the genus Streptomyces with which this strain has 98.0-99.2 % 16S rRNA gene sequence similarity. Therefore, it is concluded that strain MK-45T represents a novel species of the genus Streptomyces, for which the name Streptomycescyaneochromogenes sp. nov. is proposed, with MK-45T (=CICC 11045T=KCTC 49099T) as the type strain.

Streptomyces manganisoli sp. nov., a novel actinomycete isolated from manganese-contaminated soil.

A novel actinomycete isolate, designated strain MK44T, was isolated from a Manganese-polluted soil sample collected near Xiangtan Manganese Mine, South Central China and subjected to a polyphasic taxonomic characterization. Comparison of 16S rRNA gene sequences showed that strain MK44T was a member of the genus Streptomyces and most closely related to Streptomyces specialis JCM 16611T (97.9 %) and Streptomyces mayteni JCM 16957T (97.4 %). The DNA-DNA relatedness between strain MK44T and the above two related type species were 30.9±0.3 and 29.9±3.5 %, respectively, values which are far lower than the 70 % threshold for the delineation of a novel prokaryotic species. Furthermore, the results of physiological, biochemical and chemotaxonomic tests allowed further phenotypic differentiation. Therefore, it is concluded that strain MK44T represents a novel species of the genus Streptomyces, for which the name Streptomyces manganisoli sp. nov. is proposed. The type strain is MK44T (=GDMCC 4.137T=KCTC 39920T).

Updated incidence rates and risk factors of esophageal cancer in Nan'ao Island, a coastal high-risk area in southern China.

Esophageal cancer (EC) is one of the most common cancers in China. The purpose of this study was to investigate the updated incidence rates and risk factors of EC in Nan'ao Island, where the EC incidence rate was chronically the highest in southern China. To calculate the annual incidence rate, data on 338 EC cases from Nan'ao Cancer Registry system diagnosed during 2005-2011 were collected. A case-control study was conducted to explore the EC risk factors. One hundred twenty-five alive EC patients diagnosed during 2005-2011 and 250 controls were enrolled into the case-control study. A pre-test questionnaire on demography, dietary factors, drinking water treatment, and behavioral factors was applied to collect information of all participants. The average EC incidence rates during 2005-2011 were 66.09/105, 94.62/105, 36.83/105 for both genders, males and females, respectively, in Nan'ao Island. The EC incidence rate in males was 2.40- to 4.55-fold higher than that in females in the period from 2006 to 2011 (P < 0.05). Considering the onset age, males tend to be much younger than females and reached peak incidence rate at a younger age (P < 0.05). Drinking water treatment by filter (odds ratio [OR] = 0.28, 95% confidence interval [95% CI] = 0.13-0.58) and fruit consumption (OR = 0.55, 95% CI = 0.32-0.94) reduced the risk for EC. On the contrary, the pickled vegetables consumption (OR = 2.64, 95% CI = 1.46-4.76) and liquor drinking (OR = 2.32, 95% CI = 1.21-4.44) increased the risk for EC. These results may be of importance for future research on EC etiology and prevention strategies.

Adaptive evolution of interleukin-3 (IL3), a gene associated with brain volume variation in general human populations.

Greatly expanded brain volume is one of the most characteristic traits that distinguish humans from other primates. Recent studies have revealed genes responsible for the dramatically enlarged human brain size (i.e., the microcephaly genes), and it has been well documented that many microcephaly genes have undergone accelerated evolution along the human lineage. In addition to being far larger than other primates, human brain volume is also highly variable in general populations. However, the genetic basis underlying human brain volume variation remains elusive and it is not known whether genes regulating human brain volume variation also have experienced positive selection. We have previously shown that genetic variants (near the IL3 gene) on 5q33 were significantly associated with brain volume in Chinese population. Here, we provide further evidence that support the significant association of genetic variants on 5q33 with brain volume. Bioinformatic analyses suggested that rs31480 is likely to be the causal variant among the studied SNPs. Molecular evolutionary analyses suggested that IL3 might have undergone positive selection in primates and humans. Neutrality tests further revealed signatures of positive selection of IL3 in Han Chinese and Europeans. Finally, extended haplotype homozygosity (EHH) and relative EHH analyses showed that the C allele of SNP rs31480 might have experienced recent positive selection in Han Chinese. Our results suggest that IL3 is an important genetic regulator for human brain volume variation and implied that IL3 might have experienced weak or modest positive selection in the evolutionary history of humans, which may be due to its contribution to human brain volume.

New clustered regularly interspaced short palindromic repeat locus spacer pair typing method based on the newly incorporated spacer for Salmonella enterica.

A clustered regularly interspaced short palindromic repeat (CRISPR) typing method has recently been developed and used for typing and subtyping of Salmonella spp., but it is complicated and labor intensive because it has to analyze all spacers in two CRISPR loci. Here, we developed a more convenient and efficient method, namely, CRISPR locus spacer pair typing (CLSPT), which only needs to analyze the two newly incorporated spacers adjoining the leader array in the two CRISPR loci. We analyzed a CRISPR array of 82 strains belonging to 21 Salmonella serovars isolated from humans in different areas of China by using this new method. We also retrieved the newly incorporated spacers in each CRISPR locus of 537 Salmonella isolates which have definite serotypes in the Pasteur Institute's CRISPR Database to evaluate this method. Our findings showed that this new CLSPT method presents a high level of consistency (kappa = 0.9872, Matthew's correlation coefficient = 0.9712) with the results of traditional serotyping, and thus, it can also be used to predict serotypes of Salmonella spp. Moreover, this new method has a considerable discriminatory power (discriminatory index [DI] = 0.8145), comparable to those of multilocus sequence typing (DI = 0.8088) and conventional CRISPR typing (DI = 0.8684). Because CLSPT only costs about $5 to $10 per isolate, it is a much cheaper and more attractive method for subtyping of Salmonella isolates. In conclusion, this new method will provide considerable advantages over other molecular subtyping methods, and it may become a valuable epidemiologic tool for the surveillance of Salmonella infections.

Frequencies of insecticide resistance mutations detected by the amplicon sequencing in Plutella xylostella (Lepidoptera: Plutellidae) and Spodoptera exigua (Lepidoptera: Noctuidae) from China.

The globally prevalent pests, Diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae) and Beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae), pose significant threats to cruciferous vegetables. They have rapidly developed resistance to a wide range of insecticides, leading to significant yield losses and increased control expenses. In this study, we have established an efficient approach utilizing amplicon sequencing to detect the frequency of 15 target resistance mutant sites in 6 molecular targets, acetylcholinesterase 1 (ACE1), chitin synthase 1 (CHS1), the γ-aminobutyric acid receptor (GABAR), glutamate-gated chloride channel (GluCl), voltage-gated sodium channels (NaV), and ryanodine receptor (RyR) in P. xylostella and the frequency of 11 mutations in 5 molecular targets (except GluCl) in S. exigua in China. Our findings indicate that P. xylostella exhibits remarkably high frequency (over 88.67%) in pyrethroid resistance-related mutations T929I and L1014F of NaV. In S. exigua, the frequencies of L659F mutation were ranging from 41.92% to 74.89%. In addition, the organophosphorus resistance-related mutations A298S and G324A of ACE1 were detected at frequencies ranging from 34.29% to 75.66%, and these 2 mutations occurred simultaneously (from 29.22% to 65.79%) in P. xylostella. An interannual variation in mutation frequency from 2019 to 2021 was found for P. xylostella in HNCS. The frequency of A298S and G324A mutations steadily increased while the frequency of G4946E and I4790M mutations continuously decreased. These results unveil a worrisome scenario of multiple resistance sites in these 2 pests in China and provide valuable insights for the practical application of pesticides in the field.