ObjectiveTo investigate the gene expression changes in normal and degeneration lumbar intervertebral disc in humans, providing information for clinical. MethodsThe PCR products of 4096 human genes were spotted onto a ...ObjectiveTo investigate the gene expression changes in normal and degeneration lumbar intervertebral disc in humans, providing information for clinical. MethodsThe PCR products of 4096 human genes were spotted onto a kind of chemical-material-coated-glass slides. The total RNAs were isolated from the tissues. Both the mRNAs from the degeneration and normal lumbar intervertebral disc in humans were reversely transcribed to the cDNAs, which used as the hybridization probes with the incorporations of fluorescent dUTP. The mixed probes were then hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and analyzed with computer image analysis. ResultsAmong the 4096 targets, there were 706 genes whose expression levels differed between the degeneration and normal lumbar intervertebral disc in all cases, comprising 298 up-regulated and 358 down-regulated ones. ConclusionDNA microarray technology is an effective technique in screening for differently expressed genes between the degeneration and normal lumbar intervertebral disc. Cell apoptosis plays an important role in the process of lumbar intervertebral disc degeneration.展开更多
【Aim】Insect diapause is a critical eco-physiological adaptation for seasonal survival,but little is known about the molecular mechanisms of summer and winter diapause of Delia antiqua.Here,our main goal is to identi...【Aim】Insect diapause is a critical eco-physiological adaptation for seasonal survival,but little is known about the molecular mechanisms of summer and winter diapause of Delia antiqua.Here,our main goal is to identify candidate genes linked to pupal diapause of D.antiqua.【Methods】A straightforward in silico approach was performed to predict the secreted diapause-related proteins based on the RNA-seq and digital gene expression datasets.Amino acid sequence similarity and basic physiochemical characteristics analysis were conducted.Gene expression levels of 12 selected genes from RNA-seq data were further validated in non-diapause pupae,and different developmental stages of summer and winter diapause pupae by qRT-PCR.【Results】A total of 38 putative secreted proteins were differentially regulated between the summer and winter diapause pupae of D.antiqua.Gene Ontology analysis revealed differentially expressed genes to be associated with odorant binding,chitin and lipid metabolic processes,innate immunity and development,etc.Among them,regulation of lipid mobilization and chitin-related metabolism was found to be important during diapause development.qRT-PCR analyses showed that the expression profiles of the 12 selected genes were in good agreement with results from RNA-seq(R=0.862,P=0.001).【Conclusion】This study establishes a robust pipeline for the discovery of diapause-related genes by secretome mining based on the high-throughput sequencing.展开更多
The paper presented an implement method for a dynamic gene expression display software based on the GTK. This method established the dynamic presentation system of gene expression which according to gene expression da...The paper presented an implement method for a dynamic gene expression display software based on the GTK. This method established the dynamic presentation system of gene expression which according to gene expression data from gene chip hybridize at different time, adopted a linearity combination model and Pearson correlation coefficient algorithm. The system described the gene expression changes in graphic form, the gene expression changes with time and the changes in characteristics of the gene expression, also the changes in relations of the gene expression and regulation relationships among genes. The system also provided an integrated platform for analysis on gene chips data, especially for the research on the network ofgene regulation.展开更多
Abiotic stress is a major limiting factor to crop productivity,and heat stress is one of the important elements for reduced crop production.Plants respond to heat stress at molecular and cellular levels as well as phy...Abiotic stress is a major limiting factor to crop productivity,and heat stress is one of the important elements for reduced crop production.Plants respond to heat stress at molecular and cellular levels as well as physiological level.Heat stress alters expression patterns of numerous genes in plants.展开更多
Fiber cell initiation is a complex process involving many pathways,including phytohormones and components for transcriptional and posttranscriptional regulation.Here we report expression
Nonspecific lipid transfer proteins(ns LTPs)are widely distributed in the plant kingdom and are involved in various stress responses.To clarify the function of ns LTP genes,IbLTP1 and IbLTP2 were cloned by PCR technol...Nonspecific lipid transfer proteins(ns LTPs)are widely distributed in the plant kingdom and are involved in various stress responses.To clarify the function of ns LTP genes,IbLTP1 and IbLTP2 were cloned by PCR technology,and the sequence structures,conserved domains,and evolutionary relationships were analyzed.Sequences of c DNAs and genomic genes showed that neither gene had introns,but both had several homologous isoforms.IbLTP1 and IbLTP2 encode proteins of 114 and 94 amino acid residues respectively,without any Trp.These proteins contain a signal peptide at the N-terminal and have conserved domains of ns LTP1 and ns LTP2,respectively.The expression patterns and expression differences of IbLTP1 and IbLTP2 in different tissues and under stress were determined byreal-time RT-PCR.Results showed that IbLTP1 and IbLTP2 had higher relative expression levels in young leaves and stems,respectively,and were highly induced under sodium chloride(Na Cl)stress.The coding sequences of IbLTP1 and IbLTP2 were cloned into expression vector p ET32 a and expressed in Escherichia coli BL21(DE3),respectively.The maximal OD_(600)values of strains harboring p ET32a-IbLTP1 and p ET32a-IbLTP2 were higher than those of the p ET32 a transformed strain under Na Cl stress.展开更多
Background： The SWEET （Sugars will eventually be exported transporters） gene family plays multiple roles in plant physiological activities and development process. It participates in reproductive development and in...Background： The SWEET （Sugars will eventually be exported transporters） gene family plays multiple roles in plant physiological activities and development process. It participates in reproductive development and in the process of sugar transport and absorption, plant senescence and stress responses and plant-pathogen interaction. However, thecomprehensive analysis of SWEET genes has not been reported in cotton. Results： In this study, we identified 22, 31, 55 and 60 SWEETgenes from the sequenced genomes of Gossypium orboreum, G. rairnondii, G. hirsutum and G. borbadense, respectively. Phylogenetic tree analysis showed that the SWEET genes could be divided into four groups, which were further classified into 14 sub-clades. Further analysis of chromosomal location, synteny analysis and gene duplication suggested that the orthologs showed a good collinearity and segmental duplication events played a crucial role in the expansion of the family in cotton. Specific MtN3_slv domains were highly conserved between Arabidopsis and cotton by exon-intron organization and motif analysis. In addition, the expression pattern in different tissues indicated that the duplicated genes in cotton might have acquired new functions as a result of sub-functionalization or neo-functionalization. The expression pattern of SWEET genes showed that the different genes were induced by diverse stresses. The identification and functional analysis of SWEET genes in cotton may provide more candidate genes for genetic modification. Conclusion： SWEET genes were classified into four clades in cotton. The expression patterns suggested that the duplicated genes might have experienced a functional divergence. This work provides insights into the evolution of SWEETgenes and more candidates for specific genetic modification, which will be useful in future research.展开更多
To understand the use of real-time reverse transcription-polymerase chain reaction （real-time RT-PCR） for detecting the relative abundance of mRNA, the expression of a tobacco ferrltin gene （NtFer1） was detected b...To understand the use of real-time reverse transcription-polymerase chain reaction （real-time RT-PCR） for detecting the relative abundance of mRNA, the expression of a tobacco ferrltin gene （NtFer1） was detected by Northern blot and real-time RT-PCR. The results indicated that both of the two methods were able to detect mRNA expression of NtFer1 cleady and similady, namely NtFer1 expression was responsive to iron-ovedoad, and the abundance of NtFer1 mRNA was greatly increased after iron loaded for 6 h. To compare the effect and sensitivity of two methods, results revealed that Northern blot need 30 μg of total RNA and at least 3 days for the total protocol performance, whereas real-time RT-PCR only need 2 μg of total RNA and 1.5 h. The real-time RT-PCR is rather sensitive and effective than Northern blot. Real-time RT-PCR analysis can be used to rapidly detect the relative abundance of mRNA expression instead of Northern blot analysis.展开更多
Specific primers for the MC1R gene of alpacas(GenBank EU1358800) were designed to amplify the cDNA sequence using RT-PCR to seek variation in the sequence and explore the relationship between the expression level of M...Specific primers for the MC1R gene of alpacas(GenBank EU1358800) were designed to amplify the cDNA sequence using RT-PCR to seek variation in the sequence and explore the relationship between the expression level of MC1R gene and alpaca coat color.The MC1R gene from white alpaca was cloned successfully and sequence analysis verified that the MC1R gene,encoding 317 amino acids,was 1081 bp in length.Compared with the existing sequence in GenBank,sequence identity was 99.9%and 7 mutations were found.Primers,designed from the sequence obtained,were used to assess the relative expression of MC1R in alpacas of different coat color using QRT-PCR and SPSS 13.0 software.Relative expression of MC1R in the skin of brown alpacas was 4.32 times higher than that in white alpacas after normalization with GAPDH(P【0.01),indicating that MC1R expression may be related to coat color of alpacas.展开更多
Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein Psb...Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein PsbX has been identified in PSII,which is associated with the oxygen-evolving complex.The expression of PsbX gene protein is regulated by light.PsbX’s central role involves the regulation of PSII,facilitating the binding of quinone molecules to the Qb(PsbA)site,and it additionally plays a crucial role in optimizing the efficiency of photosynthesis.Despite these insights,a comprehensive understanding of the PsbX gene’s functions has remained elusive.Results In this study,we identified ten PsbX genes in Gossypium hirsutum L.The phylogenetic analysis results showed that 40 genes from nine species were classified into one clade.The resulting sequence logos exhibited substantial conservation across the N and C terminals at multiple sites among all Gossypium species.Furthermore,the ortholo-gous/paralogous,Ka/Ks ratio revealed that cotton PsbX genes subjected to positive as well as purifying selection pressure might lead to limited divergence,which resulted in the whole genome and segmental duplication.The expression patterns of GhPsbX genes exhibited variations across specific tissues,as indicated by the analysis.Moreover,the expression of GhPsbX genes could potentially be regulated in response to salt,intense light,and drought stresses.Therefore,GhPsbX genes may play a significant role in the modulation of photosynthesis under adverse abiotic conditions.Conclusion We examined the structure and function of PsbX gene family very first by using comparative genom-ics and systems biology approaches in cotton.It seems that PsbX gene family plays a vital role during the growth and development of cotton under stress conditions.Collectively,the results of this study provide basic information to unveil the molecular and physiological function of PsbX genes of cotton plants.展开更多
The capsid protein precursor （P1）, which plays a major role for the generation of polypeptides of swine vesicular disease virus （SVDV）, was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and e...The capsid protein precursor （P1）, which plays a major role for the generation of polypeptides of swine vesicular disease virus （SVDV）, was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and expressed in the mammalian cell line PK15 through the retroviral expression system. The activity of recombinant protein to induce immune response was evaluated in guinea pigs. IFA and Western Blot were used to detect the recombinant protein expression. The results showed that the recombinant protein could be recognized by SVDV positive serum, and animal test showed SVDV-specific antibodies. All of those results indicate that a retroviral-based vaccine carrying the capsid protein precursor （P1） of SVD is able to be expressed in the eukaryotic cell and elicites strong SVDV-specific immune responses in guinea pigs.展开更多
Objective:Metabolic dysfunction-associated steatohepatitis(MASH),a progressive subtype of metabolic dysfunction-associated steatotic liver disease(MASLD),is characterized by hepatic steatosis,lobular inflammation,and ...Objective:Metabolic dysfunction-associated steatohepatitis(MASH),a progressive subtype of metabolic dysfunction-associated steatotic liver disease(MASLD),is characterized by hepatic steatosis,lobular inflammation,and hepatocyte ballooning,and may further progress to liver fibrosis and cirrhosis.Lectin-like oxidized low-density lipoprotein receptor-1(LOX-1),a member of the scavenger receptor family,recognizes and binds oxidized low-density lipoprotein.This study aims to investigate the role of LOX-1 in MASH progression.Methods:LOX-1 expression in MASLD mouse liver was analyzed using Gene Expression Omnibus(GEO)datasets.Immunofluorescence staining was performed to detect LOX-1 and alpha-smooth muscle actin(α-SMA)levels and co-localization in fibrotic liver tissues and LX-2 cells.LOX-1 knockout(Lox-1^(−/−))mice were generated using CRISPR/caspase-9(Cas9)and genotyped by PCR and Sanger sequencing.Wild-type(WT)and Lox-1^(−/−)mice were randomized into control and Western diet model groups.Serum and liver samples were collected for alanine aminotransferase(ALT)and aspartate aminotransferase(AST)measurement by biochemical kits,liver structure evaluation by hematoxylin and eosin(HE)staining,collagen deposition by Masson staining,lipid accumulation by Oil Red O staining,and fibrotic marker gene expression by real-time quantitative PCR(RT-qPCR).Network pharmacology and search tool for the retrieval of interacting genes/proteins(STRING)-based protein-protein interaction(PPI)with Gene Ontology(GO)enrichment were used to predict downstream targets and pathways.Results:The results from the GEO datasets GSE30552 and GSE40041 indicated LOX-1 mRNA was upregulated in high fat diet(HFD)and bile duct ligation(BDL)mouse models(both P<0.001).LOX-1 and α-SMA levels were elevated in fibrotic liver tissues.Lox-1^(−/−)mice were successfully established.Biochemical tests showed that serum AST and ALT levels were significantly elevated in WT mice fed a Western diet(both P<0.001),and these levels decreased after LOX-1 knockout(both P<0.05).HE staining revealed that WT mice on the Western diet exhibited marked hepatocellular ballooning degeneration,steatosis,inflammatory cell infiltration,and periportal fibroplasia,which were significantly ameliorated by LOX-1 knockout.Masson staining demonstrated increased blue-stained collagen fibers in the liver tissues of WT mice fed the Western diet compared with controldiet mice,and LOX-1 knockout inhibited collagen fiber deposition(all P<0.05).RT‑qPCR results showed that hepatic mRNA levels of Acta2,Col1a1,and Timp1 were significantly increased in Western diet-fed mice,and LOX-1 knockout reduced the expression of these fibrogenic marker genes.Oil Red O staining indicated that hepatocytes in WT mice fed the Western diet were notably enlarged,displayed macrovesicular steatosis,and exhibited diffusely distributed red lipid droplets,whereas LOX-1 knockout alleviated hepatic lipid accumulation(both P<0.001).RT‑qPCR results further demonstrated that knockdown of LOX-1 reduced Acta2,Col1a1,and Timp1 mRNA levels in LX‑2 cells(all P<0.05).Immunofluorescence analysis revealed co‑localization of LOX-1 and α‑SMA in LX‑2 cells,and LOX-1 silencing suppressed α‑SMA expression.Network pharmacology suggested LOX-1 may promote MASH via lipid and cholesterol metabolism networks.Conclusion:LOX-1 gene knockout ameliorates Western diet-induced MASH in mice and may serve as a potential therapeutic target.展开更多
Gene expression profiling at early stages(0～2 DPA) of fiber development in Gossypium hirsutum identified a number of transcription factors which were down regulated in fiberless mutants relative to wild type controls...Gene expression profiling at early stages(0～2 DPA) of fiber development in Gossypium hirsutum identified a number of transcription factors which were down regulated in fiberless mutants relative to wild type controls and which could play a role in controlling early fiber development.Chief among these was GhMYB25,a Mixta-like MYB gene.Transgenic GhMYB25-silenced cotton展开更多
Salinity resistance and differential gene expression associated with salinity in cotton germplasm were studied,because of the lar ge scale area of salinityin China,and its significant negative effect s on the cotton p...Salinity resistance and differential gene expression associated with salinity in cotton germplasm were studied,because of the lar ge scale area of salinityin China,and its significant negative effect s on the cotton production.The salinity-resisted genes and their differential expression were studied under the stress of NaCl on cotton.There were found,under the NaCl stress,1644 genes differentially ex-pressed fro m the salinity-sensitive cotton and only 817 genes differe ntially expressed from the salinity re siste d cotton.展开更多
Cotton(Goss y pium hirsutum L.)is the leading fiber crop and one of the mainstays of the econo-my in the world.Cotton fibers,as the main product of cotton plants,are unicellular,linear structures derived from the epid...Cotton(Goss y pium hirsutum L.)is the leading fiber crop and one of the mainstays of the econo-my in the world.Cotton fibers,as the main product of cotton plants,are unicellular,linear structures derived from the epidermis of the ovule.Cotton fiber develop ment consists of fo ur discrete yet over-lapping developmentalstages:initiation,elongation,secondary wall deposition,and maturation.Therefore,it is regard as an ideal experimental model for studying plant cell elo ngation,cellulose syn-thesis,and cell wall deposition.Furt hermore,the fiber quantity and quality are established during fi-ber development.展开更多
Tomato(Solanum lycopersicum L.)is a thermophilic vegetable crop,but sensitive to high temperature stress,especially under the greenhouse conditions.Due to global climate changes,heat stress has now become a great thre...Tomato(Solanum lycopersicum L.)is a thermophilic vegetable crop,but sensitive to high temperature stress,especially under the greenhouse conditions.Due to global climate changes,heat stress has now become a great threat to tomato production and fruit quality.Many studies have been conducted to determine the functions of genes in tomato responsive to abiotic and biotic stresses,but transcriptomic information on heat stress responses of tomato fruit is still limited.To investigate heat stress associated genes in tomato fruit,a cDNA library was constructed using fruit harvested from tomato cv.P19-9 plants grown under 42℃for 0,1,2 and4 h and the expression profiles of heat stress responsive genes in tomato fruit were analyzed through RNA-seq.A total of 632224558 clean high quality paired-end reads were obtained and then mapped to reference genome for RNA-seq analysis.After quality control analysis,alignment analysis and transcript assembly,a total of 55457 RNA transcripts were obtained with functional annotations.Overall,6869 differentially expressed genes(DEGs)were identified with a significant response to one or more of the three heat stress treatment times.Based on GO enrichment analysis,22 genes potentially involved in tomato thermo-tolerance were selected and validated for their expressions through qPCR.The expression profile of tomato fruit genes obtained in this study could shed light on the mechanism and gene editing breeding projects for tomato thermo-tolerance.These findings could also benefit improvement of harvest and storage of tomato in greenhouse.展开更多
Background: Cotton is an important commercial crop for being a valuable source of natural fiber.Its production has undergone a sharp decline because of abiotic stresses,etc.Drought is one of the major abiotic stress c...Background: Cotton is an important commercial crop for being a valuable source of natural fiber.Its production has undergone a sharp decline because of abiotic stresses,etc.Drought is one of the major abiotic stress causing significant yield losses in cotton.However,plants have evolved self-defense mechanisms to cope abiotic factors like drought,salt,cold,etc.The evolution of stress responsive transcription factors such as the trihelix,a nodule-inception-like protein(NLP),and the late embryogenesis abundant proteins have shown positive response in the resistance improvement to several abiotic stresses.Results: Genome wide identification and characterization of the effects of Light-Harvesting Chloro a/b binding(LHC)genes were carried out in cotton under drought stress conditions.A hundred and nine proteins encoded by the LHC genes were found in the cotton genome,with 55,27,and 27 genes found to be distributed in Gossypium hirsutum,G.arboreum,and G.raimondii,respectively.The proteins encoded by the genes were unevenly distributed on various chromosomes.The Ka/Ks(Non-synonymous substitution rate/Synonymous substitution rate)values were less than one,an indication of negative selection of the gene family.Differential expressions of genes showed that majority of the genes are being highly upregulated in the roots as compared with leaves and stem tissues.Most genes were found to be highly expressed in MR-85,a relative drought tolerant germplasm.Conclusion: The results provide proofs of the possible role of the LHC genes in improving drought stress tolerance,and can be explored by cotton breeders in releasing a more drought tolerant cotton varieties.展开更多
One of the impediments in the genetic improvement of cotton fiber is the paucity of information about genes associated with fiber development.Availability of chromosome arm substitution line CS-
Cotton,the most important natural fiber crop in the world,is a mainstay in China's economy.However,for over two decades,cotton yields both in China and U.S.have been at a plateau.One reason for this plateau is the...Cotton,the most important natural fiber crop in the world,is a mainstay in China's economy.However,for over two decades,cotton yields both in China and U.S.have been at a plateau.One reason for this plateau is the limitation of current cotton germplasm.Cotton fibers are single cells re"sulting from elongated cells of the ovule epidermis.IAA regulates both plant growth and differentia-tion,and it has important roles in cotton fiber develop ment.To evaluate plant hormone biosynthetic genes for genetic engineering to mo dify cotton fiber,iaaM,a auxin biosynthetic gene,was linked to two ovule-specific(Ag15 and Lefsm1).展开更多
In order to screen the genes controlling watermelon rind color and luster, the experiment was carried out with yellow watermelon skin mutants as tester and green wild type watermelon as control, and transcriptome sequ...In order to screen the genes controlling watermelon rind color and luster, the experiment was carried out with yellow watermelon skin mutants as tester and green wild type watermelon as control, and transcriptome sequencing and bioinformatics analysis were done. The results show that 34.27 Gb clean data were got by transcriptome sequencing. There are 261 differentially expressed genes among Y_1_vs_G_1, Y_2_vs_G_2 and Y_3_vs_G_3. The pathways contenting most differentially expressed genes are plant hormone signal transduction pathway, phenylpropanoid biosynthesis pathway, photosynthesis pathway, starch and sucrose metabolism pathway. 9-cis-epoxycarotenoid dioxygenase(Cla002942), alcohol dehydrogenase(Cla004992), photosystem Ⅰ reaction center subunit Ⅲ, chloroplastic(precursor)(Cla009181), long-chain acyl coenzyme A synthetase(Cla017341), threonine dehydratase biosynthetic(Cla018352) candidates genes were screened out.展开更多
文摘ObjectiveTo investigate the gene expression changes in normal and degeneration lumbar intervertebral disc in humans, providing information for clinical. MethodsThe PCR products of 4096 human genes were spotted onto a kind of chemical-material-coated-glass slides. The total RNAs were isolated from the tissues. Both the mRNAs from the degeneration and normal lumbar intervertebral disc in humans were reversely transcribed to the cDNAs, which used as the hybridization probes with the incorporations of fluorescent dUTP. The mixed probes were then hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and analyzed with computer image analysis. ResultsAmong the 4096 targets, there were 706 genes whose expression levels differed between the degeneration and normal lumbar intervertebral disc in all cases, comprising 298 up-regulated and 358 down-regulated ones. ConclusionDNA microarray technology is an effective technique in screening for differently expressed genes between the degeneration and normal lumbar intervertebral disc. Cell apoptosis plays an important role in the process of lumbar intervertebral disc degeneration.
文摘【Aim】Insect diapause is a critical eco-physiological adaptation for seasonal survival,but little is known about the molecular mechanisms of summer and winter diapause of Delia antiqua.Here,our main goal is to identify candidate genes linked to pupal diapause of D.antiqua.【Methods】A straightforward in silico approach was performed to predict the secreted diapause-related proteins based on the RNA-seq and digital gene expression datasets.Amino acid sequence similarity and basic physiochemical characteristics analysis were conducted.Gene expression levels of 12 selected genes from RNA-seq data were further validated in non-diapause pupae,and different developmental stages of summer and winter diapause pupae by qRT-PCR.【Results】A total of 38 putative secreted proteins were differentially regulated between the summer and winter diapause pupae of D.antiqua.Gene Ontology analysis revealed differentially expressed genes to be associated with odorant binding,chitin and lipid metabolic processes,innate immunity and development,etc.Among them,regulation of lipid mobilization and chitin-related metabolism was found to be important during diapause development.qRT-PCR analyses showed that the expression profiles of the 12 selected genes were in good agreement with results from RNA-seq(R=0.862,P=0.001).【Conclusion】This study establishes a robust pipeline for the discovery of diapause-related genes by secretome mining based on the high-throughput sequencing.
文摘The paper presented an implement method for a dynamic gene expression display software based on the GTK. This method established the dynamic presentation system of gene expression which according to gene expression data from gene chip hybridize at different time, adopted a linearity combination model and Pearson correlation coefficient algorithm. The system described the gene expression changes in graphic form, the gene expression changes with time and the changes in characteristics of the gene expression, also the changes in relations of the gene expression and regulation relationships among genes. The system also provided an integrated platform for analysis on gene chips data, especially for the research on the network ofgene regulation.
文摘Abiotic stress is a major limiting factor to crop productivity,and heat stress is one of the important elements for reduced crop production.Plants respond to heat stress at molecular and cellular levels as well as physiological level.Heat stress alters expression patterns of numerous genes in plants.
文摘Fiber cell initiation is a complex process involving many pathways,including phytohormones and components for transcriptional and posttranscriptional regulation.Here we report expression
基金supported by grants from the National Science and Technology Pillar Program of China(2007BAD78B03)the“Eleventh-Five”Key Project of Sichuan ProvinceChina(07SG111-003-1)
文摘Nonspecific lipid transfer proteins(ns LTPs)are widely distributed in the plant kingdom and are involved in various stress responses.To clarify the function of ns LTP genes,IbLTP1 and IbLTP2 were cloned by PCR technology,and the sequence structures,conserved domains,and evolutionary relationships were analyzed.Sequences of c DNAs and genomic genes showed that neither gene had introns,but both had several homologous isoforms.IbLTP1 and IbLTP2 encode proteins of 114 and 94 amino acid residues respectively,without any Trp.These proteins contain a signal peptide at the N-terminal and have conserved domains of ns LTP1 and ns LTP2,respectively.The expression patterns and expression differences of IbLTP1 and IbLTP2 in different tissues and under stress were determined byreal-time RT-PCR.Results showed that IbLTP1 and IbLTP2 had higher relative expression levels in young leaves and stems,respectively,and were highly induced under sodium chloride(Na Cl)stress.The coding sequences of IbLTP1 and IbLTP2 were cloned into expression vector p ET32 a and expressed in Escherichia coli BL21(DE3),respectively.The maximal OD_(600)values of strains harboring p ET32a-IbLTP1 and p ET32a-IbLTP2 were higher than those of the p ET32 a transformed strain under Na Cl stress.
基金supported by the The National Key ResearchDevelopment Program of China(2016YFD0101400,2017YFD0101600)
文摘Background： The SWEET （Sugars will eventually be exported transporters） gene family plays multiple roles in plant physiological activities and development process. It participates in reproductive development and in the process of sugar transport and absorption, plant senescence and stress responses and plant-pathogen interaction. However, thecomprehensive analysis of SWEET genes has not been reported in cotton. Results： In this study, we identified 22, 31, 55 and 60 SWEETgenes from the sequenced genomes of Gossypium orboreum, G. rairnondii, G. hirsutum and G. borbadense, respectively. Phylogenetic tree analysis showed that the SWEET genes could be divided into four groups, which were further classified into 14 sub-clades. Further analysis of chromosomal location, synteny analysis and gene duplication suggested that the orthologs showed a good collinearity and segmental duplication events played a crucial role in the expansion of the family in cotton. Specific MtN3_slv domains were highly conserved between Arabidopsis and cotton by exon-intron organization and motif analysis. In addition, the expression pattern in different tissues indicated that the duplicated genes in cotton might have acquired new functions as a result of sub-functionalization or neo-functionalization. The expression pattern of SWEET genes showed that the different genes were induced by diverse stresses. The identification and functional analysis of SWEET genes in cotton may provide more candidate genes for genetic modification. Conclusion： SWEET genes were classified into four clades in cotton. The expression patterns suggested that the duplicated genes might have experienced a functional divergence. This work provides insights into the evolution of SWEETgenes and more candidates for specific genetic modification, which will be useful in future research.
基金Supported in Part by the Key Project of Chinese Ministry of Education (106065) Heilongjiang Provincial Natural ScienceFoundation (C200533)
文摘To understand the use of real-time reverse transcription-polymerase chain reaction （real-time RT-PCR） for detecting the relative abundance of mRNA, the expression of a tobacco ferrltin gene （NtFer1） was detected by Northern blot and real-time RT-PCR. The results indicated that both of the two methods were able to detect mRNA expression of NtFer1 cleady and similady, namely NtFer1 expression was responsive to iron-ovedoad, and the abundance of NtFer1 mRNA was greatly increased after iron loaded for 6 h. To compare the effect and sensitivity of two methods, results revealed that Northern blot need 30 μg of total RNA and at least 3 days for the total protocol performance, whereas real-time RT-PCR only need 2 μg of total RNA and 1.5 h. The real-time RT-PCR is rather sensitive and effective than Northern blot. Real-time RT-PCR analysis can be used to rapidly detect the relative abundance of mRNA expression instead of Northern blot analysis.
基金supported by the National Natural Science Foundation of China(No.30501070)Shanxi Natural Science Foundation(No.20041099)President Foundation of Agricultural University of Hebei (BS2007023)
文摘Specific primers for the MC1R gene of alpacas(GenBank EU1358800) were designed to amplify the cDNA sequence using RT-PCR to seek variation in the sequence and explore the relationship between the expression level of MC1R gene and alpaca coat color.The MC1R gene from white alpaca was cloned successfully and sequence analysis verified that the MC1R gene,encoding 317 amino acids,was 1081 bp in length.Compared with the existing sequence in GenBank,sequence identity was 99.9%and 7 mutations were found.Primers,designed from the sequence obtained,were used to assess the relative expression of MC1R in alpacas of different coat color using QRT-PCR and SPSS 13.0 software.Relative expression of MC1R in the skin of brown alpacas was 4.32 times higher than that in white alpacas after normalization with GAPDH(P【0.01),indicating that MC1R expression may be related to coat color of alpacas.
基金supported by National Natural Science Foundation of China(32060466)Chinese Academy of Agricultural Sciences。
文摘Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein PsbX has been identified in PSII,which is associated with the oxygen-evolving complex.The expression of PsbX gene protein is regulated by light.PsbX’s central role involves the regulation of PSII,facilitating the binding of quinone molecules to the Qb(PsbA)site,and it additionally plays a crucial role in optimizing the efficiency of photosynthesis.Despite these insights,a comprehensive understanding of the PsbX gene’s functions has remained elusive.Results In this study,we identified ten PsbX genes in Gossypium hirsutum L.The phylogenetic analysis results showed that 40 genes from nine species were classified into one clade.The resulting sequence logos exhibited substantial conservation across the N and C terminals at multiple sites among all Gossypium species.Furthermore,the ortholo-gous/paralogous,Ka/Ks ratio revealed that cotton PsbX genes subjected to positive as well as purifying selection pressure might lead to limited divergence,which resulted in the whole genome and segmental duplication.The expression patterns of GhPsbX genes exhibited variations across specific tissues,as indicated by the analysis.Moreover,the expression of GhPsbX genes could potentially be regulated in response to salt,intense light,and drought stresses.Therefore,GhPsbX genes may play a significant role in the modulation of photosynthesis under adverse abiotic conditions.Conclusion We examined the structure and function of PsbX gene family very first by using comparative genom-ics and systems biology approaches in cotton.It seems that PsbX gene family plays a vital role during the growth and development of cotton under stress conditions.Collectively,the results of this study provide basic information to unveil the molecular and physiological function of PsbX genes of cotton plants.
基金Supported by Key Technology R&D Programme (2006BAD06A03)
文摘The capsid protein precursor （P1）, which plays a major role for the generation of polypeptides of swine vesicular disease virus （SVDV）, was cloned from SVDV HK/70 strain into the retroviral vector pBABE puro and expressed in the mammalian cell line PK15 through the retroviral expression system. The activity of recombinant protein to induce immune response was evaluated in guinea pigs. IFA and Western Blot were used to detect the recombinant protein expression. The results showed that the recombinant protein could be recognized by SVDV positive serum, and animal test showed SVDV-specific antibodies. All of those results indicate that a retroviral-based vaccine carrying the capsid protein precursor （P1） of SVD is able to be expressed in the eukaryotic cell and elicites strong SVDV-specific immune responses in guinea pigs.
基金supported by the Natural Science Foundation of Hunan Province,China(211142095031)。
文摘Objective:Metabolic dysfunction-associated steatohepatitis(MASH),a progressive subtype of metabolic dysfunction-associated steatotic liver disease(MASLD),is characterized by hepatic steatosis,lobular inflammation,and hepatocyte ballooning,and may further progress to liver fibrosis and cirrhosis.Lectin-like oxidized low-density lipoprotein receptor-1(LOX-1),a member of the scavenger receptor family,recognizes and binds oxidized low-density lipoprotein.This study aims to investigate the role of LOX-1 in MASH progression.Methods:LOX-1 expression in MASLD mouse liver was analyzed using Gene Expression Omnibus(GEO)datasets.Immunofluorescence staining was performed to detect LOX-1 and alpha-smooth muscle actin(α-SMA)levels and co-localization in fibrotic liver tissues and LX-2 cells.LOX-1 knockout(Lox-1^(−/−))mice were generated using CRISPR/caspase-9(Cas9)and genotyped by PCR and Sanger sequencing.Wild-type(WT)and Lox-1^(−/−)mice were randomized into control and Western diet model groups.Serum and liver samples were collected for alanine aminotransferase(ALT)and aspartate aminotransferase(AST)measurement by biochemical kits,liver structure evaluation by hematoxylin and eosin(HE)staining,collagen deposition by Masson staining,lipid accumulation by Oil Red O staining,and fibrotic marker gene expression by real-time quantitative PCR(RT-qPCR).Network pharmacology and search tool for the retrieval of interacting genes/proteins(STRING)-based protein-protein interaction(PPI)with Gene Ontology(GO)enrichment were used to predict downstream targets and pathways.Results:The results from the GEO datasets GSE30552 and GSE40041 indicated LOX-1 mRNA was upregulated in high fat diet(HFD)and bile duct ligation(BDL)mouse models(both P<0.001).LOX-1 and α-SMA levels were elevated in fibrotic liver tissues.Lox-1^(−/−)mice were successfully established.Biochemical tests showed that serum AST and ALT levels were significantly elevated in WT mice fed a Western diet(both P<0.001),and these levels decreased after LOX-1 knockout(both P<0.05).HE staining revealed that WT mice on the Western diet exhibited marked hepatocellular ballooning degeneration,steatosis,inflammatory cell infiltration,and periportal fibroplasia,which were significantly ameliorated by LOX-1 knockout.Masson staining demonstrated increased blue-stained collagen fibers in the liver tissues of WT mice fed the Western diet compared with controldiet mice,and LOX-1 knockout inhibited collagen fiber deposition(all P<0.05).RT‑qPCR results showed that hepatic mRNA levels of Acta2,Col1a1,and Timp1 were significantly increased in Western diet-fed mice,and LOX-1 knockout reduced the expression of these fibrogenic marker genes.Oil Red O staining indicated that hepatocytes in WT mice fed the Western diet were notably enlarged,displayed macrovesicular steatosis,and exhibited diffusely distributed red lipid droplets,whereas LOX-1 knockout alleviated hepatic lipid accumulation(both P<0.001).RT‑qPCR results further demonstrated that knockdown of LOX-1 reduced Acta2,Col1a1,and Timp1 mRNA levels in LX‑2 cells(all P<0.05).Immunofluorescence analysis revealed co‑localization of LOX-1 and α‑SMA in LX‑2 cells,and LOX-1 silencing suppressed α‑SMA expression.Network pharmacology suggested LOX-1 may promote MASH via lipid and cholesterol metabolism networks.Conclusion:LOX-1 gene knockout ameliorates Western diet-induced MASH in mice and may serve as a potential therapeutic target.
文摘Gene expression profiling at early stages(0～2 DPA) of fiber development in Gossypium hirsutum identified a number of transcription factors which were down regulated in fiberless mutants relative to wild type controls and which could play a role in controlling early fiber development.Chief among these was GhMYB25,a Mixta-like MYB gene.Transgenic GhMYB25-silenced cotton
文摘Salinity resistance and differential gene expression associated with salinity in cotton germplasm were studied,because of the lar ge scale area of salinityin China,and its significant negative effect s on the cotton production.The salinity-resisted genes and their differential expression were studied under the stress of NaCl on cotton.There were found,under the NaCl stress,1644 genes differentially ex-pressed fro m the salinity-sensitive cotton and only 817 genes differe ntially expressed from the salinity re siste d cotton.
基金This work was supported by the National Natural Science Foundation of China(No 30370904and No 30671258)the National High Technology Research and Development Program(863 project)of China(No 2006AA10Z121)the Program for New Century Excellent Talents in University(No NCET-07-0712)
文摘Cotton(Goss y pium hirsutum L.)is the leading fiber crop and one of the mainstays of the econo-my in the world.Cotton fibers,as the main product of cotton plants,are unicellular,linear structures derived from the epidermis of the ovule.Cotton fiber develop ment consists of fo ur discrete yet over-lapping developmentalstages:initiation,elongation,secondary wall deposition,and maturation.Therefore,it is regard as an ideal experimental model for studying plant cell elo ngation,cellulose syn-thesis,and cell wall deposition.Furt hermore,the fiber quantity and quality are established during fi-ber development.
文摘Tomato(Solanum lycopersicum L.)is a thermophilic vegetable crop,but sensitive to high temperature stress,especially under the greenhouse conditions.Due to global climate changes,heat stress has now become a great threat to tomato production and fruit quality.Many studies have been conducted to determine the functions of genes in tomato responsive to abiotic and biotic stresses,but transcriptomic information on heat stress responses of tomato fruit is still limited.To investigate heat stress associated genes in tomato fruit,a cDNA library was constructed using fruit harvested from tomato cv.P19-9 plants grown under 42℃for 0,1,2 and4 h and the expression profiles of heat stress responsive genes in tomato fruit were analyzed through RNA-seq.A total of 632224558 clean high quality paired-end reads were obtained and then mapped to reference genome for RNA-seq analysis.After quality control analysis,alignment analysis and transcript assembly,a total of 55457 RNA transcripts were obtained with functional annotations.Overall,6869 differentially expressed genes(DEGs)were identified with a significant response to one or more of the three heat stress treatment times.Based on GO enrichment analysis,22 genes potentially involved in tomato thermo-tolerance were selected and validated for their expressions through qPCR.The expression profile of tomato fruit genes obtained in this study could shed light on the mechanism and gene editing breeding projects for tomato thermo-tolerance.These findings could also benefit improvement of harvest and storage of tomato in greenhouse.
基金This research was funded by the National Natural Science Foundation of China,grant number 31621005,31530053,31671745The National Key R&D Program of China(2021YFE0101200),PSF/CRP/18thProtocol(07).
文摘Background: Cotton is an important commercial crop for being a valuable source of natural fiber.Its production has undergone a sharp decline because of abiotic stresses,etc.Drought is one of the major abiotic stress causing significant yield losses in cotton.However,plants have evolved self-defense mechanisms to cope abiotic factors like drought,salt,cold,etc.The evolution of stress responsive transcription factors such as the trihelix,a nodule-inception-like protein(NLP),and the late embryogenesis abundant proteins have shown positive response in the resistance improvement to several abiotic stresses.Results: Genome wide identification and characterization of the effects of Light-Harvesting Chloro a/b binding(LHC)genes were carried out in cotton under drought stress conditions.A hundred and nine proteins encoded by the LHC genes were found in the cotton genome,with 55,27,and 27 genes found to be distributed in Gossypium hirsutum,G.arboreum,and G.raimondii,respectively.The proteins encoded by the genes were unevenly distributed on various chromosomes.The Ka/Ks(Non-synonymous substitution rate/Synonymous substitution rate)values were less than one,an indication of negative selection of the gene family.Differential expressions of genes showed that majority of the genes are being highly upregulated in the roots as compared with leaves and stem tissues.Most genes were found to be highly expressed in MR-85,a relative drought tolerant germplasm.Conclusion: The results provide proofs of the possible role of the LHC genes in improving drought stress tolerance,and can be explored by cotton breeders in releasing a more drought tolerant cotton varieties.
文摘One of the impediments in the genetic improvement of cotton fiber is the paucity of information about genes associated with fiber development.Availability of chromosome arm substitution line CS-
基金This work is supported by the Major State Basic Research Development Program of China(2004CB117300)the National Natural Science Foundation of China(30530490)
文摘Cotton,the most important natural fiber crop in the world,is a mainstay in China's economy.However,for over two decades,cotton yields both in China and U.S.have been at a plateau.One reason for this plateau is the limitation of current cotton germplasm.Cotton fibers are single cells re"sulting from elongated cells of the ovule epidermis.IAA regulates both plant growth and differentia-tion,and it has important roles in cotton fiber develop ment.To evaluate plant hormone biosynthetic genes for genetic engineering to mo dify cotton fiber,iaaM,a auxin biosynthetic gene,was linked to two ovule-specific(Ag15 and Lefsm1).
基金Project(31260476)supported by the National Natural Science Foundation of China
文摘In order to screen the genes controlling watermelon rind color and luster, the experiment was carried out with yellow watermelon skin mutants as tester and green wild type watermelon as control, and transcriptome sequencing and bioinformatics analysis were done. The results show that 34.27 Gb clean data were got by transcriptome sequencing. There are 261 differentially expressed genes among Y_1_vs_G_1, Y_2_vs_G_2 and Y_3_vs_G_3. The pathways contenting most differentially expressed genes are plant hormone signal transduction pathway, phenylpropanoid biosynthesis pathway, photosynthesis pathway, starch and sucrose metabolism pathway. 9-cis-epoxycarotenoid dioxygenase(Cla002942), alcohol dehydrogenase(Cla004992), photosystem Ⅰ reaction center subunit Ⅲ, chloroplastic(precursor)(Cla009181), long-chain acyl coenzyme A synthetase(Cla017341), threonine dehydratase biosynthetic(Cla018352) candidates genes were screened out.
