Coptis chinensis Franch.,also named Chinese goldthread is a member of Ranunculaceae in the order Ranunculales and represents an important lineage of early eudicots with traditional medicinal value.In our study,by usin...Coptis chinensis Franch.,also named Chinese goldthread is a member of Ranunculaceae in the order Ranunculales and represents an important lineage of early eudicots with traditional medicinal value.In our study,by using syntenic analysis combined with phylogenomic analysis of C.chinensis and four other representative genomes from basal and core eudicots,we confirmed that the WGD event in C.chinensis was shared by Aquilegia coerulea and Papaver somniferum L.and quickly occurred after Ranunculales diverged from other eudicots,likely a Ranunculales common tetraploidization(RCT).The synonymous nucleotide substitutions at synonymous sites distribution of syntenic blocks across these genomes showed that the evolutionary rate of the P.somniferum genome is faster than that of the C.chinensis genome by approximately 13.7%,possibly due to Papaveraceaes having an additional special tetraploidization event(PST).After Ks correction,the RCT dated to 115—130 million years ago(MYA),which was close to the divergence of Ranunculaceaes and Papaveraceaes approximately115.45—130.51 MYA.Moreover,we identified homologous genes related to polyploidization and speciation and constructed multiple sequence alignments with different reference genomes.Notably,the event-related subgenomes in the basal genomes all showed genomic fractionation bias,suggesting a likely allopolyploid nature of the RCT,PST and T-Alpha and T-Beta events in Tetracentron sinense.In addition,we detected that the sixteen P450 subfamilies were markedly expanded in the genomes of Ranunculales,and most of them were related to the RCT and PST events.We constructed a new platform for Early Eudicot Comparative Genomic Research(http://www.cgrpoee.top/index.html)to store more information.In summary,our findings support the WGD of C.chinensis shared by Ranunculales,which is likely an allotetraploidization event.This present effort offered new insights into the evolution of key polyploidization events and the genes related to secondary metabolites during the diversification of early eudicots.展开更多
Understanding how summer warming influences the parent and daughter shoot production in a perennial clonal grass is vital for comprehending the response of grassland productivity to global warming.Here,we conducted a s...Understanding how summer warming influences the parent and daughter shoot production in a perennial clonal grass is vital for comprehending the response of grassland productivity to global warming.Here,we conducted a simulated experiment using potted Leymus chinensis,to study the relationship between the photosynthetic activ-ity of parent shoots and the production of daughter shoots under a whole(90 days)summer warming scenario(+3°C).The results showed that the biomass of parents and buds decreased by 25.52%and 33.45%,respectively,under warming conditions.The reduction in parent shoot biomass due to warming directly resulted from decreased leaf area(18.03%),chlorophyll a(18.27%),chlorophyll b(29.21%)content,as well as a reduction in net photosynthetic rate(7.32%)and the maximum quantum efficiency of photosystem II(PSII)photochemistry(4.29%).The decline in daughter shoot biomass was linked to a decrease in daughter shoot number(33.33%)by warming.However,the number of belowground buds increased by 46.43%.The results indicated that long-term summer warming reduces biomass accumulation in parent shoot by increasing both limitation of stoma and non-stoma.Consequently,the parent shoot allocates relatively more biomass to the belowground organs to maintain the survival and growth of buds.Overall,buds,as a potential aboveground population,could remedy for the cur-rent loss of parent shoot density by increasing the number of future daughter shoots if summer warming subsides.展开更多
The Chinese tree shrew(Tupaia belangeri chinensis),a member of the mammalian order Scandentia,exhibits considerable similarities with primates,including humans,in aspects of its nervous,immune,and metabolic systems.Th...The Chinese tree shrew(Tupaia belangeri chinensis),a member of the mammalian order Scandentia,exhibits considerable similarities with primates,including humans,in aspects of its nervous,immune,and metabolic systems.These similarities have established the tree shrew as a promising experimental model for biomedical research on cancer,infectious diseases,metabolic disorders,and mental health conditions.Herein,we used metatranscriptomic sequencing to analyze plasma,as well as oral and anal swab samples,from 105 healthy asymptomatic tree shrews to identify the presence of potential zoonotic viruses.In total,eight mammalian viruses with complete genomes were identified,belonging to six viral families,including Flaviviridae,Hepeviridae,Parvovirinae,Picornaviridae,Sedoreoviridae,and Spinareoviridae.Notably,the presence of rotavirus was recorded in tree shrews for the first time.Three viruses-hepacivirus 1,parvovirus,and picornavirus-exhibited low genetic similarity(<70%)with previously reported viruses at the whole-genome scale,indicating novelty.Conversely,three other viruses-hepacivirus 2,hepatovirus A and hepevirus-exhibited high similarity(>94%)to known viral strains.Phylogenetic analyses also revealed that the rotavirus and mammalian orthoreovirus identified in this study may be novel reassortants.These findings provide insights into the diverse viral spectrum present in captive Chinese tree shrews,highlighting the necessity for further research into their potential for crossspecies transmission.展开更多
As highly social animals,Indo-Pacific humpback dolphins(Sousa chinensis)exhibit community differentiation.Nevertheless,our understanding of the external and internal factors influencing these dynamics,as well as their...As highly social animals,Indo-Pacific humpback dolphins(Sousa chinensis)exhibit community differentiation.Nevertheless,our understanding of the external and internal factors influencing these dynamics,as well as their spatiotemporal variations,is still limited.In the present study,variations in the social structure of an endangered Indo-Pacific humpback dolphin population in Xiamen Bay,China,were monitored over two distinct periods(2007–2010 and 2017–2019)to analyze the effects of habitat utilization and the composition of individuals within the population.In both periods,the population demonstrated a strikingly similar pattern of social differentiation,characterized by the division of individuals into two main clusters and one small cluster.Spatially,the two primary clusters occupied the eastern and western waters,respectively,although the core distribution area of the eastern cluster shifted further eastward between the two periods.Despite this distribution shift,the temporal stability of the social structure and inter-associations within the eastern cluster remained unaffected.A subset of 16individuals observed in both periods,comprising 51.6%and 43.2%of the population in each respective period,emerged as a foundational element of the social structure and may be responsible for sustaining social structure stability,especially during the 2007–2010 period.These observations suggest that the composition of dominant individuals,an internal factor,had a more substantial influence on the formation of the social network than changes in habitat use,an external factor.Consequently,the study proposes distinct conservation measures tailored to each of the two main clusters.展开更多
As a traditional Chinese herbal medicine,Schisandrae Chinensis Fructus(SC)has been used in medicine and food industry due to its health care and therapeutic effects.Over the past 20 years,the use of SC and its active ...As a traditional Chinese herbal medicine,Schisandrae Chinensis Fructus(SC)has been used in medicine and food industry due to its health care and therapeutic effects.Over the past 20 years,the use of SC and its active ingredient lignans in the prevention and treatment of liver diseases has been increasing,and their hepatoprotective effects has increased the interest of the public and academia.Therefore,in the present work,we first determined the effectiveness of SC in the treatment of liver diseases such as metabolic associated fatty liver disease,alcoholic liver disease,cholestatic liver disease and acute liver injury.Subsequently,the pharmacological effects and molecular mechanisms of lignans,the active components of SC,for liver disease treatment were comprehensively summarized for the first time.The results showed that the lignans in SC could achieve hepatoprotective effects by regulating lipid metabolism,anti-fibrosis,anti-inflammation,anti-oxidation,anti-tumor and regulating bile acid metabolism.The mechanism mainly involved adenosine 5’-monophosphate-activated protein kinase,endoplasmic reticulum stress,sterol regulatory element binding protein 1c,autophagy,transforming growth factor-β,mitogen-activated protein kinase,microRNA,nuclear factor kappa-B,nuclear factor erythroid-2-related factor 2,heat shock proteins and pregnane X receptor signaling pathways.These results can lay a scientific foundation for the development of hepatoprotective drugs or functional foods from SC/lignans.展开更多
The resource of Fenneropenaeus chinensis has declined sharply due to excessive fishing intensity,ecological changes and diseases.In order to supplement the fishing yield and restore resources of F.chinensis,the releva...The resource of Fenneropenaeus chinensis has declined sharply due to excessive fishing intensity,ecological changes and diseases.In order to supplement the fishing yield and restore resources of F.chinensis,the relevant authorities have carried out the activities of stock enhancement and releasing.It can increase biomass and recover resources.However,compared with increasing biomass,there were still few reports on its effect on the recovery of resources.Resource recovery is a process related to whether the released individuals can form a reproductive population.Up to now,there has been a lack of evidence whether the released F.chinensis can complete the entire life history,and form reproduction population.In this study,gravid female shrimp after spawning migration were captured from coastal waters of Haiyang,Qingdao,and Yellow Sea.After identifying parentage relationships using simple sequence repeat(SSR)and mtDNA haplotype,it was finally confirmed that there were eight released individuals in the recapture samples.It was confirmed for the first time that at least part of the released F.chinensis can complete overwintering and reproductive migration,and maintain the migration habits as their wild counterparts.Therefore,we infered that the released shrimp can reproduce under natural conditions,these F.chinensis can form reproductive populations theoretically if without human intervention.These results indicated that enhancenment and release activities have a positive effect on resource recovery.展开更多
BACKGROUND Prostate cancer(PCa)has high morbidity and mortality rates in elderly men.With a history of thousands of years,traditional Chinese medicine derived from insects could be an important source for developing c...BACKGROUND Prostate cancer(PCa)has high morbidity and mortality rates in elderly men.With a history of thousands of years,traditional Chinese medicine derived from insects could be an important source for developing cancer-targeted drugs to prevent tumorigenesis,enhance therapeutic effects,and reduce the risk of recurrence and metastasis.Multiple studies have shown that Coridius chinensis(Cc)has anticancer effects.AIM To elucidate the mechanism of action of Cc against PCa via network pharma-cology and molecular docking.METHODS Potential targets for Cc and PCa were predicted using ChemDraw 19.0 software,the PharmMapper database and the GeneCards database.Then,the STRING database was used to construct the protein–protein interaction network.Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment and molecular docking analyses were subsequently conducted to identify the key targets,active ingredients and pathways involved.RESULTS GO and KEGG analyses indicated that the PI3K-Akt signalling pathway was the critical pathway(P value<1.0×10-8).Multiple targeting ingredients that can affect multiple pathways in PCa have been identified in Cc.Seven active compounds(asponguanosines A,asponguanine B,asponguanine C,aspong-pyrazine A,N-acetyldopamine,aspongadenine B and aspongpyrazine B)were selected for molecular docking with 9 potential targets,and the results revealed that aspongpyrazine A and asponguanosine A are the main components by which Cc affects PCa(affinity<-5 kcal/mol,hydrogen bonding),but more studies are needed.CONCLUSION We used network pharmacology to predict the bioactive components and important targets of Cc for the treatment of PCa,supporting the development of Cc as a natural anticancer agent.展开更多
The purpose of this project is used for exploring the mechanism of Callistephus chinensis in the treatment of diabetes by network pharmacology and molecular docking methods.The target of Callistephus chinensis was obt...The purpose of this project is used for exploring the mechanism of Callistephus chinensis in the treatment of diabetes by network pharmacology and molecular docking methods.The target of Callistephus chinensis was obtained from SwissTargetPrediction database,while the target related to diabetes was obtained from GeneCards and OMIM databases.The target was added in String database to build the protein interaction network.GO biological process enrichment analysis and KEGG pathway enrichment analysis were carried out by Metascape software,then the target-pathway network was constructed.Molecular docking was carried out in Discovery Studio 2016 Client software to verify the binding force of Callistephus chinensis flavonoid compounds with key targets.In this study,10 potential active components were selected from the flavonoid monomer compounds of Callistephus chinensis.1847 biological processes(BP),126 cell compositions(CC)and 256 molecular functions(MF)were obtained by GO enrichment analysis;a total of 194 pathways were involved in KEGG enrichment analysis of 192 cross targets.Network analysis showed that quercetin was the main active component of flavonoids in the treatment of diabetes,AKT1,TNF,VEGFA,EGFR,SRC and other related signals were in relation to the treatment of diabetes.This study showed that Callistephus chinensis flavonoid compounds play a role in the treatment of diabetes by regulating multi-target and multi-pathway.展开更多
To clarify the taxonomic status of Phlebotomus chinensb, the morphological characters of Ph. chinensis from Shaanxi, Henan, Sichuan and Gansu Provinces, and Ph. wui and Ph. longiductus from Xinjiang were examined and ...To clarify the taxonomic status of Phlebotomus chinensb, the morphological characters of Ph. chinensis from Shaanxi, Henan, Sichuan and Gansu Provinces, and Ph. wui and Ph. longiductus from Xinjiang were examined and the rDNA-ITS2 region and mtDNA Cyt b gene sequences were analyzed simultaneously. It was found that morphological variation was very low in Ph. chinensis among the different localities; the wings of adult samples distributed in high-altitude areas were merely longer. The rDNA-ITS2 and mtDNA Cyt b sequences had an intraspecific variability. The relationships among species in the genus Phlebotomus can be inferred by molecular markers. These results suggest that there is an obvious genetic divergence at the molecular level among the Ph. chinensis populations in China.展开更多
This is the first report on a polysaccharide(RCJ2-Ib)isolated from Rosa Chinensis Flos.RCJ2-Ib was obtained through the extraction with water,precipitation with ethanol,separation with DEAE-52 column and purification ...This is the first report on a polysaccharide(RCJ2-Ib)isolated from Rosa Chinensis Flos.RCJ2-Ib was obtained through the extraction with water,precipitation with ethanol,separation with DEAE-52 column and purification with DEAE-Sepharose Fast Flow column and Sephadex G100 column.GC,FT-IR and NMR analyses revealed that RCJ2-Ib(3.3 k Da)was a 1,4-linked polymannuronic acid containing substantialβ-Danomers units.The anticoagulant effect of RCJ2-Ib evaluated by using rabbit ear venous blood and an acute blood stasis rat model showed that RCJ2-Ib had obvious anticoagulant activity in regulating endogenous and exogenous coagulation pathways and reducing serum thromboxane B2 and endothelin-1.In addition,RCJ2-Ib could also increase the number of Lactobacillus and Escherichia coli.As a result,RCJ2-Ib has the potential to inhibit thrombosis and maintain the intestinal environment.展开更多
Organic amendment is a promising,in situ phytostabilization approach to alleviate the phytotoxic effects of heavy metal contaminated soils.The aim of this study was to evaluate the feasibility of cow manure(CM)and i...Organic amendment is a promising,in situ phytostabilization approach to alleviate the phytotoxic effects of heavy metal contaminated soils.The aim of this study was to evaluate the feasibility of cow manure(CM)and its derived biochar(CMB)as a soil amendment on cadmium(Cd)availability and accumulation in low and high Cd-accumulating cultivars of Brassica chinensis L.grown in an acidic red soil.CM and CMB were applied to Cd-contaminated acidic red soil at the rates of 0,3.0and 6.0%(w/w).Application of CMB was significantly more effective than that of CM,as it reduced the availability of Cd in soil by 34.3–69.9%and its bioaccumulation in the low Cd accumulator,Aijiaoheiye 333,by 51.2 and 67.4%,respectively.The addition of CMB significantly increased the extractability and accumulation of trace metals(Zn,Mn,Fe,and Cu)by plants and improved plant biomass production.CMB application,combined with utilizing low Cd accumulating cultivars represents a new,sustainable strategy to alleviate the toxic effects on Cd and improve food safety.展开更多
Eight microsatellite markers were used to analyze genetic diversity, level of inbreeding, and effective population size of spawner and recaptured populations of Chinese shrimp(Fenneropenaeus chinensis) during stock ...Eight microsatellite markers were used to analyze genetic diversity, level of inbreeding, and effective population size of spawner and recaptured populations of Chinese shrimp(Fenneropenaeus chinensis) during stock enhancement in the Bohai Bay in 2013. A total of 254 and 238 alleles were identified in the spawner and recaptured populations, respectively, and the numbers of alleles(N_a) were 8–63 and 6–60, respectively. The numbers of effective alleles(N_e) were 2.52–21.60 and 2.67–20.72, respectively. The polymorphism information content ranged from 0.529 to 0.952. The observed heterozygosity(H_o) values(0.638–0.910 and 0.712–0.927) were lower than the expected heterozygosity(H_e) values(0.603–0.954 and 0.625–0.952), which indicated that the two populations possessed a rich genetic diversity. In 16 tests(2 populations×8 loci), 13 tests deviated from the HardyWeinberg equilibrium. F_(is) values were positive at seven loci and the inbreeding coefficients(F) of the two populations estimated by trio ML were 13.234% and 11.603%, suggesting that there was a relatively high degree of inbreeding. A certain level of inbreeding depression had occurred in the Chinese shrimp population. F_(st) values ranged from 0 to 0.059, with a mean of 0.028, displaying a low level of genetic differentiation in the two populations. Effective population sizes(3 060.2 and 3 842.8) were higher than the minimum number suggested for retaining the evolutionary potential to adapt to new environmental conditions. For enhancement activity in 2014,the ideal number of captured shrimp spawners should have ranged from 7 686 to 19 214 to maintain genetic diversity and effective population size. Further strategies to adjust the balance of economic cost, fishing effort and ideal number of shrimp spawners to maintain a satisfactory effective population size for ensuring the sustainability of Chinese shrimp are proposed.展开更多
Glutathione S-transferases(GSTs)are a superfamily of multifunction enzymes involved in the regulation of redox homeostasis and innate immune responses against various pathogenic infections in marine invertebrates.In t...Glutathione S-transferases(GSTs)are a superfamily of multifunction enzymes involved in the regulation of redox homeostasis and innate immune responses against various pathogenic infections in marine invertebrates.In the present study,a delta class GST gene(designated as FcδGST)was cloned from Fenneropenaeus chinensis using rapid amplification of c DNA ends(RACE)technology.The complete cDNA sequence of FcδGST was 780 bp in length,which includes a 27-bp 5′non-coding region(UTR),a 117-bp 3′UTR,a 636-bp open reading frame(ORF),and a polyadenylate signal site(AATAAA)presented at the upstream of poly A tail.The FcδGST gene encoded 211 amino acids peptide,including a GST_N domain and a GST_C domain,and exhibited high similarity with previously reported delta GSTs.The predicted molecular mass of FcδGST protein was 23.39 kDa,and its theoretical isoelectric point(pI)was 5.34.The FcδGST mRNA transcripts were ubiquitously expressed in all the tested tissues,with the highest expression level in hemocytes and hepatopancreas.During the stimulation of Vibrio anguillarum or white spot syndrome virus(WSSV),the m RNA expression of FcδGST in hemocytes and hepatopancreas revealed significant up-regulation.The purified recombinant FcδGST protein(designated as rFcδGST)exhibited specific catalytic activity against 1-chloro-2,4-dinitrobenzene(CDNB)substrate with relatively low stable enzymatic activities.These results indicated that FcδGST was a fragile but typical novel delta class GST member and potentially involved in the innate immune responses of F.chinensis.展开更多
[Objectives] The research aimed to evaluate impacts of diseases on quality of Paris polyphylla var. chinensis. [Methods]Ultraviolet spectrophotometry was used to determine and contrast content of total saponins in roo...[Objectives] The research aimed to evaluate impacts of diseases on quality of Paris polyphylla var. chinensis. [Methods]Ultraviolet spectrophotometry was used to determine and contrast content of total saponins in root,stem and leaf from healthy and diseased plants. Infrared characteristic absorption peaks of healthy and diseased plants were found by infrared fingerprint method for comparative analysis.[Results]Total saponins content in root sample J4 of healthy plant reached 4. 89% and was the highest,while it was 0. 13% in stem sample J11 of healthy plant and was the lowest. Total saponins content in root sample B1 of diseased plant was 1. 68% and was the highest,while it was 0. 1% in stem sample B7 of diseased plant and was the lowest. In healthy and diseased plants,total saponins content in root was significantly higher than that in stem and leaf. Content of total saponins in root from healthy plant was significantly higher than that from diseased plant,and there was little difference in stem and leaf of healthy and diseased plants. Root sample Jg of healthy plant had obvious C-O characteristic vibration of polysaccharides and glycosides and absorption peak of O-H bending vibration in steroidal saponin near 1 160. 02 cm-1,while diseased plant had no obvious absorption peak. Near 861. 97 and 768. 07 cm-1,root sample Jg of healthy plant had significant C-C stretching vibration in sugar ring and characteristic absorption peak of characteristic vibration of steroidal saponin,while root sample Bg of diseased plant had no obvious characteristic peak. [Conclusions] Diseases significantly affected composition and content of total saponins and steroidal saponins from P. polyphylla var. chinensis,further affecting quality of P. polyphylla var. chinensis.展开更多
To test the patterns of the root morphology and architecture indexes of Tamarix chinensis in response to water and salt changes in the two media of the groundwater and soil,three-year-old T.chinensis seedlings were ch...To test the patterns of the root morphology and architecture indexes of Tamarix chinensis in response to water and salt changes in the two media of the groundwater and soil,three-year-old T.chinensis seedlings were chosen as the research object.Groundwater with four salinity levels was created,and three groundwater level(GL)were applied for each salinity treatment to measure the root growth and architecture indexes.In the fresh water and brackish water treatments,the topological index(TI)of the T.chinensis roots was close to 0.5,and the root architecture was close to a dichotomous branching pattern.In the saline water and saltwater treatments,the TI of the T.chinensis roots was large and close to 1.0,and the root architecture was close to a herringbone-like branching pattern.Under different GLs and salinities,the total root length was significantly greater than the internal link length,the external link length was greater than the internal link length,and the root system showed an outward expansion strategy.The treatment with fresh water and a GL of 1.5 m was the most suitable for T.chinensis root growth,while the root growth of T.chinensis was the worst in the treatment with saline water and a GL of 0.3 m.T.chinensis can adapt to the changes in soil water and salt by regulating the growth and morphological characteristics of the root system.T.chinensis can adapt to high-salt environments by reducing its root branching and to water deficiencies by expanding the distribution and absorption area of the root system.展开更多
The effect of genotypic diversity on the age structure and bud bank of the Leymus chinensis population has not yet been demonstrated.This experiment was designed to study the age structure of the tiller,rhizome,and bu...The effect of genotypic diversity on the age structure and bud bank of the Leymus chinensis population has not yet been demonstrated.This experiment was designed to study the age structure of the tiller,rhizome,and bud of L.chinensis at genotypic diversity levels of 1,2,4,8,and 12.A total of 64 communities were established in this experiment,following the principle of randomized block experimental design.The results indicated that genotypic diversity had a significant or obviously significant effect on the cumulative length of rhizomes of 2a and 4a,dry matter accumulation of rhizomes of 1a and 2a,tiller number of 1a,tiller productivity of 3a,but had no significant effect age structure of buds.As the levels of genotypic diversity increased,the proportion of the cumulative length of rhizomes with 4a decreased and then increased,and dry matter accumulation of rhizomes with 1a and 2a gradually increased while the number of tillers with 1a gradually decreased,when the gradient of genotypic diversity increased.At the 1,2,4,8,and 12 genotype diversity levels,the number of tillers,the cumulative length of rhizomes,and dry matter accumulation in rhizomes and buds were all age structures of expanding type.However,tiller productivity was an expanding or stable age structure at levels of 1,2,4,8 genotypic diversity,while it was a declining age structure at the 12 genotype diversity gradient.An appropriate genotypic diversity is conducive to maintaining the growth and stability of the age structure of L.chinensis population,but too high a gradient of genotypic diversity can have a negative impact on the population age structure.This study provided that an appropriate number of genotypic diversity contributes to the stability of the population.展开更多
Although Vibrio parahaemolyticus has become the most common pathogen in fresh and frozen seafood,its virulence changes have often been ignored during the processing of seafood.To investigate these potential risks,we u...Although Vibrio parahaemolyticus has become the most common pathogen in fresh and frozen seafood,its virulence changes have often been ignored during the processing of seafood.To investigate these potential risks,we used frozen Penaeus chinensis as examples,and the most virulent factors of V.parahaemolyticus,including amounts,viable but nonculturable(VBNC)status,toxins TDH and TRH,and virulence genes tdh and trh,were determined.Bacterial quantities were signifi cantly reduced during drain and sprinkling phases,but caused by different factors.By SYTO9 and PI staining showed that washing was the main reason for the bacterial reduction at the drain phase,while the strain entering VBNC state was another reason at sprinkling phase.Their hemolysis toxicity,produced by TDH and TRH,became stronger after inoculation on shrimp,and could be detected throughout the process.Moreover,tdh and trh also exhibited trends similar to that of the hemolysis toxicity test.tdh was almost to a two-fold expression level during ice-glazing phase,while trh only express at a low level,less than half of the expression level before inoculation.These results demonstrated that the strains were not dead during freezing process,but became VBNC cells,which still produced and accumulated toxins,especially TDH,the most virulent factor.展开更多
基金funded by the National Natural Science Foundation of China(Grant Nos.32170236 and 31501333)the Natural Science Foundation of Hebei Province(Grant No.C2020209064)the Youth Foundation of Educational Committee of Hebei Province(Grant No.QN2020139)。
文摘Coptis chinensis Franch.,also named Chinese goldthread is a member of Ranunculaceae in the order Ranunculales and represents an important lineage of early eudicots with traditional medicinal value.In our study,by using syntenic analysis combined with phylogenomic analysis of C.chinensis and four other representative genomes from basal and core eudicots,we confirmed that the WGD event in C.chinensis was shared by Aquilegia coerulea and Papaver somniferum L.and quickly occurred after Ranunculales diverged from other eudicots,likely a Ranunculales common tetraploidization(RCT).The synonymous nucleotide substitutions at synonymous sites distribution of syntenic blocks across these genomes showed that the evolutionary rate of the P.somniferum genome is faster than that of the C.chinensis genome by approximately 13.7%,possibly due to Papaveraceaes having an additional special tetraploidization event(PST).After Ks correction,the RCT dated to 115—130 million years ago(MYA),which was close to the divergence of Ranunculaceaes and Papaveraceaes approximately115.45—130.51 MYA.Moreover,we identified homologous genes related to polyploidization and speciation and constructed multiple sequence alignments with different reference genomes.Notably,the event-related subgenomes in the basal genomes all showed genomic fractionation bias,suggesting a likely allopolyploid nature of the RCT,PST and T-Alpha and T-Beta events in Tetracentron sinense.In addition,we detected that the sixteen P450 subfamilies were markedly expanded in the genomes of Ranunculales,and most of them were related to the RCT and PST events.We constructed a new platform for Early Eudicot Comparative Genomic Research(http://www.cgrpoee.top/index.html)to store more information.In summary,our findings support the WGD of C.chinensis shared by Ranunculales,which is likely an allotetraploidization event.This present effort offered new insights into the evolution of key polyploidization events and the genes related to secondary metabolites during the diversification of early eudicots.
基金funded by the NSFC(32371669)the Science and Technology Talent Project for Distinguished Young Scholars of Jilin Province(20240602009RC)+1 种基金the NSF of Jilin Province(20240101207JC)the Scientific Research Project of the Department of Education,Jilin Province(JJKH20230687KJ).
文摘Understanding how summer warming influences the parent and daughter shoot production in a perennial clonal grass is vital for comprehending the response of grassland productivity to global warming.Here,we conducted a simulated experiment using potted Leymus chinensis,to study the relationship between the photosynthetic activ-ity of parent shoots and the production of daughter shoots under a whole(90 days)summer warming scenario(+3°C).The results showed that the biomass of parents and buds decreased by 25.52%and 33.45%,respectively,under warming conditions.The reduction in parent shoot biomass due to warming directly resulted from decreased leaf area(18.03%),chlorophyll a(18.27%),chlorophyll b(29.21%)content,as well as a reduction in net photosynthetic rate(7.32%)and the maximum quantum efficiency of photosystem II(PSII)photochemistry(4.29%).The decline in daughter shoot biomass was linked to a decrease in daughter shoot number(33.33%)by warming.However,the number of belowground buds increased by 46.43%.The results indicated that long-term summer warming reduces biomass accumulation in parent shoot by increasing both limitation of stoma and non-stoma.Consequently,the parent shoot allocates relatively more biomass to the belowground organs to maintain the survival and growth of buds.Overall,buds,as a potential aboveground population,could remedy for the cur-rent loss of parent shoot density by increasing the number of future daughter shoots if summer warming subsides.
基金supported by the National Key R&D Program of China (2021YFC2300900,2021YFC2301300)Academic Promotion Programme of Shandong First Medical University (2019QL006)+2 种基金Natural Science Foundation of Shandong Province (ZR2020QH274)Yunnan Key Research and Development Program (202103AQ100001,202102AA310055)Key Program of Chinese Academy of Sciences (KJZD-SW-L11)。
文摘The Chinese tree shrew(Tupaia belangeri chinensis),a member of the mammalian order Scandentia,exhibits considerable similarities with primates,including humans,in aspects of its nervous,immune,and metabolic systems.These similarities have established the tree shrew as a promising experimental model for biomedical research on cancer,infectious diseases,metabolic disorders,and mental health conditions.Herein,we used metatranscriptomic sequencing to analyze plasma,as well as oral and anal swab samples,from 105 healthy asymptomatic tree shrews to identify the presence of potential zoonotic viruses.In total,eight mammalian viruses with complete genomes were identified,belonging to six viral families,including Flaviviridae,Hepeviridae,Parvovirinae,Picornaviridae,Sedoreoviridae,and Spinareoviridae.Notably,the presence of rotavirus was recorded in tree shrews for the first time.Three viruses-hepacivirus 1,parvovirus,and picornavirus-exhibited low genetic similarity(<70%)with previously reported viruses at the whole-genome scale,indicating novelty.Conversely,three other viruses-hepacivirus 2,hepatovirus A and hepevirus-exhibited high similarity(>94%)to known viral strains.Phylogenetic analyses also revealed that the rotavirus and mammalian orthoreovirus identified in this study may be novel reassortants.These findings provide insights into the diverse viral spectrum present in captive Chinese tree shrews,highlighting the necessity for further research into their potential for crossspecies transmission.
基金supported by the National Natural Science Foundation of China (32030011,31630071)National Key Research and Development Program of China (2022YFF1301600)Priority Academic Program Development of Jiangsu Higher Education Institutions。
文摘As highly social animals,Indo-Pacific humpback dolphins(Sousa chinensis)exhibit community differentiation.Nevertheless,our understanding of the external and internal factors influencing these dynamics,as well as their spatiotemporal variations,is still limited.In the present study,variations in the social structure of an endangered Indo-Pacific humpback dolphin population in Xiamen Bay,China,were monitored over two distinct periods(2007–2010 and 2017–2019)to analyze the effects of habitat utilization and the composition of individuals within the population.In both periods,the population demonstrated a strikingly similar pattern of social differentiation,characterized by the division of individuals into two main clusters and one small cluster.Spatially,the two primary clusters occupied the eastern and western waters,respectively,although the core distribution area of the eastern cluster shifted further eastward between the two periods.Despite this distribution shift,the temporal stability of the social structure and inter-associations within the eastern cluster remained unaffected.A subset of 16individuals observed in both periods,comprising 51.6%and 43.2%of the population in each respective period,emerged as a foundational element of the social structure and may be responsible for sustaining social structure stability,especially during the 2007–2010 period.These observations suggest that the composition of dominant individuals,an internal factor,had a more substantial influence on the formation of the social network than changes in habitat use,an external factor.Consequently,the study proposes distinct conservation measures tailored to each of the two main clusters.
基金supported by National Natural Science Foundation of China(81891012,U19A2010,81630101)Sichuan Province Science and Technology Support Program(2021JDRC0041)Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-D-202209).
文摘As a traditional Chinese herbal medicine,Schisandrae Chinensis Fructus(SC)has been used in medicine and food industry due to its health care and therapeutic effects.Over the past 20 years,the use of SC and its active ingredient lignans in the prevention and treatment of liver diseases has been increasing,and their hepatoprotective effects has increased the interest of the public and academia.Therefore,in the present work,we first determined the effectiveness of SC in the treatment of liver diseases such as metabolic associated fatty liver disease,alcoholic liver disease,cholestatic liver disease and acute liver injury.Subsequently,the pharmacological effects and molecular mechanisms of lignans,the active components of SC,for liver disease treatment were comprehensively summarized for the first time.The results showed that the lignans in SC could achieve hepatoprotective effects by regulating lipid metabolism,anti-fibrosis,anti-inflammation,anti-oxidation,anti-tumor and regulating bile acid metabolism.The mechanism mainly involved adenosine 5’-monophosphate-activated protein kinase,endoplasmic reticulum stress,sterol regulatory element binding protein 1c,autophagy,transforming growth factor-β,mitogen-activated protein kinase,microRNA,nuclear factor kappa-B,nuclear factor erythroid-2-related factor 2,heat shock proteins and pregnane X receptor signaling pathways.These results can lay a scientific foundation for the development of hepatoprotective drugs or functional foods from SC/lignans.
基金Laoshan Laboratory under contract No.LSKJ202203803。
文摘The resource of Fenneropenaeus chinensis has declined sharply due to excessive fishing intensity,ecological changes and diseases.In order to supplement the fishing yield and restore resources of F.chinensis,the relevant authorities have carried out the activities of stock enhancement and releasing.It can increase biomass and recover resources.However,compared with increasing biomass,there were still few reports on its effect on the recovery of resources.Resource recovery is a process related to whether the released individuals can form a reproductive population.Up to now,there has been a lack of evidence whether the released F.chinensis can complete the entire life history,and form reproduction population.In this study,gravid female shrimp after spawning migration were captured from coastal waters of Haiyang,Qingdao,and Yellow Sea.After identifying parentage relationships using simple sequence repeat(SSR)and mtDNA haplotype,it was finally confirmed that there were eight released individuals in the recapture samples.It was confirmed for the first time that at least part of the released F.chinensis can complete overwintering and reproductive migration,and maintain the migration habits as their wild counterparts.Therefore,we infered that the released shrimp can reproduce under natural conditions,these F.chinensis can form reproductive populations theoretically if without human intervention.These results indicated that enhancenment and release activities have a positive effect on resource recovery.
基金the Major Project of Science and Technology Foundation of Guizhou Provincial Health Commission,No.gzwkj2023-579Zunyi Medical University Innovation Project,No.S202310661123.
文摘BACKGROUND Prostate cancer(PCa)has high morbidity and mortality rates in elderly men.With a history of thousands of years,traditional Chinese medicine derived from insects could be an important source for developing cancer-targeted drugs to prevent tumorigenesis,enhance therapeutic effects,and reduce the risk of recurrence and metastasis.Multiple studies have shown that Coridius chinensis(Cc)has anticancer effects.AIM To elucidate the mechanism of action of Cc against PCa via network pharma-cology and molecular docking.METHODS Potential targets for Cc and PCa were predicted using ChemDraw 19.0 software,the PharmMapper database and the GeneCards database.Then,the STRING database was used to construct the protein–protein interaction network.Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment and molecular docking analyses were subsequently conducted to identify the key targets,active ingredients and pathways involved.RESULTS GO and KEGG analyses indicated that the PI3K-Akt signalling pathway was the critical pathway(P value<1.0×10-8).Multiple targeting ingredients that can affect multiple pathways in PCa have been identified in Cc.Seven active compounds(asponguanosines A,asponguanine B,asponguanine C,aspong-pyrazine A,N-acetyldopamine,aspongadenine B and aspongpyrazine B)were selected for molecular docking with 9 potential targets,and the results revealed that aspongpyrazine A and asponguanosine A are the main components by which Cc affects PCa(affinity<-5 kcal/mol,hydrogen bonding),but more studies are needed.CONCLUSION We used network pharmacology to predict the bioactive components and important targets of Cc for the treatment of PCa,supporting the development of Cc as a natural anticancer agent.
文摘The purpose of this project is used for exploring the mechanism of Callistephus chinensis in the treatment of diabetes by network pharmacology and molecular docking methods.The target of Callistephus chinensis was obtained from SwissTargetPrediction database,while the target related to diabetes was obtained from GeneCards and OMIM databases.The target was added in String database to build the protein interaction network.GO biological process enrichment analysis and KEGG pathway enrichment analysis were carried out by Metascape software,then the target-pathway network was constructed.Molecular docking was carried out in Discovery Studio 2016 Client software to verify the binding force of Callistephus chinensis flavonoid compounds with key targets.In this study,10 potential active components were selected from the flavonoid monomer compounds of Callistephus chinensis.1847 biological processes(BP),126 cell compositions(CC)and 256 molecular functions(MF)were obtained by GO enrichment analysis;a total of 194 pathways were involved in KEGG enrichment analysis of 192 cross targets.Network analysis showed that quercetin was the main active component of flavonoids in the treatment of diabetes,AKT1,TNF,VEGFA,EGFR,SRC and other related signals were in relation to the treatment of diabetes.This study showed that Callistephus chinensis flavonoid compounds play a role in the treatment of diabetes by regulating multi-target and multi-pathway.
基金supported by the Chinese National Natural Science Foundations(No.30771869)
文摘To clarify the taxonomic status of Phlebotomus chinensb, the morphological characters of Ph. chinensis from Shaanxi, Henan, Sichuan and Gansu Provinces, and Ph. wui and Ph. longiductus from Xinjiang were examined and the rDNA-ITS2 region and mtDNA Cyt b gene sequences were analyzed simultaneously. It was found that morphological variation was very low in Ph. chinensis among the different localities; the wings of adult samples distributed in high-altitude areas were merely longer. The rDNA-ITS2 and mtDNA Cyt b sequences had an intraspecific variability. The relationships among species in the genus Phlebotomus can be inferred by molecular markers. These results suggest that there is an obvious genetic divergence at the molecular level among the Ph. chinensis populations in China.
基金supported by Research on Precision Nutrition and Health Food,Department of Science and Technology of Henan Province(CXJD2021006)Key Project in Science and Technology Agency of Henan Province(202102110283 and 202102110149)。
文摘This is the first report on a polysaccharide(RCJ2-Ib)isolated from Rosa Chinensis Flos.RCJ2-Ib was obtained through the extraction with water,precipitation with ethanol,separation with DEAE-52 column and purification with DEAE-Sepharose Fast Flow column and Sephadex G100 column.GC,FT-IR and NMR analyses revealed that RCJ2-Ib(3.3 k Da)was a 1,4-linked polymannuronic acid containing substantialβ-Danomers units.The anticoagulant effect of RCJ2-Ib evaluated by using rabbit ear venous blood and an acute blood stasis rat model showed that RCJ2-Ib had obvious anticoagulant activity in regulating endogenous and exogenous coagulation pathways and reducing serum thromboxane B2 and endothelin-1.In addition,RCJ2-Ib could also increase the number of Lactobacillus and Escherichia coli.As a result,RCJ2-Ib has the potential to inhibit thrombosis and maintain the intestinal environment.
基金financially supported by Ministry of Science and Technology of China (2012AA100405)Zhejiang University Innovative Research Funds, China (2015FZA6008)
文摘Organic amendment is a promising,in situ phytostabilization approach to alleviate the phytotoxic effects of heavy metal contaminated soils.The aim of this study was to evaluate the feasibility of cow manure(CM)and its derived biochar(CMB)as a soil amendment on cadmium(Cd)availability and accumulation in low and high Cd-accumulating cultivars of Brassica chinensis L.grown in an acidic red soil.CM and CMB were applied to Cd-contaminated acidic red soil at the rates of 0,3.0and 6.0%(w/w).Application of CMB was significantly more effective than that of CM,as it reduced the availability of Cd in soil by 34.3–69.9%and its bioaccumulation in the low Cd accumulator,Aijiaoheiye 333,by 51.2 and 67.4%,respectively.The addition of CMB significantly increased the extractability and accumulation of trace metals(Zn,Mn,Fe,and Cu)by plants and improved plant biomass production.CMB application,combined with utilizing low Cd accumulating cultivars represents a new,sustainable strategy to alleviate the toxic effects on Cd and improve food safety.
基金The National Basic Research Program of China(973 Program)Adaptive Response of Fishery Species to Environmental Changes and Their Effects on Population Dynamics under contract No.2015CB453303the International Science&Technology Cooperation Program of China under contract No.2013DFA31410
文摘Eight microsatellite markers were used to analyze genetic diversity, level of inbreeding, and effective population size of spawner and recaptured populations of Chinese shrimp(Fenneropenaeus chinensis) during stock enhancement in the Bohai Bay in 2013. A total of 254 and 238 alleles were identified in the spawner and recaptured populations, respectively, and the numbers of alleles(N_a) were 8–63 and 6–60, respectively. The numbers of effective alleles(N_e) were 2.52–21.60 and 2.67–20.72, respectively. The polymorphism information content ranged from 0.529 to 0.952. The observed heterozygosity(H_o) values(0.638–0.910 and 0.712–0.927) were lower than the expected heterozygosity(H_e) values(0.603–0.954 and 0.625–0.952), which indicated that the two populations possessed a rich genetic diversity. In 16 tests(2 populations×8 loci), 13 tests deviated from the HardyWeinberg equilibrium. F_(is) values were positive at seven loci and the inbreeding coefficients(F) of the two populations estimated by trio ML were 13.234% and 11.603%, suggesting that there was a relatively high degree of inbreeding. A certain level of inbreeding depression had occurred in the Chinese shrimp population. F_(st) values ranged from 0 to 0.059, with a mean of 0.028, displaying a low level of genetic differentiation in the two populations. Effective population sizes(3 060.2 and 3 842.8) were higher than the minimum number suggested for retaining the evolutionary potential to adapt to new environmental conditions. For enhancement activity in 2014,the ideal number of captured shrimp spawners should have ranged from 7 686 to 19 214 to maintain genetic diversity and effective population size. Further strategies to adjust the balance of economic cost, fishing effort and ideal number of shrimp spawners to maintain a satisfactory effective population size for ensuring the sustainability of Chinese shrimp are proposed.
基金Supported by the Project of Sanya Yazhou Bay Science and Technology City Management Foundation (No.SKJC-KJ-2019KY01)the Startup Fund of Young Talents Project of Ocean University of China。
文摘Glutathione S-transferases(GSTs)are a superfamily of multifunction enzymes involved in the regulation of redox homeostasis and innate immune responses against various pathogenic infections in marine invertebrates.In the present study,a delta class GST gene(designated as FcδGST)was cloned from Fenneropenaeus chinensis using rapid amplification of c DNA ends(RACE)technology.The complete cDNA sequence of FcδGST was 780 bp in length,which includes a 27-bp 5′non-coding region(UTR),a 117-bp 3′UTR,a 636-bp open reading frame(ORF),and a polyadenylate signal site(AATAAA)presented at the upstream of poly A tail.The FcδGST gene encoded 211 amino acids peptide,including a GST_N domain and a GST_C domain,and exhibited high similarity with previously reported delta GSTs.The predicted molecular mass of FcδGST protein was 23.39 kDa,and its theoretical isoelectric point(pI)was 5.34.The FcδGST mRNA transcripts were ubiquitously expressed in all the tested tissues,with the highest expression level in hemocytes and hepatopancreas.During the stimulation of Vibrio anguillarum or white spot syndrome virus(WSSV),the m RNA expression of FcδGST in hemocytes and hepatopancreas revealed significant up-regulation.The purified recombinant FcδGST protein(designated as rFcδGST)exhibited specific catalytic activity against 1-chloro-2,4-dinitrobenzene(CDNB)substrate with relatively low stable enzymatic activities.These results indicated that FcδGST was a fragile but typical novel delta class GST member and potentially involved in the innate immune responses of F.chinensis.
基金Supported by the National Key R&D Program(2018YFC1708005)the Sichuan Science and Technology Plan Project(2018JY0111)the Innovating Research Program of Postgraduates of Southwest Minzu University in 2018(cx2018sz83)
文摘[Objectives] The research aimed to evaluate impacts of diseases on quality of Paris polyphylla var. chinensis. [Methods]Ultraviolet spectrophotometry was used to determine and contrast content of total saponins in root,stem and leaf from healthy and diseased plants. Infrared characteristic absorption peaks of healthy and diseased plants were found by infrared fingerprint method for comparative analysis.[Results]Total saponins content in root sample J4 of healthy plant reached 4. 89% and was the highest,while it was 0. 13% in stem sample J11 of healthy plant and was the lowest. Total saponins content in root sample B1 of diseased plant was 1. 68% and was the highest,while it was 0. 1% in stem sample B7 of diseased plant and was the lowest. In healthy and diseased plants,total saponins content in root was significantly higher than that in stem and leaf. Content of total saponins in root from healthy plant was significantly higher than that from diseased plant,and there was little difference in stem and leaf of healthy and diseased plants. Root sample Jg of healthy plant had obvious C-O characteristic vibration of polysaccharides and glycosides and absorption peak of O-H bending vibration in steroidal saponin near 1 160. 02 cm-1,while diseased plant had no obvious absorption peak. Near 861. 97 and 768. 07 cm-1,root sample Jg of healthy plant had significant C-C stretching vibration in sugar ring and characteristic absorption peak of characteristic vibration of steroidal saponin,while root sample Bg of diseased plant had no obvious characteristic peak. [Conclusions] Diseases significantly affected composition and content of total saponins and steroidal saponins from P. polyphylla var. chinensis,further affecting quality of P. polyphylla var. chinensis.
基金financially supported by the Joint Funds of the National Natural Science Foundation of China(U2006215)the National Natural Science Foundation of China(31770761)+2 种基金the Shandong Key Laboratory of Coastal Environmental Processes,YICCAS(2019SDHADKFJJ16)the Natural Science Foundation of Shangdong Province(ZR2020QD003)Taishan Scholars Program of Shandong Province,China(TSQN201909152)。
文摘To test the patterns of the root morphology and architecture indexes of Tamarix chinensis in response to water and salt changes in the two media of the groundwater and soil,three-year-old T.chinensis seedlings were chosen as the research object.Groundwater with four salinity levels was created,and three groundwater level(GL)were applied for each salinity treatment to measure the root growth and architecture indexes.In the fresh water and brackish water treatments,the topological index(TI)of the T.chinensis roots was close to 0.5,and the root architecture was close to a dichotomous branching pattern.In the saline water and saltwater treatments,the TI of the T.chinensis roots was large and close to 1.0,and the root architecture was close to a herringbone-like branching pattern.Under different GLs and salinities,the total root length was significantly greater than the internal link length,the external link length was greater than the internal link length,and the root system showed an outward expansion strategy.The treatment with fresh water and a GL of 1.5 m was the most suitable for T.chinensis root growth,while the root growth of T.chinensis was the worst in the treatment with saline water and a GL of 0.3 m.T.chinensis can adapt to the changes in soil water and salt by regulating the growth and morphological characteristics of the root system.T.chinensis can adapt to high-salt environments by reducing its root branching and to water deficiencies by expanding the distribution and absorption area of the root system.
基金supported by the National Natural Science Foundation of China(32071860).
文摘The effect of genotypic diversity on the age structure and bud bank of the Leymus chinensis population has not yet been demonstrated.This experiment was designed to study the age structure of the tiller,rhizome,and bud of L.chinensis at genotypic diversity levels of 1,2,4,8,and 12.A total of 64 communities were established in this experiment,following the principle of randomized block experimental design.The results indicated that genotypic diversity had a significant or obviously significant effect on the cumulative length of rhizomes of 2a and 4a,dry matter accumulation of rhizomes of 1a and 2a,tiller number of 1a,tiller productivity of 3a,but had no significant effect age structure of buds.As the levels of genotypic diversity increased,the proportion of the cumulative length of rhizomes with 4a decreased and then increased,and dry matter accumulation of rhizomes with 1a and 2a gradually increased while the number of tillers with 1a gradually decreased,when the gradient of genotypic diversity increased.At the 1,2,4,8,and 12 genotype diversity levels,the number of tillers,the cumulative length of rhizomes,and dry matter accumulation in rhizomes and buds were all age structures of expanding type.However,tiller productivity was an expanding or stable age structure at levels of 1,2,4,8 genotypic diversity,while it was a declining age structure at the 12 genotype diversity gradient.An appropriate genotypic diversity is conducive to maintaining the growth and stability of the age structure of L.chinensis population,but too high a gradient of genotypic diversity can have a negative impact on the population age structure.This study provided that an appropriate number of genotypic diversity contributes to the stability of the population.
基金funded by grants from the National Key Research and Development Program of China[2018YFC1602201]Key Research and Development Program of Shaanxi Province[2021NY-158]+1 种基金the National Natural Science Foundation of China[31671780]the Agricultural Product Quality and Safety Risk Assessment Foundation of the Ministry of Agriculture and Rural Affairs of the People’s Republic of China[GJFP2020002,GJFP2020003].
文摘Although Vibrio parahaemolyticus has become the most common pathogen in fresh and frozen seafood,its virulence changes have often been ignored during the processing of seafood.To investigate these potential risks,we used frozen Penaeus chinensis as examples,and the most virulent factors of V.parahaemolyticus,including amounts,viable but nonculturable(VBNC)status,toxins TDH and TRH,and virulence genes tdh and trh,were determined.Bacterial quantities were signifi cantly reduced during drain and sprinkling phases,but caused by different factors.By SYTO9 and PI staining showed that washing was the main reason for the bacterial reduction at the drain phase,while the strain entering VBNC state was another reason at sprinkling phase.Their hemolysis toxicity,produced by TDH and TRH,became stronger after inoculation on shrimp,and could be detected throughout the process.Moreover,tdh and trh also exhibited trends similar to that of the hemolysis toxicity test.tdh was almost to a two-fold expression level during ice-glazing phase,while trh only express at a low level,less than half of the expression level before inoculation.These results demonstrated that the strains were not dead during freezing process,but became VBNC cells,which still produced and accumulated toxins,especially TDH,the most virulent factor.