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The consecutive monoculture obstacle is a major problem in the field of Rehmannia glutinosa( R. glutinosa),has severely declined the yield and quality of R. glutinosa. Here,using hi TAIL-PCR and RACE techniques,we have cloned the full-length transcript( 1 573 bp) of Unigene 29334＿All screened by DGE as a consecutive monoculture obstacle response gene of R. glutinosa. Based on ORF Finder prediction,all ORFs detected in the full-length transcript were less than 300 nt,which suggested that the above transcript was confirmed to be a long non-coding RNA( LncRNA). With alignment in R. glutinosa transcriptome,this LncRNA was partially homologous to alanine glyoxylate transaminase 2 gene( Rg AGT2),which was named LncRNA-RgATG2. To further explore the function of LncRNA-RgAGT2,we have examined expression patterns of LncRNA-RgAGT2 and Rg AGT2 at five critical development stages( seedling,elongation,pre-expanding,mid-expanding,late-expanding) in the first and second year replanting of R. glutinosa,respectively. The results indicated that LncRNA-RgAGT2,as a potential regulator,is possible to play a vital role in Rg AGT2 expression regulation. Meanwhile,LncRNA-RgAGT2 has presented significant variation in all development stages of R. glutinosa,which could be used as a " diagnostic label" to assess consecutive monoculture obstacle. This study,for the first time,showed that LncRNA was responsible for the response and regulation of consecutive monoculture obstacle,which would be a powerful supplement to reveal the molecular mechanisms of consecutive monoculture obstacle of R. glutinosa.
Subject(s)
Cloning, Molecular , Gene Expression , RNA, Long Noncoding , Rehmannia , TranscriptomeABSTRACT
Objective To detect the SNR and geometric accuracy of MRI based on American College of Radiology (ACR)standards.Methods The SNR and geometric accuracy of Siemens Skyra 3.0T,Siemens Trio 3.0T and GE Excite HD 1.5T MRI were measured with ACR phantom,and the detection results were calculated according to the standards.Results The SNR values of Siemens Skyra 3.0T,Siemens Trio 3.0T and GE Excite HD 1.5T MRI were 589.98,438.50 and 277.12 respectively.Siemens Skyra 3.0T MRI had the values of geometric accuracy being-1.93％ at X direction,-3.20％ at Y direction and 0.68％ at Z direction,Siemens Trio 3.0T MRI had the value of geometric accuracy being-0.87％ at X direction,-2.33％ at Y direction and 1.49％ at Z direction,GE Excite HD 1.5T MRI had the values of geometric accuracy being 0.20％ at X direction,-1.53％ at Y direction and 1.69％ at Z direction.Conclusion The detection of the SNR and geometric accuracy of MRI can effectively guarantee the image quality.
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Objective To test the spatial resolution and average glandular dose (AGD) of full field digital mammography system to obtain high-resolution and contrast breast X-ray images with the lowest dose.Methods The M12 breast X-ray performance test phantom was placed on the breast support center position,then line-pair card was put on the M12 phantom and fixed groove,and exposure was carried out with auto exposure control,28 kV tube voltage and common anode filtration combination so as to detect the spatial resolution.Half-value layer (HVL) was measured by Fluke TNT12000 ray detection tools,2 mm-thickness plate and aluminum sheets with different thicknesses,and then AGD was calculated accordingly.Results The spatial resolutions were all higher than 7 lp/mm and proved to meet the quality standard,which had the vertical spatial resolution being 8 lp/mm,the maximum value of the lateral spatial resolution being 10 lp/mm and the minimum value being 8 lp/mm.AGD was lower than 2 mGy and then proved qualified which was limited within 0.60 and 0.61 mGy.Conclusion The test of spatial resolution and AGD of full digital mammography system can assess the performance and radiation dose,reflect the performance of equipment,ensure image quality and lower radiation dose.
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Objective To detect the SNR and geometric accuracy of MRI based on American College of Radiology (ACR)standards.Methods The SNR and geometric accuracy of Siemens Skyra 3.0T,Siemens Trio 3.0T and GE Excite HD 1.5T MRI were measured with ACR phantom,and the detection results were calculated according to the standards.Results The SNR values of Siemens Skyra 3.0T,Siemens Trio 3.0T and GE Excite HD 1.5T MRI were 589.98,438.50 and 277.12 respectively.Siemens Skyra 3.0T MRI had the values of geometric accuracy being-1.93％ at X direction,-3.20％ at Y direction and 0.68％ at Z direction,Siemens Trio 3.0T MRI had the value of geometric accuracy being-0.87％ at X direction,-2.33％ at Y direction and 1.49％ at Z direction,GE Excite HD 1.5T MRI had the values of geometric accuracy being 0.20％ at X direction,-1.53％ at Y direction and 1.69％ at Z direction.Conclusion The detection of the SNR and geometric accuracy of MRI can effectively guarantee the image quality.
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Objective To test the spatial resolution and average glandular dose (AGD) of full field digital mammography system to obtain high-resolution and contrast breast X-ray images with the lowest dose.Methods The M12 breast X-ray performance test phantom was placed on the breast support center position,then line-pair card was put on the M12 phantom and fixed groove,and exposure was carried out with auto exposure control,28 kV tube voltage and common anode filtration combination so as to detect the spatial resolution.Half-value layer (HVL) was measured by Fluke TNT12000 ray detection tools,2 mm-thickness plate and aluminum sheets with different thicknesses,and then AGD was calculated accordingly.Results The spatial resolutions were all higher than 7 lp/mm and proved to meet the quality standard,which had the vertical spatial resolution being 8 lp/mm,the maximum value of the lateral spatial resolution being 10 lp/mm and the minimum value being 8 lp/mm.AGD was lower than 2 mGy and then proved qualified which was limited within 0.60 and 0.61 mGy.Conclusion The test of spatial resolution and AGD of full digital mammography system can assess the performance and radiation dose,reflect the performance of equipment,ensure image quality and lower radiation dose.
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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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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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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>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
Compound SIPI5047 was synthesize by using piperazine as starting material in five reaction steps, and its central none-opioid analgesic activity was studied. Its analgesic activity, pharmacological mechanism, action type and drug dependence were well studied in vivo and in vitro. The results show that SIPI5047 has potent analgesic activities in vivo, which is quite similar to morphine and also much more powerful than paracetamol. SIPI5047 has no efficacy to reduce fever or inflammation, but has an obvious action on central nervous system. SIPI5057 has no apparent affinity with the mu-receptor and it is an antagonist that acts on the polyamine site of the NMDA receptor. SIPI5057 appears no drug dependence. SIPI5047 is a novel central none-opioid analgesic agent and more worthy of further research as a new drug candidate.