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HMGR, 3-hydroxy-3-methylglutaryl-CoA reductase, is a major rate-limiting enzyme in mevalonate (MVA) pathway for isoprenoids and subsequent tanshinone biosynthesis in the Chinese traditional bulk herbal medicine Danshen, Salvia miltiorrhiza, mainly for cardiovascular disorders. In this paper, the genomic SmHMGR genes of 38 cultivated populations of S. miltiorrhiza collected in China were for the first time sequenced to reveal the genetic diversity and phylogeny. The SmHMGR gene was shown to be intron-free, 1650~1659 bp in complete CDS with the majority being 1656 bp, and two unique populations (W-FJLY-V-1 and W-SCHY-W-1) being 1659 and 1650 bp respectively. A total of 103 SNP variation sites were detected with a variation rate of 6.22%, most of which occurred in S. miltiorrhiza f. alba population W-SCHY-W-1; a total of 25 amino acid variation sites were found, of which 19 was in W-SCHY-W-1. The same four populations, W-SCHY-W-1, V-HBAG-V-1, V-JLCC-V-1 and S-NM-V-1 could be discriminated from the remaining 34 by both the SNP fingerprints and the deduced amino acid variation sites. Other or composite DNA markers are needed for better identification. The SmHMGR gene of white flower S. miltiorrhiza f. alba population W-SCHY-W-1 is especially rich in variations and worthy of further studies. Phylogenetic trees based on both the gene and the deduced amino acid sequences showed a very similar two-clade topological structure. This research enriched the content and the genetic means for the molecular identification, genetic diversity and phylogenetic studies of the cultivated S. miltiorrhiza populations, and laid a solid foundation for further related and in-depth investigations. [ABSTRACT FROM AUTHOR]

Human papillomavirus (HPV) can cause condyloma acuminatum and cervical cancer. Some mutations of these viruses are closely related to the persistent infection of cervical cancer and are ideal cancer vaccine targets. Several databases have been developed to collect HPV sequences, but no HPV mutation database has been published. This paper reports a Chinese HPV mutation database (HPVMD-C), which contains 149 HPV genotypes, 468 HPV mutations, 3409 protein sequences, 4727 domains and 236 epitopes. We analyzed the mutation distribution among HPV genotypes, domains and epitopes. We designed a visualization tool to display these mutations, domains and epitopes and provided more detailed information about the disease, region and related literature. We also proposed an HPV genotype prediction tool, which can predict HPV carcinogenic or non-carcinogenic risk genotypes. We expect that HPVMD-C will complement the existing database and provide valuable resources for HPV vaccine research and cervical cancer treatment. HPVMD-C is freely available at Database URL : http://bioinfo.zstu.edu.cn/hpv. [ABSTRACT FROM AUTHOR]

Background: Pebrine, caused by microsporidium Nosema bombycis, is a devastating disease that causes serious economic damages to the sericulture industry. Studies on development of therapeutic and diagnostic options for managing pebrine in silkworms are very limited. Methionine aminopeptidase type 2 (MetAP2) of microsporidia is an essential gene for their survival and has been exploited as the cellular target of drugs such as fumagillin and its analogues in several microsporidia spp., including Nosema of honeybees. Methods: In the present study, using molecular and bioinformatics tools, we performed in-depth characterization and phylogenetic analyses of MetAP2 of Nosema bombycis isolated from Guangdong province of China. Results: The full length of MetAP2 gene sequence of Nosema bombycis (Guangdong isolate) was found to be 1278 base pairs (bp), including an open reading frame of 1,077 bp, encoding a total of 358 amino acids. The bioinformatics analyses predicted the presence of typical alpha-helix structural elements, and absence of transmembrane domains and signal peptides. Additionally, other characteristics of a stable protein were also predicted. The homology-based 3D models of MetAP2 of Nosema bombycis (Guangdong isolate) with high accuracy and reliability were developed. The MetAP2 protein was expressed and purified. The observed molecular weight of MetAP2 protein was found to be ~43-45 kDa. The phylogenetic analyses showed that MetAP2 gene and amino acids sequences of Nosema bombycis (Guangdong isolate) shared a close evolutionary relationship with Nosema spp. of wild silkworms, but it was divergent from microsporidian spp. of other insects, Aspergillus spp., Saccharomyces cerevisiae, and higher animals including humans. These analyses indicated that the conservation and evolutionary relationships of MetAP2 are closely linked to the species relationships. Conclusion: This study provides solid foundational information that could be helpful in optimization and development of diagnostic and treatment options for managing the threat of Nosema bombycis infection in sericulture industry of China. [ABSTRACT FROM AUTHOR]

Introduction: The escalating occurrence of infectious disease outbreaks in humans and animals necessitates innovative, effective, and integrated research to better comprehend their transmission and dynamics. Viral infection in livestock has led to profound economic losses globally. Pneumonia is the prevalent cause of death in sheep. However, very few studies exist regarding virus-related pathogens in sheep. Metagenomics sequencing technologies in livestock research hold significant potential to elucidate these contingencies and enhance our understanding. Methods: Therefore, this study aims to characterize respiratory viromes in paired nasal swabs from Inner Mongolian feedlot sheep in China using metaviromic sequencing. Through deep sequencing, de novo assembly, and similarity searches using translated protein sequences, several previously uncharacterized and known viruses were identified in this study. Results: Among these discoveries, a novel Bovine Rhinitis B Virus (BRBV) (BRBVsheep) strain was serendipitously detected in the nasal swabs of domestic sheep (Ovis aries). To facilitate further molecular epidemiological studies, the entire genome of BRBV-sheep was also determined. Owing to the unique sequence characteristics and phylogenetic position of BRBV-sheep, genetically distinct lineages of BRBV in sheep may exist. A TaqMan-based qRT-PCR assay targeting the 3D polymerase gene was developed and used to screen 592 clinical sheep specimens. The results showed that 44.59% of the samples (264/592) were positive. These findings suggest that BRBV sheep are widespread among Inner Mongolian herds. Conclusion: This discovery marks the initial identification of BRBV in sheep within Inner Mongolia, China. These findings contribute to our understanding of the epidemiology and genetic evolution of BRBV. Recognizing the presence of BRBV in sheep informs strategies for disease management and surveillance and the potential development of targeted interventions to control its spread. [ABSTRACT FROM AUTHOR]

Background: High-risk human papillomavirus (HR-HPV) infection is an important factor for the development of cervical cancer. HPV18 is the second most common HR-HPV after HPV16. Methods: In this study, MEGA11 software was used to analyze the variation and phylogenetic tree of HPV18 E6-E7 and L1 genes. The selective pressure to E6, E7 and L1 genes was estimated using pamlX. In addition, the B cell epitopes of L1 amino acid sequences and T cell epitopes of E6-E7 amino acid sequences in HPV18 were predicted by ABCpred server and IEDB website, respectively. Results: A total of 9 single nucleotide variants were found in E6-E7 sequences, of which 2 were nonsynonymous variants and 7 were synonymous variants. Twenty single nucleotide variants were identified in L1 sequence, including 11 nonsynonymous variants and 9 synonymous variants. Phylogenetic analysis showed that E6-E7 and L1 sequences were all distributed in A lineage. In HPV18 E6, E7 and L1 sequences, no positively selected site was found. The nonconservative substitution R545C in L1 affected hypothetical B cell epitope. Two nonconservative substitutions, S82A in E6, and R53Q in E7, impacted multiple hypothetical T cell epitopes. Conclusion: The sequence variation data of HPV18 may lay a foundation for the virus diagnosis, further study of cervical cancer and vaccine design in central China. [ABSTRACT FROM AUTHOR]

Brucella BP26 proves to be a highly immunogenic antigen with excellent specificity in brucellosis detection. In China, the authorized use of the Bp26-deleted vaccine M5ΔBP26 for preventing small ruminant brucellosis highlights the importance of developing accurate detection methods targeting BP26, particularly for the diagnosis of differentiation between infected and vaccinated animals (DIVA). Using the traditional mouse hybridoma technique, we successfully obtained 12 monoclonal antibodies (mAbs) targeting BP26. The efficacy of these mAbs in detecting various animal brucellosis cases using the competitive ELISA method was evaluated. Among them, only the E10 mAb exhibited significant efficiency, being inhibited by 100, 97.62, and 100% of brucellosis-positive sera from cattle, small ruminants, and canines, respectively. The E10-based competitive enzyme-linked immunosorbent assay (cELISA) outperformed the BP26-based indirect enzyme-linked immunosorbent assay (iELISA) in accuracy, particularly for cattle and small ruminant brucellosis, with cELISA sensitivity reaching 97.62% compared to 64.29% for iELISA for small ruminants. Although cELISA showed slightly lower specificity than iELISA, it still maintained high accuracy in canine brucellosis detection. The epitope of mAb E10 was identified in the amino acid sequence QPIYVYPDDKNNLKEPTITGY, suggesting its potential as a diagnostic antigen for brucellosis. In conclusion, the E10-based cELISA presents an effective means of detecting animal brucellosis, particularly significant for DIVA diagnosis in China, where the BP26-mutant vaccine is widely used. [ABSTRACT FROM AUTHOR]

The red Chinese sand pear (Pyrus pyrifolia Nakai) is native to China and exhibits a unique fruit coloration pattern. Flavonoid 3′-hydroxylase (F3′H) catalyzes the hydroxylation of flavonoids, which subsequently determines the components of anthocyanins and the color of plant organs. Two genes encoding flavonoid 3′-hydroxylase (F3′H), PpF3′HI and PpF3′HII, have been identified in red Chinese sand pears. The coding regions for PpF3′HI and PpF3′HII were 1542 and 1536 bp in length, respectively. PpF3′HI shared 95% of its amino acid sequence identity with PpF3′HII, and a highly conserved P450 superfamily domain was found both in PpF3′HI and in PpF3′HII. Phylogenetic analysis showed that PpF3′HI and PpF3′HII clustered with MdF3′HI and MdF3′HII, respectively. PpF3′H genes were highly expressed in anthocyanin-enriched tissues such as young leaves, and transcription of PpF3′H genes corresponded to anthocyanin biosynthesis during the developmental stages, bagging treatment, and postharvest UV-B/visible irradiation treatment. A Y1H assay showed that PpMYB10 and PpHY5 could interact with the −419 bp to 0 bp and −746 bp to −396 bp fragments of the PpF3′HI promoter region, respectively. Understanding the mechanism of flavonoid hydroxylation patterns will, in turn, promote the development of new technologies for modifying flavonoid and anthocyanin composition in fruits. [ABSTRACT FROM AUTHOR]

[Objectives] The paper aimed to identify and characterize the etiological agent of the gout which many goose flocks suffered from in the severe outbreaks of gout in 4 to 16-day-old goslings which had caused about 2% to 20% mortality in the Eastern China since 2017,bringing huge economic losses to the goose flocks.[Methods] Tissue samples from diseased goslings were inoculated into goose embryos to isolate pathogens,and the isolates were identified by PCR detection,genome sequencing and phylogenetic analysis. Positive allantoic fluid was inoculated into goose embryonic kidney epithelial cells to determinate virus titer. Goslings(Yangzhou white goose)were inoculated with the virus as an animal reproduction experiment,and observing anatomical and histopathological changes,detecting virus shedding and distribution in clocal sample and organ samples were detected by PCR,and serum uric acid was measured by ELISA.[Results] A virus designated goose astrovirus(GAstV)JSHA was isolated from allantoic fluid of goose embryos following three serial passages. Phylogenetic analysis indicated that the JSHA strain had more than 99% identity with GAstV strains(SD01,SDPY,GD)isolated from geese with gout while it was highly divergent from all other avastroviruses(only 27.3% to 57.0%)according to the amino acid sequence of the full-length ORF2. Cytopathic changes including rounding,clumping and detachment of cells from the surface were found in goose embryonic kidney epithelial cells with positive allantoic fluid,and the virus titer was 104.25 TCID50·mL-1. Animal reproduction experiment showed that the virus caused the mortality of 25%(5/20),the reduction of body weight,pale and swollen kidney,visceral gout and the increase of serum uric acid. Histopathological changes were observed in the kidneys,livers and spleens,including necrosis of renal tubular,liver and spleen,the deposition of urates and inflammatory infiltration. Accordingly,the virus load was higher in kidney,liver and spleen than that in other tissues. Virus shedding of goslings was detected in cloacal swabs at 3 days post infection,with a shedding peak from 6-9 d post infection.[Conclusions] GAstV JSHA is the main cause of gout in gosling. [ABSTRACT FROM AUTHOR]

[Objectives] The single-stranded DNA binding protein Whirly(WHY), a transcription factor, exists in plants widely and plays an important role in plant leaf senescence. This paper aimed to clone CsWHY1, a single-stranded DNA binding protein gene in cucumber, and to analyze its expression characteristics to provide reference for its functional research on cucumber leaf senescence. [Methods] The full length cDNA of CsWHY1 was cloned with North China cucumber ‘9930’ ‘Changchunmici’ and ‘Xintaimici’ as experimental materials. A series of bioinformatics software were used to speculate nucleotide sequence and protein function of CsWHY1. In addition, the subcellular localization was conducted by onion epidermal cells transient expression system, and gene expression pattern of CsWHY1 was investigated by RT-qPCR in different tissues of 3 cucumber varieties. [Results] The ORF of CsWHY1 gene was 831 bp, encoding 276 amino acids. The relative molecular mass of CsWHY1, a hydrophilic protein, was 30.729×103 with the theoretical isoelectric point(pI) of 9.00. Amino acid sequence analysis revealed that CsWHY1 protein had a conserved Whirly domain, and it was highly homologous in dicotyledonous and monocotyledonous plants and had the closest genetic relationship with melon. Subcellular localization analysis showed that CsWHY1 was located in the plastids of onion epidermal cell. RT-qPCR analysis showed that CsWHY1 expressed in different tissues of three cucumber varieties, with the highest expression level in leaves and the lower expression in roots and stems, and the expression level of CsWHY1 in aged leaves was significantly higher than that in mature leaves. [Conclusions] The expression of CsWHY1 was tissue-specific. CsWHY1 had a high expression level in senescent leaves and participated in the senescence process of leaves in cucumber. [ABSTRACT FROM AUTHOR]

Background: Citrus tristeza virus (CTV) is one of the most serious threats to the citrus industry, and is present in both wild and cultivated citrus. The origin and dispersal patterns of CTV is still poorly understood in China. Methods: In this study, 524 CTV suspected citrus samples from China were collected, including 354 cultivated citrus samples and 174 wild citrus samples. Finally, 126 CTV coat protein sequences were obtained with time-stamped from 10 citrus origins in China. Bayesian phylodynamic inference were performed for CTV origin and dispersal patterns study in China. Result: We found that CTV was mainly distributed in southern and coastal areas of China. The substitution rate of CTV was 4.70 × 10− 4 subs/site/year (95% credibility interval: 1.10 × 10− 4 subs/site/year ~ 9.10 × 10− 4 subs/site/year), with a slight increasing trend in CTV populations between 1990 and 2006. The CTV isolates in China shared a most common recent ancestor around 1875 (95% credibility interval: 1676.57 ~ 1961.02). The CTV in China was originated from wild citrus in Hunan and Jiangxi, and then spread from the wild citrus to cultivated citrus in the growing regions of Sichuan, Chongqing, Hubei, Fujian, Zhejiang, Guangxi and Guangdong provinces. Conclusions: This study has proved that CTV in China was originated from wild citrus in Hunan and Jiangxi. The spatial-temporal distribution and dispersal patterns has uncovered the population and pandemic history of CTV, providing hints toward a better understanding of the spread and origin of CTV in China. [ABSTRACT FROM AUTHOR]

Gypsophila huashanensis Y. W. Tsui & D. Q. Lu (Caryophyllaceae) is an endemic herb species to the Qinling Mountains in China. In this study, we characterized its whole plastid genome using the Illumina sequencing platform. The complete plastid genome of G. huashanensis is 152,457 bp in length, including a large single-copy DNA region of 83,476 bp, a small single-copy DNA region of 17,345 bp, and a pair of inverted repeat DNA sequences of 25,818 bp. The genome contains 130 genes comprising 85 protein-coding genes, 37 tRNA genes, and eight rRNA genes. Evolutionary analysis showed that the non-coding regions of Caryophyllaceae exhibit a higher level of divergence than the exon regions. Gene site selection analysis suggested that 11 coding protein genes (accD, atpF, ndhA, ndhB, petB, petD, rpoCl, rpoC2, rps16, ycfl, and ycf2) have some sites under protein sequence evolution. Phylogenetic analysis showed that G. huashanensis is most closely related to the congeneric species G. oldhamiana. These results are very useful for studying phylogenetic evolution and species divergence in the family Caryophyllaceae. [ABSTRACT FROM AUTHOR]

Goose astrovirus genotype 2 (GoAstV2), as a contagious pathogen of fatal visceral gout in goslings, has been widely distributed in major goose-producing regions in China since 2017, leading to significant economic losses to the Chinese goose industry. In this study, a novel goose astrovirus (GoAstV-SDHZ) was isolated and identified from laying hens with nephritis for the first time. Phylogenetic analysis of the sequenced genome of ORF2 gene revealed that the GoAstV-SDHZ strain clustered into Group 2 GoAstVs and shared the highest identity with the other representative GoAstV2 in the nucleotide (ORF1a: 97.2–99.7%; ORF1b: 98.4–99.8%; ORF2: 97.2–99.9%) and amino acid sequence (ORF1a: 98.2–100%; ORF1b: 98.8–99.6%; ORF2: 97.4–99.6%). In summary, this study provides the first evidence of the GoAstV2 infection in Chinese laying hens, which raises potential threat to the poultry industry in China. [ABSTRACT FROM AUTHOR]

Among the most important transcription factors in plants, the v-myb avian myeloblastosis viral oncogene homolog (MYB) regulates the expression network of response genes under stresses such as fungal infection. In China, the canker disease Valsa mali threatens the survival of Malus sieversii, an ancestor of cultivated apples. Using the M. sieversii genome, we identified 457 MsMYB and 128 R2R3-MsMYB genes that were randomly distributed across 17 chromosomes. Based on protein sequence and structure, the R2R3-MsMYB genes were phylogenetically divided into 29 categories, and 26 conserved motifs were identified. We further predicted cis-elements in the 2000-kb promoter region of R2R3-MsMYBs based on the genome. Transcriptome analysis of M. sieversii under V. mali infection showed that 27 R2R3-MsMYBs were significantly differentially expressed, indicating their key role in the response to V. mali infection. Using transient transformation, MsMYB14, MsMYB24, MsMYB39, MsMYB78, and MsMYB108, which were strongly induced by V. mali infection, were functionally identified. Among the five MsMYBs, MsMYB14 and MsMYB78 were both important in enhancing resistance to diseases, whereas MsMYB24 inhibited resistance. Based on the results of this study, we gained a better understanding of the MsMYB transcription factor family and laid the foundation for a future research program on disease prevention strategies in M. sieversii. [ABSTRACT FROM AUTHOR]

The first complete mitochondrial genome of the freshwater goby Rhinogobius davidi was determined by high-throughput sequencing. This genome was 16,627 bp in length and consisted of 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and 2 non-coding control regions. Phylogenetic analysis based on the amino acid sequences of 13 mitochondrial protein-coding genes from R. davidi and 23 relatives suggested that R. davidi had a close mitogenome relationship with Rhinogobius giurinus. This complete genome of R. davidi will provide basal molecular data for future studies on taxonomy, comparative genomics, and adaptive evolution in Rhinogobius. [ABSTRACT FROM AUTHOR]

African swine fever virus (ASFV) is a highly infectious viral pathogen that endangers the global pig industry, and no effective vaccine is available thus far. The CD2v protein is a glycoprotein on the outer envelope of ASFV, which mediates the transmission of the virus in the blood and recognition of the virus serotype, playing an important role in ASFV vaccine development and disease prevention. Here, we generated two specific monoclonal antibodies (mAbs), 6C11 and 8F12 (subtype IgG1/kappa-type), against the ASFV CD2v extracellular domain (CD2v-ex, GenBank: MK128995.1, 1–588 bp) and characterized their specificity. Peptide scanning technology was used to identify the epitopes recognized by mAbs 6C11 and 8F12. As a result, two novel B cell epitopes, 38DINGVSWN45 and 134GTNTNIY140, were defined. Amino acid sequence alignment showed that the defined epitopes were conserved in all referenced ASFV strains from various regions of China including the highly pathogenic, epidemic strain, Georgia2007/1 (NC_044959.2), with the same noted substitutions compared to the four foreign ASFV wild-type strains. This study provides important reference values for the design and development of an ASFV vaccine and useful biological materials for the functional study of the CD2v protein by deletion analysis. [ABSTRACT FROM AUTHOR]

Rice false smut (RFS), caused by Ustilaginoidea virens, is an important fungal disease of rice. In China, Methyl Benzimidazole Carbamates (MBCs), including carbendazim, are common fungicides used to control RFS and other rice diseases. In this study, resistance of U. virens to carbendazim was monitored for three consecutive years during 2018 to 2020. A total of 321 U. virens isolates collected from Jiangsu Province of China were tested for their sensitivity to carbendazim on PSA. The concentration at which mycelial growth is inhibited by 50% (EC50) of the carbendazim-sensitive isolates was 0.11 to 1.38 µg/mL, with a mean EC50 value of 0.66 μg/mL. High level of resistance to carbendazim was detected in 14 out of 321 isolates. The resistance was stable but associated with a fitness penalty. There was a statistically significant and moderate negative correlation (r= −0.74, p < 0.001) in sensitivity between carbendazim and diethofencarb. Analysis of the U. virens genome revealed two potential MBC targets, Uvβ1Tub and Uvβ2Tub, that putatively encode β-tubulin gene. The two β-tubulin genes in U. virens share 78% amino acid sequence identity, but their function in MBC sensitivity has been unclear. Both genes were identified and sequenced from U. virens sensitive and resistant isolates. It is known that mutations in the β2-tubulin gene have been shown to confer resistance to carbendazim in other fungi. However, no mutation was found in the Uvβ2Tub gene in either resistant or sensitive isolates. Variations including point mutations, non-sense mutations, codon mutations, and frameshift mutations were found in the Uvβ1Tub gene from the 14 carbendazim-resistant isolates, which have not been reported in other fungi before. Thus, these results indicated that variations of Uvβ1Tub result in the resistance to carbendazim in field isolates of Ustilaginoidea virens. [ABSTRACT FROM AUTHOR]

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Pheretima vulgaris has been prescribed for the treatment of cardiovascular diseases in China for several hundred years in the form of dried powder in the clinic. However, the peptides with the potential antithrombotic activity of this source have never been reported. The total active proteins from Pheretima vulgaris were hydrolyzed by eight different commercial proteases and the alcalase hydrolysate showed the strongest thrombolytic activity. Four original thrombolytic peptides were isolated and characterized using bioactivity‐directed fractionation of the active hydrolysate. The amino acid sequences were identified as HEPLPEP (m/z 818.40076), EYPLPEP (m/z 844.39648), LGEPSVP (m/z 698.39648), and LLAPP (m/z 510.28043) by nanoLC‐ESI‐Orbitrap mass spectrometry with PEAKS software. HEPLPEP and EYPLPEP, containing the common ‐PLPEP residue, showed superior thrombolytic activity in plasmin assay and fibrinogen‐thrombin time assay. This research confirmed that Pheretima vulgaris was a potential source of active peptides with thrombolytic activities and provided novel candidates for the thrombolytic agents. Practical applications: Thrombosis has become the leading cause of mortality as it was the common underlying pathology of cardiovascular diseases, such as ischemic heart disease, and stroke. The demand for thrombolytics has increased gradually as the incidence trends of thrombosis‐related diseases raise with the aging of the population. Four novel thrombolytic peptides were characterized from Pheretima vulgaris proteins hydrolysates, among which HEPLPEP and EYPLPEP could prevent the formation of thrombus and degrade existing thrombus in vitro. These peptides are promising to be meritorious templates for developing thrombolytic agents. The structure–function relationship of peptides resulting from the presence of specific residues in these sequences may contribute to extending the knowledge about their thrombolytic activity, which may be useful in designing novel thrombolytic agents. The present research based on a bioactivity‐directed isolation strategy could also be applied to other animal‐derived traditional Chinese medicines with proteins or peptides as their function basis. [ABSTRACT FROM AUTHOR]

Silene conoidea L. is an annual troublesome broadleaf weed in winter wheat fields in China. In recent years, field applications of tribenuron-methyl have been ineffective in controlling S. conoidea in Hebei Province, China. The aim of this study was to determine the molecular basis of tribenuron-methyl resistance in S. conoidea. Whole-plant response assays revealed that the resistant population (R) exhibited a higher level of resistance (382.3-fold) to tribenuron-methyl. The R population also showed high cross-resistance to other acetolactate synthase (ALS) inhibitors, including imazethapyr, bispyribac-sodium and florasulam. However, the R population could be controlled by the field-recommended rates of bentazone, MCPA, fluroxypyr, carfentrazone-ethyl and bromoxynil. In vitro ALS activity assays indicated that the tribenuron-methyl I50 value for the R population was 18.5 times higher than those for the susceptible population (S). ALS gene sequencing revealed an amino acid mutation, Trp-574-Leu, in the R population. Pretreatment with the P450 inhibitor malathion indicated that the R population might have cytochrome P450-mediated metabolic resistance. These results suggest that the Trp-574-Leu mutation and P450-mediated enhanced metabolism coexist in S. conoidea to generate tribenuron-methyl resistance. This is the first time that target-site and non-target-site resistance to tribenuron-methyl has been reported in S. conoidea. [ABSTRACT FROM AUTHOR]

Background: Porcine epidemic diarrhea virus (PEDV), an enteric coronavirus, has become the major causative agent of acute gastroenteritis in piglets since 2010 in China. Results: In the current study, 91 complete spike (S) gene sequences were obtained from PEDV positive samples collected from 17 provinces in China from March 2020 to March 2021. A phylogenetic analysis showed that 92.3% (84 out of 91) of the identified strains belonged to GII subtype, while 7.7% (7 out of 91) were categorized as S-INDEL like strains and grouped within GI-c clade. Based on a recombination analysis, six of S-INDEL like strains were recombinant strains originated from S-INDEL strain FR/001/2014 and virulent strain AJ1102. In addition, PEDV variant strains (CH/GDMM/202012, CH/GXDX/202010 et al) carrying novel insertions (360QGRKS364 and 1278VDVF1281) in the S protein were observed. Furthermore, the deduced amino acid sequences for the S protein showed that multiple amino acid substitutions in the antigenic epitopes in comparison with the vaccine strains. Conclusions: In conclusion, these data provide novel molecular evidence on the epidemiology and molecular diversity of PEDV in 2020–2021. This information may help design a strategy for controlling and preventing the prevalence of PEDV variant strains in China. [ABSTRACT FROM AUTHOR]

Simple Summary: The rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), is indigenous to the United States and has become a significant invasive agricultural pest in China. In this study, we identified and cloned one trehalase gene (LoTRE1) encoding a soluble protein in L. oryzophilus and compared the relative expression levels of LoTRE1 in different tissues. The purified LoTRE1 protein was obtained using a prokaryotic expression system, and its enzymatic properties were explored. Amino acid sequence homology modeling of LoTRE1 and molecular docking between the LoTRE1 protein and substrate trehalose were simulated, which further provided a theoretical basis for revealing the role of LoTRE1 in the degradation mechanism of trehalose. In addition, the LoTRE1 double-stranded RNA (dsRNA) was synthesized in vitro, and its RNAi effect in L. oryzophilus was detected via feeding. The results suggested that LoTRE1 played a vital role in L. oryzophilus development, which could be useful for providing information for insect pest control in the future. Trehalase is the only enzyme known for the irreversible splitting of trehalose and plays a major role in insect growth and development. In this report, we describe a basic study of the trehalase gene fragment encoding a soluble trehalase from Lissorhoptrus oryzophilus (LoTRE1). Sequence alignment and phylogenetic analysis suggested that LoTRE1 was similar to some known insect trehalases and belongs to the Coleoptera trehalase group. Additionally, LoTRE1 was expressed mainly in the fat body. Purified protein was obtained using heterologous expression of LoTRE1 in Escherichia coli, and the recombinant protein exhibited the ability to decompose trehalose. Enzyme–substrate docking indicated the potential involvement of other residues in the catalytic activity, in addition to Asp 333. Moreover, feeding of adults on LoTRE1 dsRNA silenced the transcription of LoTRE1 and thereby reduced the activity of trehalase and increased the trehalose content; it also led to a 12% death rate. This study reveals essential molecular features of trehalase and offers insights into the structural aspects of this enzyme, which might be related to its function. Taken together, the findings demonstrate that LoTRE1 is indispensable for adults of this pest and provide a new target for the control of L. oryzophilus. [ABSTRACT FROM AUTHOR]

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

During a surveillance study to monitor porcine epidemic diarrohoea virus and transmissible gastroenteritis virus in India, a total of 1043 swine samples including faeces (n = 264) and clotted blood (n = 779) were collected and tested. Five samples (four faecal and one serum) showed cytopathic effects in Vero cells. Transmission electron microscopy of infectious cell supernatant revealed the presence of two types of virions. Next-generation sequencing (de novo) allowed the complete genome sequence of mammalian orthorubulavirus 5 (MRuV5; 15246 bp) and that of all 10 gene segments of mammalian orthoreovirus to be determined. Genetic analysis of MRuV5 revealed grouping of the Indian MRuV5 with isolates from various mammalian species in South Korea and China, sharing more than 99% nucleotide sequence identity. The deduced amino acid sequences of the HN, NP, and F genes of MRuV5 isolates showed three (92L, 111R, 447H), two (86S, 121S), and two (139T, 246T) amino acid substitutions, respectively, compared to previously reported virus strains. Phylogenic analysis based on S1 gene sequences showed the Indian MRV isolates to be clustered in lineage IV of MRV type 3, with the highest nucleotide sequence identity (97.73%) to MRV3 strain ZJ2013, isolated from pigs in China. The protein encoded by the MRV3 S1 gene was found to contain the amino acid residues 198-204NLAIRLP, 249I, 340D, and 419E, which are known to be involved in sialic acid binding and neurotropism. This is the first report of co-isolation and whole-genomic characterisation of MRuV5 and MRV3 in domestic pigs in India. The present study lays a foundation for further surveillance studies and continuous monitoring of the emergence and spread of evolving viruses that might have pathogenic potential in animal and human hosts. [ABSTRACT FROM AUTHOR]

Poleroviruses are positive-sense, single-stranded viruses. In this study, we describe the identification of a novel polerovirus isolated from soybean displaying curled leaves. The complete viral genome sequence was identified using high-throughput sequencing and confirmed using rapid amplification of cDNA ends (RACE), RT-PCR and Sanger sequencing. Its genome organization is typical of the members of genus Polerovirus, containing seven putative open reading frames (ORFs). The full genome is composed of single-stranded RNA of 5822 nucleotides in length, with the highest nucleotide sequence identity (79.07% with 63% coverage) for cowpea polerovirus 2 (CPPV2). Amino acid sequence identities of the protein products between the virus and its relatives are below the threshold determined by the International Committee of Taxonomy of Viruses (ICTV) for species demarcation, and this strongly supports this virus' status as a novel species, for which the name soybean chlorotic leafroll virus (SbCLRV) is proposed. Recombination analysis identified a recombination event in the ORF5 of the 3' portion in the genome. Phylogenetic analyses of the genome and encoded protein sequences revealed that the new virus is closely related to phasey bean mild yellows virus, CPPV2 and siratro latent polerovirus. Subsequently, we demonstrated the infectivity of SbCLRV in Nicotiana benthamiana via infectious cDNA clone generation and agroinoculation. [ABSTRACT FROM AUTHOR]

The incidence of cervical cancer is closely related to high‐risk human papillomavirus (HR‐HPV). Women in Jingzhou had relatively high susceptibility to HPV‐51, whose ratio was 9.61% (456/4743) among HR‐HPV‐positive samples and ranked fifth in all analyzed HR‐HPV types. In this study, variations and phylogenetic trees of HPV‐51 E6−E7 and L1 sequences were analyzed by MEGA‐X. The selective pressure was estimated using PAML. The B‐cell epitope of L1 amino acid sequences and T‐cell epitope of E6 and E7 amino acid sequences were further predicted by ABCpred server and IEDB website, respectively. In the E6−E7 sequences 14 single nucleotide variants occurred, among which 4 were nonsynonymous variants and 10 were synonymous variants. A total of 41 single nucleotide variants were identified in the L1 sequences, including 10 nonsynonymous variants and 31 synonymous variants. All the isolates of both E6−E7 and L1 were classified into the A variant lineage. In HPV‐51 E6−E7 and L1 sequences, no positively selected site was found. Two nonconservative substitutions, H119Y and N176S in L1, affected multiple hypothetical B‐cell epitopes. Three nonconservative substitutions, T86P, S100L in E6, and F29L in E7, affected multiple hypothetical T‐cell epitopes. Elucidation of the HR‐HPV prevalence characteristics and genetic variations of HPV‐51 in central China may contribute to future investigations of diagnostic probes, therapeutic or preventative vaccines with wider coverage. Highlights: This is the first study on the genetic variation of E6, E7, and L1 genes of human papillomavirus 51 in Hubei Province, China.Compared with the results of other geological locations, some new variants of E6, E7, and L1 genes of human papillomavirus 51 are found in this study.Phylogenetic analysis shows E6, E7, and L1 gene sequences of human papillomavirus 51 in central China are mostly distributed in the A lineage.This study provides information for the investigation of human papillomavirus epidemic characteristics. The discovery of new variants of human papillomavirus 51 may lay the basis for the development of the virus diagnosis and further study of cervical cancer in central China. [ABSTRACT FROM AUTHOR]

Infectious laryngotracheitis virus (ILTV) causes severe respiratory disease in chickens and results in huge economic losses in the poultry industry worldwide. To correlate the genomic difference with the replication and pathogenicity, phenotypes of three ILTVs isolated from chickens in China from 2016 to 2018 were sequenced by high-throughput sequencing. Based on the entire genome, the isolates GD2018 and SH2017 shared 99.9% nucleotide homology, while the isolate SH2016 shared 99.7% nucleotide homology with GD2018 and SH2017, respectively. Each virus genome contained 82 ORFs encoding 77 kinds of protein, 31 of which share the same amino acid sequence in the three viruses. GD2018 and SH2017 shared 57 proteins with the same amino acid sequence, while SH2016 shared 42 and 41 proteins with the amino acid sequences of GD2018 and SH2017, respectively. SH2016 propagated efficiently in allantoic fluid and on chorioallantoic membranes (CAMs) of SPF chicken embryo eggs, while GD2018 and SH2017 proliferated well only on CAMs. GD2018 propagated most efficiently on CAMs and LMH cells among three isolates. SH2016 caused serious clinical symptoms, while GD2018 and SH2017 caused mild and moderate clinical symptoms in chickens, although the sero of the chickens infected with those three isolates were all positive for anti-ILTV antibody at 14 and 21 days after challenge. Three ILTVs with high genetic homology showed significant differences in the replication in different culture systems and the pathogenicity of chickens, providing basic materials for studying the key determinants of pathogenicity of ILTV. [ABSTRACT FROM AUTHOR]

Dendroctonus armandi (Tsai and Li) (Coleoptera: Curculionidae: Scolytinae) is considered to be the most destructive forest pest in the Qinling and Bashan Mountains of China. Low winter temperatures limit insect's populations, distribution, activity, and development. Insects have developed different strategies such as freeze-tolerance and freeze-avoidance to survive in low temperature conditions. In the present study, we used gene cloning, real-time polymerase chain reaction (PCR), RNA interference (RNAi), and heterologous expression to study the function of the D. armandi antifreeze protein gene (DaAFP). We cloned the 800 bp full-length cDNA encoding 228 amino acids of DaAFP and analyzed its structure using bioinformatics analysis. The DaAFP amino acid sequence exhibited 24–86% similarity with other insect species. The expression of DaAFP was high in January and in the larvae, head, and midgut of D. armandi. In addition, the expression of DaAFP increased with decreasing temperature and increasing exposure time. RNAi analysis also demonstrated that AFP plays an important role in the cold tolerance of overwintering larvae. The thermal hysteresis and antifreeze activity assay of DaAFP and its mutants indicated that the more regular the DaAFP threonine-cystine-threonine (TXT) motif, the stronger the antifreeze activity. These results suggest that DaAFP plays an essential role as a biological cryoprotectant in overwintering D. armandi larvae and provides a theoretical basis for new pest control methods. [ABSTRACT FROM AUTHOR]

Abstract: In past years, spider venoms have attracted increasing attention due to their extraordinary chemical and pharmacological diversity. The recently popularized proteomic method highly improved our ability to analyze the proteins in the venom. However, the lack of information about isolated venom proteins sequences dramatically limits the ability to confidently identify venom proteins. In the present paper, the venom from Araneus ventricosus was analyzed using two complementary approaches: 2-DE/Shotgun-LC-MS/MS coupled to MASCOT search and 2-DE/Shotgun-LC-MS/MS coupled to manual de novo sequencing followed by local venom protein database (LVPD) search. The LVPD was constructed with toxin-like protein sequences obtained from the analysis of cDNA library from A. ventricosus venom glands. Our results indicate that a total of 130 toxin-like protein sequences were unambiguously identified by manual de novo sequencing coupled to LVPD search, accounting for 86.67% of all toxin-like proteins in LVPD. Thus manual de novo sequencing coupled to LVPD search was proved an extremely effective approach for the analysis of venom proteins. In addition, the approach displays impeccable advantage in validating mutant positions of isoforms from the same toxin-like family. Intriguingly, methyl esterifcation of glutamic acid was discovered for the first time in animal venom proteins by manual de novo sequencing. [Copyright &y& Elsevier]

The complete genome sequence of a virus from lily (Lilium lancifolium Thunb.) growing in Huoshan County, Anhui Province, China, was determined. The whole genome consists of 9558 nucleotides, excluding the poly(A) tail, and encodes a 3061-amino-acid polyprotein (GenBank number ON365558) typical of potyviruses. This is the first complete genome sequence of iris potyvirus B (IPB), for which only a partial sequence from Iris domestica was reported previously. Comparative analysis of this genome sequence with those of closely related potyviruses identified nine cleavage sites and the conserved motifs typical of potyviruses. The complete polyprotein ORF shares 73.6% nucleotide and 81.6% amino acid sequence identity with that of iris potyvirus A (IPA, GenBank number MH898493). Phylogenetic analysis showed that IPB is related to IPA and clusters in a group with lily yellow mosaic virus (LYMV). This is the first report of IPB infecting lily plants. [ABSTRACT FROM AUTHOR]

Fowl adenovirus serotype 4 (FAdV-4), the causative agent of hepatitis-hydropericardium syndrome (HHS), distributed widely in the poultry farms in China. Hexon is one of the major capsid proteins associated with viral species or serotypes. However, the epitopes of Hexon protein remain largely unknown. In this study, a monoclonal antibody (mAb) specific to Hexon protein of FAdV-4, designated as 3G8, was generated. Subsequently, the linear peptide recognized by 3G8 was mapped and identified as 213AYGAYVK219 using a series of overlapping peptides generated from Hexon protein. Amino acid sequence analysis revealed that the epitope recognized by 3G8 was highly conserved across all the FAdVs. The epitope was immunogenic and could be recognized by FAdV-4 positive chicken serum samples. These findings will enrich our knowledge regarding the epitope on Hexon and provide valuable information for further characterization of the antigenicity of Hexon protein. [ABSTRACT FROM AUTHOR]

Mitochondrial genomes provide new insights that help elucidating biological features, genetic evolution, and classification of protozoans. Theileria uilenbergi (T. uilenbergi), transmitted by Haemaphysalis qinghaiensis and H. longicornis, is considered as highly pathogenic to sheep and goats in China. This study reports and outlines features of its mitochondrial genome. The T. uilenbergi mitochondrial genome is a linear monomeric molecule of 6.0 kb length, which encodes three protein-coding genes named cytochrome c oxidase I (cox1), cytochrome b (cob), and cytochrome c oxidase III (cox3), as well as six large subunit (LSU) rRNA gene fragments, and ends in terminal inverted repeats (TIRs). The array structure and organization of the mitochondrial genome of T. uilenbergi is identical to that of T. parva. Phylogenetic analysis based on the amino acid sequences of cox1, cob, and cox3 genes suggests that T. uilenbergi is distantly related to the group of transforming Theileria species such as T. parva. This study contributes to a comprehensive understanding of the phylogeny and evolution of the mitochondrial genome of piroplasms and provides useful information of diagnostic marker for T. uilenbergi. [ABSTRACT FROM AUTHOR]

Larix olgensis is one of the main coniferous tree species in northeastern China and has excellent timber properties and strong tolerance to stress. Thirteen HD-Zip family genes with a complete CDS region were identified on the basis of cambium transcriptome data from Larix olgensis. All 13 genes were analyzed via bioinformatics by their conserved domain protein sequence and amino acid composition, including their physicochemical properties and protein structure. The spatiotemporal expression and abiotic stress responses of these genes were analyzed by real-time quantitative PCR. The results showed that the 13 HD-Zip genes of Larix olgensis were expressed in the roots, stems, and leaves at different stages. The expression of three of these genes (LoHDZ2, LoHDZ11, LoHDZ13) was highest in nonlignified roots, indicating that they might be related to the secondary growth of Larix olgensis; in addition, three genes (LoHDZ5, LoHDZ9, LoHDZ10) were highly expressed in partially and completely lignified stems and leaves. These 13 genes were expressed specifically under drought stress. The expression of two of them (LoHDZ1, LoHDZ5) was obviously upregulated, and the expression of 6 genes (LoHDZ2, LoHDZ3, LoHDZ4, LoHDZ8, LoHDZ10, LoHDZ13) was significantly downregulated. The expression trends indicate that these genes could be involved in drought stress. The expression of all 13 genes was downregulated when the plants were treated with 0.2 M NaCl for 96 h, indicating that these genes are inhibited by salt stress. Overall, the results have significant implications for the study of the gene function of members of the LoHD-Zip transcription factor family. [ABSTRACT FROM AUTHOR]

Chicken infectious anemia (CIA), caused by chicken anemia virus (CAV), is an important immunosuppressive disease that seriously threatens the global poultry industry. Here, we isolated and identified 30 new CAV strains from CAV-positive flocks. The VP1 genes of these strains were sequenced and analyzed at the nucleotide and amino acid levels and were found to have very similar nucleotide sequences (> 97% identity); however, they showed 93.9–100.0% sequence identity to the VP1 genes of 55 reference strains. Furthermore, alignment of the deduced amino acid sequences revealed some unique mutations. Phylogenetic analysis indicated the division of VP1 amino acid sequences into two groups (A and B) and four subgroups (A1, A2, A3 and A4). Interestingly, 22 of the newly isolated strains and some Asian reference strains belonged to the A1 group, whereas the remaining eight new isolates belonged to the A3 group. To evaluate the pathogenicity of the epidemic CAV strains from China, the representative strains CAV-JL16/8901 and CAV-HeN19/3001 and the reference strain Cux-1 were selected for animal experiments. Chickens infected with the isolates and reference strain all showed thymus atrophy and bone marrow yellowing. The mortality rates for CAV-JL16/8901, CAV-HeN19/3001, and the reference strain was 30%, 20%, and 0%, respectively, indicating that the epidemic strains pose a more serious threat to chickens. We not only analyzed the molecular evolution of the epidemic strains but also showed for the first time that the epidemic strains in China are more pathogenic than reference strain Cux-1. Effective measures should be established to prevent the spread of CIA in China. [ABSTRACT FROM AUTHOR]

Bovine herpesvirus type 1 (BoHV-1) is one of the most critical pathogens in cattle and is prevalent in China. BoHV-1 is divided into two gene types, BoHV-1.1 and 1.2, which are further differentiated into two subtypes, BoHV-1.2a and 1.2b. However, the phylogenetic analysis of BoHV-1 isolates has not been reported in China. To perform a molecular epidemiological survey based on isolates from cattle in China, 102 lung tissue samples of calves under ten months of age with respiratory disease (BRD) that died from 2016 to 2019 in China were used to isolate BoHV-1 with Madin-Darby bovine kidney (MDBK) cells. Part of the BoHV-1 isolates were applied to the phylogenetic analysis based on the region of the glycoprotein C (gC) gene of BoHV-1. Thirty BoHV-1 isolates were obtained, and the gC gene of 13 isolates was amplified by polymerase chain reaction (PCR) methods and sequenced. The result of the phylogenetic analysis according to the 451-nucleotide portion of the gC gene found that all of 13 isolates belonged to the BoHV-1.2b gene subtype, but these isolates had located two different phylogenetic tree branches. The gC gene sequence homology of isolates in group1 was higher with a reference strain of BoHV-1.2b EVI14 up to 98.0–100%, while in group 2, this was higher with reference strain BoHV-1.2b B589 up to 97.8–99.8%. The deduced amino acid sequence of gC from isolates in group 2 had two amino acid mutations with interference strain BoHV-1.2b K22 or BoHV-1.1 COOPER. The cytopathic effects (CPEs) of BoHV-1 isolates in group 2 were ulcered on the centration like a volcano on MDBK cell, and different from traditional CPEs of BoHV-1. Overall, BoHV-1.2b seems to be the primary strain of BoHV-1 in cattle in China and is also a critical cause of BRD. These BoHV-1.2b isolates had significant genetic variations. • The paper firstly reported the molecular epidemiology study of BoHV-1 strains from naturally infected cattle in China. • Bovine herpesvirus type 1.2b is prevalent gene type of BoHV-1 in cattle with respiratory disease in China. • Bovine herpesvirus type 1.2b isolates had significant genetic variations, further divided into two groups. • The cytopathic effects (CPEs) of BoHV-1 isolates in group 2 were different from traditional BoHV-1 CPEs. [ABSTRACT FROM AUTHOR]

Family of Gastromyzontidae originates from China and Southeast Asia, which is widely distributed in the southern part of the Yangtze River. In this study, we sequenced seven fish complete mitogenomes from three of all 18 genera in the Gastromyzontidae on the Illumina HiSeq platform. Lengths of seven complete mitochondrial genome sequences varied from 16,554 to 16,574 bp, with an AT bias of 53.6–54.9%. Maximum-likelihood phylogenetic tree was reconstructed using the connected protein sequences of 13 protein-coding genes of 34 species of fish. The phylogeny revealed that the genera Gastromyzon, Sewellia, and Erromyzon were monophyletic, and other genera Beaufortia, Vanmanenia, Formosania, and Pseudogastromyzon were paraphyletic or polyphyletic. Phylogeny corrected the taxonomic status of Metahomaloptera omeiensis, which should be categorized under the family of Gastromyzontidae rather than Balitoridae, denoting the significance of this study to the overall fisheries research. [ABSTRACT FROM AUTHOR]

A bacterial strain designated NYYP31T was isolated from the leaves of an annual halophytes, Suaeda corniculata Bunge, collected from the southern edge of the Gurbantunggut desert, north-west China. Strain NYYP31T was Gram-staining negative, strictly aerobic, rod-shaped, non-motile, and non-spore-forming. Growth was observed at 4–42 °C, at pH 5.0–10.0, in the presence of up to 8% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences and coding sequences of 92 protein clusters showed that strain NYYP31T should be assigned to the genus Sphingobacterium. 16S rRNA gene sequence similarity analysis showed that strain NYYP31T was most closely related to the type strain of Sphingobacterium daejeonense (97.9%) and Sphingobacterium lactis (97.7%). The predominant isoprenoid quinone was MK-7. The major fatty acids were identified as iso-C15:0, iso-C17:0 3-OH and summed feature 3 (C16:1ω7c and/or C16:1ω6c). The polar lipids were phosphatidylethanolamine, two unidentified phospholipids, three unidentified lipids, three unidentified amino phospholipids, and two unidentified glycolipids. The genomic DNA G + C content was 36.4 mol%. The average nucleotide identity (ANI) values for strain NYYP31T to the type strains of S. daejeonense and S. lactis were 77.9 and 74.1%, respectively, which were below the cut-off level (95–96%) for species delineation. Based on the above results, strain NYYP31T represents a novel species of the genus Sphingobacterium, for which the name Sphingobacterium endophyticum sp. nov. is proposed. The type strain is NYYP31T (= CGMCC 1.16979T = NBRC 114258T). [ABSTRACT FROM AUTHOR]

Background: Aleutian mink disease parvovirus (AMDV) causes Aleutian mink disease (AMD), which is a serious infectious disease of mink. The aim of this study was to get a better understanding of the molecular epidemiology of AMDV in northeast China to control and prevent AMD from further spreading. This study for the first time isolated AMDV from fecal swab samples of mink in China. Results: A total of 157/291 (54.0%) of the fecal swab samples were positive for AMDV. Of these, 23 AMDV positive samples were randomly selected for sequence alignment and phylogenetic analysis based on the acquired partial fragments of VP2 gene with the hypervariable region. Comparative DNA sequence analysis of 23 AMDV isolates with a reference nonpathogenic (AMDV-G) strain revealed 8.3% difference in partial VP2 nucleotide sequences. Amino acid alignment indicated the presence of several genetic variants, as well as one single amino acid residue deletion. The most concentrated area of variation was located in the hypervariable region of VP2 protein. According to phylogenetic analysis, the Chinese AMDV strains and the other reference AMDV strains from different countries clustered into three groups (clades A, B and C). Most of the newly sequenced strains were found to form a Chinese-specific group, which solely consisted of Chinese AMDV strains. Conclusion: These findings indicated that a high genetic diversity was found in Chinese AMDV strains and the virus distribution were not dependent on geographical origin. Both local and imported AMDV positive species were prevalent in the Chinese mink farming population. The genetic evidence of AMDV variety and epidemic isolates have importance in mink farming practice. [ABSTRACT FROM AUTHOR]

Background: Babesiosis, a tick-borne disease caused by protozoans of the genus Babesia, is widespread in subtropical and tropical countries. Mitochondria are essential organelles that are responsible for energy transduction and metabolism, calcium homeostasis and cell signaling. Mitochondrial genomes could provide new insights to help elucidate and investigate the biological features, genetic evolution and classification of the protozoans. Nevertheless, there are limited data on the mitochondrial genomes of ovine Babesia spp. in China. Methods: Herein, we sequenced, assembled and annotated the mitochondrial genomes of six ovine Babesia isolates; analyzed the genome size, gene content, genome structure and cytochrome b (cytb) amino acid sequences and performed comparative mitochondrial genomics and phylogenomic analyses among apicomplexan parasites. Results: The mitochondrial genomes range from 5767 to 5946 bp in length with a linear form and contain three protein-encoding genes, cytochrome c oxidase subunit 1 (cox1), cytochrome c oxidase subunit 3 (cox3) and cytb, six large subunit rRNA genes (LSU) and two terminal inverted repeats (TIR) on both ends. The cytb gene sequence analysis indicated the binding site of anti-Babesia drugs that targeted the cytochrome bc1 complex. Babesia microti and Babesia rodhaini have a dual flip-flop inversion of 184–1082 bp, whereas other Babesia spp. and Theileria spp. have one pair of TIRs, 25–1563 bp. Phylogenetic analysis indicated that the six ovine Babesia isolates were divided into two clades, Babesia sp. and Babesia motasi. Babesia motasi isolates were further separated into two small clades (B. motasi Hebei/Ningxian and B. motasi Tianzhu/Lintan). Conclusions: The data provided new insights into the taxonomic relationships and drug targets of apicomplexan parasites. [ABSTRACT FROM AUTHOR]

Background: Ethylene Responsive Factor (ERF) is involved in various processes of plant development and stress responses. In wheat, several ERFs have been identified and their roles in mediating biotic or abiotic stresses have been elucidated. However, their effects on wheat plant architecture and yield-related traits remain poorly studied. Results: In this study, TaERF8, a new member of the ERF family, was isolated in wheat (Triticum aestivum L.). Three homoeologous TaERF8 genes, TaERF8-2A, TaERF8-2B and TaERF8-2D (named according to sub-genomic origin), were cloned from the common wheat cultivar Chinese Spring. The three homoeologs showed highly similar protein sequences, with identical AP2 domain. Whereas homoeologs sequence polymorphism analysis allowed the establishment of ten, two and three haplotypes, respectively. Expression analysis revealed that TaERF8s were constitutively expressed through entire wheat developmental stages. Analysis of related agronomic traits of TaERF8-2B overexpressing transgenic lines showed that TaERF8-2B plays a role in regulating plant architecture and yield-related traits. Association analysis between TaERF8-2B haplotypes (Hap-2B-1 and Hap-2B-2) and agronomic traits showed that TaERF8-2B was associated with plant height, heading date and 1000 kernel weight (TKW). The TaERF8-2B haplotypes distribution analysis revealed that Hap-2B-2 frequency increased in domesticated emmer wheat and modern varieties, being predominant in five major China wheat producing zones. Conclusion: These results indicated that TaERF8s are differentially involved in the regulation of wheat growth and development. Haplotype Hap-2B-2 was favored during domestication and in Chinese wheat breeding. Unveiling that the here described molecular marker TaERF8-2B-InDel could be used for marker-assisted selection, plant architecture and TKW improvement in wheat breeding. [ABSTRACT FROM AUTHOR]

Background: This study aims to investigate the T-cell receptor (TCR) repertoire in patients with acute coronary syndrome (ACS).Methods: The TCR repertoires of 9 unstable angina patients (UA), 14 acute myocardial infarction patients (AMI) and 9 normal coronary artery (NCA) patients were profiled using high-throughput sequencing (HTS). The clonal diversity of the TCR repertoires in different groups was analyzed, as well as the frequencies of variable (V), diversity (D) and joining(J) gene segments.Results: ACS patients including UA and AMI, showed reduced TCRβ diversity than NCA patients. ACS patients presented higher levels of clonal expansion. The clonotype overlap of complementarity determining region 3(CDR3) was significantly varied between different groups. A total of 10 V genes and 1 J gene were differently utilized between ACS and NCA patients. We identified some shared CDR3 amino acid sequences that were presented in ACS but not in NCA patients.Conclusions: This study revealed the distinct TCR repertoires in patients with ACS and demonstrated the presence of disease associated T-cell clonotypes. These findings suggested a role of T cells in ACS and provided a new way to explore the mechanisms of ACS. [ABSTRACT FROM AUTHOR]

BACKGROUND: Early blight caused by Alternaria spp. is amongst the most important diseases in potato. Demethylation inhibitor (DMI) fungicides are widely used to control the disease but long‐term use may decrease its control efficacy due to fungicide resistance. This study investigated the occurrence of difenoconazole resistance in Alternaria spp. and molecular resistant mechanisms. RESULTS: EC50 values of 160 isolates to difenoconazole ranged from 0.026 μg mL−1 to 15.506 μg mL−1 and the frequency of difenoconazole sensitivity formed a non‐normal distribution curve with a major and a minor peak. Isolates with EC50 values of 4.121 and 5.461 μg mL−1 were not controlled effectively at fungicide doses of 50 and 100 μg mL−1. Cross‐resistance was observed between DMI fungicides difenoconazole and propiconazole, but not between difenoconazole and other fungicide groups, including boscalid, iprodione, or carbendazim. The CYP51gene was 1673 bp encoding 525 amino acids in length and contained two introns. All sensitive and resistant isolates had the identical amino acid sequence of CYP51, with the exception of one resistant isolate carrying a mutation of R511W. A 6 bp insertion in the upstream region was observed in half of the resistant isolates. In the absence of propiconazole, the relative expression of CYP51 was not significantly different in sensitive and resistant isolates. In the presence of difenoconazole, expression of CYP51 gene was induced significantly in the DMI‐resistant isolates but not in the sensitive ones. CONCLUSION: Induced expression of CYP51 in resistant isolates exposed to difenoconazole is an important determinant for DMI resistance in potato pathogens Alternaria sect. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

In order to identify accurately the circular object contained in the ceramic pot excavated from Jurou Li's grave of the Jin dynasty (1115–1234 ce) in Xi'an, Shaanxi, China, mass spectrometry was applied to determine the amino acid sequences of the residual proteins extracted from the sample, after preliminary starch grain analysis. The sequences were searched against a standard protein sequence database. The proteins extracted were identified as originating from domesticated barley (Hordeum vulgare), soybean (Glycine max), and fermentative microorganisms (Kluyveromyces lactis, Lipomyces starkeyi, Wickerhamomyces ciferrii, Nadsonia fulvescens and Zygosaccharomyces rouxii). These findings indicate that the extremely degraded object in the ceramic pot was made from barley by fermentation with the addition of soy sauce, providing direct evidence of culinary culture in the Jin dynasty. [ABSTRACT FROM AUTHOR]

Background: Artemisia weed pollen allergy is important in the northern hemisphere. While over 350 species of this genus have been recorded, there has been no full investigation into whether different species may affect the allergen diagnosis and treatment. This study aimed to evaluate the variations in amino acid sequences and the content of major allergens, and how these affect specific IgE binding capacity in representative Artemisia species. Methods: Six representative Artemisia species from China and Artemisia vulgaris from Europe were used to determine allergen amino acid sequences by transcriptome, gene sequencing and mass spectrometry of the purified allergen component proteins. Sandwich ELISAs were developed and applied for Art v 1, Art v 2 and Art v 3 allergen quantification in different species. Aqueous pollen extracts and purified allergen components were used to assess IgE binding by ELISA and ImmunoCAP with mugwort allergic patient serum pools and individual sera from five areas in China. Results: The Art v 1 and Art v 2 homologous allergen sequences in the seven Artemisia species were highly conserved. Art v 3 type allergens in A. annua and A. sieversiana were more divergent compared to A. argyi and A. vulgaris. The allergen content of Art v 1 group in the seven extracts ranged from 3.4% to 7.1%, that of Art v 2 from 1.0% to 3.6%, and Art v 3 from 0.3% to 10.5%. The highest IgE binding potency for most Chinese Artemisia allergy patients was with A. annua pollen extract, followed by A. vulgaris and A. argyi, with A. sieversiana significantly lower. Natural Art v 1‐3 isoallergens from different species have almost equivalent IgE binding capacity in Artemisia allergic patients from China. Conclusion and clinical relevance: There was high sequence similarity but different content of the three group allergens from different Artemisia species. Choice of Artemisia annua and A. argyi pollen source for diagnosis and immunotherapy is recommended in China. [ABSTRACT FROM AUTHOR]

Background: Grain weight is an important yield component. Selection of advanced lines with heavy grains show high grain sink potentials and strong sink activity, which is an increasingly important objective in wheat breeding programs. Rice OsGS3 has been identified as a major quantitative trait locus for both grain weight and grain size. However, allelic variation of GS3 has not been characterized previously in hexaploid wheat. Results: We cloned 2445, 2393, and 2409 bp sequences of the homologs TaGS3-4A, TaGS3-7A, and TaGS3-7D in wheat 'Changzhi 6406', a cultivar that shows high grain weight. The TaGS3 genes each contained five exons and four introns, and encoded a deduced protein of 170, 169, and 169 amino acids, respectively. Phylogenetic analysis of plant GS3 protein sequences revealed GS3 to be a monocotyledon-specific gene and the GS3 proteins were resolved into three classes. The length of the atypical Gγ domain and the cysteine-rich region was conserved within each class and not conserved between classes. A single-nucleotide polymorphism in the fifth exon (at position 1907) of TaGS3-7A leads to an amino acid change (ALA/THR) and showed different frequencies in two pools of Chinese wheat accessions representing extremes in grain weight. Association analysis indicated that the TaGS3-7A-A allele was associated with higher grain weight in the natural population. The TaGS3-7A-A allele was favoured in global modern wheat cultivars but the allelic frequency varied among different wheat-production regions of China, which indicated that this allele is of potential utility to improve wheat grain weight in certain wheat-production areas of China. Conclusions: The novel molecular information on wheat GS3 homologs and the KASP functional marker designed in this study may be useful in marker-assisted breeding for genetic improvement of wheat. [ABSTRACT FROM AUTHOR]

Aims: Isolating culturable bacteria associated with sea cucumber (Apostichopus japonicus) and investigating their potential bioactivities are important approaches to discover natural marine products. Methods and Results: A total of 161 isolates were obtained from sea cucumber collected along the Weihai coast of the North Yellow Sea, China. Identification and phylogenetic analysis of 61 isolates were conducted by 16S rRNA gene sequencing. The isolates belonged to 13 genera from 10 families in three phyla, including Firmicutes, Actinobacteria and Proteobacteria. The antimicrobial activities of all strains were determined using six indicator strains. Of the 161 isolates, 93 showed antibacterial activities against at least one of the indicator strains. The 26 strains with the strongest inhibitory effects were selected for screening the biosynthetic gene clusters of polyketide synthase (PKS‐I, ‐II) and nonribosomal peptide synthetase (NRPS). Phylogenetic trees based on the amino acid sequences of the PKS or NRPS genes were constructed. Eleven strains with PKS genes and four strains with NRPS genes were detected. Conclusions: The data reveal the diversity of culturable bacteria associated with A. japonicus. Most strains showed broad‐spectrum antimicrobial activities, and some strains with antimicrobial activities possessed PKS and NRPS genes. Significance and Impact of the Study: The results suggest that culturable bacteria associated with A. japonicus may act as a promising source of bioactive substances. [ABSTRACT FROM AUTHOR]

Porcine circovirus type 2 (PCV2) is the cause of postweaning multisystemic wasting syndrome (PMWS), which encompasses several distinct symptoms in pigs. PCV2 infection and clinical incidence of PMWS have increased in recent years, possibly due to shifts in viral populations and mutations. In this study, we identified PVC2 strains currently afflicting pig populations in mainland China, because this is a prerequisite for developing a specific vaccine to control the spread of PMWS. We collected 235 tissue samples from 16 provinces between 2014 and 2016. Of these, 152 samples were positive for PCV2. We compared the sequences we obtained for the PVC2 capsid gene, ORF2, to those of the Chinese PCV2 sequences deposited in GenBank between 2002 and 2016 (n = 648). Phylogenetic analyses demonstrated that the PCV2d genotype was the most prevalent strain in the sample population included in GenBank and among the positive samples from this study. We also found one PCV2c strain among the GenBank sequences. Furthermore, PCV2a-2F was the predominant genotype in the PCV2a cluster. Amino acid sequence comparisons demonstrated 70.8-100% identity within PCV ORF2 and several consistent mutations in ORF2. More interestingly, six isolates were classified as recombinant strains. Cumulatively, this study represents the first comprehensive description of PCV2 strains distribution, including recent samples, in Chinese porcine populations. We demonstrate the existence of high genetic variability among PVC2 strains and the ability of this virus to rapidly evolve. [ABSTRACT FROM AUTHOR]

The genome of HAdV-B14p1 strain BJ430, isolated from a six-month-old baby diagnosed with bronchial pneumonia at the Beijing Children’s Hospital in December 2010, was sequenced, analyzed, and compared with reference adenovirus genome sequences archived in GenBank. This genome is 34,762 bp in length, remarkably presenting 99.9% identity with the genome from HAdV14p1 strain 303600, which was isolated in the USA (2006). Even more remarkable, it is 99.7% identical with the HAdV-B14p (prototype “de Wit” strain) genome, isolated from The Netherlands in 1955. The patient and its parents presumably had no or limited contact with persons from the USA and Ireland, both of which reported outbreaks of the re-emergent virus HAdV-14p1 recently. These genome data, its analysis, and this report provide a reference for any additional HAdV-B14 outbreak in China and provide the basis for the development of adenovirus vaccines and molecular pathogen surveillance protocols in high-risk areas. [ABSTRACT FROM AUTHOR]

Ureteropelvic junction obstruction (UPJO) is the most common congenital anomaly of the urinary tract. Evidence has shown that BMP4 and Id2 play crucial roles in nephrogenesis, alterations of which may cause ureteral developmental anomalies. Here, we directly sequenced the coding sequences in BMP4 and Id2 genes of 108 unrelated Chinese patients with ureteropelvic junction stenosis. One missense mutation c.485G> A (p.R162Q) in BMP4 and two synonymous mutations (c.1167T> C in BMP4 and c.108A> G in Id2) were detected in three cases. None of these variations were present in the 150 normal controls. Comparative amino acid sequence alignments of BMP4 in humans and other vertebrate orthologs show that p.R162 located to a highly conserved amino acid residue. Moreover, computational analysis predicted that R162Q probably infect the function of BMP4 protein. Conclusion: The mutation c.485G> A in BMP4 might be one of the causes of human UPJO. Further functional studies are required to validate the association between this variation and UPJO. Otherwise, Id2 mutations do not seem to be involved in this disease. [ABSTRACT FROM AUTHOR]

Metagenomic DNA libraries constructed from the Dagong Ancient Brine Well were screened for genes with Na+/H+ antiporter activity on the antiporter-deficient Escherichia coli KNabc strain. One clone with a stable Na+-resistant phenotype was obtained and its Na+/H+ antiporter gene was sequenced and designated as m- nha. The deduced amino acid sequence of M-Nha protein consists of 523 residues with a calculated molecular weight of 58 147 Da and a pI of 5.50, which is homologous with NhaH from Halobacillus dabanensis D-8T (92%) and Halobacillus aidingensis AD-6T (86%), and with Nhe2 from Bacillus sp. NRRL B-14911 (64%). It had a hydropathy profile with 10 putative transmembrane domains and a long carboxyl terminal hydrophilic tail of 140 amino acid residues, similar to Nhap from Synechocystis sp. and Aphanothece halophytica, as well as NhaG from Bacillus subtilis. The m- nha gene in the antiporter-negative mutant E. coli KNabc conferred resistance to Na+ and the ability to grow under alkaline conditions. The difference in amino acid sequence and the putative secondary structure suggested that the m- nha isolated from the Dagong Ancient Brine Well in this study was a novel Na+/H+ antiporter gene. [ABSTRACT FROM AUTHOR]

Background: Pentastomiasis is a rare parasitic infection of humans. Pentastomids are dioecious obligate parasites requiring multiple hosts to complete their lifecycle. Despite their worm-like appearance, they are commonly placed into a separate sub-class of the subphylum Crustacea, phylum Arthropoda. However, their systematic position is not uncontested and historically, they have been considered as a separate phylum. Methodology/Principal Findings: An appraisal of Armillifer agkistrodontis was performed in terms of morphology and genetic identification after its lifecycle had been established in a multi-host model, i.e., mice and rats as intermediate hosts, and snakes (Agkistrodon acutus and Python molurus) as definitive hosts. Different stages of the parasite, including eggs, larvae and adults, were isolated and examined morphologically using light and electron microscopes. Phylogenetic and cluster analysis were also undertaken, focusing on the 18S rRNA and the Cox1 gene. The time for lifecycle completion was about 14 months, including 4 months for the development of eggs to infectious larvae in the intermediate host and 10 months for infectious larvae to mature in the final host. The main morphological difference between A. armillatus and Linguatula serrata is the number of abdominal annuli. Based on the 18S rRNA sequence, the shortest hereditary distance was found between A. agkistrodontis and Raillietiella spp. The highest degree of homology in the Cox 1 nucleic acid sequences and predicted amino acid sequences was found between A. agkistrodontis and A. armillatus. Conclusion: This is the first time that a multi-host model of the entire lifecycle of A. agkistrodontis has been established. Morphologic and genetic analyses supported the notion that pentastomids should be placed into the phylum Arthropoda. Author Summary: Little information is currently available on the lifecycle and morphology of pentastomids, a new zoonotic parasite in China. The lifecycle of Armillifer agkistrodontis was established in multiple hosts, i.e., an intermediate host and a definitive host, and the parasite examined in terms of morphology and genetic relationship with other species. The time required for the completion of an entire lifecycle was about 14 months. The main morphological difference between A. armillatus and L. serrata is the number of abdominal annuli. The genetic data supported the notion that pentastomids belong to the phylum Arthropoda. Based on the 18S rRNA sequence, the shortest hereditary distance was found between A. agkistrodontis and Raillietiella spp. The highest similarity in the Cox 1 nucleic acid sequences was found between A. agkistrodontis and A. armillatus. The established multi-host model provides a possible approach to confirm suspected infections and offers an opportunity to further study this parasite. [ABSTRACT FROM AUTHOR]