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In recent years, the increase in the number of cases of postural asphyxia has gradually attracted the attention and discussion of forensic scientists domestically and internationally, but a systematic, comprehensive and recognized expert consensus and identification standard has not been established at home and abroad. This paper reviews the case characteristics, occurrence, mechanism of death, and identification criteria of postural asphyxia, to provide reference for future research.
Subject(s)
Humans , Asphyxia/etiology , Forensic Medicine , Forensic PathologyABSTRACT
Objective To study the relationship between material mechanics and bone material parameters of rat skulls and their correlation with age by examination of the parameters. Methods Forty-eight healthy male SD rats were divided into 2, 4, 6, 8, 17, 26, 52 and 104 week groups according to their age. Each group had six rats. The right cranium was compressed by KD Ⅱ-0.2 microcomputer controlled electronic universal testing machine, and material mechanics parameters (ultimate load, compression strength and compression modulus) were measured, then the skull slices were cut off and scanned by Micro-CT system to detect bone material parameters (skull thickness, bone mineral density, bone volume, and trabecular thickness). Results The differences in ultimate load, compression strength and compression modulus among all groups had statistical significance (P<0.05), and were positively correlated with age within 26 weeks (P<0.05). The differences in skull thickness, bone mineral density, bone volume and trabecular thickness among all groups had statistical significance (P<0.05), and were positively correlated with age within 52 weeks (P<0.05). All material mechanics parameters were positively correlated with bone material parameters (P<0.05). Conclusion There is a positive correlation between bone material parameters (skull thickness, bone mineral density, bone volume, trabecular thickness), material mechanics parameter (skull ultimate load, compression strength, compression modulus) and age in a certain range, which can be used to infer age.
Subject(s)
Animals , Male , Rats , Biomechanical Phenomena , Bone Density , Rats, Sprague-Dawley , Skull/diagnostic imagingABSTRACT
Objective:To explore the composition characteristics of rhizosphere soil under <italic>Rehmannia glutinosa-Zea mays</italic> intercropping model,and screen out special signal substances in rhizosphere soil of <italic>R. glutinosa</italic> under intercropping <italic>Z. mays</italic>, so as to provide the basis for the study of allelopathic substances in continuous cropping obstacle of <italic>R. glutinosa</italic>. Method:In this experiment,rhizosphere soils of <italic>R. glutinosa</italic> under <italic>Z. mays </italic>intercropping and <italic>R. glutinosa </italic>single cropping models in July,August,September and October were taken as the research objects, and the volatile organic compounds in ethyl acetate fraction were analyzed by gas chromatography-mass spectrometry (GC-MS). Principal component analysis (PCA), hierachical cluster analysis (HCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) analysis were performed on the data by SIMCA 14.1 to screen out potential differences in volatile organic compounds between the two models. Result:The types of volatile organic compounds in intercropping and single cropping models were mainly hydrocarbons, alcohols, esters, ketones, amides, acids and other substances. Specifically, the average relative contents of hydrocarbons,esters and amides in intercropping model were 58.46%,32.15% and 5.42% respectively,while the relative contents of hydrocarbons,esters and amides in single cropping model were 37.27%,36.11% and 21.13%. The results of PCA and HCA showed that the characteristics of volatile organic compounds in the ethyl acetate fraction of rhizosphere soil under intercropping and single cropping models could be clearly divided into two categories,the screening results of potential differential components based on OPLS-DA analysis indicated that various components, such as dibutyl phthalate,(<italic>Z</italic>)-9-oleamide,<italic>β</italic>-caryophyllene,dioctyl iso-phthalate, phthalate (2-propylamyl) diester, <italic>n</italic>-hexadecane,octodecane, <italic>n</italic>-heneicosane, were screened from rhizosphere soil under the two models. Conclusion:The <italic>R. glutinosa-Z. mays</italic> intercropping model has certain effects on the volatile organic compounds in the rhizosphere soil of <italic>R. glutinosa</italic>,and the effect of the selected components on the growth and quality characteristics of <italic>R. glutinosa</italic> still need to be further studied.
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NRT1 family proteins play an important roles for absorbing and transporting of nitrate in different plants. In order to identify the NRT1 family genes of Rehmannia glutinosa, this study used 11 NRT1 homologous proteins of Arabidopsis as probe sequences and aligned with the transcriptome data of R. glutinosa by using NCBI BLASTN software. Resulting there were 18 NRT1 proteins were identified in R. glutinosa. On basis of this, a series of the molecular characteristics of R. glutinosa NRT1 proteins including the conserved domains, the transmembrane structure, the subcellular location and phylogenetic features were in detail analyzed. At same time, it were systematically analyzed that the temporal and spatial expression patterns and characteristics of R. glutinosa NRT1 family genes in response to different stress factors. The results indicated that 18 R. glutinosa NRT1 family genes with the length of coding region from 1 260 bp to 1 806 bp, encoded proteins ranging from 419 to 601 amino acids, and all of they owned the domains of typical peptide transporter with 7 to 12 transmembrane domains. These R. glutinosa NRT1 family proteins mostly were found to locate on cellular plasma membrane, and belonged to the hydrophobic proteins. Furthermore, the evolutionary analysis found that the 18 R. glutinosa NRT1 protein family could be divided into two subfamilies, of which 14 NRT1 family genes might occur the positive selection, and 4 genes occur the passivation selection during the evolution process of R. glutinosa. In addition the expression analysis showed that 18 R. glutinosa NRT1 family genes have the distinct expression patterns in different tissues of R. glutinosa, and their expression levels were also obvious difference in response to various stress. These findings infield that 18 R. glutinosa NRT1 family proteins might have obviously different functional roles in nitrate transport of R. glutinosa. In conclusion, this study lays a solid theoretical foundation for clarifying the absorption and transport molecular mechanism of N element during R. glutinosa growth and development, and at same time for deeply studying the molecular function of R. glutinosa NRT1 proteins in absorption and transport of nitrate.
Subject(s)
Anion Transport Proteins , Membrane Transport Proteins , Nitrates , Phylogeny , Plant Proteins/metabolism , Rehmannia/genetics , TranscriptomeABSTRACT
The present study analyzed the effects of planting density on the development, quality, and gene transcription characte-ristics of Rehmannia glutinosa using 85-5 and J9 as materials with three planting densities of 5 000, 25 000, and 50 000 plants/Mu(1 Mu≈667 m~2). The agronomic characteristics of leaves and tuberous roots, the content of catalpol and acteoside, and the changes of gene expression were determined. The results showed that the leaf size, the diameter of tuberous root, leaf biomass, tuberous root number, and tuberous root biomass per plant at low density were significantly higher than those of medium and high densities. The content of catalpol and acteoside in leaves was higher at high density. The content of catalpol in tuberous roots was higher at low density, and the change trend was similar to that in leaves, while the content of acteoside in tuberous roots was higher at high density. Transcriptome analysis found that about 1/2 of the expansin genes could change regularly in response to density treatment, which was rela-ted to the development of tuberous roots. The change trend of the gene expression of multiple catalytic enzymes involved in the biosynthesis of catalpol and acteoside was consistent with that of their content, which was presumedly involved in the accumulation and regulation of density-responsive medicinal components. Based on the analysis of the development, medicinal components, and gene expression characteristics of R. glutinosa at different densities, this study is expected to provide an important basis for regulating the quality and yield of medicinal materials of R. glutinosa by managing the planting density.
Subject(s)
Gene Expression Profiling , Plant Leaves/genetics , Plant Roots/genetics , Rehmannia/genetics , Transcription, GeneticABSTRACT
Objective To study the relations of the dural thickness and the biomechanical parameters with postmortem interval (PMI) of human cadavers, and to explore the feasibility of the two indexes used for PMI estimation. Methods Dural samples were collected at different postmortem intervals of 6 h, 12 h, 18 h, 24 h, 30 h, 36 h, 48 h, 60 h, 72 h, 84 h and 96 h, then fabricated into 4.0 cm×1.0 cm A and B test specimens. The thickness of the A test specimen was measured, and the biomechanical parameters, such as ultimate load, maximum force deformation, tensile strength, elastic modulus and fracture force, were measured. The regression equations of thickness, biomechanical parameters and PMI were fitted respectively, and the difference between the predicted value and actual measured value of PMI was verified by the verification group. The B test specimen was fixed with 10% neutral formaldehyde solution, then tissue sections were prepared to observe its morphological changes. Results From 6 h to 96 h after death, the dural thickness decreased gradually, the collagen fibers gradually changed from clear arrangement to mutual fusion, and the number of nuclei decreased gradually. The dural thickness, ultimate load, tensile strength, elastic modulus and fracture force decreased sequentially, among which the dural thickness, ultimate load, elastic modulus and fracture force had a correlation with PMI (P<0.05). In the return test, the difference between the predicted value and actual measured value of PMI in the verification group had no statistical significance (P>0.05). Conclusion The dural thickness, ultimate load, elastic modulus and fracture force change sequentially from 6 h to 96 h after death. The regression equation established by the relationship between the changes and PMI can be used for PMI estimation.
Subject(s)
Humans , Autopsy , Cadaver , Fractures, Bone , Postmortem Changes , Time FactorsABSTRACT
Objective: To establish a chemical constitution-disease target-metabolic signaling pathway network of Leonurus japonicus, and explore the mechanism of multi-component, multi-target and multi-pathway of L. japonicus for the treatment of dysmenorrhea caused by cold coagulation and blood stasis. Methods: TCMSP database and Swiss Target Prediction server were used to obtain the chemical components and action targets of L. japonicus. Combined with DrugBank, DisGeNET, TTD and other databases, the action targets of L. japonicus for treating dysmenorrhea symptoms wtih cold coagulation and blood stasis were obtained. Through DAVID's website, GO function enrichment analysis and KEGG pathway analysis were conducted on the targets, and cytoscape 3.6.0 was used to construct the network diagram of the active component-dysmenorrhea target-metabolic signaling pathway of L. japonicus, to explore the mechanism of action of L. japonicus in treating dysmenorrhea with cold coagulation and blood stasis. Furthermore, Western blotting experiments were conducted to verify the effect of L. japonicus on the expression of PTGS1 and PTGS2 proteins in the uterine tissues of the model rats with cold coagulation and blood stasis dysmenorrhea. Results: Eight potential active ingredients of L. japonicus were obtained, including 22 dysmenorrhea related disease targets, main targets PTGS1, PTGS2, ALOX5, PLAG2A, AKR1C3, etc. A total of 71 GO items were obtained by GO functional enrichment analysis (P < 0.05), including 46 biological process (BP) items, four cell component (CC) items, and 21 molecular function (MF) items. There were 22 possible targets for the treatment of dysmenorrhea, and seven signaling pathways were obtained through KEGG pathway enrichment and screening (P < 0.05), involving arachidonic acid metabolism, linoleic acid metabolism, steroid hormone biosynthesis, etc. L. japonicus significantly increased the protein expression levels of PTGS1 and PTGS2 in the uterus of the model rats (P < 0.05), which confirmed some of the predicted results of network pharmacology. Conclusion: L. japonicus may treat dysmenorrhea with cold coagulation and blood stasis by acting on arachidonic acid metabolism pathway.
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Spatholobi Caulis, the vine stem of Spatholobus suberectus and widely used in China and Southeast Asian nations, has the effects on nourishing the blood and promoting blood flow, regulating menstruation and relieving pain, and invigorating the nerves. Through consulting the herbal textual and local chronicles, we summarized the original textual research and medicinal evolution on Spatholobi Caulis to analyze the changes of varieties in different historical periods. Further, the major active ingredient in Spatholobi Caulis was discussed. According to the literature to date, 60 flavonoids compounds have been isolated and could be divided into isoflavones, dihydroflavones, flavanols, dihydroflavonols, procyaninides, chalcones, pterocarpans, isoflavanols, isoflavanones and aurone according to their molecular structures. These indicative ingredients in Spatholobi Caulis showed pharmacological activities on regulation of the blood system, anti-tumor, anti-oxidation, anti-virus, anti-bacteria and inhibition of melanin deposition. This review will provide reference and basis for the sustainable use of resources and industry development on Spatholobi Caulis.
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Calcium signaling plays a critical role in response to various abiotic and biotic stresses in plants. Preliminarily evidence showed that calcium signaling perceived and transduced the harmful signaling generated from continuous cropping stress in R. glutinosa. To investigate the roles of calcium signaling in continuous cropping injury formation, the key genes involved in calcium signaling transduction were identified in R. glutinosa transcriptome through bioinformatic methods. Furthermore, the calcium ion concentration in both normal and continuous cropping R. glutinosa root cells were measured by potassium pyroantimonate precipitation and calcium fluorescence method. As a result, a set of 84 calcium signaling-related genes, including 5 CaMs, 12 CBLs, 21 CDPKs, 21 CIPKs, 16 CMLs, and 9 CRKs were captured in R. glutinosa transcriptome. The analysis of expression profile in continuous cropping compared to normal growth R. glutinosa indicated that continuous cropping stress significantly increased the expression of calcium signaling-related genes in continuous cropping R. glutinosa. At the same time, the abundance levels of 12 calcium signaling-related genes quantified by qPCR further validated the high expression of calcium signaling-related genes presented in continous cropping R. glutinosa. In addition, the continuous cropping condition significantly promoted the accumulation of intracellular calcium ions in R. glutinosa based on two methods of potassium pyroantimonate precipitation and calcium fluorescence. This study verified the possible roles of calcium signaling in the formation of continuous cropping injury on molecular and cellular level, which lays a solid foundation for illuminating formation mechanism of continuous cropping injury on molecular level.
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Although consecutive monoculture problems have been studied for many years, no effective treatments are currently available. The complexity of systems triggered the formation of consecutive monoculture problems was one major cause. This paper elaborated the physiological and ecological mechanisms of consecutive monoculture problem formation based on the interaction relationship among multiple factors presented in the rhizosphere soil of consecutive monoculture plants. At same time, in this paper the multiple interactions among cultivated medicinal plants, autotoxic allelochemicals and rhizosphere microbial were proposed to be most important causes that derived the formation of consecutive monoculture problem. The paper also highlighted the advantage of 'omics' technologies integrating plant functional genomics and metabolomics as well as microbial macro-omics in understanding the multiple factor interaction under a particular ecological environment. Additionally, taking R. glutinosa as an example, the paper reviewed the molecular mechanism for the formation of R. glutinosa consecutive monoculture problem from the perspective of the accumulation of allelopathic autotoxins, the rhizosphere microecology catastrophe and theresponding of consecutive monoculture plants. Simultaneously, the roles of mutilple 'omics' technologies in comprehending these formation mechanism were described in detail. This paper provides finally a new insight to solve systematically the mechanism of consecutive monoculture problem formation on molecular level.
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This paper clarified in detail the definition, characteristics of allelopathy and its association with consecutive monoculture problem.Most of studies have indicated that it is critical to parse the formation mechanisms of consecutive monoculture problem that identification of allelochemicals and verification of its function. Here, we proposed a new approach to separate and identify the allelochemical group precisely and effectively, in which the "knock-out/in" methods of targeting ingredients in the model of medicinal effect identification and quality control were applied. This method will contribute to deep understanding plant allelopathy, and provide theoretical basis and technical support for alleviating consecutive monoculture problems simultaneously.
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The efficacy of Rehmannia glutinosa which as a large quantity of traditional Chinese medicine is significant. However, the land must be given up after one season of R. glutinosa cultivation or replanted after a period of 8-10 years because of the severe continuous cropping obstacles. MicroRNAs is a class of endogenous non-coding small RNAs, which participate in regulation of physiological activities by target mRNA cleavage or translational repression in plants. In recent years,studies on the role of miRNAs in plants have made significant progresses,especially in medicinal plants.MiRNAs from some different medicinal plant species have been identified with regulatory effects.When plants are exposed to environmental stress, miRNAs act on stress-related genes and initiate stress-resistance mechanisms in the body against adverse factors. R. glutinosa is also a kind of environmental stress. It is conducive to deciphering the molecular mechanism of continuous cropping obstacles for us by researching miRNAs. This article reviews the production of miRNAs, mechanism, research approaches and characteristics of resisting the environmental stresses in plants, the development trends and future prospect of R. glutinosa miRNAs research.
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Using cDNA from Rehmannia glutinosa leaf as template, a 972 bp fragment of expansin gene which containing a 762 bp ORF that encoded 253 amino acids, was cloned, named RgEXPA10, which GenBank accession number for this gene is KF011918. A 1 207 bp genomic sequence of RgEXPA10 was amplified by PCR with leaf DNA as template, sequencing analysis revealed that three exons and two introns in RgEXPA10 genomic sequence, and which GenBank accession number is KF011919. Molecular and bioinformatic analyses indicated that RgEXPA10 protein have DPBB_1 and Pollen_allerg_1 domain, also including a 26 aa nuclear localization signal and a 19 aa transmembrane region. Phylogenetic analysis revealed that RgEXPA10 showed the highest homology with AtEXPA8 among the 26 α-expansins in Arabidopsis thaliana. However, the RgEXPA10 indicated the highest homology with the expansin from Solanum lycopersicum among 22 plant species. Expression patterns using qRT-PCR analysis showed that RgEXPA10 mainly expressed in unfolded leaf, followed by the tuberous root at stage of expanding period, and rarely expressed in senescing leaf. And RgEXPA10 showed higher expression level in tuberous root at 60 and 90 days after emergence. The transcription level of RgEXPA10 significantly reduced under all the three stresses including continuous cropping conditions, salinity and waterlogging. This study will lay foundations for molecular function in development and regulation of different stresses for R. glutinosa.
Subject(s)
Amino Acid Sequence , Cloning, Molecular , DNA, Complementary , Gene Expression Regulation, Plant , Genes, Plant , Molecular Sequence Data , Phylogeny , Plant Leaves , Plant Proteins , Genetics , Plant Roots , Rehmannia , GeneticsABSTRACT
In order to study the development characteristics of Rehmannia glutinosa tuberous root expansion and reveal the regulation mechanism of the genes related to hormones in this process, R. glutinosa "wen-85" was used as the experimental material in this study. R. glutinosa tuberous roots of different developmental stages were collected to observe phenotype and tissue morphology using resin semi-thin sections method. The genes related to hormone biosynthesis and response were chosen from the transcriptome of R. glutinosa, which was previously constructed by our laboratory, their expression levels at different development stages were measured by real-time quantitative PCR. The results showed that the root development could be divided into six stages: seeding, elongation, pre-expanding, mid-expanding, late-expanding and maturity stage. The anatomic characteristics indicated that the fission of secondary cambium initiated the tuberous root expansion, and the continuous and rapid division of secondary cambium and accessory cambium kept the sustained and rapid expansion of tuberous root. In addition, a large number oleoplasts were observed in root on the semi-thin and ultra-thin section. The quantitative analysis suggested that the genes related to biosynthesis and response of the IAA, CK, ABA,ethylene, JA and EB were up-regulated expressed, meanwhile, GA synthesis and response genes were down-regulated expressed and the genes of GA negative regulation factors were up-regulated expressed. The maximum levels of most genes expression occurred in the elongation and pre-expansion stage, indicating these two stages were the key periods to the formation and development of tuberous roots. Oleoplasts might be the essential cytological basis for the formation and storage of the unique medicinal components in R. glutinosa. The results of the study are helpful for explanation of development and the molecular regulation mechanism of the tuberous root in R. glutinosa.
Subject(s)
Gene Expression Regulation, Developmental , Genetics , Gene Expression Regulation, Plant , Genetics , Lipid Droplets , Metabolism , Microscopy, Electron, Transmission , Plant Growth Regulators , Pharmacology , Plant Proteins , Genetics , Metabolism , Plant Roots , Genetics , Metabolism , Rehmannia , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Time FactorsABSTRACT
To clone and analyze a member of the Auxin/indole-3-acetic acid (Aux/IAA) gene family, RgIAA1, from Rehmannia glutinosa. The transcriptional EST database of R. glutinosa was used to clone the new Aux/IAA gene by cDNA probe of AtIAA14. Bioinformatics was applied to analyze the sequence characteristics of RgIAA1 protein and construct phylogenetiC trees. Quantitative RT-PCR has been applied to detect the transcription level of RgIAA1 in seven tissues as well as in leaves under three stresses. The results showed that, the cDNA sequence of RgIAA1 contains 903 bp was obtained. The open reading frame (ORF) of RgIAA1 was 681 bp encoding 226 amino acids, which has typical structural domains and characteristic sequence of Aux/IAA family proteins. RgIAA1 showed the highest expression level in unfolded leaf, followed by the stem. And the expression of RglAA1 was quickly decreased with leaf growing up. The transcription level increased under continuous cropping conditions while it reduced both in salinity and waterlogging stresses. RgIAA1, an Aux/IAA gene from R. glutinosa has been obtained for the first time, which can lay the foundation for further studies about its molecular function in development and responses to stress.
Subject(s)
Amino Acid Sequence , Cloning, Molecular , Gene Expression Regulation, Plant , Indoleacetic Acids , Metabolism , Molecular Sequence Data , Organ Specificity , Phylogeny , Plant Proteins , Chemistry , Genetics , Rehmannia , Classification , Genetics , Physiology , Stress, Physiological , GeneticsABSTRACT
Based on the early transcriptome and digital differentially expressed profiling library construction in consecutive monoculture (two-year culturing) Rehmannia glutinosa, we screened and chose the twelve differentially expressed protein genes which might be related with calcium signal system. The spatiotemporal expression of these genes was measured by the real-time quantitative PCR, and the relative expression values of the genes related with calcium signal system in different development stages and tissues of normal growth (one-year culturing) and succession cropping of R. glutinosa (two-year culturing) was elaborated in detail. In addition, disposed succession cropping of R. glutinosa was treated with different levels of calcium signal blocking agents in order to verify the mode of action of calcium signal system on consecutive monoculture problem in R. glutinosa. Among the twelve genes, two calcium channels away from the cytoplasm were down-regulated expressed, while the ten calcium channels toward the cytoplasm were up-regulated expressed. The results implied that succession cropping caused calcium ions flowing from endoplasmic reticulum to cytoplasm. While the key genes in calcium signal respond components such as CBL, CBP, CIBP, PLC, etc. were down-regulated expressed significantly in succession cropping of R. glutinosa which were disposed with calcium signal blocking agents, the extent of the damage was relieved, and approached the normal growth (one-year culturing) level. This result strongly showed that calcium signal system participated in the perceiving, conducting and magnifying processes of succession cropping obstacles of R. glutinosa.
Subject(s)
Calcium , Metabolism , Calcium Signaling , Gene Expression Regulation, Developmental , Plant Proteins , Genetics , Metabolism , Rehmannia , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the expression and clinical significance of Semaphorin4D (Sema4D) mRNA in peripheral blood lymphocyte, Sema4D on platelet surface, soluble Sema4D (sSema4D) in plasma in patients with cerebral infarction.</p><p><b>METHODS</b>Taking 299 patients with cerebral infarction as the case group while 195 healthy adults as the control group. The mRNA expression of Sema4D was detected by Real-time PCR, and Sema4D expression on platelet by flow cytometry, sSema4D by ELISA. Then, the expression of Sema4D on platelet surface and the concentration of sSema4D in plasma of the 195 selected patients following 2 weeks' treatment were tested.</p><p><b>RESULTS</b>The expression of Sema4D mRNA significantly increased in the case group \[(2.23, 2.66)×10(4) IU/ml\] than in the control group \[(0.49, 0.53)×10(4)IU/ml\] (P < 0.01). The level of Sema4D on platelet surface in the case group (191.62 ± 46.56) significantly decreased than in the control group (303.33 ± 112.66) (P < 0.01). But the concentration of sSema4D in plasma in the case group \[(1.34 ± 0.56) µg/L\] was obviously higher than in the control group \[(0.61 ± 0.31) µg/L\] (P < 0.01). The expression of Sema4D on platelet was obviously relevant with the concentration of sSema4D in plasma in the case group with the correlation coefficient as 0.328 (P < 0.01). The expression of Sema4D on platelet obviously peaked up following 2 weeks' routine therapy in the case group, which was close to that in the control group. Meanwhile the concentration of sSema4D in plasma was downward corrected to the normal in the case group.</p><p><b>CONCLUSION</b>The increased expressions and plasma levels, and reduced expressions on platelet of Sema4D in acute period, which returned to normal 2 weeks after treatment in the case group may be related to the occurrence of acute cerebral infarction, reflecting the development process of cerebral infarction.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Antigens, CD , Blood , Metabolism , Blood Platelets , Metabolism , Case-Control Studies , Cerebral Infarction , Blood , Lymphocytes , Metabolism , Semaphorins , Blood , MetabolismABSTRACT
<p><b>OBJECTIVE</b>Based on previous study, authors used the suppression subtractive hybridization (SSH) technique to construct the forward and reverse subtractive cDNA libraries of consecutive monoulture Rehmannia glutinosa. Five genes related with consecutive monoculture problem of R. glutinosa were chosen from the each of two subtractive libraries. And their spatiotemporal expression was measured in order to explore the functions in consecutive monoculture problem of R. glutinosa.</p><p><b>METHOD</b>Using the real-time quantitative PCR, we tested the relative expression values of the genes in different development stages and tissues of normal growth (one-year culturing) and consecutive monoculture (two-year culturing) R. glutinosa.</p><p><b>RESULT</b>The five genes (calcium-dependent protein kinase, s-adenosyl-methionine synthetase, Aminocyclopropane-1-carboxylate oxidase, methyltransferase, calpain), which were chosen from the forward library had high expression in consecutive monoculture R. glutinosa, especially in root, and were hardly expression in normal growth R. glutinosa. On the contrary, the other five genes (RNA-dependent RNA polymerase, RNA replicase, DNA-directed RNA polymerase IIa, cyclin D, RNA binding protein) chosen from the reverse library had high expression in one-year R. glutinosa, but were down regulated or shut down in consecutive monoculture R. glutinosa.</p><p><b>CONCLUSION</b>The key genes, which regulate inessential metabolism parthway (such as cyclin D, DNA-directed RNA polymerase IIa), were restrained or shut down in consecutive monoculture R. glutinosa. Calcium and ethylene signaling might played key roles in the formation of consecutive monoculture problem, resulting in disturbing normal metabolic process and syndrome of disease in R. glutinosa appeared in turn.</p>
Subject(s)
Cell Culture Techniques , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Gene Library , Plant Proteins , Genetics , Metabolism , Rehmannia , Genetics , MetabolismABSTRACT
This paper introduced the work principle, execution, use and major characteristics of the remote hydraulic pressure control injection. Using Pascal principle, it is more accurate, convenient, cheap and safe. It could be used in all the fields of the medicine.