Plant anatomy is patterned early during leaf development which suggests studying the spatial–temporal transcriptomes of primordia will help identify critical regulative and functional genes.We successfully isolated t...Plant anatomy is patterned early during leaf development which suggests studying the spatial–temporal transcriptomes of primordia will help identify critical regulative and functional genes.We successfully isolated the leaf primordia tissues from the C3grass rice and the C4grass foxtail millet by laser capture microdissection(LCM)and studied the gene expression throughout leaf developmental stages.Our data analysis uncovered the conserved expression patterns of certain gene clusters both in rice and foxtail millet during leaf development.We revealed genes and transcription factors involved in vein formation,stomatal development,and suberin accumulation.We identified 79 candidate genes associated with functional regulation of C4anatomy formation.Screening phenotype of the candidate genes revealed that knock-out of a putative polar auxin transport related gene NAL1 resulted significantly reduced veinal space in rice leaf.Our present work provides a foundation for future analyses of genes with novel functions in grasses and their role in leaf development,in particular the role in leaves with a contrasting C3vs.C4biosynthetic pathway.展开更多
Betalain has received increased attention because of its high nutritional value and crucial physiological functions.Based on the elucidation of its core biosynthetic pathway,betalain can be produced in additional plan...Betalain has received increased attention because of its high nutritional value and crucial physiological functions.Based on the elucidation of its core biosynthetic pathway,betalain can be produced in additional plants by metabolic engineering.Synthesis of betalain in carrot(Daucus carota L.)can improve its nutritional quality and economic value by extracting betalain from the fleshy root,non-edible part,and processing residue of carrot.In this study,two different constructs,namely,pYB:mCD(AomelOS,BvCYP76AD1S,and BvDODA1S)and p YB:CDD(BvCYP76AD1S,BvDODA1S,and MjcDOPA5GTS),were introduced into carrot for betanin synthesis by Agrobacterium-mediated transformation.Betanin can be synthetized in both transgenic calli,and p YB:m CD-transgenic callus can be used to produce betacyanin by suspension culture.However,pYB:mCD-transgenic seedlings can synthetize betanin only by tyrosine feeding.The p YB:CDD-transgenic lines can synthetize betanin in whole plants.The betanin content in fleshy root of pYB:CDD-transgenic carrot was(63.4±9)μg·g^(-1)fresh weight according to quantitative analysis.These betanin-producing carrot plant materials can be used to synthesize betanin for industrial application or consumption as dietary sources.展开更多
Porcine reproductive and respiratory syndrome(PRRS)is a globally prevalent contagious disease caused by the positive-strand RNA PRRS virus(PRRSV),resulting in substantial economic losses in the swine industry.Modifyin...Porcine reproductive and respiratory syndrome(PRRS)is a globally prevalent contagious disease caused by the positive-strand RNA PRRS virus(PRRSV),resulting in substantial economic losses in the swine industry.Modifying the CD163 SRCR5 domain,either through deletion or substitution,can eff1ectively confer resistance to PRRSV infection in pigs.However,large fragment modifications in pigs inevitably raise concerns about potential adverse effects on growth performance.Reducing the impact of genetic modifications on normal physiological functions is a promising direction for developing PRRSV-resistant pigs.In the current study,we identified a specific functional amino acid in CD163 that influences PRRSV proliferation.Viral infection experiments conducted on Marc145 and PK-15CD163 cells illustrated that the mE535G or corresponding pE529G mutations markedly inhibited highly pathogenic PRRSV(HP-PRRSV)proliferation by preventing viral binding and entry.Furthermore,individual viral challenge tests revealed that pigs with the E529G mutation had viral loads two orders of magnitude lower than wild-type(WT)pigs,confirming effective resistance to HP-PRRSV.Examination of the physiological indicators and scavenger function of CD163 verified no significant differences between the WT and E529G pigs.These findings suggest that E529G pigs can be used for breeding PRRSV-resistant pigs,providing novel insights into controlling future PRRSV outbreaks.展开更多
Objective:To determine the genetic diversity of Plasmodium(P.)knowlesi isolates from Sabah,Malaysian Borneo and Peninsular Malaysia,targeting the S-type SSU rRNA gene and including aspects of natural selection and hap...Objective:To determine the genetic diversity of Plasmodium(P.)knowlesi isolates from Sabah,Malaysian Borneo and Peninsular Malaysia,targeting the S-type SSU rRNA gene and including aspects of natural selection and haplotype.Methods:Thirty-nine blood samples infected with P.knowlesi were collected in Sabah,Malaysian Borneo and Peninsular Malaysia.The S-type SSU rRNA gene was amplified using polymerase chain reaction,cloned into a vector,and sequenced.The natural selection and haplotype of the S-type SSU rRNA gene sequences were determined using DnaSP v6 and illustrated using NETWORK v10.This study's 39 S-type SSU rRNA sequences and eight sequences from the Genbank database were subjected to phylogenetic analysis using MEGA 11.Results:Overall,the phylogenetic analysis showed no evidence of a geographical cluster of P.knowlesi isolates from different areas in Malaysia based on the S-type SSU rRNA gene sequences.The S-type SSU rRNA gene sequences were relatively conserved and with a purifying effect.Haplotype sharing of the S-type SSU rRNA gene was observed between the P.knowlesi isolates in Sabah,Malaysian Borneo,but not between Sabah,Malaysian Borneo and Peninsular Malaysia.Conclusions:This study suggests that the S-type SSU rRNA gene of P.knowlesi isolates in Sabah,Malaysian Borneo,and Peninsular Malaysia has fewer polymorphic sites,representing the conservation of the gene.These features make the S-type SSU rRNA gene suitable for comparative studies,such as determining the evolutionary relationships and common ancestry among P.knowlesi species.展开更多
Florets are the basic structural units of spikelets,and their morphogenesis determines the yield and quality of rice grains.However,whether and how pseudouridine-5’-phosphate glycosidase participates in rice spikelet...Florets are the basic structural units of spikelets,and their morphogenesis determines the yield and quality of rice grains.However,whether and how pseudouridine-5’-phosphate glycosidase participates in rice spikelet development remains an open question.In this study,we identified a novel gene,OsPPG,which encodes a peroxisome-localized pseudouridine-5’-phosphate glycosidase and regulates the development of rice spikelets.osppg mutants exhibited abnormal sterile lemma,lemma,palea,lodicule,stamens,and pistils;male sterility;shorter panicles;and reduced plant height.OsPPG was found to regulate several OsMADS genes,thereby affecting the morphogenesis of rice spikelets.Furthermore,metabolomics revealed that the OsPPG gene was involved in the decomposition of pseudouridine via the pyrimidine metabolism pathway and may affect the jasmonic acid signaling pathway.These results suggest that OsPPG is a key regulator of rice spikelet development.展开更多
Alfalfa(Medicago sativa.L.)is a globally significant autotetraploid legume forage crop.However,despite its importance,establishing efficient gene editing systems for cultivated alfalfa remains a formidable challenge.I...Alfalfa(Medicago sativa.L.)is a globally significant autotetraploid legume forage crop.However,despite its importance,establishing efficient gene editing systems for cultivated alfalfa remains a formidable challenge.In this study,we pioneered the development of a highly effective ultrasonic-assisted leaf disc transformation system for Gongnong 1 alfalfa,a variety widely cultivated in Northeast China.Subsequently,we created a single transcript CRISPR/Cas9(CRISPR_2.0)toolkit,incorporating multiplex gRNAs,designed for gene editing in Gongnong 1.Both Cas9 and gRNA scaffolds were under the control of the Arabidopsis ubiquitin-10 promoter,a widely employed polymeraseⅡconstitutive promoter known for strong transgene expression in dicots.To assess the toolkit’s efficiency,we targeted PALM1,a gene associated with a recognizable multifoliate phenotype.Utilizing the CRISPR_2.0 toolkit,we directed PALM1 editing at two sites in the wild-type Gongnong 1.Results indicated a 35.1%occurrence of editing events all in target 2 alleles,while no mutations were detected at target 1 in the transgenic-positive lines.To explore more efficient sgRNAs,we developed a rapid,reliable screening system based on Agrobacterium rhizogenes-mediated hairy root transformation,incorporating the visible reporter MtLAP1.This screening system demonstrated that most purple visible hairy roots underwent gene editing.Notably,sgRNA3,with an 83.0%editing efficiency,was selected using the visible hairy root system.As anticipated,tetra-allelic homozygous palm1 mutations exhibited a clear multifoliate phenotype.These palm1 lines demonstrated an average crude protein yield increase of 21.5%compared to trifoliolate alfalfa.Our findings highlight the modified CRISPR_2.0 system as a highly efficient and robust gene editing tool for autotetraploid alfalfa.展开更多
The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of t...The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.展开更多
The overuse of nitrogen(N)fertilizer in fields has increased production costs and raised environmental concerns.Increasing the N use efficiency(NUE)of rice varieties is crucial for sustainable agriculture.Here we repo...The overuse of nitrogen(N)fertilizer in fields has increased production costs and raised environmental concerns.Increasing the N use efficiency(NUE)of rice varieties is crucial for sustainable agriculture.Here we report the cloning and characterization of OsNPF3.1,a gene that controls rice NUE.An amino acid mutation in the OsNPF3.1 coding region caused different NUEs in wild and cultivated rice.OsNPF3.1,which is expressed mainly in the aerial parts of rice,also affects rice plant height,heading date,and thousand-grain weight.The OsNPF3.1 protein is located in the plasma membrane.When OsNPF3.1 was subjected to artificial selection,two naturally varying loci were associated with NUE,of which OsNPF3.1Chr6_8741040differed between indica and japonica rice.OsNPF3.1 can be used as a new target gene for breeding rice varieties with high NUE.展开更多
Despite increasing knowledge of barnyardgrass(Echinochloa crus-galli) interference with rice, relatively little is known how endophytes improve the ability of rice against barnyardgrass stress. Here, we provided a det...Despite increasing knowledge of barnyardgrass(Echinochloa crus-galli) interference with rice, relatively little is known how endophytes improve the ability of rice against barnyardgrass stress. Here, we provided a detailed temporal characterization of rice root-associated microbiomes during co-cultivation with barnyardgrass and a comparison with the microbiomes of weed-free rice plants. Alpha diversity analysis indicated that barnyardgrass had the opposite effects on endophytic bacteria and fungi in rice roots, in terms of the community diversity, richness and coverage at the rice seedling stage. Principal coordinate analysis showed that barnyardgrass had only a minor effect on the community composition of endophytes in rice roots at the rice seedling stage, but showed a significant and maximum interference at the heading stage. Rice recruited many endophytes to resist biotic stress from barnyardgrass, especially for fungi. PICRUSt(phylogenetic investigation of communities by reconstruction of unobserved states) predictive analysis indicated that 23 metabolic pathways of bacteria were overrepresented in rice. In addition, the main trophic mode of fungi was pathotroph according to FUNGuild analysis. A positive correlation between bacteria and fungi in rice roots was found via network analysis. Anaeromyxobacter, Azospira and Pseudolabrys were the vital bacteria, Phaeosphaeria and Funneliformis were the dominant fungi in maintaining the stability of the ecological network. These results provided data and a theoretical basis for the in-depth understanding of what role endophytes play in rice resistance to barnyardgrass stress and will have implications on improving the resistance of rice against biotic stress using root microbiota.展开更多
Objective:To investigate the cardioprotective effect of naringenin against isoproterenol(ISO)-induced cardiotoxicity in rats.Methods:Rats were divided into five groups:the normal group,the ISO group(85 mg/kg b.w.);the...Objective:To investigate the cardioprotective effect of naringenin against isoproterenol(ISO)-induced cardiotoxicity in rats.Methods:Rats were divided into five groups:the normal group,the ISO group(85 mg/kg b.w.);the ISO+naringenin(50 mg/kg b.w.)group,the ISO+naringenin(100 mg/kg b.w.)group and the ISO+propranolol(10 mg/kg b.w.)group.Plasma creatine kinase-MB(CK-MB),cardiac troponin T,lactate dehydrogenase,brain natriuretic peptide(BNP),and IL-10,as well as cardiac transforming growth factor-β1(TGF-β1),vascular endothelial growth factor(VEGF)and malondialdehyde(MDA)were examined.In addition,NLRP3 and mRNA-208a expressions were evaluated by RT-PCR analysis.Histopathological examination was also performed to assess cardiac damages.Results:Naringenin treatment significantly decreased plasma lactate dehydrogenase,CK-MB,cardiac troponin T,BNP,and IL-10,as well as cardiac TGF-β1,VEGF,and MDA while increasing p-Akt and superoxide dismutase in ISO-administered rats.It also reduced NLRP3 and mRNA-208a gene expression levels.Furthermore,naringenin improved ISO-induced cardiac damage.Conclusions:Naringenin attenuates myocardial dysfunction in ISO-treated rats by decreasing oxidative stress and increasing cardiac endogenous antioxidant system,which may be modulated partly by improvement of NLRP3 and mRNA-208a gene expression.展开更多
Wheat is one of the major food crops in the world.Functional validation of the genes in increasing the grain yield of wheat by genetic engineering is essential for feeding the ever-growing global population.This study...Wheat is one of the major food crops in the world.Functional validation of the genes in increasing the grain yield of wheat by genetic engineering is essential for feeding the ever-growing global population.This study investigated the role of ABP7,a bHLH transcription factor from maize involved in kernel development,in regulating grain yield-related traits in transgenic wheat.Molecular characterization showed that transgenic lines HB123 and HB287 contained multicopy integration of ABP7 in the genome with higher transgene expression.At the same time,QB205 was a transgenic event of single copy insertion with no significant difference in ABP7 expression compared to wild-type(WT) plants.Phenotyping under field conditions showed that ABP7 over-expressing transgenic lines HB123 and HB287 exhibited improved grain yield-related traits(e.g.,grain number per spike,grain weight per spike,thousand-grain weight,grain length,and grain width) and increased grain yield per plot,compared to WT plants,whereas line QB205 did not.In addition,total chlorophyll,chlorophyll a,chlorophyll b,and total soluble sugars were largely increased in the flag leaves of both HB123and HB287 transgenic lines compared to the WT.These results strongly suggest that ABP7 positively regulates yieldrelated traits and plot grain yield in transgenic wheat.Consequently,ABP7 can be utilized in wheat breeding for grain yield improvement.展开更多
The genetic pathways of rice seedling growth have a major impact on seedling emergence from soil and development.In this study,we identified a new bHLH transcription factor,BEAR1,from rice RNAi mutant library.Both the...The genetic pathways of rice seedling growth have a major impact on seedling emergence from soil and development.In this study,we identified a new bHLH transcription factor,BEAR1,from rice RNAi mutant library.Both the BEAR1-RNAi and bear1 CRISPR mutants had accelerated seedling growth.Histological section of leaves showed accelerated development of lacuna and vascular bundles in bear1 mutant.GUS staining revealed that BEAR1 was highly expressed in coleoptiles and leaves at seedling stage.Expression analysis of gibberellin(GA)biosynthesis and metabolic genes and content determination of active GAs indicated that the expression of GA biosynthesis genes,especially OsKS4 and OsCPS2,were upregulated and the GAs content were significantly increased in bear1,which correlated with the seedling phenotype of bear1 mutant.Molecular and biochemical assays revealed that BEAR1 directly binds to the promoter of OsKS4,thereby repressing its expression.Haplotypes analysis showed clear differentiation in indica and japonica rice varieties,and a strong correlation with seedling height.These findings provide novel insights into the regulation of seedling growth in rice.展开更多
Foxtail millet(Setaria italica)is an important C4 model crop;however,due to its high-density planting and high stature,lodging at the filling stage resulted in a serious reduction in yield and quality.Therefore,it is ...Foxtail millet(Setaria italica)is an important C4 model crop;however,due to its high-density planting and high stature,lodging at the filling stage resulted in a serious reduction in yield and quality.Therefore,it is imperative to identify and deploy the genes controlling foxtail millet plant height.In this study,we used a semi-dwarf line 263A and an elite high-stalk breeding variety,Chuang 29 to construct an F2 population to identify dwarf genes.We performed transcriptome analysis(RNA-seq)using internode tissues sampled at three jointing stages of 263A and Chuang 29,as well as bulk segregant analysis(BSA)on their F2 population.A total of 8918 differentially expressed genes(DEGs)were obtained from RNA-seq analysis,and GO analysis showed that DEGs were enriched in functions such as‘‘gibberellin metabolic process”and‘‘oxidoreductase activity”,which have previously been shown to be associated with plant height.A total 593 mutated genes were screened by BSA-seq method.One hundred and seventy-six out of the 593 mutated genes showed differential expression levels between the two parental lines,and seven genes not only showed differential expression in two or three internode tissues but also showed high genomic variation in coding regions,which indicated they play a crucial role in plant height determination.Among them,we found a gibberellin biosynthesis related GA20 oxidase gene(Seita.5G404900),which had a single-base at the third exon,leading to the frameshift mutation at 263A.Cleaved amplified polymorphic sequence assay and association analysis proved the single-base in Seita.5G404900 co-segregated with dwarf phenotype in two independent F2 populations planted in entirely different environments.Taken together,the candidate genes identified in this study will help to elucidate the genetic basis of foxtail millet plant height,and the molecular marker will be useful for marker-assisted dwarf breeding.展开更多
Barley(Hordeum vulgare L.)is an important economic crop for food,feed and industrial raw materials.In the present research,112 barley landraces from the Shanghai region were genotyped using genotyping-by-sequencing(GB...Barley(Hordeum vulgare L.)is an important economic crop for food,feed and industrial raw materials.In the present research,112 barley landraces from the Shanghai region were genotyped using genotyping-by-sequencing(GBS),and the genetic diversity and population structure were analyzed.The results showed that 210,268 Single Nucleotide Polymorphisms(SNPs)were present in total,and the average poly-morphism information content(PIC)was 0.1642.Genetic diversity and population structure analyses suggested that these barley landraces were differentiated and could be divided into three sub-groups,with morphological traits of row-type and adherence of the hulls the main distinguishing factors between groups.Genotypes with similar or duplicated names were also investigated according to their genetic backgrounds and seed appearances.This study provided valuable information on barley landraces from the Shanghai region,and showed that all these barley landraces should be protected and used for future breeding programs.展开更多
Colorectal carcinoma(CRC)is one such dispersed cancer globally and also prominent one in causing cancer-based death.Conventionally,pathologists execute CRC diagnosis through visible scrutinizing under the microscope t...Colorectal carcinoma(CRC)is one such dispersed cancer globally and also prominent one in causing cancer-based death.Conventionally,pathologists execute CRC diagnosis through visible scrutinizing under the microscope the resected tissue samples,stained and fixed through Haematoxylin and Eosin(H&E).The advancement of graphical processing systems has resulted in high potentiality for deep learning(DL)techniques in interpretating visual anatomy from high resolution medical images.This study develops a slime mould algorithm with deep transfer learning enabled colorectal cancer detection and classification(SMADTL-CCDC)algorithm.The presented SMADTL-CCDC technique intends to appropriately recognize the occurrence of colorectal cancer.To accomplish this,the SMADTLCCDC model initially undergoes pre-processing to improve the input image quality.In addition,a dense-EfficientNet technique was employed to extract feature vectors from the pre-processed images.Moreover,SMA with Discrete Hopfield neural network(DHNN)method was applied for the recognition and classification of colorectal cancer.The utilization of SMA assists in appropriately selecting the parameters involved in the DHNN approach.A wide range of experiments was implemented on benchmark datasets to assess the classification performance.A comprehensive comparative study highlighted the better performance of the SMADTL-CDC model over the recent approaches.展开更多
A healthy balanced diet and a healthy lifestyle are very closely linked.Whichever the biological link is,it is overwhelming to understand.Modifications in how food is served,divided up,and supervised,such as the intro...A healthy balanced diet and a healthy lifestyle are very closely linked.Whichever the biological link is,it is overwhelming to understand.Modifications in how food is served,divided up,and supervised,such as the introduction of nutritional hygiene standards,food handling practices,and the entry of macro and micronutrients,have had a big impact on human health in the last few decades.Growing evidence indicates that our gut microbiota may affect our health in ways that are at least in part influenced by our diet and the ingredients used in the preparation of our food and drinks,as well as other factors.As a new problem,this one is getting a lot of attention,but it would be hard to figure out how the gut microbiota and nutrition molecules work together and how they work in certain situations.Genetic analysis,metagenomic characterization,configuration analysis of foodstuffs,and the shift to digital health information have provided massive amounts of data that might be useful in tackling this problem.Machine learning and deep learning methods will be employed extensively as part of this research in order to blend complicated data frames and extract crucial information that will be capable of exposing and grasping the incredibly delicate links that prevail between diet,gut microbiome,and overall wellbeing.Nutrition,well-being,and gut microorganisms are a few subjects covered in this field.It takes into account not only databases and high-speed technology,but also virtual machine problem-solving skills,intangible assets,and laws.This is how it works:Computer vision,data mining,and analytics are all discussed extensively in this study piece.We also point out limitations in existing methodologies and new situations that discovered in the context of current scientific knowledge in the decades to come.We also provide background on"bioinformatics"algorithms;recent developments may seem to herald a revolution in clinical research,pushing traditional techniques to the sidelines.Furthermore,their true potential rests in their ability to work in conjunction with,rather than as a substitute for,traditional research hypotheses and procedures.When new metadata propositions are made by focusing on easily understandable frameworks,they will always need to be rigorously validated and brought into question.Because of the huge datasets available,assumption analysis may be used to complement rather than a substitute for more conventional concept-driven scientific investigation.It is only by employing all of us that we will all increase the quality of evidence-based practice.展开更多
Chimeric plants composed of green and albino tissues have great ornamental value.To unveil the functional genes responsible for albino phenotypes in chimeric plants,we inspected the complete plastid genomes(plastomes)...Chimeric plants composed of green and albino tissues have great ornamental value.To unveil the functional genes responsible for albino phenotypes in chimeric plants,we inspected the complete plastid genomes(plastomes)in green and albino leaf tissues from 23 ornamental chimeric plants belonging to 20 species,including monocots,dicots,and gymnosperms.In nine chimeric plants,plastomes were identical between green and albino tissues.Meanwhile,another 14 chimeric plants were heteroplasmic,showing a mutation between green and albino tissues.We identified 14 different point mutations in eight functional plastid genes related to plastid-encoded RNA polymerase(rpo)or photosystems which caused albinism in the chimeric plants.Among them,12 were deleterious mutations in the target genes,in which early termination appeared due to small deletion-mediated frameshift or single nucleotide substitution.Another was single nucleotide substitution in an intron of the ycf3 and the other was a missense mutation in coding region of the rpoC2 gene.We inspected chlorophyll structure,protein functional model of the rpoC2,and expression levels of the related genes in green and albino tissues of Reynoutria japonica.A single amino acid change,histidine-to-proline substitution,in the rpoC2 protein may destabilize the peripheral helix of plastid-encoded RNA polymerase,impairing the biosynthesis of the photosynthesis system in the albino tissue of R.japonica chimera plant.展开更多
Aim: To detect the expression of hepatitis B virus (HBV) genes (HB S and C genes) in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fer...Aim: To detect the expression of hepatitis B virus (HBV) genes (HB S and C genes) in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization (IVF) technique. Methods: Human sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method. Polymerase chain reaction (PCR) was used to detect HB S and pre-Core/Core (pre-C/C) coding genes both in one- and two-cell embryos. Reverse transcription-PCR (RT-PCR) analysis was used to study the expression of the two genes. Fluorescence in situ hybridization (FISH) analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome. Results: Both HB S and pre-C/C coding genes are present and transcribed in one- and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences. Conclusion: Sperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells. These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line.展开更多
Objective:To detect the in vitro total phenolics,flavonoids contents and antioxidant activity of essential oil,various organic extracts from the leaves of tropical medicinal plant Tetrastigma from Sabah.Methods:The dr...Objective:To detect the in vitro total phenolics,flavonoids contents and antioxidant activity of essential oil,various organic extracts from the leaves of tropical medicinal plant Tetrastigma from Sabah.Methods:The dry powder leaves of Tetrastigma were extracted with different organic solvent such as hexane.ethyl acetate,chloroform,butanol and aqueous methanol.The total phenolic and total flavonoids contents of the essential oil and various organic extracts such as hexane,ethyl acetate,chloroform,butanol and aqueous ethanol were determined by Folin-Ciocalteu method and the assayed antioxidant activity was determined in vitro models such as antioxidant capacity by radical scavenging activity usingα,α-diphenyl-β-picrylhydrazyl (DPPH) method.Results:The total phenolic contents of the essential oil and different extracts as gallic acid equivalents were found to be highest in methanol extract(386.22 mg/g) followed by ethyl acetate(190.89 mg/g).chloroform(175.89 mg/g).hexane(173.44 mg/g).and Imtanol extract (131.72 mg/g) and the phenolic contents not detected in essential oil.The antioxidant capacity of the essential oil and different extracts as ascorbic acid standard was in the order of methanol extract】ethyl acetate extract】chloroform】butanol】hexane extract also the antioxidant activity was not detected in essential oil.Conclusions:The findings show that the extent of antioxidant activity of the essential oil and all extracts are in accordance with the amount of phenolics present in that extract.Leaves of Tetrastigma being rich in phenolics may provide a good source of antioxidant.展开更多
基金supported by the National Key Research and Development Program of China(NKRDP)(2022YFF1001700)the Agricultural Science and Technology Innovation Program(2020YFE0202300)the National Natural Science Foundation of China(31871313)。
文摘Plant anatomy is patterned early during leaf development which suggests studying the spatial–temporal transcriptomes of primordia will help identify critical regulative and functional genes.We successfully isolated the leaf primordia tissues from the C3grass rice and the C4grass foxtail millet by laser capture microdissection(LCM)and studied the gene expression throughout leaf developmental stages.Our data analysis uncovered the conserved expression patterns of certain gene clusters both in rice and foxtail millet during leaf development.We revealed genes and transcription factors involved in vein formation,stomatal development,and suberin accumulation.We identified 79 candidate genes associated with functional regulation of C4anatomy formation.Screening phenotype of the candidate genes revealed that knock-out of a putative polar auxin transport related gene NAL1 resulted significantly reduced veinal space in rice leaf.Our present work provides a foundation for future analyses of genes with novel functions in grasses and their role in leaf development,in particular the role in leaves with a contrasting C3vs.C4biosynthetic pathway.
基金supported by National Natural Science Foundation of China(Grant No.32072563)Key Research and Development Projects of Ningxia Hui Autonomous Region(Grant No.2022BBF02008)+2 种基金Key Project Fund of the Shanghai Municipal Committee of Agriculture(Grant Nos.2021-02-08-00-12-F00795,Chuangzi 20181e5)Shanghai Academic Technology Research Leader(Grant No.19XD1432300)Priority Academic Program Development of Jiangsu Higher Education Institutions Project(PAPD)。
文摘Betalain has received increased attention because of its high nutritional value and crucial physiological functions.Based on the elucidation of its core biosynthetic pathway,betalain can be produced in additional plants by metabolic engineering.Synthesis of betalain in carrot(Daucus carota L.)can improve its nutritional quality and economic value by extracting betalain from the fleshy root,non-edible part,and processing residue of carrot.In this study,two different constructs,namely,pYB:mCD(AomelOS,BvCYP76AD1S,and BvDODA1S)and p YB:CDD(BvCYP76AD1S,BvDODA1S,and MjcDOPA5GTS),were introduced into carrot for betanin synthesis by Agrobacterium-mediated transformation.Betanin can be synthetized in both transgenic calli,and p YB:m CD-transgenic callus can be used to produce betacyanin by suspension culture.However,pYB:mCD-transgenic seedlings can synthetize betanin only by tyrosine feeding.The p YB:CDD-transgenic lines can synthetize betanin in whole plants.The betanin content in fleshy root of pYB:CDD-transgenic carrot was(63.4±9)μg·g^(-1)fresh weight according to quantitative analysis.These betanin-producing carrot plant materials can be used to synthesize betanin for industrial application or consumption as dietary sources.
基金Major Scientific and Technological Projects in Agricultural Biological Breeding of China(2023ZD0404302)Youth Program of National Natural Science Foundation of China(32202754)。
文摘Porcine reproductive and respiratory syndrome(PRRS)is a globally prevalent contagious disease caused by the positive-strand RNA PRRS virus(PRRSV),resulting in substantial economic losses in the swine industry.Modifying the CD163 SRCR5 domain,either through deletion or substitution,can eff1ectively confer resistance to PRRSV infection in pigs.However,large fragment modifications in pigs inevitably raise concerns about potential adverse effects on growth performance.Reducing the impact of genetic modifications on normal physiological functions is a promising direction for developing PRRSV-resistant pigs.In the current study,we identified a specific functional amino acid in CD163 that influences PRRSV proliferation.Viral infection experiments conducted on Marc145 and PK-15CD163 cells illustrated that the mE535G or corresponding pE529G mutations markedly inhibited highly pathogenic PRRSV(HP-PRRSV)proliferation by preventing viral binding and entry.Furthermore,individual viral challenge tests revealed that pigs with the E529G mutation had viral loads two orders of magnitude lower than wild-type(WT)pigs,confirming effective resistance to HP-PRRSV.Examination of the physiological indicators and scavenger function of CD163 verified no significant differences between the WT and E529G pigs.These findings suggest that E529G pigs can be used for breeding PRRSV-resistant pigs,providing novel insights into controlling future PRRSV outbreaks.
基金This study was supported by the Ministry of Higher Education,Malaysia(FRGS0322-SG-1/2013)Universiti Malaysia Sabah(GUG0521-2/2020).
文摘Objective:To determine the genetic diversity of Plasmodium(P.)knowlesi isolates from Sabah,Malaysian Borneo and Peninsular Malaysia,targeting the S-type SSU rRNA gene and including aspects of natural selection and haplotype.Methods:Thirty-nine blood samples infected with P.knowlesi were collected in Sabah,Malaysian Borneo and Peninsular Malaysia.The S-type SSU rRNA gene was amplified using polymerase chain reaction,cloned into a vector,and sequenced.The natural selection and haplotype of the S-type SSU rRNA gene sequences were determined using DnaSP v6 and illustrated using NETWORK v10.This study's 39 S-type SSU rRNA sequences and eight sequences from the Genbank database were subjected to phylogenetic analysis using MEGA 11.Results:Overall,the phylogenetic analysis showed no evidence of a geographical cluster of P.knowlesi isolates from different areas in Malaysia based on the S-type SSU rRNA gene sequences.The S-type SSU rRNA gene sequences were relatively conserved and with a purifying effect.Haplotype sharing of the S-type SSU rRNA gene was observed between the P.knowlesi isolates in Sabah,Malaysian Borneo,but not between Sabah,Malaysian Borneo and Peninsular Malaysia.Conclusions:This study suggests that the S-type SSU rRNA gene of P.knowlesi isolates in Sabah,Malaysian Borneo,and Peninsular Malaysia has fewer polymorphic sites,representing the conservation of the gene.These features make the S-type SSU rRNA gene suitable for comparative studies,such as determining the evolutionary relationships and common ancestry among P.knowlesi species.
基金partially supported by the Key Program of the National Natural Science of Fujian Province(2022J02010)Science and Technology Project of State Administration of Science,Technology and Industry for National Defense,Seed Innovation and Industrialization Project of Fujian Province(zycxny2021003)Fujian Province Public Welfare Scientific Research Program(2016R1018-8)。
文摘Florets are the basic structural units of spikelets,and their morphogenesis determines the yield and quality of rice grains.However,whether and how pseudouridine-5’-phosphate glycosidase participates in rice spikelet development remains an open question.In this study,we identified a novel gene,OsPPG,which encodes a peroxisome-localized pseudouridine-5’-phosphate glycosidase and regulates the development of rice spikelets.osppg mutants exhibited abnormal sterile lemma,lemma,palea,lodicule,stamens,and pistils;male sterility;shorter panicles;and reduced plant height.OsPPG was found to regulate several OsMADS genes,thereby affecting the morphogenesis of rice spikelets.Furthermore,metabolomics revealed that the OsPPG gene was involved in the decomposition of pseudouridine via the pyrimidine metabolism pathway and may affect the jasmonic acid signaling pathway.These results suggest that OsPPG is a key regulator of rice spikelet development.
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA26030301)Hohhot Key R&D Project(2023-JBGSS-1),the National Natural Science Foundation of China(U23A200206,32071864,32325035)+1 种基金the Taishan Scholar Program of Shandong(to Chunxiang Fu)the Shandong Provincial Natural Science Foundation(ZR202210270038)。
文摘Alfalfa(Medicago sativa.L.)is a globally significant autotetraploid legume forage crop.However,despite its importance,establishing efficient gene editing systems for cultivated alfalfa remains a formidable challenge.In this study,we pioneered the development of a highly effective ultrasonic-assisted leaf disc transformation system for Gongnong 1 alfalfa,a variety widely cultivated in Northeast China.Subsequently,we created a single transcript CRISPR/Cas9(CRISPR_2.0)toolkit,incorporating multiplex gRNAs,designed for gene editing in Gongnong 1.Both Cas9 and gRNA scaffolds were under the control of the Arabidopsis ubiquitin-10 promoter,a widely employed polymeraseⅡconstitutive promoter known for strong transgene expression in dicots.To assess the toolkit’s efficiency,we targeted PALM1,a gene associated with a recognizable multifoliate phenotype.Utilizing the CRISPR_2.0 toolkit,we directed PALM1 editing at two sites in the wild-type Gongnong 1.Results indicated a 35.1%occurrence of editing events all in target 2 alleles,while no mutations were detected at target 1 in the transgenic-positive lines.To explore more efficient sgRNAs,we developed a rapid,reliable screening system based on Agrobacterium rhizogenes-mediated hairy root transformation,incorporating the visible reporter MtLAP1.This screening system demonstrated that most purple visible hairy roots underwent gene editing.Notably,sgRNA3,with an 83.0%editing efficiency,was selected using the visible hairy root system.As anticipated,tetra-allelic homozygous palm1 mutations exhibited a clear multifoliate phenotype.These palm1 lines demonstrated an average crude protein yield increase of 21.5%compared to trifoliolate alfalfa.Our findings highlight the modified CRISPR_2.0 system as a highly efficient and robust gene editing tool for autotetraploid alfalfa.
基金supported by the National Natural Science Foundation of China(32001532 and 31860411)the National Key Research and Development Program of China,(2022YFF1000020)+1 种基金Hunan Seed Industry Innovation Project(2021NK1012)the Yunnan Tobacco Company Project(2020530000241009)。
文摘The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.
基金supported by the National Natural Science Foundation of China(32060476 and 31860371)Guangxi Department of Science and Technology(AA22068087-4)+3 种基金Guangxi Natural Science Foundation of China(2015GXNSFAA139054,2018GXNSFAA138124,and 2020GXNSFAA259041)Guangxi Ministry of Science and Technology(AB21238009)Special Fund of Local Science and Technology Development for the Central Guidance(ZY21195034)Guangxi Academy of Agricultural Sciences(2021JM04,2021JM49,2021YT030,QN-25,and QN-35)。
文摘The overuse of nitrogen(N)fertilizer in fields has increased production costs and raised environmental concerns.Increasing the N use efficiency(NUE)of rice varieties is crucial for sustainable agriculture.Here we report the cloning and characterization of OsNPF3.1,a gene that controls rice NUE.An amino acid mutation in the OsNPF3.1 coding region caused different NUEs in wild and cultivated rice.OsNPF3.1,which is expressed mainly in the aerial parts of rice,also affects rice plant height,heading date,and thousand-grain weight.The OsNPF3.1 protein is located in the plasma membrane.When OsNPF3.1 was subjected to artificial selection,two naturally varying loci were associated with NUE,of which OsNPF3.1Chr6_8741040differed between indica and japonica rice.OsNPF3.1 can be used as a new target gene for breeding rice varieties with high NUE.
基金supported by the National Natural Science Foundation of China(Grant No.31701803)Changsha Natural Science Foundation,China(Grant No.kq2202336)the Special Project of Hunan Innovative Province Construction,China(Grant No.S2021ZCKPZT0004)。
文摘Despite increasing knowledge of barnyardgrass(Echinochloa crus-galli) interference with rice, relatively little is known how endophytes improve the ability of rice against barnyardgrass stress. Here, we provided a detailed temporal characterization of rice root-associated microbiomes during co-cultivation with barnyardgrass and a comparison with the microbiomes of weed-free rice plants. Alpha diversity analysis indicated that barnyardgrass had the opposite effects on endophytic bacteria and fungi in rice roots, in terms of the community diversity, richness and coverage at the rice seedling stage. Principal coordinate analysis showed that barnyardgrass had only a minor effect on the community composition of endophytes in rice roots at the rice seedling stage, but showed a significant and maximum interference at the heading stage. Rice recruited many endophytes to resist biotic stress from barnyardgrass, especially for fungi. PICRUSt(phylogenetic investigation of communities by reconstruction of unobserved states) predictive analysis indicated that 23 metabolic pathways of bacteria were overrepresented in rice. In addition, the main trophic mode of fungi was pathotroph according to FUNGuild analysis. A positive correlation between bacteria and fungi in rice roots was found via network analysis. Anaeromyxobacter, Azospira and Pseudolabrys were the vital bacteria, Phaeosphaeria and Funneliformis were the dominant fungi in maintaining the stability of the ecological network. These results provided data and a theoretical basis for the in-depth understanding of what role endophytes play in rice resistance to barnyardgrass stress and will have implications on improving the resistance of rice against biotic stress using root microbiota.
文摘Objective:To investigate the cardioprotective effect of naringenin against isoproterenol(ISO)-induced cardiotoxicity in rats.Methods:Rats were divided into five groups:the normal group,the ISO group(85 mg/kg b.w.);the ISO+naringenin(50 mg/kg b.w.)group,the ISO+naringenin(100 mg/kg b.w.)group and the ISO+propranolol(10 mg/kg b.w.)group.Plasma creatine kinase-MB(CK-MB),cardiac troponin T,lactate dehydrogenase,brain natriuretic peptide(BNP),and IL-10,as well as cardiac transforming growth factor-β1(TGF-β1),vascular endothelial growth factor(VEGF)and malondialdehyde(MDA)were examined.In addition,NLRP3 and mRNA-208a expressions were evaluated by RT-PCR analysis.Histopathological examination was also performed to assess cardiac damages.Results:Naringenin treatment significantly decreased plasma lactate dehydrogenase,CK-MB,cardiac troponin T,BNP,and IL-10,as well as cardiac TGF-β1,VEGF,and MDA while increasing p-Akt and superoxide dismutase in ISO-administered rats.It also reduced NLRP3 and mRNA-208a gene expression levels.Furthermore,naringenin improved ISO-induced cardiac damage.Conclusions:Naringenin attenuates myocardial dysfunction in ISO-treated rats by decreasing oxidative stress and increasing cardiac endogenous antioxidant system,which may be modulated partly by improvement of NLRP3 and mRNA-208a gene expression.
文摘Wheat is one of the major food crops in the world.Functional validation of the genes in increasing the grain yield of wheat by genetic engineering is essential for feeding the ever-growing global population.This study investigated the role of ABP7,a bHLH transcription factor from maize involved in kernel development,in regulating grain yield-related traits in transgenic wheat.Molecular characterization showed that transgenic lines HB123 and HB287 contained multicopy integration of ABP7 in the genome with higher transgene expression.At the same time,QB205 was a transgenic event of single copy insertion with no significant difference in ABP7 expression compared to wild-type(WT) plants.Phenotyping under field conditions showed that ABP7 over-expressing transgenic lines HB123 and HB287 exhibited improved grain yield-related traits(e.g.,grain number per spike,grain weight per spike,thousand-grain weight,grain length,and grain width) and increased grain yield per plot,compared to WT plants,whereas line QB205 did not.In addition,total chlorophyll,chlorophyll a,chlorophyll b,and total soluble sugars were largely increased in the flag leaves of both HB123and HB287 transgenic lines compared to the WT.These results strongly suggest that ABP7 positively regulates yieldrelated traits and plot grain yield in transgenic wheat.Consequently,ABP7 can be utilized in wheat breeding for grain yield improvement.
基金supported by the National Natural Science Foundation of China(32071932)Scientific Research Foundation for Scholars of HZNU(2019QDL015)the Natural Science Foundation of Zhejiang province(LQ22C130001)。
文摘The genetic pathways of rice seedling growth have a major impact on seedling emergence from soil and development.In this study,we identified a new bHLH transcription factor,BEAR1,from rice RNAi mutant library.Both the BEAR1-RNAi and bear1 CRISPR mutants had accelerated seedling growth.Histological section of leaves showed accelerated development of lacuna and vascular bundles in bear1 mutant.GUS staining revealed that BEAR1 was highly expressed in coleoptiles and leaves at seedling stage.Expression analysis of gibberellin(GA)biosynthesis and metabolic genes and content determination of active GAs indicated that the expression of GA biosynthesis genes,especially OsKS4 and OsCPS2,were upregulated and the GAs content were significantly increased in bear1,which correlated with the seedling phenotype of bear1 mutant.Molecular and biochemical assays revealed that BEAR1 directly binds to the promoter of OsKS4,thereby repressing its expression.Haplotypes analysis showed clear differentiation in indica and japonica rice varieties,and a strong correlation with seedling height.These findings provide novel insights into the regulation of seedling growth in rice.
基金supported by the National Key Research and Development Program of China (2018YFD1000702/ 2018YFD1000700)the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural SciencesOperating Expenses for Basic Scientific Research of Institute of Crop Science, Chinese Academy of Agricultural Sciences
文摘Foxtail millet(Setaria italica)is an important C4 model crop;however,due to its high-density planting and high stature,lodging at the filling stage resulted in a serious reduction in yield and quality.Therefore,it is imperative to identify and deploy the genes controlling foxtail millet plant height.In this study,we used a semi-dwarf line 263A and an elite high-stalk breeding variety,Chuang 29 to construct an F2 population to identify dwarf genes.We performed transcriptome analysis(RNA-seq)using internode tissues sampled at three jointing stages of 263A and Chuang 29,as well as bulk segregant analysis(BSA)on their F2 population.A total of 8918 differentially expressed genes(DEGs)were obtained from RNA-seq analysis,and GO analysis showed that DEGs were enriched in functions such as‘‘gibberellin metabolic process”and‘‘oxidoreductase activity”,which have previously been shown to be associated with plant height.A total 593 mutated genes were screened by BSA-seq method.One hundred and seventy-six out of the 593 mutated genes showed differential expression levels between the two parental lines,and seven genes not only showed differential expression in two or three internode tissues but also showed high genomic variation in coding regions,which indicated they play a crucial role in plant height determination.Among them,we found a gibberellin biosynthesis related GA20 oxidase gene(Seita.5G404900),which had a single-base at the third exon,leading to the frameshift mutation at 263A.Cleaved amplified polymorphic sequence assay and association analysis proved the single-base in Seita.5G404900 co-segregated with dwarf phenotype in two independent F2 populations planted in entirely different environments.Taken together,the candidate genes identified in this study will help to elucidate the genetic basis of foxtail millet plant height,and the molecular marker will be useful for marker-assisted dwarf breeding.
基金funded by the Shanghai Agriculture Applied Technology Development Program(Grant No.2019-02-08-00-08-F01109)the Climbing Plan(Grant No.PG22211)and the Ear-Marked Fund for CARS(Grant No.CARS-05-01A-02)N.G.H.is supported at Rothamsted Research by the Biotechnology and Biological Sciences Research Council(BBSRC)via the Designing Future Wheat Programme(BB/P016855/1).
文摘Barley(Hordeum vulgare L.)is an important economic crop for food,feed and industrial raw materials.In the present research,112 barley landraces from the Shanghai region were genotyped using genotyping-by-sequencing(GBS),and the genetic diversity and population structure were analyzed.The results showed that 210,268 Single Nucleotide Polymorphisms(SNPs)were present in total,and the average poly-morphism information content(PIC)was 0.1642.Genetic diversity and population structure analyses suggested that these barley landraces were differentiated and could be divided into three sub-groups,with morphological traits of row-type and adherence of the hulls the main distinguishing factors between groups.Genotypes with similar or duplicated names were also investigated according to their genetic backgrounds and seed appearances.This study provided valuable information on barley landraces from the Shanghai region,and showed that all these barley landraces should be protected and used for future breeding programs.
基金the Deanship of Scientific Research(DSR),King Abdulaziz University,Jeddah,under grant No.(FP-000-000-1441).
文摘Colorectal carcinoma(CRC)is one such dispersed cancer globally and also prominent one in causing cancer-based death.Conventionally,pathologists execute CRC diagnosis through visible scrutinizing under the microscope the resected tissue samples,stained and fixed through Haematoxylin and Eosin(H&E).The advancement of graphical processing systems has resulted in high potentiality for deep learning(DL)techniques in interpretating visual anatomy from high resolution medical images.This study develops a slime mould algorithm with deep transfer learning enabled colorectal cancer detection and classification(SMADTL-CCDC)algorithm.The presented SMADTL-CCDC technique intends to appropriately recognize the occurrence of colorectal cancer.To accomplish this,the SMADTLCCDC model initially undergoes pre-processing to improve the input image quality.In addition,a dense-EfficientNet technique was employed to extract feature vectors from the pre-processed images.Moreover,SMA with Discrete Hopfield neural network(DHNN)method was applied for the recognition and classification of colorectal cancer.The utilization of SMA assists in appropriately selecting the parameters involved in the DHNN approach.A wide range of experiments was implemented on benchmark datasets to assess the classification performance.A comprehensive comparative study highlighted the better performance of the SMADTL-CDC model over the recent approaches.
文摘A healthy balanced diet and a healthy lifestyle are very closely linked.Whichever the biological link is,it is overwhelming to understand.Modifications in how food is served,divided up,and supervised,such as the introduction of nutritional hygiene standards,food handling practices,and the entry of macro and micronutrients,have had a big impact on human health in the last few decades.Growing evidence indicates that our gut microbiota may affect our health in ways that are at least in part influenced by our diet and the ingredients used in the preparation of our food and drinks,as well as other factors.As a new problem,this one is getting a lot of attention,but it would be hard to figure out how the gut microbiota and nutrition molecules work together and how they work in certain situations.Genetic analysis,metagenomic characterization,configuration analysis of foodstuffs,and the shift to digital health information have provided massive amounts of data that might be useful in tackling this problem.Machine learning and deep learning methods will be employed extensively as part of this research in order to blend complicated data frames and extract crucial information that will be capable of exposing and grasping the incredibly delicate links that prevail between diet,gut microbiome,and overall wellbeing.Nutrition,well-being,and gut microorganisms are a few subjects covered in this field.It takes into account not only databases and high-speed technology,but also virtual machine problem-solving skills,intangible assets,and laws.This is how it works:Computer vision,data mining,and analytics are all discussed extensively in this study piece.We also point out limitations in existing methodologies and new situations that discovered in the context of current scientific knowledge in the decades to come.We also provide background on"bioinformatics"algorithms;recent developments may seem to herald a revolution in clinical research,pushing traditional techniques to the sidelines.Furthermore,their true potential rests in their ability to work in conjunction with,rather than as a substitute for,traditional research hypotheses and procedures.When new metadata propositions are made by focusing on easily understandable frameworks,they will always need to be rigorously validated and brought into question.Because of the huge datasets available,assumption analysis may be used to complement rather than a substitute for more conventional concept-driven scientific investigation.It is only by employing all of us that we will all increase the quality of evidence-based practice.
基金This work was supported by the Bio&Medical Technology Development Program of the NRF,MSIP,Republic of Korea(grant no.NRF-2015M3A9A5030733)grants from the Nuclear R&D Program of the Ministry of Science and ICT(MSIT)and the research program of KAERI,Republic of Korea.
文摘Chimeric plants composed of green and albino tissues have great ornamental value.To unveil the functional genes responsible for albino phenotypes in chimeric plants,we inspected the complete plastid genomes(plastomes)in green and albino leaf tissues from 23 ornamental chimeric plants belonging to 20 species,including monocots,dicots,and gymnosperms.In nine chimeric plants,plastomes were identical between green and albino tissues.Meanwhile,another 14 chimeric plants were heteroplasmic,showing a mutation between green and albino tissues.We identified 14 different point mutations in eight functional plastid genes related to plastid-encoded RNA polymerase(rpo)or photosystems which caused albinism in the chimeric plants.Among them,12 were deleterious mutations in the target genes,in which early termination appeared due to small deletion-mediated frameshift or single nucleotide substitution.Another was single nucleotide substitution in an intron of the ycf3 and the other was a missense mutation in coding region of the rpoC2 gene.We inspected chlorophyll structure,protein functional model of the rpoC2,and expression levels of the related genes in green and albino tissues of Reynoutria japonica.A single amino acid change,histidine-to-proline substitution,in the rpoC2 protein may destabilize the peripheral helix of plastid-encoded RNA polymerase,impairing the biosynthesis of the photosynthesis system in the albino tissue of R.japonica chimera plant.
基金Acknowledgment This work was supported by a grant from the National Nature Science Foundation of China (No. 39970374). The authors wish to thank Professor Yi-Pong Hu, Second Military Medical University of China, for his kindness in providing us the recombinant plasmid (pBR322-HBV). We wish to thank Mr. Michael Talion of Shantou University Medical College, English Language Training Section for his assistance in proofreading this manuscript. We gratefully acknowledge the support of the leaders of Shantou University Medical College.
文摘Aim: To detect the expression of hepatitis B virus (HBV) genes (HB S and C genes) in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization (IVF) technique. Methods: Human sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method. Polymerase chain reaction (PCR) was used to detect HB S and pre-Core/Core (pre-C/C) coding genes both in one- and two-cell embryos. Reverse transcription-PCR (RT-PCR) analysis was used to study the expression of the two genes. Fluorescence in situ hybridization (FISH) analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome. Results: Both HB S and pre-C/C coding genes are present and transcribed in one- and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences. Conclusion: Sperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells. These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line.
基金Biotechnology Research Institute,University Malaysia Sabah,Malaysia for financial support of this entire work
文摘Objective:To detect the in vitro total phenolics,flavonoids contents and antioxidant activity of essential oil,various organic extracts from the leaves of tropical medicinal plant Tetrastigma from Sabah.Methods:The dry powder leaves of Tetrastigma were extracted with different organic solvent such as hexane.ethyl acetate,chloroform,butanol and aqueous methanol.The total phenolic and total flavonoids contents of the essential oil and various organic extracts such as hexane,ethyl acetate,chloroform,butanol and aqueous ethanol were determined by Folin-Ciocalteu method and the assayed antioxidant activity was determined in vitro models such as antioxidant capacity by radical scavenging activity usingα,α-diphenyl-β-picrylhydrazyl (DPPH) method.Results:The total phenolic contents of the essential oil and different extracts as gallic acid equivalents were found to be highest in methanol extract(386.22 mg/g) followed by ethyl acetate(190.89 mg/g).chloroform(175.89 mg/g).hexane(173.44 mg/g).and Imtanol extract (131.72 mg/g) and the phenolic contents not detected in essential oil.The antioxidant capacity of the essential oil and different extracts as ascorbic acid standard was in the order of methanol extract】ethyl acetate extract】chloroform】butanol】hexane extract also the antioxidant activity was not detected in essential oil.Conclusions:The findings show that the extent of antioxidant activity of the essential oil and all extracts are in accordance with the amount of phenolics present in that extract.Leaves of Tetrastigma being rich in phenolics may provide a good source of antioxidant.