Agarwood is the resinous heartwood of Aquilaria species.However,low yields and high costs of existing stimulation methods have led to the need for new techniques to produce agarwood rapidly and effectively.We develope...Agarwood is the resinous heartwood of Aquilaria species.However,low yields and high costs of existing stimulation methods have led to the need for new techniques to produce agarwood rapidly and effectively.We developed a biological agarwood-inducing technique(Agar-Bit)that produces high yields and quality within 6 months.To better understand agarwood formation by Agar-Bit,dynamic gene expressions of key synthetases pathways of sesquiterpenes and chalcone-related enzymes at different times were determined after both Agar-Bit and the traditional burning chisel drilling(BCD)stimulation on Aquilaria sinensis trees.The qRT-PCR results show that some characteristic synthase genes were expressed at greatly different levels and times compared with the controls.For the Agar-Bit technology,main changes were after the 3rd or 5th month,while BCD expression clearly changed at the 5th month.Essential oils and total chromone contents were simultaneously determined.In the Agar-Bit group,both were higher and similar to natural levels.The Agar-Bit methodology is a new option for producing agarwood as demonstrated by genetic and chemical aspects.The differences in gene expression within 6 months for both groups indicates that the mechanisms of the two methods are different.These findings provide information on genetic variation during the process of agarwood formation.展开更多
Camptotheac acuminata Decne is a unique tree species in China with an important secondary metabolite,camptothecin(CPT),used in the treatment of cancer.Nitrogen(N)is an important element that affects plant growth and t...Camptotheac acuminata Decne is a unique tree species in China with an important secondary metabolite,camptothecin(CPT),used in the treatment of cancer.Nitrogen(N)is an important element that affects plant growth and the accumulation of CPT.Reports on the effect of N on CPT synthesis from a genetic perspective are scarce.To explore the effects of different N sources and levels on CPT synthesis in C.acuminata,two-year-old seedlings were fertilized with different concentrations of pure ammonium sulphate,source of ammonium N(NH4+-N),and potassium nitrate for nitrate N(NO3--N).Concentrations of 2.5,5,7.5,and10 g pot-1 NH4+-N and NO3--N were used.The results showed that 7.5 g NH4+-N and NO3--N treatments were best for growth and fresh weight of leaves.Compared with the other treatments,the CPT content,tryptophan synthase and tryptophan decarboxylase activities,and expression of the CaTSB and CaTDCl genes under the 2.5 g NH4+-N and NO3--N treatments peaked significantly at 30 days.However,the expression of CaTDC2 surpassed that of the other two genes at 60 days.Therefore,compared with NH4+-N source,the NO3--N source was more beneficial for growth,and NO3--N was better for CPT yield.Consequently,leaves of C.acuminata treated with 2.5 g NO3--N could be harvested after 30 days to obtain maximum CPT content.CaTDC1 is more closely linked to CPT synthesis.The results of this study improved the production of CPT in C.acuminata via fertilization.展开更多
Objective. To explore the role of insulin-like growth factor 1 (IGF-1)gene expression abnormality in neurotrophic causes of diabetic peripheral neurophathy.Methods. Diabetes was induced in Sprague Dawley rats by allox...Objective. To explore the role of insulin-like growth factor 1 (IGF-1)gene expression abnormality in neurotrophic causes of diabetic peripheral neurophathy.Methods. Diabetes was induced in Sprague Dawley rats by alloxan. The parameters were measured as follows: IGF-1 mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR); IGF-1 peptide by enzyme-linked immunosorbent assay (ELISA); electrophysiological parameters of nerves by evoked electromyogram; morphometric evaluation of sciatic nerves under light microscope and transmission electron microscope.Results. During early diabetic stage, IGF-1 mRNA [(0.430±0.031)vs. (0.370±0.016), P ＜0.01,(0.430 ± 0.031 ) vs. (0.280 ± 0.010) , P ＜0.001, respectively], IGF - 1 peptide contents [ (38.44 ± 3.60)ng/mgvs. (30.06±2.41) ng/mg, P ＜0.01, (38.44±3.6) ng/mgvs. (3.71 +2.70) ng/mg, P ＜0.001,respectively] in sciatic nerve tissue reduced in diabetic rats with hyperglycemia and varied with severity of diabetic state when compared with non-diabetic control rats, and further gradually down-regulated in the diabetic rats with duration of diabetes [IGF-1 mRNA (0. 320 ± 0. 021) ～ (0. 230 + 0. 060); IGF-1 peptide (28.80 ± 3.30) ～(19. 51 + 1.80)ng/mg]. Furthermore, they correlated with nerve functional (sensory nerve conduction velocity:r = 0. 741, P ＜0. 001; amplitude ofevokedpotential: r = 0. 716, P ＜0. 001, respectively)andstructuralabnormality (axonal areas r = 0. 81, P ＜ 0. 001 ) of sciatic nerve. No difference was found in the above parameters between diabetic rats with euglycemia and non-diabetic control group.Conclusion. IGF-1 gene expression in tissues was down-regulated from early diabetic stage, and varied with the severity and duration of diabetic state. The decrement in IGF-1 level might contribute to the initiation and development of diabetic neuropathy via autocrine or paracrine pathway.展开更多
A method for transient gene expression was developed for western white pine(WWP,Pinus monticola Dougl.ex D.Don)using reporter gene uidA encodingβ-glucuronidase(GUS).GUS was transiently expressed in cross sections of ...A method for transient gene expression was developed for western white pine(WWP,Pinus monticola Dougl.ex D.Don)using reporter gene uidA encodingβ-glucuronidase(GUS).GUS was transiently expressed in cross sections of primary and secondary needles,cotyledons,and current and second year stems of WWP via vacuum-infiltration with Agrobacterium tumefaciens.Histochemical assays of cross sections of secondary needles showed stronger blue color indicating GUS expression at day 1 and 2 than on other days post agroinfiltration(dpa).GUS activity expressed inside WWP cells was confirmed using light microscopy.In fluorometric assays,GUS expression was high at 1 dpa and lasted until 4 dpa in detached secondary needles,while similarly high expression levels only lasted until 2 dpa in attached secondary needles then dropped significantly.Although the length of GUS-staining zones varied among different WWP organs and between growth and dormant seasons,all tested WWP tissues using the protocol had high levels of transient GUS expression.Thus,heterologous candidate genes or endogenous silencing can be expressed in various WWP tissues or organs using this agroinfiltration approach.The current protocol for efficient transient gene expression will aid functional genomics study of WWP and its pathogens and related conifer species.展开更多
As an irreplaceable dietary constituent,lipids play a vital important role in health,but their effects on aging process and longevity are still not well known yet.In this paper,the metabolic profiling and gene express...As an irreplaceable dietary constituent,lipids play a vital important role in health,but their effects on aging process and longevity are still not well known yet.In this paper,the metabolic profiling and gene expression levels of Caenorhabditis elegans were investigated to explore the effects of different edible oils on senescence and lifespan.The results showed that teaseed oil(TO)could prolong the life expectancy and slow down the aging process of C.elegans.Compared to the control group,the intake of lard oil(LO)and TO increased the expression levels of genes related to inhibition of protein aggregation(akt-1,daf-16,hsf-1,hsp-16.2)and lipid metabolism(daf-7,daf-1,mdt-15,lipl-4,fat-5,fat-6,fat-7),with a more significant alteration in TO group.Metabolomics revealed that palm oil can upregulated plenty of fatty acids(palmitic acid,stearic acid,tetracosanoic acid),together with some amino acids(tryptophan,L-aspartate,L-valine)and carbohydrate(D-glucose),while the trend was opposite in TO group.Besides,moderate-to-strong correlations were found between differential metabolites and changed genes.In general,this paper claimed that TO could prolong lifespan and slow down aging process via regulating the lipids,amino acids and carbohydrates metabolism.展开更多
This paper deals with the reflectance estimation model issue to improve the estimation accuracy. We propose a model containing two core procedures: dimensionality reduction and model mining. First, the dimensionality ...This paper deals with the reflectance estimation model issue to improve the estimation accuracy. We propose a model containing two core procedures: dimensionality reduction and model mining. First, the dimensionality reduction algorithm of hyperspectral data based on dependence degree(DRNDDD) is proposed to reduce the redundant hyperspectral band. DRND-DD solves the selection of suitable hyperspectral band via rough set theory. Furthermore, to improve the computation speed and accuracy of the model, based on DRND-DD, this paper proposes reflectance estimation model mining of leaf nitrogen concentration(LNC) for hyperspectral data by using hybrid gene expression programming(REMLNC-HGEP). Experimental results on three datasets demonstrate that the DRND-DD algorithm can obtain good results with a very short running time compared with principal component analysis(PCA), singular value decomposition(SVD), a dimensionality reduction algorithm based on the positive region(AR-PR) and a dimensionality reduction algorithm based on a discernable matrix(ARDM), and REMLNC-HGEP has low average time-consumption, high model mining success ratio and estimation accuracy. It was concluded that the REMLNC-HGEP performs better than the regression methods.展开更多
Objective:To establish a systematic framework for selecting the best clustering algorithm and provide an evaluation method for clustering analyses of gene expression data. Methods: Based on data structure (internal in...Objective:To establish a systematic framework for selecting the best clustering algorithm and provide an evaluation method for clustering analyses of gene expression data. Methods: Based on data structure (internal information) and function classification (external information), the evaluation of gene expression data analyses were carried out by using 2 approaches. Firstly, to assess the predictive power of clusteringalgorithms, Entropy was introduced to measure the consistency between the clustering results from different algorithms and the known and validated functional classifications. Secondly, a modified method of figure of merit (adjust-FOM) was used as internal assessment method. In this method, one clustering algorithm was used to analyze all data but one experimental condition, the remaining condition was used to assess the predictive power of the resulting clusters. This method was applied on 3 gene expression data sets (2 from the Lyer's Serum Data Sets, and 1 from the Ferea's Saccharomyces Cerevisiae Data Set). Results: A method based on entropy and figure of merit (FOM) was proposed to explore the results of the 3 data sets obtained by 6 different algorithms, SOM and Fuzzy clustering methods were confirmed to possess the highest ability to cluster. Conclusion: A method based on entropy is firstly brought forward to evaluate clustering analyses.Different results are attained in evaluating same data set due to different function classification. According to the curves of adjust_FOM and Entropy_FOM, SOM and Fuzzy clustering methods show the highest ability to cluster on the 3 data sets.展开更多
Objective: To analyze the differential expression genes (DEGs) between Barrett’s esophagus (BE) and normal esophagus mucosa and explore the target genes related to the development and progression of BE. Methods: The ...Objective: To analyze the differential expression genes (DEGs) between Barrett’s esophagus (BE) and normal esophagus mucosa and explore the target genes related to the development and progression of BE. Methods: The total RNAs of matched BE and normal esophagus mucosa of BE patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of BE and normal esophagus mucosa were hybridized with Agilent oligomicroarray (30 968 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. Results: (1) The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2) There were 142 up-regulated genes and 284 down-regulated genes among 2-fold DEGs. Conclusion: Microarray-based studies are feasible in endoscopically obtained tissues. Many BE-associated genes are screened by the high-throughput gene chip. The development and progression of BE is a complicated process involving multiple genes and multiple procedures, and functional study of these genes may help to identify the key genes or pathways involved in the pathogenesis and development of BE.展开更多
Objective: To investigate the role of cytokines IL-2, IL-4, IL-10, IL-12, and IFN-γ in pathogenesis of acute graft-versus-host disease (aGVHD) after allogeneic peripheral blood stem cell transplantation (allo-PBSCT)....Objective: To investigate the role of cytokines IL-2, IL-4, IL-10, IL-12, and IFN-γ in pathogenesis of acute graft-versus-host disease (aGVHD) after allogeneic peripheral blood stem cell transplantation (allo-PBSCT). Methods: Forty-two patients undergoing allo-PBSCT were included in this study. Reverse-transcriptase polymerase chain reaction (RT-PCR) was used to analyze gene expression of cytokines IL-2, IL-4, IL-10, IL-12, and IFN-γ. Results: All patients achieved engraftment, 18 patients developed grade ⅠGVHD, 6 patients developed grade Ⅱ-Ⅳ GVHD. The gene expression of IL-2, IL-12, and IFN-γ increased, the gene expression of IL-4 and IL-10 decreased. Conclusion: Cytokines IL-2, IL-12, and IFN-γ lead to a positive regulation of the development in human aGVHD, and IL-4 and IL-10 play negative regulatory roles.展开更多
Glutathione-S-transferase(GST,EC2.5.1.18)multifunctional protease is important for detoxification,defense against biotic and abiotic stresses,and secondary metabolic material transport for plant growth and development...Glutathione-S-transferase(GST,EC2.5.1.18)multifunctional protease is important for detoxification,defense against biotic and abiotic stresses,and secondary metabolic material transport for plant growth and development.In this study,71 members of the BpGST family were identified from the entire Betula platyphylla Suk.genome.Most of the members encode proteins with amino acid lengths ranging from 101 to 875 and were localized to the cytoplasm by a prediction.BpGSTs can be divided into seven subfamilies,with a majority of birch U and F subfamily members according to gene structure,conserved motifs and evolutionary analysis.GST family genes showed collinearity with 22 genes in Oryza sativa L.,and three genes in Arabidopsis thaliana;promoter cis-acting elements predicted that the GST gene family is functional in growth,hormone regulation,and abiotic stress response.Most members of the F subfamily of GST(BpGSTFs)were expressed in roots,stems,leaves,and petioles,with the most expression observed in leaves.On the basis of the expression profiles of F subfamily genes(BpGSTF1 to BpGSTF13)during salt,mannitol and ABA stress,BpGSTF proteins seem to have multiple functions depending on the type of abiotic stress;for instance,BpGSTs may function at different times during abiotic stress.This study enhances understanding of the GST gene family and provides a basis for further exploration of their function in birch.展开更多
Germin and Germin-like protein （GLP） have various proposed roles in plant developmental stages and stress- related processes. A novel GLP cDNA clone was isolated from a cDNA library of Tamarix hispida. ThGLP, coded ...Germin and Germin-like protein （GLP） have various proposed roles in plant developmental stages and stress- related processes. A novel GLP cDNA clone was isolated from a cDNA library of Tamarix hispida. ThGLP, coded 225aa, possesses conserved motif of plant germin and Germin-like protein. ThGLP belongs to true germin subfamily through phylogenetic analyses. Gene expression profiles in roots and leaves were evaluated using real-time quantitative RT-PCR. The results show that the gene was highly induced by drought, salt, low temperature, CdCl2 and abscisic acid treatments. Our results demonstrate that the ThGLP gene is expressed in leaves and roots, is involved in different abiotic stress re-sponses and controlled by an ABA-dependent signaling pathway.展开更多
Objective To investigate the mutation and expression of tumor suppressor gene-PTEN mRNA and explore their roles in tumorigenesis and progression of ovarian cancer. Methods Mutated exon 5 of PTEN gene was examined in n...Objective To investigate the mutation and expression of tumor suppressor gene-PTEN mRNA and explore their roles in tumorigenesis and progression of ovarian cancer. Methods Mutated exon 5 of PTEN gene was examined in normal ovary(n = 5), ovarian cyst (n =5), ovarian borderline tumor (n = 9), epithelial ovarian cancer(n = 60), and ovarian cancer cell line (n = 1)by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). mRNA expression of PTEN gene was evaluated in corresponding tissues and cell line by reverse transcription polymerase chain reaction(RT-PCR). The mutation and mRNA expression of PTEN gene were compared with clini-copathological features of ovarian cancer. Results Mutated exon 5 of PTEN gene was detected only in 5(7.1%)cases of epithelial ovarian cancer. mRNA expression level of PTEN gene in ovarian borderline tumor or ovarian cancer was lower than that in normal ovary or ovarian cyst(P < 0.05). The level of PTEN gene mRNA expression was negatively correlated with clinicopathological staging of ovarian cancer, whereas positively correlated with histological differentiation (P < 0.05). mRNA expression level of PTEN gene in ovarian endometrioid cancer was significantly lower than that in ovarian serous or mucinous cancer (P < 0.05=. Conclusions Mutation of PTEN gene occurs in ovarian cancer. Down-regulated expression of PTEN is probably an important molecular event in tumorigenesis of ovarian cancer. Abnormal expression of PTEN gene is involved in progression of ovarian cancer. Reduced expression of PTEN gene is closely associated with tumorigenesis and pathobiological behaviors of ovarian endometrioid cancer.展开更多
Objective. The experiment was designed to study the association of cerebral ischemia and reperfusion with endothelin- 1 (ET- 1) gene expression of rat brains and time-dependent changes of ET- 1 gene expression during...Objective. The experiment was designed to study the association of cerebral ischemia and reperfusion with endothelin- 1 (ET- 1) gene expression of rat brains and time-dependent changes of ET- 1 gene expression during cerebral ischemia.Materials and methods. Thirty- three male SD rats were divided into dot blot hybridization(n = 27) and in silu hybridization groups(n= 6). The focal cerebral ischemia and reperfusion models were made with suture embolism of middle cerebral artery. Dot blot hybridization groups were redivided into control and ischemic subgroups (ischemia for 0. 5 , 1 , 1. 5 , 3 , 6 , 12 , 24 , 48 and 72 h respectively). In situ hybridization groups were redivided into ischemia and reperfusion groups. After 24 h ischemia and 24 h reperfusion,ET1 gene expressions were investigated with in situ hybridization and the resuhs were analyzed with IBAS 2000 Image Analysis System.Results. Dot blot hybridization showed that ET-1 mRNA of cerebral cortex and caudate- putamen was increased at 6 h of ischemia and reached peak at 24 h (3. 9 and 3. 7 fold respectively) ,and at 72 h of ischemia it remained at high levels(3. 5 and 2. 1 fold respectively). In silu hybridization showed that the levels of ET- 1 mRNA of cerebral cortex and caudate-putamen were also markedly increased both in 24 h ischemia and 24 h reperfusion groups (P<0. 01 , P<0. 05 respectively) .Conclusions. ET-1 gene expression in focal ischemic brain tissue were markedly and progressively increased during cerebral ischemia and reperfusion and downregulation of ET- 1 gene expression may be a new approach to the treatment of ischemic cerebrovascular diseases.展开更多
Objective To study the expression and switching of Thl/Th2 cytokines gene in human gliomas and its effects on occurring and developing of human gliomas. Methods Interleukin（IL）-2 and interferon-3, represent Thl typ...Objective To study the expression and switching of Thl/Th2 cytokines gene in human gliomas and its effects on occurring and developing of human gliomas. Methods Interleukin（IL）-2 and interferon-3, represent Thl type cytokines. IL-4, IL-6, IL-10, and IL-13 represent Th2 type cytokines. The gene expressions of Th1/Th2 cytokines in human glioma cells, glioma infiltrating lymphocytes, and glioma cell lines were detected by reverse transcription polymerase chain reaction （RT-PCR）. The biological activity of cytokines in the supematant of glioma cell lines was assayed by enzyme-linked immunosorbent assay （ELISA） method. Results The total positive rates of Th1 and Th2 type cytokines gene in human glioma cells were 14.77% and 75%. The total positive rates of Th1 and Th2 type cytokines gene in glioma infiltrating lymphocytes were 22.73% and 68.17%. There was obviously predominant expression of Th2 type cytokines in human glioma tissues, glioma infiltrating lymphocytes, and glioma cell lines. There was no unbalanced expression of Th1/Th2 cytokines in normal brain tissues. Conclusion There is a predominant expression of Th2 type cytokines in human glioma cells. The switching of Th1/Th2 cytokines gene may play an important role in the occurring and developing of human gliomas.展开更多
Basic Leu zipper ( bZIP ) genes play pivotal, versatile roles in abiotic or biotic stress responses and in other biological processes. Knowledge on the evolutionary relationships and patterns of gene expression of bZI...Basic Leu zipper ( bZIP ) genes play pivotal, versatile roles in abiotic or biotic stress responses and in other biological processes. Knowledge on the evolutionary relationships and patterns of gene expression of bZIP family members in woody plants, however, has been limited. Here we identified and characterized 47 BpbZIP genes across the silver birch ( Betula pendula Roth.) genome. With reference to bZIP classifications for Arabidopsis thaliana , all BpbZIP proteins clustered among 10 groups in phylogeny. The bZIP domains were divided into five patterns based on intron positions and splicing phases. A total of 24 conserved motifs were detected in BpbZIPs with high group specificity. We also analyzed the protein structure of the BpAREB/ABF/ ABI5 subfamily, the most important subfamily in the bZIP family. Expression analyses demonstrated that BpbZIP genes were widely involved in abscisic acid, salt, drought, and heat stress responses, with BpbZIP07/ABF4 and BpbZIP21/ABF2 most highly expressed. Our results on genome-wide identification, evolutionary relationships, gene structure, and motif and promoter element identification for BpbZIP family members in silver birch provide a comprehensive understanding of bZIP transcription factors in birch and will lead to a deeper understanding of their evolution and potential biological functions.展开更多
O6-methylguanine-DNA methyltransferase (MGMT) gene expression in 6 Mer+ (HeLa S3, SMMC-7721,SGC-7901, B-239, AGZY83-a, and Cc80 1) and 2 Mer- (SHG-44 , and HeLa MR) human tumor cell lines was examined. Southern blot ...O6-methylguanine-DNA methyltransferase (MGMT) gene expression in 6 Mer+ (HeLa S3, SMMC-7721,SGC-7901, B-239, AGZY83-a, and Cc80 1) and 2 Mer- (SHG-44 , and HeLa MR) human tumor cell lines was examined. Southern blot analysis revealed no deletion, amplification, or rearrangement of the MGMT gene in these cell lines. However ,～ 1. 0 kb transcripts were detected in the 6 Mer+ cell lines but not in the 2 Mer-cell lines by Northern blot analysis. Furthermore,a rough correlation between MGMT activity and mRNA level in these cell lines was observed. These results suggest that transcriptional regulation of the MGMT gene is the molecular basis of the absence of MGMT activity in Mer-cell lines.展开更多
This research was designed to assess the changes in anthocyanin content in grape skins of Vitis amurensis and to explore m RNA transcriptions of 11 structural genes（PAL,CHS3, CHI1, F3H2, F30 H, F3050 H, DFR, LDOX, UF...This research was designed to assess the changes in anthocyanin content in grape skins of Vitis amurensis and to explore m RNA transcriptions of 11 structural genes（PAL,CHS3, CHI1, F3H2, F30 H, F3050 H, DFR, LDOX, UFGT,OMT and GST） related to anthocyanin biosynthesis during grape berry development, by the use of HPLC-MS/MS and real-time Q-PCR analysis. Accumulation of anthocyanins began at veraison, continued throughout the later berry development and reached a peak at maturity. Veraison is the time when the berries turn from green to purple. Expression of PAL, CHI1, and LDOX were up-regulated from 2 to4 weeks after flowering（WAF）, down-regulated from6 WAF to veraison, whereas DFR was up-regulated at8 WAF, and then up-regulated from veraison to maturity.CHS3, F3050 H, UFGT, GST, and OMT were down-regulated from 2 WAF to veraison, and then up-regulated from veraison to maturity. The transcriptional expressions of the11 structural genes also showed positive correlations with the anthocyanin content from veraison to maturity. Positive correlations were also observed between OMT transcriptional level and the content of methoxyl-anthocyanins, and between F3050 H transcriptional level and the content of delphinidin anthocyanins. F3H2 and F30 H expression was up-regulated at 2 WAF. F3H2 expression was down-regulated from 4 WAF to veraison and then up-regulated again from veraison to maturity. F30 H expression was down-regulated at 4 WAF and then up-regulated again from 6 WAF to maturity. F30 H transcriptional level was correlated positively with the cyanidin anthocyanin concentration from veraison to maturity. These results indicate that the onset of anthocyanin synthesis during berry development coincides with a coordinated increase in the expression of a number of genes in the anthocyanin biosynthetic pathway.展开更多
Expansins are cell-wall-loosening proteins that have multiple roles during plant development and stress- related processes. In this study, a novel expansin gene PwEXP2 was cloned by rapid amplification of cDNA ends ba...Expansins are cell-wall-loosening proteins that have multiple roles during plant development and stress- related processes. In this study, a novel expansin gene PwEXP2 was cloned by rapid amplification of cDNA ends based on the cDNA library of Picea wilsonii and EST fragment of PwEXP2. It was found that PwEXP2 coded 253 amino acids, and putative signal peptides exist at the N-terminal, followed by 8 cysteines, a HFD （His-Phe-Asp） conserved domain, and 4 tryptophan residues at the C-terminal. PwEXP2 was located in cytoplasm and nucleus when transformed in an onion epidermal cell. Quantitative real-time PCR assays showed that PwEXP2 was expressed in various tissues with a relatively high level in needles and low level in mature pollen. The expression level of PwEXP2 dramatically increased after seed germination. Gene expression profiles in abiotic stresses showed that PwEXP2 was induced by high temperature and osmotic stress but not involved in ABA-dependent signaling path- way. These results display the important roles of the PwEXP2 in plant development and multiple adversity stresses.展开更多
Taxol(Paclitaxel),an important anticancer drug,is derived at very low yields from Taxus(yew)species that grow very slowly.In the present study,thirteen genes that encode enzymes involved in Taxol biosynthesis in Taxus...Taxol(Paclitaxel),an important anticancer drug,is derived at very low yields from Taxus(yew)species that grow very slowly.In the present study,thirteen genes that encode enzymes involved in Taxol biosynthesis in Taxus spp.were analyzed with bioinformatics methods,and their expression levels in different tissues and after cold and hormone treatments were also analyzed.The results indicated that many cis-elements related to abiotic stresses and hormones were found in the promoter sequences of the 8 genes involved in Taxol biosynthesis.Moreover,the 13 enzymes encoded by the target genes were located in different organelles and had many phosphorylation sites in the response proteins.The 13 genes were expressed highly either in roots or in stems,with lower transcripts in needles,and they were highly expressed after treatment with cold,gibberellin,methyl jasmonate or coronatine,consistent with predictions based on the bioinformatics analysis.These results suggest that the factors such as hormones and abiotic stresses stimulate taxane biosynthesis in yews,providing an important way to sustainably generate taxanes from yew trees or their cell cultures to improve Taxol yields.展开更多
Nicotinamide treatment of plants and plant cell cultures has been shown to promote defense and decrease levels of DNA methylation.In the present study,we used RNA-seq technology to study overall changes in gene expres...Nicotinamide treatment of plants and plant cell cultures has been shown to promote defense and decrease levels of DNA methylation.In the present study,we used RNA-seq technology to study overall changes in gene expression induced in roots of 3-month-old spruce(Picea abies)seedlings grown from nicotinamide-treated seeds to examine the molecular mechanisms underlying the defense promotion.Approximately 350 genes were identified as differentially expressed in roots after the seed treatment.Stress response genes,including transcription factors MYB77 and LHY and two chitinase enzymes,were generally upregulated,whereas genes thought to be involved in epigenetic regulation such as DDM1,known to promote DNA methylation,were present at high frequency among the downregulated genes.Across all samples,the expression of downregulated epigenetic-related genes was highly correlated with the nicotinamide treatment,indicating a common regulation.Our results support an earlier hypothesis regarding a potential role of nicotinamide as a defense-signal mediator.展开更多
基金supported by Grant Number NDRC2011-51 from the National Development and Reform Commission(NDRC),the Office of New High-Tech Industrial Developmentsupported by Grant Number18A36002 from Key project of institutions of colleges and universities in Henan province,Henan Education Department。
文摘Agarwood is the resinous heartwood of Aquilaria species.However,low yields and high costs of existing stimulation methods have led to the need for new techniques to produce agarwood rapidly and effectively.We developed a biological agarwood-inducing technique(Agar-Bit)that produces high yields and quality within 6 months.To better understand agarwood formation by Agar-Bit,dynamic gene expressions of key synthetases pathways of sesquiterpenes and chalcone-related enzymes at different times were determined after both Agar-Bit and the traditional burning chisel drilling(BCD)stimulation on Aquilaria sinensis trees.The qRT-PCR results show that some characteristic synthase genes were expressed at greatly different levels and times compared with the controls.For the Agar-Bit technology,main changes were after the 3rd or 5th month,while BCD expression clearly changed at the 5th month.Essential oils and total chromone contents were simultaneously determined.In the Agar-Bit group,both were higher and similar to natural levels.The Agar-Bit methodology is a new option for producing agarwood as demonstrated by genetic and chemical aspects.The differences in gene expression within 6 months for both groups indicates that the mechanisms of the two methods are different.These findings provide information on genetic variation during the process of agarwood formation.
基金This research was sponsored by the 13th Five-Year-Plan National Projects for Camptotheca acuminata Decne Effi cient Cultivation Technology Research of China(2017YFD0600706).
文摘Camptotheac acuminata Decne is a unique tree species in China with an important secondary metabolite,camptothecin(CPT),used in the treatment of cancer.Nitrogen(N)is an important element that affects plant growth and the accumulation of CPT.Reports on the effect of N on CPT synthesis from a genetic perspective are scarce.To explore the effects of different N sources and levels on CPT synthesis in C.acuminata,two-year-old seedlings were fertilized with different concentrations of pure ammonium sulphate,source of ammonium N(NH4+-N),and potassium nitrate for nitrate N(NO3--N).Concentrations of 2.5,5,7.5,and10 g pot-1 NH4+-N and NO3--N were used.The results showed that 7.5 g NH4+-N and NO3--N treatments were best for growth and fresh weight of leaves.Compared with the other treatments,the CPT content,tryptophan synthase and tryptophan decarboxylase activities,and expression of the CaTSB and CaTDCl genes under the 2.5 g NH4+-N and NO3--N treatments peaked significantly at 30 days.However,the expression of CaTDC2 surpassed that of the other two genes at 60 days.Therefore,compared with NH4+-N source,the NO3--N source was more beneficial for growth,and NO3--N was better for CPT yield.Consequently,leaves of C.acuminata treated with 2.5 g NO3--N could be harvested after 30 days to obtain maximum CPT content.CaTDC1 is more closely linked to CPT synthesis.The results of this study improved the production of CPT in C.acuminata via fertilization.
基金This work was supported by the National Natural Sciences Foun-dation of China.This work was originally published in Chinese Journal of Internal Medicine (2001 40(2): 93-7)in Chinese.
文摘Objective. To explore the role of insulin-like growth factor 1 (IGF-1)gene expression abnormality in neurotrophic causes of diabetic peripheral neurophathy.Methods. Diabetes was induced in Sprague Dawley rats by alloxan. The parameters were measured as follows: IGF-1 mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR); IGF-1 peptide by enzyme-linked immunosorbent assay (ELISA); electrophysiological parameters of nerves by evoked electromyogram; morphometric evaluation of sciatic nerves under light microscope and transmission electron microscope.Results. During early diabetic stage, IGF-1 mRNA [(0.430±0.031)vs. (0.370±0.016), P ＜0.01,(0.430 ± 0.031 ) vs. (0.280 ± 0.010) , P ＜0.001, respectively], IGF - 1 peptide contents [ (38.44 ± 3.60)ng/mgvs. (30.06±2.41) ng/mg, P ＜0.01, (38.44±3.6) ng/mgvs. (3.71 +2.70) ng/mg, P ＜0.001,respectively] in sciatic nerve tissue reduced in diabetic rats with hyperglycemia and varied with severity of diabetic state when compared with non-diabetic control rats, and further gradually down-regulated in the diabetic rats with duration of diabetes [IGF-1 mRNA (0. 320 ± 0. 021) ～ (0. 230 + 0. 060); IGF-1 peptide (28.80 ± 3.30) ～(19. 51 + 1.80)ng/mg]. Furthermore, they correlated with nerve functional (sensory nerve conduction velocity:r = 0. 741, P ＜0. 001; amplitude ofevokedpotential: r = 0. 716, P ＜0. 001, respectively)andstructuralabnormality (axonal areas r = 0. 81, P ＜ 0. 001 ) of sciatic nerve. No difference was found in the above parameters between diabetic rats with euglycemia and non-diabetic control group.Conclusion. IGF-1 gene expression in tissues was down-regulated from early diabetic stage, and varied with the severity and duration of diabetic state. The decrement in IGF-1 level might contribute to the initiation and development of diabetic neuropathy via autocrine or paracrine pathway.
文摘A method for transient gene expression was developed for western white pine(WWP,Pinus monticola Dougl.ex D.Don)using reporter gene uidA encodingβ-glucuronidase(GUS).GUS was transiently expressed in cross sections of primary and secondary needles,cotyledons,and current and second year stems of WWP via vacuum-infiltration with Agrobacterium tumefaciens.Histochemical assays of cross sections of secondary needles showed stronger blue color indicating GUS expression at day 1 and 2 than on other days post agroinfiltration(dpa).GUS activity expressed inside WWP cells was confirmed using light microscopy.In fluorometric assays,GUS expression was high at 1 dpa and lasted until 4 dpa in detached secondary needles,while similarly high expression levels only lasted until 2 dpa in attached secondary needles then dropped significantly.Although the length of GUS-staining zones varied among different WWP organs and between growth and dormant seasons,all tested WWP tissues using the protocol had high levels of transient GUS expression.Thus,heterologous candidate genes or endogenous silencing can be expressed in various WWP tissues or organs using this agroinfiltration approach.The current protocol for efficient transient gene expression will aid functional genomics study of WWP and its pathogens and related conifer species.
基金supported by National Key R&D Program of China(2019YFC1606205)Natural Science Foundation of China(32172136)Jiangsu distinguished professor project and Talent plan of Taihu Lake.
文摘As an irreplaceable dietary constituent,lipids play a vital important role in health,but their effects on aging process and longevity are still not well known yet.In this paper,the metabolic profiling and gene expression levels of Caenorhabditis elegans were investigated to explore the effects of different edible oils on senescence and lifespan.The results showed that teaseed oil(TO)could prolong the life expectancy and slow down the aging process of C.elegans.Compared to the control group,the intake of lard oil(LO)and TO increased the expression levels of genes related to inhibition of protein aggregation(akt-1,daf-16,hsf-1,hsp-16.2)and lipid metabolism(daf-7,daf-1,mdt-15,lipl-4,fat-5,fat-6,fat-7),with a more significant alteration in TO group.Metabolomics revealed that palm oil can upregulated plenty of fatty acids(palmitic acid,stearic acid,tetracosanoic acid),together with some amino acids(tryptophan,L-aspartate,L-valine)and carbohydrate(D-glucose),while the trend was opposite in TO group.Besides,moderate-to-strong correlations were found between differential metabolites and changed genes.In general,this paper claimed that TO could prolong lifespan and slow down aging process via regulating the lipids,amino acids and carbohydrates metabolism.
基金supported in part by the National Natural Science Foundation of China (11&zd167, 51507084, 61572262)NSF of Jiangsu Province (BK20141427)+2 种基金NUPT (NY214097)Open research fund of Key Lab of Broadband Wireless Communication and Sensor Network Technology (NUPT), Ministry of Education (NYKL201507)Qinlan Project of Jiangsu Province and the General Project of National Natural Science Found of China under Grant 41471300
文摘This paper deals with the reflectance estimation model issue to improve the estimation accuracy. We propose a model containing two core procedures: dimensionality reduction and model mining. First, the dimensionality reduction algorithm of hyperspectral data based on dependence degree(DRNDDD) is proposed to reduce the redundant hyperspectral band. DRND-DD solves the selection of suitable hyperspectral band via rough set theory. Furthermore, to improve the computation speed and accuracy of the model, based on DRND-DD, this paper proposes reflectance estimation model mining of leaf nitrogen concentration(LNC) for hyperspectral data by using hybrid gene expression programming(REMLNC-HGEP). Experimental results on three datasets demonstrate that the DRND-DD algorithm can obtain good results with a very short running time compared with principal component analysis(PCA), singular value decomposition(SVD), a dimensionality reduction algorithm based on the positive region(AR-PR) and a dimensionality reduction algorithm based on a discernable matrix(ARDM), and REMLNC-HGEP has low average time-consumption, high model mining success ratio and estimation accuracy. It was concluded that the REMLNC-HGEP performs better than the regression methods.
文摘Objective:To establish a systematic framework for selecting the best clustering algorithm and provide an evaluation method for clustering analyses of gene expression data. Methods: Based on data structure (internal information) and function classification (external information), the evaluation of gene expression data analyses were carried out by using 2 approaches. Firstly, to assess the predictive power of clusteringalgorithms, Entropy was introduced to measure the consistency between the clustering results from different algorithms and the known and validated functional classifications. Secondly, a modified method of figure of merit (adjust-FOM) was used as internal assessment method. In this method, one clustering algorithm was used to analyze all data but one experimental condition, the remaining condition was used to assess the predictive power of the resulting clusters. This method was applied on 3 gene expression data sets (2 from the Lyer's Serum Data Sets, and 1 from the Ferea's Saccharomyces Cerevisiae Data Set). Results: A method based on entropy and figure of merit (FOM) was proposed to explore the results of the 3 data sets obtained by 6 different algorithms, SOM and Fuzzy clustering methods were confirmed to possess the highest ability to cluster. Conclusion: A method based on entropy is firstly brought forward to evaluate clustering analyses.Different results are attained in evaluating same data set due to different function classification. According to the curves of adjust_FOM and Entropy_FOM, SOM and Fuzzy clustering methods show the highest ability to cluster on the 3 data sets.
文摘Objective: To analyze the differential expression genes (DEGs) between Barrett’s esophagus (BE) and normal esophagus mucosa and explore the target genes related to the development and progression of BE. Methods: The total RNAs of matched BE and normal esophagus mucosa of BE patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of BE and normal esophagus mucosa were hybridized with Agilent oligomicroarray (30 968 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. Results: (1) The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2) There were 142 up-regulated genes and 284 down-regulated genes among 2-fold DEGs. Conclusion: Microarray-based studies are feasible in endoscopically obtained tissues. Many BE-associated genes are screened by the high-throughput gene chip. The development and progression of BE is a complicated process involving multiple genes and multiple procedures, and functional study of these genes may help to identify the key genes or pathways involved in the pathogenesis and development of BE.
文摘Objective: To investigate the role of cytokines IL-2, IL-4, IL-10, IL-12, and IFN-γ in pathogenesis of acute graft-versus-host disease (aGVHD) after allogeneic peripheral blood stem cell transplantation (allo-PBSCT). Methods: Forty-two patients undergoing allo-PBSCT were included in this study. Reverse-transcriptase polymerase chain reaction (RT-PCR) was used to analyze gene expression of cytokines IL-2, IL-4, IL-10, IL-12, and IFN-γ. Results: All patients achieved engraftment, 18 patients developed grade ⅠGVHD, 6 patients developed grade Ⅱ-Ⅳ GVHD. The gene expression of IL-2, IL-12, and IFN-γ increased, the gene expression of IL-4 and IL-10 decreased. Conclusion: Cytokines IL-2, IL-12, and IFN-γ lead to a positive regulation of the development in human aGVHD, and IL-4 and IL-10 play negative regulatory roles.
基金supported by the National Key Research and Development Program of China(No.2021YFD2200304)FundamentalResearch Funds for the Central Universities(2572022DQ08)the National Natural Science Foundation of China(No32171738).
文摘Glutathione-S-transferase(GST,EC2.5.1.18)multifunctional protease is important for detoxification,defense against biotic and abiotic stresses,and secondary metabolic material transport for plant growth and development.In this study,71 members of the BpGST family were identified from the entire Betula platyphylla Suk.genome.Most of the members encode proteins with amino acid lengths ranging from 101 to 875 and were localized to the cytoplasm by a prediction.BpGSTs can be divided into seven subfamilies,with a majority of birch U and F subfamily members according to gene structure,conserved motifs and evolutionary analysis.GST family genes showed collinearity with 22 genes in Oryza sativa L.,and three genes in Arabidopsis thaliana;promoter cis-acting elements predicted that the GST gene family is functional in growth,hormone regulation,and abiotic stress response.Most members of the F subfamily of GST(BpGSTFs)were expressed in roots,stems,leaves,and petioles,with the most expression observed in leaves.On the basis of the expression profiles of F subfamily genes(BpGSTF1 to BpGSTF13)during salt,mannitol and ABA stress,BpGSTF proteins seem to have multiple functions depending on the type of abiotic stress;for instance,BpGSTs may function at different times during abiotic stress.This study enhances understanding of the GST gene family and provides a basis for further exploration of their function in birch.
基金This study was supported by national natural science foundation (Grant No. 30972386)Central university basic scientific business specific foundation (Grant No. DL09BA22)Genetically modified organisms breeding major projects (Grant No.2009ZX08009-098B)
文摘Germin and Germin-like protein （GLP） have various proposed roles in plant developmental stages and stress- related processes. A novel GLP cDNA clone was isolated from a cDNA library of Tamarix hispida. ThGLP, coded 225aa, possesses conserved motif of plant germin and Germin-like protein. ThGLP belongs to true germin subfamily through phylogenetic analyses. Gene expression profiles in roots and leaves were evaluated using real-time quantitative RT-PCR. The results show that the gene was highly induced by drought, salt, low temperature, CdCl2 and abscisic acid treatments. Our results demonstrate that the ThGLP gene is expressed in leaves and roots, is involved in different abiotic stress re-sponses and controlled by an ABA-dependent signaling pathway.
文摘Objective To investigate the mutation and expression of tumor suppressor gene-PTEN mRNA and explore their roles in tumorigenesis and progression of ovarian cancer. Methods Mutated exon 5 of PTEN gene was examined in normal ovary(n = 5), ovarian cyst (n =5), ovarian borderline tumor (n = 9), epithelial ovarian cancer(n = 60), and ovarian cancer cell line (n = 1)by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). mRNA expression of PTEN gene was evaluated in corresponding tissues and cell line by reverse transcription polymerase chain reaction(RT-PCR). The mutation and mRNA expression of PTEN gene were compared with clini-copathological features of ovarian cancer. Results Mutated exon 5 of PTEN gene was detected only in 5(7.1%)cases of epithelial ovarian cancer. mRNA expression level of PTEN gene in ovarian borderline tumor or ovarian cancer was lower than that in normal ovary or ovarian cyst(P < 0.05). The level of PTEN gene mRNA expression was negatively correlated with clinicopathological staging of ovarian cancer, whereas positively correlated with histological differentiation (P < 0.05). mRNA expression level of PTEN gene in ovarian endometrioid cancer was significantly lower than that in ovarian serous or mucinous cancer (P < 0.05=. Conclusions Mutation of PTEN gene occurs in ovarian cancer. Down-regulated expression of PTEN is probably an important molecular event in tumorigenesis of ovarian cancer. Abnormal expression of PTEN gene is involved in progression of ovarian cancer. Reduced expression of PTEN gene is closely associated with tumorigenesis and pathobiological behaviors of ovarian endometrioid cancer.
文摘Objective. The experiment was designed to study the association of cerebral ischemia and reperfusion with endothelin- 1 (ET- 1) gene expression of rat brains and time-dependent changes of ET- 1 gene expression during cerebral ischemia.Materials and methods. Thirty- three male SD rats were divided into dot blot hybridization(n = 27) and in silu hybridization groups(n= 6). The focal cerebral ischemia and reperfusion models were made with suture embolism of middle cerebral artery. Dot blot hybridization groups were redivided into control and ischemic subgroups (ischemia for 0. 5 , 1 , 1. 5 , 3 , 6 , 12 , 24 , 48 and 72 h respectively). In situ hybridization groups were redivided into ischemia and reperfusion groups. After 24 h ischemia and 24 h reperfusion,ET1 gene expressions were investigated with in situ hybridization and the resuhs were analyzed with IBAS 2000 Image Analysis System.Results. Dot blot hybridization showed that ET-1 mRNA of cerebral cortex and caudate- putamen was increased at 6 h of ischemia and reached peak at 24 h (3. 9 and 3. 7 fold respectively) ,and at 72 h of ischemia it remained at high levels(3. 5 and 2. 1 fold respectively). In silu hybridization showed that the levels of ET- 1 mRNA of cerebral cortex and caudate-putamen were also markedly increased both in 24 h ischemia and 24 h reperfusion groups (P<0. 01 , P<0. 05 respectively) .Conclusions. ET-1 gene expression in focal ischemic brain tissue were markedly and progressively increased during cerebral ischemia and reperfusion and downregulation of ET- 1 gene expression may be a new approach to the treatment of ischemic cerebrovascular diseases.
文摘Objective To study the expression and switching of Thl/Th2 cytokines gene in human gliomas and its effects on occurring and developing of human gliomas. Methods Interleukin（IL）-2 and interferon-3, represent Thl type cytokines. IL-4, IL-6, IL-10, and IL-13 represent Th2 type cytokines. The gene expressions of Th1/Th2 cytokines in human glioma cells, glioma infiltrating lymphocytes, and glioma cell lines were detected by reverse transcription polymerase chain reaction （RT-PCR）. The biological activity of cytokines in the supematant of glioma cell lines was assayed by enzyme-linked immunosorbent assay （ELISA） method. Results The total positive rates of Th1 and Th2 type cytokines gene in human glioma cells were 14.77% and 75%. The total positive rates of Th1 and Th2 type cytokines gene in glioma infiltrating lymphocytes were 22.73% and 68.17%. There was obviously predominant expression of Th2 type cytokines in human glioma tissues, glioma infiltrating lymphocytes, and glioma cell lines. There was no unbalanced expression of Th1/Th2 cytokines in normal brain tissues. Conclusion There is a predominant expression of Th2 type cytokines in human glioma cells. The switching of Th1/Th2 cytokines gene may play an important role in the occurring and developing of human gliomas.
基金The work was supported by the Fundamental Research Funds for the Central Universities of China(2572018CL04)the InnovationProject of State Key Laboratory of Tree Genetics and Breeding(2020A02)the Heilongjiang Touyan Innovation Team Program(Tree Genetics andBreeding Innovation Team).
文摘Basic Leu zipper ( bZIP ) genes play pivotal, versatile roles in abiotic or biotic stress responses and in other biological processes. Knowledge on the evolutionary relationships and patterns of gene expression of bZIP family members in woody plants, however, has been limited. Here we identified and characterized 47 BpbZIP genes across the silver birch ( Betula pendula Roth.) genome. With reference to bZIP classifications for Arabidopsis thaliana , all BpbZIP proteins clustered among 10 groups in phylogeny. The bZIP domains were divided into five patterns based on intron positions and splicing phases. A total of 24 conserved motifs were detected in BpbZIPs with high group specificity. We also analyzed the protein structure of the BpAREB/ABF/ ABI5 subfamily, the most important subfamily in the bZIP family. Expression analyses demonstrated that BpbZIP genes were widely involved in abscisic acid, salt, drought, and heat stress responses, with BpbZIP07/ABF4 and BpbZIP21/ABF2 most highly expressed. Our results on genome-wide identification, evolutionary relationships, gene structure, and motif and promoter element identification for BpbZIP family members in silver birch provide a comprehensive understanding of bZIP transcription factors in birch and will lead to a deeper understanding of their evolution and potential biological functions.
文摘O6-methylguanine-DNA methyltransferase (MGMT) gene expression in 6 Mer+ (HeLa S3, SMMC-7721,SGC-7901, B-239, AGZY83-a, and Cc80 1) and 2 Mer- (SHG-44 , and HeLa MR) human tumor cell lines was examined. Southern blot analysis revealed no deletion, amplification, or rearrangement of the MGMT gene in these cell lines. However ,～ 1. 0 kb transcripts were detected in the 6 Mer+ cell lines but not in the 2 Mer-cell lines by Northern blot analysis. Furthermore,a rough correlation between MGMT activity and mRNA level in these cell lines was observed. These results suggest that transcriptional regulation of the MGMT gene is the molecular basis of the absence of MGMT activity in Mer-cell lines.
基金supported by China Agriculture Research System(CARS-30)Jilin Agricultural Science and Technology College seed fund project(2013-903)
文摘This research was designed to assess the changes in anthocyanin content in grape skins of Vitis amurensis and to explore m RNA transcriptions of 11 structural genes（PAL,CHS3, CHI1, F3H2, F30 H, F3050 H, DFR, LDOX, UFGT,OMT and GST） related to anthocyanin biosynthesis during grape berry development, by the use of HPLC-MS/MS and real-time Q-PCR analysis. Accumulation of anthocyanins began at veraison, continued throughout the later berry development and reached a peak at maturity. Veraison is the time when the berries turn from green to purple. Expression of PAL, CHI1, and LDOX were up-regulated from 2 to4 weeks after flowering（WAF）, down-regulated from6 WAF to veraison, whereas DFR was up-regulated at8 WAF, and then up-regulated from veraison to maturity.CHS3, F3050 H, UFGT, GST, and OMT were down-regulated from 2 WAF to veraison, and then up-regulated from veraison to maturity. The transcriptional expressions of the11 structural genes also showed positive correlations with the anthocyanin content from veraison to maturity. Positive correlations were also observed between OMT transcriptional level and the content of methoxyl-anthocyanins, and between F3050 H transcriptional level and the content of delphinidin anthocyanins. F3H2 and F30 H expression was up-regulated at 2 WAF. F3H2 expression was down-regulated from 4 WAF to veraison and then up-regulated again from veraison to maturity. F30 H expression was down-regulated at 4 WAF and then up-regulated again from 6 WAF to maturity. F30 H transcriptional level was correlated positively with the cyanidin anthocyanin concentration from veraison to maturity. These results indicate that the onset of anthocyanin synthesis during berry development coincides with a coordinated increase in the expression of a number of genes in the anthocyanin biosynthetic pathway.
基金supported by National Natural Science Foundation(Grant No.31270663)a grant from Agricultural Ministry of China(Grant No.2013ZX08009003-002-004)
文摘Expansins are cell-wall-loosening proteins that have multiple roles during plant development and stress- related processes. In this study, a novel expansin gene PwEXP2 was cloned by rapid amplification of cDNA ends based on the cDNA library of Picea wilsonii and EST fragment of PwEXP2. It was found that PwEXP2 coded 253 amino acids, and putative signal peptides exist at the N-terminal, followed by 8 cysteines, a HFD （His-Phe-Asp） conserved domain, and 4 tryptophan residues at the C-terminal. PwEXP2 was located in cytoplasm and nucleus when transformed in an onion epidermal cell. Quantitative real-time PCR assays showed that PwEXP2 was expressed in various tissues with a relatively high level in needles and low level in mature pollen. The expression level of PwEXP2 dramatically increased after seed germination. Gene expression profiles in abiotic stresses showed that PwEXP2 was induced by high temperature and osmotic stress but not involved in ABA-dependent signaling path- way. These results display the important roles of the PwEXP2 in plant development and multiple adversity stresses.
基金This work was supported by National Natural Science Foundation of China(31570675)a Grant from the National Key Research and Development Program of China(2017YFD060070605)a Grant for National non-profit Research Institutions of Chinese Academy of Forestry(CAFYBB2018SY009).
文摘Taxol(Paclitaxel),an important anticancer drug,is derived at very low yields from Taxus(yew)species that grow very slowly.In the present study,thirteen genes that encode enzymes involved in Taxol biosynthesis in Taxus spp.were analyzed with bioinformatics methods,and their expression levels in different tissues and after cold and hormone treatments were also analyzed.The results indicated that many cis-elements related to abiotic stresses and hormones were found in the promoter sequences of the 8 genes involved in Taxol biosynthesis.Moreover,the 13 enzymes encoded by the target genes were located in different organelles and had many phosphorylation sites in the response proteins.The 13 genes were expressed highly either in roots or in stems,with lower transcripts in needles,and they were highly expressed after treatment with cold,gibberellin,methyl jasmonate or coronatine,consistent with predictions based on the bioinformatics analysis.These results suggest that the factors such as hormones and abiotic stresses stimulate taxane biosynthesis in yews,providing an important way to sustainably generate taxanes from yew trees or their cell cultures to improve Taxol yields.
基金supported by AForsk(https://aforsk.com)[15-416]Stiftelsen Tornspiran(http://stiftelsentornspiran.se)+2 种基金Anna och Nils Hakanssons Stiftelse(http://www.annaochnilshakanssonsstiftelse.se)Helge Ax:son Johnsons stiftelse(http://haxsonj.se/www/)[770721-0204]Magnus Bergvalls Stiftelse(http://www.magnbergvallsstiftelse.nu)[2014-00501]。
文摘Nicotinamide treatment of plants and plant cell cultures has been shown to promote defense and decrease levels of DNA methylation.In the present study,we used RNA-seq technology to study overall changes in gene expression induced in roots of 3-month-old spruce(Picea abies)seedlings grown from nicotinamide-treated seeds to examine the molecular mechanisms underlying the defense promotion.Approximately 350 genes were identified as differentially expressed in roots after the seed treatment.Stress response genes,including transcription factors MYB77 and LHY and two chitinase enzymes,were generally upregulated,whereas genes thought to be involved in epigenetic regulation such as DDM1,known to promote DNA methylation,were present at high frequency among the downregulated genes.Across all samples,the expression of downregulated epigenetic-related genes was highly correlated with the nicotinamide treatment,indicating a common regulation.Our results support an earlier hypothesis regarding a potential role of nicotinamide as a defense-signal mediator.
