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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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Objective: To determine the content of index components in different parts of Gardenia jasminoides (pericarp, seeds, whiskers), study the fingerprint, and compare the contents and compositions differences of different parts of G. jasminoides, in order to provide the theoretical basis for different efficacies of G. jasminoides pericarp and seeds, explore the exploitation and utilization values of G. jasminoides whiskers, and avoid waste of gardenia medicinal resources. Method: The contents of geniposide and crocetin Ⅰ was were determined by HPLC, the content of total iridoid glycosides was determined by ultraviolet spectrophotometry, and three index components in different parts of G. jasminoides were analyzed. HPLC fingerprints of different parts of G. jasminoides were collected, the common pattern of HPLC fingerprints of different parts of G. jasminoides of different origins and with different processing methods was established, and the similarity evaluation software was used for data analysis; comparative analysis on fingerprints of different parts of G. jasminoides was conducted. Result: Content change of index components in G. jasminoides pericarp and seeds from Henan, Fujian and Jiangxi were the same. Content of geniposide:Fujian > Henan > Jiangxi, the contents of three components in G. jasminoides pericarp from Fujian were much higher than those from Henan and Jiangxi, the contents of crocetin Ⅰ and total iridoid glycosides:Fujian > Jiangxi > Henan, the contents of total iridoid glycosides from Fujian, Jiangxi were much higher than those from Henan. The order of three index components in G. jasminoides whiskers from different origins from high to low, the content of geniposide and crocetin Ⅰ was Fujian > Jiangxi and Henan, the content of total iridoid glycosides was Fujian > Jiangxi > Henan.In the same part, there were 22 common peaks in the fingerprints of G. jasminoides pericarp, except for S13-S15, the similarity of other samples were more than 0.9;the fingerprints of G. jasminoides seeds had 22 common peaks, except for S22-S30, the similarities of other samples were more than 0.9;the fingerprints of G. jasminoides whiskers had 16 common peaks, except for S7-S9, the similarities of other samples were more than 0.9.In different parts, the fingerprints of G. jasminoides whiskers were significant different from those of pericarp and seeds, the number of peaks in G. jasminoides whiskers reduced, the order of height of peaks 2, 3, 5 of G. jasminoides from high to low were whiskers > gardenia > seeds. There was not peak X in the seeds, the height of peak X of gardenia in whiskers was higher than that in pericarp, except for the peak 17, the height of all peaks in seeds were higher than that in whiskers. Conclusion: There are significant differences in the contents of index components in G. jasminoides pericarp and seeds. The content of total glycosides in gardenia is high, suggesting that it can be used to extract total iridoid glycosides. The fingerprints can reflect the content difference and species distribution of different parts of G. jasminoides, so as to provide theoretical support for the studies for pharmacodynamic material basis of G. jasminoides and the scientificity and rationality of the separate application of G. jasminoides pericarp and seeds.
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Objective: To discuss the phenotypic character and the HPLC fingerprints of radial striations from different germplasms Rehmanniae Radix. Method: The changes in the shape and column diameter of the radial striations of Rehmanniae Radix were observed and measured in the whole growth period. Besides,the HPLC fingerprints of the root,radial and un-radial striations were established to sign the chemical quality and analyzed by principal component analysis(PCA)and systematic cluster analysis. Result: There were significantly differences and regularities in the shape and proportion of the radial striations of different germplasms Rehmanniae Radix. The fingerprints showed the consistency between different types of chemical ingredients,and the differences in chemical quality characteristics mainly lay in the content of chemical compositions and theirs relative ratio. The results of PCA indicated that active ingredients, such as acteoside,catalpol,rehmaionoside D,rehmaionoside A and leonuride, were involved in the quality expression of different parts from various germplasms of Rehmanniae Radix,but each ingredient had a distinctive contribution rate to the differential quality expression between different parts from various germplasms of Rehmanniae Radix. However,the other components involved in the differential quality expression had different contribution rates in different germplasms.The systematic cluster analysis indicated that great differences in the chemical quality between the radial striations and un-radial striations of Beijing-1,Qinhuai,Qinhuai Zheng and 1706 germplasms,but with small differences in 85-5 and Baixuan germplasms. Conclusion: There are differences in phenotypic character of the radial striations and HPLC fingerprints between different germplasms Rehmanniae Radix.
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Objective:To study HPLC fingerprints of Achyranthis Bidentatae Radix from different origins,compare different specifications in the same origin,and explore the effect of origin and specifications on the quality of Achyranthis Bidentatae Radix and relationship between the specifications and the internal quality of Achyranthis Bidentatae Radix, in order to provide basis for the identification of its origin. Method:The HPLC fingerprints of Achyranthis Bidentatae Radix from different origins and with different specifications in the same origin were collected. The similarity analysis,cluster analysis and principal component analysis were adopted to analyze the fingerprints,the differences in fingerprints of Achyranthis Bidentatae Radix from different origins and with different specifications in the same origin were compared. Result:Analysis of different origins and principal component analysis could be used to distinguish Achyranthis Bidentatae Radix from five producing areas,and the identification results of origin analysis was better than those of cluster analysis and similarity analysis. Analysis of different specifications, similarity analysis or principal component analysis could not distinguish Achyranthis Bidentatae Radix with different specifications. Conclusion:There are significant differences in chemical composition and peak height among Achyranthis Bidentatae Radix from different origins,with less differences in chemical composition and peak height of Achyranthis Bidentatae Radix with different specifications, the principal component analysis could be used to identify origins of Achyranthis Bidentatae Radix.
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Objective:To investigate and compare the fingerprints of different polarity fractions (petroleumether,chloroform,ethyl acetate,n-butanol,water) of fresh Gardeniae Fructus with different fruit shapes,and further understand the content difference and distribution of its chemical composition. Method:Gardeniae Fructus was reflux extracted by water in order to obtain the water extract; water extract 0.1 g and dissolved with 50 mL water,then it was extracted by petroleum ether,chloroform,ethyl acetate and n-butanol in turn in order to obtain the different extraction phases and the water phase. Each phase was condensed to extractum. Finally,the samples were analyzed by high performance liquid chromatography (HPLC) fingerprints and the similarity evaluation software was used for data analysis. Result:Fingerprint of chloroform fraction of water extract in gardenia from different habitats can be used to distinguish Gardeniae Fructus from Fujian,Henan and Jiangxi. The differences between the water extract of Gardeniae Fructus from Fujian and those of Henan and Jiangxi were mainly manifested in the petroleum ether fraction, and the fat-soluble components of Gardeniae Fructus were more than those of Henan and Jiangxi. The differences between the water extract of Gardeniae Fructus from Henan and those of Fujian and Jiangxi were mainly manifested in the ethyl acetate fraction,and the content of iridoid glycosides was significantly higher than that in Fujian and Henan. The differences between the water extract of gardenia from Jiangxi and those of Fujian and Henan were mainly manifested in the n-butanol fraction,organic acid peak C1 not detected. The fingerprint of chloroform fraction of water extract in Gardeniae Fructus can be used to distinguish Gardeniae Fructus of six ribs and Gardeniae Fructus of seven ribs from Fujian and Henan,and the contents of all components of Gardeniae Fructus of seven ribs were more than those in Gardeniae Fructus of six ribs. The fingerprint of petroleum ether fraction of water extract in Gardeniae Fructus can be used to distinguish Gardeniae Fructus of six ribs and Gardeniae Fructus of seven ribs from Jiangxi. The Z3 peak of Gardeniae Fructus of six ribs from Henan was obviously higher than that of Gardeniae Fructus of seven ribs. The contents of all components on chloroform and ethyl acetate fractions of water extract in Gardeniae Fructus of seven ribs were significantly higher than those of Gardeniae Fructus of six ribs. Conclusion:There are significant differences on chemical constituents and content among Gardeniae Fructus from Fujian,Henan and Jiangxi. The main difference of fingerprint between Gardeniae Fructus of six ribs and Gardeniae Fructus of seven ribs is the peak height.
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Iridoid synthase( IS),the key enzyme in the natural biosynthesis of vegetal iridoids,catalyzes the irreversible cyclization of 10-oxogeranial to epi-iridodial. In this study,we screened the Rehmannia glutinosa transcriptome data by BLASTn with Catharanthus roseus CrIS cDNA,and found four c DNA fragments with length of 1 527,1 743,1 425,1 718 bp,named RgIS1,RgIS2,RgIS3 and RgIS4,respectively. Bioinformatics analysis revealed that the four iridoid synthase genes encoding proteins with 389-392 amino acid residues,protein molecular weights were between 44. 30-44. 74 k Da,and theoretical isoelectric points were between 5. 30 and 5. 87. Subcellular localization predictions showed that the four iridoid synthase were distributed in the cytoplasm. Structure analysis revealed that R. glutinosa iridoid synthases contain six conserved short-chain dehydrogenase/reductase( SDR) motifs,and their 3 D models were composed typical dinucleotide-binding " Rossmann" folds covered by helical C-terminal extensions. Using the amino acid sequences of four R. glutinosa iridoid synthases,phylogenetic analysis was performed,the result indicated that RgIS3,CrIS and Olea europaea OeIS were grouped together,the other R. glutinosa iridoid synthases and fifteen proteins in other plants had close relationship. Real-time fluorescent quantitative PCR revealed that RgIS1 and RgIS3 highly expressed in unfold leaves,however,RgIS2 and RgIS4 highly expressed in stems and tuberous roots,respectively. RgIS3 showed higher expression levels in non-radial striations( nRS) of the two cultivars,and RgIS1 and RgIS2 had higher expression levels in nRS of QH,while RgIS4 had less expression levels in nRS of QH1. RgIS1,RgIS2 and RgIS3 were up-regulated by Me JA treatment,although the time and degree of response differed. Our findings are helpful to reveal molecular function of R. glutinosa iridoid synthases and provide a clue for studing the molecular mechanism of iridoid biosynthesis.
Subject(s)
Cloning, Molecular , Genes, Plant , Iridoids , Metabolism , Ligases , Genetics , Phylogeny , Rehmannia , GeneticsABSTRACT
The history of Rehmannia glutinosa breeding has already beyond 100 years. There are rich cultivated varieties and wild germplasm resources in R. glutinosa. However, there also exist a lot of problems, such as, the pedigree of the existing varieties is not clear, the genetic basis is narrow, backward method of germplasm enhancement and breeding. Breeding of new varieties has been unable to meet the demand of R. glutinosa production in the new era. This paper summarizes the species of Rehmannia and their distribution, the diversity of plant morphology and the quality of R. glutinosa germplasm resources, as well as the progress of R. glutinosa breeding in recent 100 years. For ensuring the orderly, effective and safe production of R. glutinosa, the authors suggest to establish the wild resources protection area and germplasm resources garden, deeply study the genetic base of quality, strengthen application of new breeding method such as mutation breeding, haploid breeding and gene editing.
Subject(s)
Plant Breeding , Plants, Medicinal , Genetics , Rehmannia , GeneticsABSTRACT
To investigate the effect of different initial processing methods on the quality of Gardenia and determine the best cooking time in gardenia processing through the determination of index components content. The contents of geniposide, crocetin Ⅰ and total iridoid glycosides in Gardenia were determined before storage, six months after storage and one year after storage. During storage, the contents of geniposide, crocetin Ⅰ and total iridoid glycosides in directly dried Gardenia were 1.68%, 0.45% and 6.45% respectively. The contents of geniposide, crocetin Ⅰ and total iridoid glycosides in Gardenia with different steaming time were 1.34%-0.5%, 0.28%-0.06% and 6.09%-1.59% respectively. The contents of geniposide, crocetin Ⅰ and total iridoid glycosides in Gardenia with different boiling time (adding alum)were 1.42%-0.41%, 0.35%-0.07% and 6.40%-1.65% respectively. The direct drying of Gardenia samples could not achieve the function of killing enzyme and protecting glycosides. The enzymes from degradation of the index components were basically destroyed after steaming time of 13 min or boiling (adding alum) time of 8 min, achieving the function of killing enzyme and protecting glycosides.
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Objective To observe the clinical efficacy of Bai Xiao moxibustion plus electroacupuncture in treating lumbar intervertebral disc herniation (LIDH). Method Ninety-six LIDH patients were randomized into a treatment group and a control group, 48 cases each. The treatment group was intervened by Bai Xiao moxibustion plus electroacupuncture, while the control group was intervened by electroacupuncture alone. Before and after the treatment, the lower back pain scores of Japanese Orthopedic Association (JOA) and Visual Analogue Scale (VAS) were evaluated, and the clinical efficacies of the two groups were compared. Result The JOA and VAS lower back pain scores were changed significantly after the treatment in both groups (P<0.05). After the treatment, the JOA and VAS lower back pain scores of the treatment group were significantly different from those of the control group (P<0.05). The pain release time was (2.95±0.59)d after the intervention in the treatment group versus (4.26±0.68)d in the control group, and the between-group difference was statistically significant (P<0.05). The total effective rate was 95.7％ in the treatment group versus 91.7％ in the control group, and the between-group difference was statistically significant (P<0.05). Conclusion Bai Xiao moxibustion plus electroacupuncture is an effective method in treating LIDH and it can reduce the lower back pain.
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An HPLC method was established to determine the contents of catalpol, acteoside, rehmaionoside A, rehmaionoside D, leonuride in three part of Rehmanni glutinosa in Beijing No.1 variety R. glutinosa during the growth period, This method, in combination with its HPLC fingerprint was used to evaluate its overall quality characteristics.The results showed that：① the content of main components of R. glutinosa varied in different growth stages ;② there was a great difference of the content of main components between theradial striations and the non-radial striations; ③ the two sections almost have the same content distribution of catalpol, acteoside and rehmaionoside D; ④the content of rehmaionoside A in non-radial striations was higher than that in radial striations,while the content of leonuride in radial striations was higher than that in non-radial striations.; ⑤the HPLC fingerprint of radial striations, non-radial striations and whole root tuber were basically identical, except for the big difference in the content of chemical components. The result of clustering displayed that the radial striations, non-radial striations, and whole root were divided into two groups. In conclusion, there was a significant difference in the quality characteristics of radial striations and non-radial striations of R. glutinosa. This research provides a reference for quality evaluation and geoherbalism of R. glutinosa.
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Tyrosine decarboxylase (TyrDC) is an important enzyme in the secondary metabolism of several plant species, and was hypothesized to play a key role in the biosynthesis of phenylethanoid glycosides. Based on the transcriptome data, we cloned the full-length cDNA (GenBank accession NO. KU640395) of RgTyDC gene from Rehmannia glutinosa, and then performed bioinformatic analysis of the sequence. Further, we detected the expression pattern in different organs and hair roots treated with four elicitors by qRT-PCR. The results showed that the full length of RgTyDC cDNA was 1 530 bp encoding 509 amino acids. The molecular weight of the putative RgTyDC protein was about 56.6 kDa and the theoretical isoelectric point was 6.25. The RgTyDC indicated the highest homology with Sesamum indicum SiTyDC and Erythranthe guttata EgTyDC, both of them were reached 88%. RgTyDC highly expressed in R. glutinosa leaf, especially in senescing leaf, and rarely expressed in tuberous root. After the treatment of SA and MeJA, the relative expression level of RgTyDC mRNA was substantially increased. The results provide a foundation for exploring the molecular function of RgTyDC involved in phenylethanoid glycosides biosynthesis.
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<p><b>OBJECTIVE</b>To study the interactions between hemoglobin (Hb) and other proteins within human erythrocytes by using the electrophoresis release test (ERT) and co-immunoprecipitation.</p><p><b>METHODS</b>First, the fresh normal adult anti-coagulated whole blood was washed to prepare packed RBCs, which were further prepared to erythrocyte suspension and hemolysate.The erythrocyte suspension and hemolysate were analyzed by the electrophoresis release test (ERT) at the same time, and then the band of HbA of erythrocyte sample (RA) and the corresponding band of hemolysate sample (HA) were cut out from the gel and were enriched by freeze-thaw method. Then, the samples were bound with hemoglobin β antibody (37-8) AC, the complexes were separated through 5%-12% SDS-PAGE followed by Q-TOF.</p><p><b>RESULTS</b>Five bands were found in the gel, each of which was treated by hemoglobin β antibody (37-8) AC, the protein bands of 16,20,22,28 and 50 kD were emerged in RA, HA and RBC lysis.The bands were identified by MS, and the results showed that these bands were hemoglobin, band 3, peroxiredoxin2 (Prx2), band 3 and β-actin, band 3 respectively.</p><p><b>CONCLUSION</b>HbA may interact with Prx2, Band 3 and β-actin, and then the complexes are formed with each other within erythrocytes.</p>
Subject(s)
Humans , Electrophoresis, Polyacrylamide Gel , Erythrocytes , Hemoglobins , Immunoprecipitation , ProteinsABSTRACT
To detect 4 MicroRNA [miRNA] in the stool samples of colorectal cancer [CRC] patients to determine whether these miRNAs could be biomarkers in CRC screening or treatment. A retrospective comparison study was carried out in the Department of Colorectal Surgery, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China from September 2009 to March 2011. We detected 4 miRNAs [miR-143, miR-145, miR-21, and miR-106a] in the stool samples of 38 CRC patients and 13 healthy individuals. Total RNA from the stool samples was extracted using the EZNA TM stool RNA kit R6828-01. The miRNA quantification was carried out using TaqMan miRNA assays and the TaqMan Gene Expression Master Mix. The expression levels of miR-143 and miR-145 in the stool of the CRC patients were lower than in those of the healthy persons [p<0.005, median of 2[-Ct]]. No statistically significant difference was found in the expression levels of both miR-21 and miR-106a between the stool of CRC patients and those of the healthy persons [p>0.05]. The detection of fecal miRNAs is a potential method for CRC diagnosis or screening. Particularly, the down-regulation of fecal miR-143 and miR-145 could be a potential marker for CRC